text
stringlengths 310
8.3k
| input_ids
listlengths 1.02k
1.02k
| length
int64 1.02k
1.02k
| source_domain
stringclasses 7
values | writing_style_average
float64 -3.46
3.36
| required_expertise_average
float64 -3.74
4.36
| facts_and_trivia_average
float64 -3.78
3.96
| educational_value_average
float64 -3.85
3.97
|
|---|---|---|---|---|---|---|---|
it would have the effect of wages increasing worldwide and potentially a convergence in wages for highly skilled people. With the advent of technology that has increased productivity manifold times, manufacturing would be based in places not where the wages are lower, but where there is an abundant supply of skilled workers, technology and innovation. If the free movement of people is restricted, employers will be forced to move operations to other countries, thus perpetuating wage disparity.
This brings us to the H-1B visa program that has a mere 65,000 visas, plus an additional 20,000 for those who have graduated with advance degrees. Due to the well publicized layoffs of US workers at companies like Disney by H-1B workers, there appears to be no appetite by Congress to increase H-1B visa numbers even though there is a dire need to do so. By continuing to limit and stifle the H-1B program, US employers will remain less competitive and will not be able to pass on the benefits to consumers. We need more H-1B visa numbers rather than less. We also need to respect H-1B workers rather than deride them, even if they work at IT consulting company, as they too wish to abide by the law and to pursue their dreams in America. The best way to reform the H-1B program is to provide more mobility to H-1B visa workers. By providing more mobility, which includes being able to obtain a green card quickly, H-1B workers will not be stuck with the employer who brought them on the H-1B visa, and this can also result in rising wages within the occupation as a whole. Mobile foreign workers will also be incentivized to start their own innovative companies in America, which in turn will result in more jobs. This is the best way to reform the H-1B visa program, rather than to further shackle it with stifling laws and regulations, labor attestations and quotas. Market forces can better control the H-1B program from abuses and distortions than labor attestations!
As we meditate over yet another July 4th weekend celebrating America's independence, we should note that the world faces a stark choice today. Should countries be more open or less open? The ideological line between left and right is blurring as another more distinct line is being drawn between open and closed nations. America was founded on principles of openness and its ability to embrace people from all over the word, but that may change if the proponents for a closed and isolated world have their way. If America becomes closed, just like Britain will likely be after Brexit, there will be fewer opportunities for businesses to sell outside national borders, and they will be further stymied and unable to grow if they cannot gain access to the best talent. Moreover, innovation will get stifled if the best people from around the world cannot cluster together to develop new products and change paradigms. Immigration is what fuels these advances, which in turn promises more growth and prosperity. Do we want to revive the industries of the past to bring back those illusory jobs, such as steel manufacturing or coal mining, after technology has already marched on, or do we want to imagine about autonomous vehicles (notwithstanding the recent Tesla car setback), nanotechnology that will automatically repair our cells and space travel through a wormhole? Brexit and xenophobia go hand in hand. Will America buck this trend in favor of immigration and innovation when it goes to the polls in November 2016?
An Eventful Thursday for Immigration Law at the Supreme Court: United States v. Texas, Mathis v. United States, and What's Next
by David Isaacson, Partner with ABIL member, Cyrus D. Mehta
On Thursday, June 23, the U.S. Supreme Court issued two decisions of significance to immigration law: a 4-4 affirmance without opinion in United States v. Texas, and a 5-3 decision in Mathis v. United States. The first, which was more obviously immigration-related, is very disappointing and has rightly received a great deal of media attention, but the second is also worth noting and is somewhat more positive.
The Court's evenly-divided decision – or one might say lack of decision – in United States v. Texas left standing the previous 2-1 decision of a panel of the Court of Appeals for the Fifth Circuit, which had uph
|
[
372,
723,
505,
278,
2779,
310,
281,
1179,
10231,
3186,
8157,
322,
19998,
263,
17221,
297,
281,
1179,
363,
10712,
2071,
24455,
2305,
29889,
2973,
278,
17623,
310,
15483,
393,
756,
11664,
3234,
2068,
25941,
3064,
29892,
12012,
3864,
723,
367,
2729,
297,
7600,
451,
988,
278,
281,
1179,
526,
5224,
29892,
541,
988,
727,
338,
385,
18666,
424,
11421,
310,
2071,
24455,
17162,
29892,
15483,
322,
24233,
362,
29889,
960,
278,
3889,
10298,
310,
2305,
338,
22078,
29892,
5703,
414,
674,
367,
11826,
304,
4337,
6931,
304,
916,
10916,
29892,
4550,
25722,
29884,
1218,
281,
482,
17508,
537,
29889,
13,
4013,
23522,
502,
304,
278,
379,
29899,
29896,
29933,
1998,
29874,
1824,
393,
756,
263,
15187,
29871,
29953,
29945,
29892,
29900,
29900,
29900,
1998,
294,
29892,
2298,
385,
5684,
29871,
29906,
29900,
29892,
29900,
29900,
29900,
363,
1906,
1058,
505,
23588,
411,
6564,
14496,
29889,
16809,
304,
278,
1532,
970,
1891,
6568,
22450,
310,
3148,
17162,
472,
14582,
763,
17944,
491,
379,
29899,
29896,
29933,
17162,
29892,
727,
5692,
304,
367,
694,
623,
300,
568,
491,
11559,
304,
7910,
379,
29899,
29896,
29933,
1998,
29874,
3694,
1584,
2466,
727,
338,
263,
2970,
817,
304,
437,
577,
29889,
2648,
3133,
292,
304,
4046,
322,
380,
361,
280,
278,
379,
29899,
29896,
29933,
1824,
29892,
3148,
5703,
414,
674,
3933,
3109,
5100,
3321,
322,
674,
451,
367,
2221,
304,
1209,
373,
278,
23633,
304,
11233,
414,
29889,
1334,
817,
901,
379,
29899,
29896,
29933,
1998,
29874,
3694,
3265,
1135,
3109,
29889,
1334,
884,
817,
304,
3390,
379,
29899,
29896,
29933,
17162,
3265,
1135,
589,
680,
963,
29892,
1584,
565,
896,
664,
472,
13315,
8799,
292,
5001,
29892,
408,
896,
2086,
6398,
304,
633,
680,
491,
278,
4307,
322,
304,
12359,
434,
1009,
12561,
29879,
297,
6813,
29889,
450,
1900,
982,
304,
11736,
278,
379,
29899,
29896,
29933,
1824,
338,
304,
3867,
901,
22458,
1793,
304,
379,
29899,
29896,
29933,
1998,
29874,
17162,
29889,
2648,
13138,
901,
22458,
1793,
29892,
607,
7805,
1641,
2221,
304,
4017,
263,
7933,
5881,
9098,
29892,
379,
29899,
29896,
29933,
17162,
674,
451,
367,
10771,
411,
278,
5703,
261,
1058,
6296,
963,
373,
278,
379,
29899,
29896,
29933,
1998,
29874,
29892,
322,
445,
508,
884,
1121,
297,
20493,
281,
1179,
2629,
278,
26818,
408,
263,
3353,
29889,
21600,
9117,
17162,
674,
884,
367,
297,
1760,
440,
1891,
304,
1369,
1009,
1914,
24233,
1230,
14582,
297,
6813,
29892,
607,
297,
2507,
674,
1121,
297,
901,
17643,
29889,
910,
338,
278,
1900,
982,
304,
11736,
278,
379,
29899,
29896,
29933,
1998,
29874,
1824,
29892,
3265,
1135,
304,
4340,
528,
547,
280,
372,
411,
380,
361,
1847,
14243,
322,
1072,
8250,
29892,
10212,
1098,
342,
800,
322,
13911,
294,
29889,
28794,
8249,
508,
2253,
2761,
278,
379,
29899,
29896,
29933,
1824,
515,
633,
6394,
322,
1320,
441,
1080,
1135,
10212,
1098,
342,
800,
29991,
13,
2887,
591,
1612,
10388,
975,
3447,
1790,
5468,
29871,
29946,
386,
4723,
355,
10894,
1218,
6813,
29915,
29879,
21820,
29892,
591,
881,
4443,
393,
278,
3186,
17240,
263,
18561,
7348,
9826,
29889,
10575,
10916,
367,
901,
1722,
470,
3109,
1722,
29973,
450,
1957,
5996,
1196,
1546,
2175,
322,
1492,
338,
1999,
1038,
292,
408,
1790,
901,
8359,
1196,
338,
1641,
12061,
1546,
1722,
322,
5764,
19079,
29889,
6813,
471,
11091,
373,
18671,
310,
1722,
2264,
322,
967,
11509,
304,
953,
13842,
2305,
515,
599,
975,
278,
1734,
29892,
541,
393,
1122,
1735,
565,
278,
410,
9340,
363,
263,
5764,
322,
23968,
3186,
505,
1009,
982,
29889,
960,
6813,
7415,
5764,
29892,
925,
763,
14933,
674,
5517,
367,
1156,
5826,
29916,
277,
29892,
727,
674,
367,
28145,
28602,
1907,
363,
5381,
267,
304,
19417,
5377,
4797,
28199,
29892,
322,
896,
674,
367,
4340,
380,
962,
1000,
322,
9368,
304,
6548,
565,
896,
2609,
11581,
2130,
304,
278,
1900,
24242,
29889,
12808,
29892,
24233,
362,
674,
679,
380,
361,
839,
565,
278,
1900,
2305,
515,
2820,
278,
3186,
2609,
9867,
4208,
304,
2693,
716,
9316,
322,
1735,
26811,
335,
1516,
29889,
1954,
29885,
16783,
338,
825,
4084,
1379,
1438,
3061,
2925,
29892,
607,
297,
2507,
27584,
901,
14321,
322,
25831,
537,
29889,
1938,
591,
864,
304,
6664,
573,
278,
6397,
2722,
310,
278,
4940,
304,
6963,
1250,
1906,
4486,
375,
706,
17643,
29892,
1316,
408,
22973,
12012,
3864,
470,
17148,
1375,
292,
29892,
1156,
15483,
756,
2307,
8575,
287,
373,
29892,
470,
437,
591,
864,
304,
14034,
1048,
28273,
681,
24413,
313,
1333,
2541,
11235,
278,
7786,
323,
267,
433,
1559,
731,
1627,
511,
23432,
866,
3049,
3002,
393,
674,
6336,
26032,
1749,
9101,
322,
2913,
9850,
1549,
263,
281,
555,
29716,
29973,
5826,
29916,
277,
322,
921,
264,
3021,
711,
423,
748,
1361,
297,
1361,
29889,
2811,
6813,
1321,
384,
445,
534,
355,
297,
7853,
310,
5198,
16783,
322,
24233,
362,
746,
372,
5771,
304,
278,
1248,
3137,
297,
3979,
29871,
29906,
29900,
29896,
29953,
29973,
13,
2744,
6864,
1319,
498,
1295,
3250,
363,
1954,
29885,
16783,
7927,
472,
278,
22569,
9245,
29901,
3303,
3900,
325,
29889,
10319,
29892,
5792,
275,
325,
29889,
3303,
3900,
29892,
322,
1724,
29915,
29879,
8084,
13,
1609,
4699,
28156,
1100,
29892,
3455,
1089,
411,
17571,
6227,
4509,
29892,
315,
29646,
360,
29889,
2191,
400,
29874,
13,
2951,
498,
1295,
3250,
29892,
5306,
29871,
29906,
29941,
29892,
278,
501,
29889,
29903,
29889,
22569,
9245,
16610,
1023,
1602,
12112,
310,
26002,
304,
5198,
16783,
4307,
29901,
263,
29871,
29946,
29899,
29946,
2756,
3568,
749,
1728,
9426,
297,
3303,
3900,
325,
29889,
10319,
29892,
322,
263,
29871,
29945,
29899,
29941,
10608,
297,
5792,
275,
325,
29889,
3303,
3900,
29889,
450,
937,
29892,
607,
471,
901,
12879,
5198,
16783,
29899,
12817,
29892,
338,
1407,
23451,
292,
322,
756,
1492,
368,
4520,
263,
2107,
5376,
310,
5745,
8570,
29892,
541,
278,
1473,
338,
884,
7088,
451,
292,
322,
338,
10579,
901,
6374,
29889,
13,
1576,
9245,
29915,
29879,
1584,
368,
29899,
4563,
2618,
10608,
785,
470,
697,
1795,
1827,
10225,
310,
10608,
785,
297,
3303,
3900,
325,
29889,
10319,
2175,
13407,
278,
3517,
29871,
29906,
29899,
29896,
10608,
310,
263,
9451,
310,
278,
9245,
310,
2401,
29872,
1338,
363,
278,
29008,
386,
12594,
3121,
29892,
607,
750,
318,
561
] | 1,024 |
CommonCrawl
| 1.114749 | 0.78191 | 0.486978 | 0.921842 |
eld District Judge Andrew Hanen's preliminary injunction against Deferred Action for Parents of Americans and Lawful Permanent Residents (DAPA) and against the related expansion of Deferred Action for Childhood Arrivals ("DACA+"). This is quite a momentous outcome to have been reached without explanation. As former Solicitor General Walter Dellinger has written: "It is hard to know what to say about an immigration opinion affecting 4.3 million people that reads, in its entirety: 'The judgment is affirmed by an equally divded Court.' Seldom have so many hopes been crushed by so few words."
It has long been customary for an evenly divided Supreme Court to affirm the judgment below without offering opinions, as was done here (and has been done since at least 1909), although this is not a custom followed by all multi-member appellate courts in the United States. The Court of Appeals for the First Circuit sitting en banc, for example, has provided opinions explaining the views of its judges when it has divided 3 to 3 in recent years and so affirmed the district courts below, as in the immigration detention case of Castaneda v. Souza, on which the First Circuit split in December 2015. If the Supreme Court were to adopt a similar custom, one would at least have the satisfaction of knowing the reasoning behind the Justices' votes. In this particular case, one might also have hoped that some of the Justices who voted to affirm the Fifth Circuit could have been convinced to change their minds by a compelling dissenting opinion that they knew they would have to confront publicly, although presumably draft opinions were circulated internally, given the long lapse of time between oral argument in the case on April 16 and the issuance of decision last week. The 4-4 deadlock, and the Supreme Court's custom of not issuing opinions in that scenario, has left those Justices who voted to affirm the Fifth Circuit in the position of being able to do so without having to explain formally and publicly why such a position is legally coherent.
While it is frustrating that the injunction in United States v. Texas was affirmed without explanation and without any precedential decision, however, this does have the benefit of leaving the door open for a different outcome in the long run. United States v. Texas could return to the Supreme Court once a 9th Justice is seated on the Court, and potentially be decided differently, in one of at least two ways.
First, as SCOTUSBlog pointed out soon after the 4-4 decision came down, the government can petition the Supreme Court to rehear the case, and ask that the petition be held until a 9th Justice is seated on the Court. Former Solicitor General Dellinger also endorsed that approach in his above-quoted post at Slate's "Supreme Court Breakfast Table". This would be one way for U.S. v. Texas to come back before the Supreme Court, potentially quite quickly after a 9th Justice is seated. Under Supreme Court Rule 44, a petition for rehearing ordinarily will only be granted both "by a majority of the Court" and "at the instance of a Justice who concurred in the judgment or decision", but it is unclear how this latter requirement could possibly be applied in the case of a 4-4 affirmance without opinion, where the Court has not issued its own judgment and there is no public record of any Justice concurring in the affirmance more than any other Justice. Thus, it appears that an ordinary majority, presumably composed of the 4 Justices who voted to reverse plus a newly arrived 9th Justice, could grant a petition for rehearing if it were still pending when a 9th Justice were confirmed.
SCOTUSBlog also noted today, however, that the Court had denied one petition in another case seeking such rehearing by a 9-member Court, which may not bode well for the rehearing possibility. Specifically, the Court denied rehearing in Hawkins v. Community Bank of Raymore, which had been the subject of an affirmance without opinion by an equally divided Court on March 22, 2016. One might think, though, that there is a significant difference between a case like U.S. v. Texas, decided just before the end of the term and affecting national policy so substantially, and a case like Hawkins, in which the Court failed 3 months earlier to reach a conclusion regarding the questions "(1) Whether "primarily and un
|
[
2495,
7457,
26817,
11571,
7169,
264,
29915,
29879,
758,
2576,
3821,
10899,
651,
2750,
897,
14373,
9123,
363,
1459,
1237,
310,
23035,
322,
7927,
1319,
349,
3504,
296,
2538,
16719,
313,
29928,
3301,
29909,
29897,
322,
2750,
278,
4475,
13184,
310,
897,
14373,
9123,
363,
10517,
6614,
826,
15081,
29879,
4852,
29928,
2477,
29909,
29974,
2564,
910,
338,
3755,
263,
3256,
681,
21957,
304,
505,
1063,
7450,
1728,
8252,
29889,
1094,
4642,
4956,
293,
2105,
4593,
10705,
360,
514,
5621,
756,
3971,
29901,
376,
3112,
338,
2898,
304,
1073,
825,
304,
1827,
1048,
385,
5198,
16783,
9426,
6602,
292,
29871,
29946,
29889,
29941,
7284,
2305,
393,
13623,
29892,
297,
967,
4152,
1017,
29901,
525,
1576,
24284,
338,
2756,
381,
2168,
491,
385,
18018,
1933,
7176,
9245,
6169,
317,
2495,
290,
505,
577,
1784,
26926,
1063,
2181,
15392,
491,
577,
2846,
3838,
1213,
13,
3112,
756,
1472,
1063,
2888,
653,
363,
385,
1584,
368,
13931,
22569,
9245,
304,
2756,
3568,
278,
24284,
2400,
1728,
27032,
26971,
29892,
408,
471,
2309,
1244,
313,
392,
756,
1063,
2309,
1951,
472,
3203,
29871,
29896,
29929,
29900,
29929,
511,
5998,
445,
338,
451,
263,
2888,
5643,
491,
599,
2473,
29899,
14242,
623,
514,
403,
28033,
297,
278,
3303,
3900,
29889,
450,
9245,
310,
2401,
29872,
1338,
363,
278,
3824,
12594,
3121,
16246,
427,
289,
4564,
29892,
363,
1342,
29892,
756,
4944,
26971,
24232,
278,
8386,
310,
967,
6577,
2710,
746,
372,
756,
13931,
29871,
29941,
304,
29871,
29941,
297,
7786,
2440,
322,
577,
2756,
381,
2168,
278,
6474,
28033,
2400,
29892,
408,
297,
278,
5198,
16783,
1439,
2509,
1206,
310,
4834,
273,
8710,
325,
29889,
9194,
1362,
29892,
373,
607,
278,
3824,
12594,
3121,
6219,
297,
5846,
29871,
29906,
29900,
29896,
29945,
29889,
960,
278,
22569,
9245,
892,
304,
9332,
263,
2788,
2888,
29892,
697,
723,
472,
3203,
505,
278,
26470,
310,
13797,
278,
24481,
5742,
278,
3387,
1575,
29915,
18952,
29889,
512,
445,
3153,
1206,
29892,
697,
1795,
884,
505,
24936,
393,
777,
310,
278,
3387,
1575,
1058,
24854,
304,
2756,
3568,
278,
29008,
386,
12594,
3121,
1033,
505,
1063,
25617,
304,
1735,
1009,
27656,
491,
263,
752,
7807,
16317,
296,
292,
9426,
393,
896,
6363,
896,
723,
505,
304,
21751,
970,
368,
29892,
5998,
2225,
24873,
18195,
26971,
892,
18342,
630,
25106,
29892,
2183,
278,
1472,
23012,
344,
310,
931,
1546,
470,
284,
2980,
297,
278,
1206,
373,
3786,
29871,
29896,
29953,
322,
278,
17759,
749,
310,
10608,
1833,
4723,
29889,
450,
29871,
29946,
29899,
29946,
7123,
908,
29892,
322,
278,
22569,
9245,
29915,
29879,
2888,
310,
451,
17759,
292,
26971,
297,
393,
10483,
29892,
756,
2175,
1906,
3387,
1575,
1058,
24854,
304,
2756,
3568,
278,
29008,
386,
12594,
3121,
297,
278,
2602,
310,
1641,
2221,
304,
437,
577,
1728,
2534,
304,
5649,
28269,
322,
970,
368,
2020,
1316,
263,
2602,
338,
2814,
635,
16165,
261,
296,
29889,
13,
8809,
488,
372,
338,
1424,
4627,
1218,
393,
278,
10899,
651,
297,
3303,
3900,
325,
29889,
10319,
471,
2756,
381,
2168,
1728,
8252,
322,
1728,
738,
9399,
2556,
10608,
29892,
3138,
29892,
445,
947,
505,
278,
14169,
310,
10124,
278,
3050,
1722,
363,
263,
1422,
21957,
297,
278,
1472,
1065,
29889,
3303,
3900,
325,
29889,
10319,
1033,
736,
304,
278,
22569,
9245,
2748,
263,
29871,
29929,
386,
17181,
338,
409,
630,
373,
278,
9245,
29892,
322,
19998,
367,
8459,
17587,
29892,
297,
697,
310,
472,
3203,
1023,
5837,
29889,
13,
6730,
29892,
408,
12314,
2891,
29965,
1744,
1188,
11520,
714,
4720,
1156,
278,
29871,
29946,
29899,
29946,
10608,
2996,
1623,
29892,
278,
5874,
508,
5697,
654,
278,
22569,
9245,
304,
337,
354,
279,
278,
1206,
29892,
322,
2244,
393,
278,
5697,
654,
367,
4934,
2745,
263,
29871,
29929,
386,
17181,
338,
409,
630,
373,
278,
9245,
29889,
3812,
261,
4956,
293,
2105,
4593,
360,
514,
5621,
884,
1095,
943,
287,
393,
2948,
297,
670,
2038,
29899,
339,
5715,
1400,
472,
317,
9632,
29915,
29879,
376,
29903,
786,
10291,
9245,
28301,
11255,
6137,
1642,
910,
723,
367,
697,
982,
363,
501,
29889,
29903,
29889,
325,
29889,
10319,
304,
2041,
1250,
1434,
278,
22569,
9245,
29892,
19998,
3755,
9098,
1156,
263,
29871,
29929,
386,
17181,
338,
409,
630,
29889,
7634,
22569,
9245,
27308,
29871,
29946,
29946,
29892,
263,
5697,
654,
363,
337,
354,
4362,
22420,
6275,
674,
871,
367,
16896,
1716,
376,
1609,
263,
13638,
310,
278,
9245,
29908,
322,
376,
271,
278,
2777,
310,
263,
17181,
1058,
378,
2764,
1127,
297,
278,
24284,
470,
10608,
613,
541,
372,
338,
20871,
920,
445,
7480,
11809,
1033,
10075,
367,
7436,
297,
278,
1206,
310,
263,
29871,
29946,
29899,
29946,
2756,
3568,
749,
1728,
9426,
29892,
988,
278,
9245,
756,
451,
16610,
967,
1914,
24284,
322,
727,
338,
694,
970,
2407,
310,
738,
17181,
3022,
1038,
292,
297,
278,
2756,
3568,
749,
901,
1135,
738,
916,
17181,
29889,
6549,
29892,
372,
5692,
393,
385,
15311,
13638,
29892,
2225,
24873,
13725,
310,
278,
29871,
29946,
3387,
1575,
1058,
24854,
304,
11837,
2298,
263,
15141,
11977,
29871,
29929,
386,
17181,
29892,
1033,
16690,
263,
5697,
654,
363,
337,
354,
4362,
565,
372,
892,
1603,
28235,
746,
263,
29871,
29929,
386,
17181,
892,
16725,
29889,
13,
7187,
2891,
29965,
1744,
1188,
884,
11682,
9826,
29892,
3138,
29892,
393,
278,
9245,
750,
17935,
697,
5697,
654,
297,
1790,
1206,
25738,
1316,
337,
354,
4362,
491,
263,
29871,
29929,
29899,
14242,
9245,
29892,
607,
1122,
451,
289,
356,
1532,
363,
278,
337,
354,
4362,
13331,
29889,
26321,
29892,
278,
9245,
17935,
337,
354,
4362,
297,
10875,
11335,
325,
29889,
19184,
10253,
310,
9596,
5514,
29892,
607,
750,
1063,
278,
4967,
310,
385,
2756,
3568,
749,
1728,
9426,
491,
385,
18018,
13931,
9245,
373,
4779,
29871,
29906,
29906,
29892,
29871,
29906,
29900,
29896,
29953,
29889,
3118,
1795,
1348,
29892,
2466,
29892,
393,
727,
338,
263,
7282,
4328,
1546,
263,
1206,
763,
501,
29889,
29903,
29889,
325,
29889,
10319,
29892,
8459,
925,
1434,
278,
1095,
310,
278,
1840,
322,
6602,
292,
4797,
8898,
577,
23228,
368,
29892,
322,
263,
1206,
763,
10875,
11335,
29892,
297,
607,
278,
9245,
5229,
29871,
29941,
7378,
8859,
304,
6159,
263,
15997,
11211,
278,
5155,
18227,
29896,
29897,
26460,
376,
9469,
6275,
322,
443
] | 1,024 |
CommonCrawl
| 0.68817 | 1.085592 | 0.248479 | 0.600564 |
conditionally liable" spousal guarantors are unambiguously excluded from being Equal Credit Opportunity Act (ECOA) "applicants" because they are not integrally part of "any aspect of a credit transaction"; and (2) whether the Federal Reserve Board has authority under the ECOA to include by regulation spousal guarantors as "applicants" to further the purposes of eliminating discrimination against married women." Without meaning to minimize the significance of discrimination against married women seeking credit, one might reasonably suggest that this sort of garden-variety issue of statutory interpretation does not call for unusual procedural measures to achieve a final, reasoned resolution to the same extent as the issue of the legitimacy of DAPA and DACA+.
Even if rehearing is not granted, it is likely that U.S. v. Texas may ultimately return to the Supreme Court, unless it is rendered moot in the meantime by Congressional enactment of comprehensive immigration reform or recission of DAPA by a hypothetical Republican President. (Hillary Clinton, the presumptive Democratic Presidential nominee, has made quite clear that she would seek to defend and expand administrative relief such as DAPA, not rescind it.) The injunction issued by Judge Hanen and upheld by the Fifth Circuit was a preliminary injunction, and the case would now ordinarily be expected to proceed to a trial on the merits, or at least some sort of further proceedings. At the conclusion of such further proceedings, Judge Hanen may then issue a permanent injunction. The grant of such a permanent injunction could be appealed back to the Fifth Circuit, and if it were again affirmed by the Fifth Circuit, the government could seek certiorari from the Supreme Court regarding that affirmance. One hopes that by the time the case worked its way back up through the Fifth Circuit to the Supreme Court in this fashion, there will be a 9th Justice seated on the Supreme Court.
There has also been some speculation that a new case regarding DAPA and DACA+ could be commenced, in a Circuit other than the Fifth Circuit, which might come out differently and create a circuit split. It may be unlikely that such other litigation, even if deemed feasible, would make it back to the Supreme Court sooner than one of the other two routes discussed above. If there is a way to launch such other litigation despite the current national scope of the injunction against DAPA and DACA+, however, it could have other benefits: David Leopold, for example, suggests in a recent blog post that further litigation might allow DAPA and DACA+ to go into effect in portions of the United States even if not nationwide. Achieving such a goal would be difficult, given the current nationwide injunction against DAPA and DACA+ issued by Judge Hanen and upheld by the Fifth Circuit, but it appears that some intelligent and creative attorneys may be looking to see if they can find a way.
Ultimately, however, it appears that the future of DAPA and DACA+ will likely depend on who (if anyone) fills the currently vacant 9th seat on the Supreme Court. In this, as in many other things, the outcome of this November's elections will be crucial.
If Hillary Clinton is elected President and the Democrats retake the majority in the Senate, the 9th Justice who would be confirmed, whether that is President Obama's nominee Chief Judge Merrick Garland of the Court of Appeals for the D.C. Circuit or a new nominee put forward by President Clinton, would likely vote to overturn the injunction against DAPA and DACA+ if and when the case returned to the Supreme Court. In the tragic event of a Donald Trump Presidency, on the other hand, the issue would be moot, since DAPA and DACA+ would be rescinded anyway. In the event that Hillary Clinton is elected President and the Republicans maintain control of the Senate, one hopes that they would not attempt to block a Supreme Court nomination indefinitely, but given the current behavior of the Senate Republican majority, one cannot be sure. Therefore, a Democratic victory in not only the Presidential election, but also a sufficient number of Senate elections to reclaim the majority (requiring a net gain of at least four seats), would give the best hope for a revival of DAPA and DACA+. If the Democr
|
[
16122,
635,
619,
519,
29908,
805,
681,
284,
1410,
279,
424,
943,
526,
443,
14727,
5794,
429,
13347,
515,
1641,
11243,
284,
24596,
277,
438,
3016,
6997,
3185,
313,
29923,
3217,
29909,
29897,
376,
932,
506,
1934,
29908,
1363,
896,
526,
451,
3990,
635,
760,
310,
376,
1384,
9565,
310,
263,
16200,
10804,
1769,
322,
313,
29906,
29897,
3692,
278,
14879,
27811,
12590,
756,
14329,
1090,
278,
382,
3217,
29909,
304,
3160,
491,
1072,
2785,
805,
681,
284,
1410,
279,
424,
943,
408,
376,
932,
506,
1934,
29908,
304,
4340,
278,
11976,
310,
10397,
1218,
2313,
5632,
3381,
2750,
8300,
5866,
1213,
13932,
6593,
304,
6260,
675,
278,
26002,
310,
2313,
5632,
3381,
2750,
8300,
5866,
25738,
16200,
29892,
697,
1795,
2769,
2197,
4368,
393,
445,
2656,
310,
16423,
29899,
5927,
3305,
2228,
310,
1002,
329,
706,
19854,
947,
451,
1246,
363,
22910,
6449,
3631,
15366,
304,
6176,
263,
2186,
29892,
2769,
287,
10104,
304,
278,
1021,
15834,
408,
278,
2228,
310,
278,
25204,
326,
4135,
310,
360,
3301,
29909,
322,
360,
2477,
29909,
29974,
29889,
13,
29923,
854,
565,
337,
354,
4362,
338,
451,
16896,
29892,
372,
338,
5517,
393,
501,
29889,
29903,
29889,
325,
29889,
10319,
1122,
18973,
736,
304,
278,
22569,
9245,
29892,
6521,
372,
338,
13751,
2730,
327,
297,
278,
6839,
603,
491,
6005,
29878,
15750,
427,
627,
358,
310,
15171,
6270,
5198,
16783,
11736,
470,
1162,
2333,
310,
360,
3301,
29909,
491,
263,
13752,
300,
936,
21178,
7178,
29889,
313,
29950,
453,
653,
2233,
16929,
29892,
278,
2225,
398,
415,
573,
19083,
7178,
616,
2245,
457,
29872,
29892,
756,
1754,
3755,
2821,
393,
1183,
723,
16508,
304,
24663,
322,
7985,
19185,
18892,
1316,
408,
360,
3301,
29909,
29892,
451,
620,
29883,
513,
372,
1846,
450,
10899,
651,
16610,
491,
26817,
7169,
264,
322,
318,
561,
2495,
491,
278,
29008,
386,
12594,
3121,
471,
263,
758,
2576,
3821,
10899,
651,
29892,
322,
278,
1206,
723,
1286,
22420,
6275,
367,
3806,
304,
8469,
304,
263,
14260,
373,
278,
2778,
1169,
29892,
470,
472,
3203,
777,
2656,
310,
4340,
8469,
886,
29889,
2180,
278,
15997,
310,
1316,
4340,
8469,
886,
29892,
26817,
7169,
264,
1122,
769,
2228,
263,
17667,
10899,
651,
29889,
450,
16690,
310,
1316,
263,
17667,
10899,
651,
1033,
367,
5929,
7943,
1250,
304,
278,
29008,
386,
12594,
3121,
29892,
322,
565,
372,
892,
1449,
2756,
381,
2168,
491,
278,
29008,
386,
12594,
3121,
29892,
278,
5874,
1033,
16508,
2284,
1611,
1306,
515,
278,
22569,
9245,
11211,
393,
2756,
3568,
749,
29889,
3118,
26926,
393,
491,
278,
931,
278,
1206,
3796,
967,
982,
1250,
701,
1549,
278,
29008,
386,
12594,
3121,
304,
278,
22569,
9245,
297,
445,
13460,
29892,
727,
674,
367,
263,
29871,
29929,
386,
17181,
409,
630,
373,
278,
22569,
9245,
29889,
13,
8439,
756,
884,
1063,
777,
1580,
2785,
393,
263,
716,
1206,
11211,
360,
3301,
29909,
322,
360,
2477,
29909,
29974,
1033,
367,
844,
9223,
29892,
297,
263,
12594,
3121,
916,
1135,
278,
29008,
386,
12594,
3121,
29892,
607,
1795,
2041,
714,
17587,
322,
1653,
263,
11369,
6219,
29889,
739,
1122,
367,
25057,
393,
1316,
916,
11872,
335,
362,
29892,
1584,
565,
316,
22580,
28326,
1821,
29892,
723,
1207,
372,
1250,
304,
278,
22569,
9245,
29548,
1135,
697,
310,
278,
916,
1023,
12049,
15648,
2038,
29889,
960,
727,
338,
263,
982,
304,
6826,
1316,
916,
11872,
335,
362,
15020,
278,
1857,
4797,
6874,
310,
278,
10899,
651,
2750,
360,
3301,
29909,
322,
360,
2477,
29909,
29974,
29892,
3138,
29892,
372,
1033,
505,
916,
23633,
29901,
4699,
23146,
29892,
363,
1342,
29892,
14661,
297,
263,
7786,
12618,
1400,
393,
4340,
11872,
335,
362,
1795,
2758,
360,
3301,
29909,
322,
360,
2477,
29909,
29974,
304,
748,
964,
2779,
297,
2011,
1080,
310,
278,
3303,
3900,
1584,
565,
451,
5233,
8157,
29889,
15542,
15387,
1316,
263,
7306,
723,
367,
5189,
29892,
2183,
278,
1857,
5233,
8157,
10899,
651,
2750,
360,
3301,
29909,
322,
360,
2477,
29909,
29974,
16610,
491,
26817,
7169,
264,
322,
318,
561,
2495,
491,
278,
29008,
386,
12594,
3121,
29892,
541,
372,
5692,
393,
777,
13052,
296,
322,
907,
1230,
1098,
272,
484,
952,
1122,
367,
3063,
304,
1074,
565,
896,
508,
1284,
263,
982,
29889,
13,
29965,
1896,
15084,
29892,
3138,
29892,
372,
5692,
393,
278,
5434,
310,
360,
3301,
29909,
322,
360,
2477,
29909,
29974,
674,
5517,
8839,
373,
1058,
313,
361,
5019,
29897,
27793,
278,
5279,
11757,
424,
29871,
29929,
386,
12949,
373,
278,
22569,
9245,
29889,
512,
445,
29892,
408,
297,
1784,
916,
2712,
29892,
278,
21957,
310,
445,
3979,
29915,
29879,
20209,
674,
367,
7618,
1455,
29889,
13,
3644,
9143,
653,
2233,
16929,
338,
11467,
7178,
322,
278,
14189,
1446,
337,
19730,
278,
13638,
297,
278,
18148,
29892,
278,
29871,
29929,
386,
17181,
1058,
723,
367,
16725,
29892,
3692,
393,
338,
7178,
4250,
3304,
29915,
29879,
2245,
457,
29872,
14546,
26817,
341,
3127,
860,
7455,
1049,
310,
278,
9245,
310,
2401,
29872,
1338,
363,
278,
360,
29889,
29907,
29889,
12594,
3121,
470,
263,
716,
2245,
457,
29872,
1925,
6375,
491,
7178,
2233,
16929,
29892,
723,
5517,
11719,
304,
975,
685,
278,
10899,
651,
2750,
360,
3301,
29909,
322,
360,
2477,
29909,
29974,
565,
322,
746,
278,
1206,
4133,
304,
278,
22569,
9245,
29889,
512,
278,
25305,
293,
1741,
310,
263,
18935,
27504,
4360,
3615,
1270,
29892,
373,
278,
916,
1361,
29892,
278,
2228,
723,
367,
2730,
327,
29892,
1951,
360,
3301,
29909,
322,
360,
2477,
29909,
29974,
723,
367,
620,
29883,
513,
287,
8763,
29889,
512,
278,
1741,
393,
9143,
653,
2233,
16929,
338,
11467,
7178,
322,
278,
8063,
550,
7344,
2761,
310,
278,
18148,
29892,
697,
26926,
393,
896,
723,
451,
4218,
304,
2908,
263,
22569,
9245,
2245,
3381,
297,
1753,
18639,
29892,
541,
2183,
278,
1857,
6030,
310,
278,
18148,
21178,
13638,
29892,
697,
2609,
367,
1854,
29889,
7857,
29892,
263,
19083,
15354,
297,
451,
871,
278,
7178,
616,
8271,
29892,
541,
884,
263,
8002,
1353,
310,
18148,
20209,
304,
1162,
8342,
278,
13638,
313,
276,
339,
8491,
263,
7787,
11581,
310,
472,
3203,
3023,
22091,
511,
723,
2367,
278,
1900,
4966,
363,
263,
6664,
2561,
310,
360,
3301,
29909,
322,
360,
2477,
29909,
29974,
29889,
960,
278,
14189
] | 1,024 |
CommonCrawl
| 0.489145 | 1.288046 | -0.048229 | 0.415892 |
ats can also regain the majority in the House of Representatives, then the issue of DAPA and DACA+ could be rendered moot in a much more pleasant way: comprehensive immigration reform, along the lines of the Senate CIR bill S.744 that was passed by the Democratic Senate with a bipartisan majority in 2013 but denied a vote in 2013 and 2014 in the Republican-controlled House of Representatives, could become law. Hopefully, many of the U.S. citizen relatives of those who would be affected by DAPA, DACA+, or comprehensive immigration reform will be motivated by these possibilities to turn out and vote in November.
Until comprehensive immigration reform becomes law or DAPA and DACA+ come into effect, however, immigration attorneys will remain on the lookout for other small pieces of good news where we can find them. The Supreme Court's decision Thursday in Mathis v. United States, while at first glance not about an immigration case at all, provided just such a piece of good news for noncitizens with certain types of criminal convictions. (Since most significant criminal convictions would have precluded applying for DAPA and DACA+, the set of noncitizens who will benefit from Mathis has very little overlap with the set of those harmed by U.S. v. Texas, so it may only be from the perspective of immigration attorneys that Thursday was something of a good news / bad news situation; hopefully I do not strike the reader as too insensitive for examining the two decisions in one blog post nonetheless.)
Mathis was primarily a sentencing case, arising under the Armed Career Criminal Act, or ACCA. That statute provides for harsher criminal sentences against those with certain sorts of prior criminal convictions. ACCA has been interpreted by the Supreme Court to provide for a "categorical approach", where what is important is what one can be certain a person has been convicted of, that is, the elements of their crime, and not other facts regarding what they may actually have done in the past.
In its use of the categorical approach, ACCA operates similarly to several provisions of the Immigration and Nationality Act (INA) regarding noncitizens with criminal convictions. As the Supreme Court explained in Moncrieffe v. Holder, 569 U.S. ____, 133 S.Ct. 1678 (2013), the categorical approach, grounded in the language of immigration statutes that ask what a noncitizen was "convicted" of, "has a long pedigree in our Nation's immigration law." Indeed, it goes back more than 100 years, at least back to the Second Circuit's decision in United States ex rel. Mylius v. Uhl, 210 F.860 (2d Cir. 1914). There are some provisions of immigration law that have been interpreted to deviate from the categorical approach, such as the $10,000 loss threshold for a fraud crime to qualify as an aggravated felony under section 101(a)(43)(M)(i) of the INA, addressed in the Supreme Court's 2009 decision in Nijhawan v. Holder, but they are the exception, not the rule.
Because both ACCA cases and many areas of immigration law rely on the categorical approach, the reasoning of ACCA cases is often found to control in immigration cases. Moncrieffe, for example, which addressed the immigration consequences of a conviction under Georgia law for possession of marijuana with intent to distribute, cited and relied upon Shepard v. United States, 544 U.S. 13 (2005), and Johnson v. United States, both ACCA cases. (The particular ACCA provision involved in Johnson was held unconstitutionally vague by the Court, but the principles behind the categorical approach were still usefully elucidated in that case.) Footnote 2 of the Mathis majority opinion specifically acknowledged the applicability of the categorical approach discussed in Mathis to immigration cases, citing Kawashima v. Holder, 565 U. S. 478, 482–483 (2012).
In both the ACCA context and the immigration context, issues have arisen regarding the application of
|
[
1446,
508,
884,
1072,
475,
278,
13638,
297,
278,
5619,
310,
28757,
29892,
769,
278,
2228,
310,
360,
3301,
29909,
322,
360,
2477,
29909,
29974,
1033,
367,
13751,
2730,
327,
297,
263,
1568,
901,
21246,
982,
29901,
15171,
6270,
5198,
16783,
11736,
29892,
3412,
278,
3454,
310,
278,
18148,
315,
8193,
11118,
317,
29889,
29955,
29946,
29946,
393,
471,
4502,
491,
278,
19083,
18148,
411,
263,
289,
27494,
21603,
13638,
297,
29871,
29906,
29900,
29896,
29941,
541,
17935,
263,
11719,
297,
29871,
29906,
29900,
29896,
29941,
322,
29871,
29906,
29900,
29896,
29946,
297,
278,
21178,
29899,
6451,
839,
5619,
310,
28757,
29892,
1033,
4953,
4307,
29889,
7963,
3730,
29892,
1784,
310,
278,
501,
29889,
29903,
29889,
14497,
264,
14576,
310,
1906,
1058,
723,
367,
15201,
491,
360,
3301,
29909,
29892,
360,
2477,
29909,
29974,
29892,
470,
15171,
6270,
5198,
16783,
11736,
674,
367,
17385,
630,
491,
1438,
24496,
304,
2507,
714,
322,
11719,
297,
3979,
29889,
13,
29965,
20233,
15171,
6270,
5198,
16783,
11736,
7415,
4307,
470,
360,
3301,
29909,
322,
360,
2477,
29909,
29974,
2041,
964,
2779,
29892,
3138,
29892,
5198,
16783,
1098,
272,
484,
952,
674,
3933,
373,
278,
1106,
449,
363,
916,
2319,
12785,
310,
1781,
9763,
988,
591,
508,
1284,
963,
29889,
450,
22569,
9245,
29915,
29879,
10608,
498,
1295,
3250,
297,
5792,
275,
325,
29889,
3303,
3900,
29892,
1550,
472,
937,
21798,
451,
1048,
385,
5198,
16783,
1206,
472,
599,
29892,
4944,
925,
1316,
263,
8424,
310,
1781,
9763,
363,
1661,
20752,
466,
575,
411,
3058,
4072,
310,
22161,
7602,
919,
1080,
29889,
313,
23036,
1556,
7282,
22161,
7602,
919,
1080,
723,
505,
758,
13347,
15399,
363,
360,
3301,
29909,
322,
360,
2477,
29909,
29974,
29892,
278,
731,
310,
1661,
20752,
466,
575,
1058,
674,
14169,
515,
5792,
275,
756,
1407,
2217,
25457,
411,
278,
731,
310,
1906,
4023,
2168,
491,
501,
29889,
29903,
29889,
325,
29889,
10319,
29892,
577,
372,
1122,
871,
367,
515,
278,
18520,
310,
5198,
16783,
1098,
272,
484,
952,
393,
498,
1295,
3250,
471,
1554,
310,
263,
1781,
9763,
847,
4319,
9763,
6434,
29936,
27581,
306,
437,
451,
21283,
278,
9591,
408,
2086,
1663,
575,
3321,
363,
4392,
2827,
278,
1023,
1602,
12112,
297,
697,
12618,
1400,
1661,
621,
6393,
1846,
13,
11309,
275,
471,
19434,
263,
2665,
16750,
1206,
29892,
564,
5921,
1090,
278,
826,
2168,
15825,
315,
28479,
3185,
29892,
470,
319,
4174,
29909,
29889,
2193,
1002,
1082,
8128,
363,
4023,
845,
261,
22161,
25260,
2750,
1906,
411,
3058,
23551,
310,
7536,
22161,
7602,
919,
1080,
29889,
319,
4174,
29909,
756,
1063,
21551,
491,
278,
22569,
9245,
304,
3867,
363,
263,
376,
29883,
20440,
936,
2948,
613,
988,
825,
338,
4100,
338,
825,
697,
508,
367,
3058,
263,
2022,
756,
1063,
7602,
18186,
310,
29892,
393,
338,
29892,
278,
3161,
310,
1009,
17268,
29892,
322,
451,
916,
17099,
11211,
825,
896,
1122,
2869,
505,
2309,
297,
278,
4940,
29889,
13,
797,
967,
671,
310,
278,
11608,
936,
2948,
29892,
319,
4174,
29909,
1751,
1078,
22829,
304,
3196,
1326,
12112,
310,
278,
1954,
29885,
16783,
322,
3086,
537,
3185,
313,
1177,
29909,
29897,
11211,
1661,
20752,
466,
575,
411,
22161,
7602,
919,
1080,
29889,
1094,
278,
22569,
9245,
10824,
297,
2598,
29883,
2546,
17615,
325,
29889,
379,
3194,
29892,
29871,
29945,
29953,
29929,
501,
29889,
29903,
29889,
903,
1649,
3383,
29871,
29896,
29941,
29941,
317,
29889,
29907,
29873,
29889,
29871,
29896,
29953,
29955,
29947,
313,
29906,
29900,
29896,
29941,
511,
278,
11608,
936,
2948,
29892,
5962,
287,
297,
278,
4086,
310,
5198,
16783,
1002,
2667,
393,
2244,
825,
263,
1661,
20752,
19642,
471,
376,
20580,
18186,
29908,
310,
29892,
376,
5349,
263,
1472,
25922,
929,
297,
1749,
22900,
29915,
29879,
5198,
16783,
4307,
1213,
14598,
29892,
372,
5771,
1250,
901,
1135,
29871,
29896,
29900,
29900,
2440,
29892,
472,
3203,
1250,
304,
278,
6440,
12594,
3121,
29915,
29879,
10608,
297,
3303,
3900,
429,
1104,
29889,
1619,
19646,
325,
29889,
501,
4415,
29892,
29871,
29906,
29896,
29900,
383,
29889,
29947,
29953,
29900,
313,
29906,
29881,
25079,
29889,
29871,
29896,
29929,
29896,
29946,
467,
1670,
526,
777,
1326,
12112,
310,
5198,
16783,
4307,
393,
505,
1063,
21551,
304,
29668,
403,
515,
278,
11608,
936,
2948,
29892,
1316,
408,
278,
395,
29896,
29900,
29892,
29900,
29900,
29900,
6410,
16897,
363,
263,
5227,
566,
17268,
304,
4021,
1598,
408,
385,
946,
3874,
29894,
630,
7995,
2592,
1090,
4004,
29871,
29896,
29900,
29896,
29898,
29874,
5033,
29946,
29941,
5033,
29924,
5033,
29875,
29897,
310,
278,
2672,
29909,
29892,
20976,
297,
278,
22569,
9245,
29915,
29879,
29871,
29906,
29900,
29900,
29929,
10608,
297,
405,
823,
29882,
1450,
273,
325,
29889,
379,
3194,
29892,
541,
896,
526,
278,
3682,
29892,
451,
278,
5751,
29889,
13,
29933,
5658,
1716,
319,
4174,
29909,
4251,
322,
1784,
10161,
310,
5198,
16783,
4307,
19104,
373,
278,
11608,
936,
2948,
29892,
278,
24481,
310,
319,
4174,
29909,
4251,
338,
4049,
1476,
304,
2761,
297,
5198,
16783,
4251,
29889,
2598,
29883,
2546,
17615,
29892,
363,
1342,
29892,
607,
20976,
278,
5198,
16783,
27721,
310,
263,
7602,
2463,
1090,
16762,
4307,
363,
17715,
310,
1766,
26323,
1648,
411,
7609,
304,
1320,
2666,
29892,
274,
1573,
322,
337,
2957,
2501,
2296,
29886,
538,
325,
29889,
3303,
3900,
29892,
29871,
29945,
29946,
29946,
501,
29889,
29903,
29889,
29871,
29896,
29941,
313,
29906,
29900,
29900,
29945,
511,
322,
11717,
325,
29889,
3303,
3900,
29892,
1716,
319,
4174,
29909,
4251,
29889,
313,
1576,
3153,
319,
4174,
29909,
25161,
9701,
297,
11717,
471,
4934,
443,
3075,
5008,
635,
25160,
491,
278,
9245,
29892,
541,
278,
18671,
5742,
278,
11608,
936,
2948,
892,
1603,
671,
3730,
560,
1682,
333,
630,
297,
393,
1206,
1846,
7527,
6812,
29871,
29906,
310,
278,
5792,
275,
13638,
9426,
10816,
24084,
3192,
278,
15576,
3097,
310,
278,
11608,
936,
2948,
15648,
297,
5792,
275,
304,
5198,
16783,
4251,
29892,
7537,
292,
476,
1450,
1161,
2946,
325,
29889,
379,
3194,
29892,
29871,
29945,
29953,
29945,
501,
29889,
317,
29889,
29871,
29946,
29955,
29947,
29892,
29871,
29946,
29947,
29906,
29994,
29946,
29947,
29941,
313,
29906,
29900,
29896,
29906,
467,
13,
797,
1716,
278,
319,
4174,
29909,
3030,
322,
278,
5198,
16783,
3030,
29892,
5626,
505,
564,
7674,
11211,
278,
2280,
310
] | 1,024 |
CommonCrawl
| 0.580846 | 1.412633 | 0.556914 | 0.835831 |
the categorical approach to what are known as "divisible" statutes. In effect, such statutes contain multiple separate crimes, and so one can look at the record of the conviction, using what is known as the "modified categorical approach" to determine which of these crimes applied. The Supreme Court clarified in Mathis that this is only to be done when the difference between the components of the statute of conviction turns on a true element, a fact on which a jury would have to agree to convict, or which a defendant would have to admit in a guilty plea. It does not apply to alternate means of commission of a crime, even if they are listed in the statute of conviction.
Mr. Mathis had been convicted multiple times of burglary under Iowa law, which covered unlawful entry into "any building, structure, [or] land, water or air vehicle." Mathis, slip op. at 5. For ACCA purposes, on the other hand, the Court had held years earlier that a conviction only counted as "burglary" if it involved unlawful entry into a building or other structure. The question thus became whether one could look at the record of Mr. Mathis's conviction to see whether he had been convicted of breaking into a building or other structure, on the one hand, which would qualify as ACCA "burglary", or breaking into a land, water or air vehicle, which would not so qualify. Iowa case law made clear that these were merely alternative means of committing a single crime, and that a jury could convict someone of burglary without agreeing on whether the defendant had burgled a building or a vehicle. The government sought nevertheless, however, to argue based on documents from Mr. Mathis's prior criminal cases that he had in fact been convicted of burglarizing a house and not a vehicle.
In a decision written by Justice Kagan and joined by Chief Justice Roberts and Justices Kennedy, Thomas, and Sotomayor, the Court held that this was not permissible. Facts on which a jury need not be unanimous are not elements of a crime, and so the Iowa burglary statute at issue was not truly divisible: it created only one crime, not many. Because the categorical approach focuses on what the defendant was convicted of doing, and not what he or she may have actually done, one cannot pick through the record of a prior case and speculate regarding whether the jury might have agreed on something that the law did not require it to agree on in order to convict.
Justice Breyer, joined by Justice Ginsburg, dissented, arguing that if the record revealed that evidence supported conviction of a defendant only pursuant to one statutory word or phrase, it should not matter whether that word or phrase was termed an element or a means. Justice Alito, writing only for himself, compared the Court's ACCA case law to a Belgian woman who had set out for Brussels and ended up in Zagreb, Croatia, by following her GPS too unquestioningly. Accusing the majority of "pointless formalism", he hypothesized a lengthy plea colloquy in which a defendant admitted to burglarizing a house at "10 Main St." in the face of lengthy questioning from the judge regarding whether this address might represent a yacht, house boat, trailer, or tent. (The hypothetical defendant's response to this last query was said to be, "No, it's made of brick. I scraped my knee on the brick.") Even in this case, he lamented, "[a]s the Court sees things, none of this would be enough."
Before addressing some of the implications of this decision for immigration purposes, I will pause to note that Justice Alito's lengthy hypothetical colloquy, humorous though it may be, strikes me as not really supporting his argument, and perhaps even as weakening it. In the real world, a judge would almost never go through such a lengthy discussion of the nature of the premises burgled if that did not affect the crime of which the defendant was to be convicted or the punishment for which the defendant was eligible. If the defendant had actually broken into a car parked right outside the garage of the house at 10 Main Street to steal money and jewelry, rather than breaking into a car parked inside the garage and the garage itself to steal the same money and jewelry, one would not expect him to start an argument with the judge, in the context of a
|
[
278,
11608,
936,
2948,
304,
825,
526,
2998,
408,
376,
4563,
275,
1821,
29908,
1002,
2667,
29889,
512,
2779,
29892,
1316,
1002,
2667,
1712,
2999,
5004,
2181,
1355,
29892,
322,
577,
697,
508,
1106,
472,
278,
2407,
310,
278,
7602,
2463,
29892,
773,
825,
338,
2998,
408,
278,
376,
1545,
2164,
11608,
936,
2948,
29908,
304,
8161,
607,
310,
1438,
2181,
1355,
7436,
29889,
450,
22569,
9245,
7542,
2164,
297,
5792,
275,
393,
445,
338,
871,
304,
367,
2309,
746,
278,
4328,
1546,
278,
7117,
310,
278,
1002,
1082,
310,
7602,
2463,
12169,
373,
263,
1565,
1543,
29892,
263,
2114,
373,
607,
263,
432,
2857,
723,
505,
304,
8661,
304,
7602,
919,
29892,
470,
607,
263,
822,
5818,
723,
505,
304,
20000,
297,
263,
27719,
5644,
29874,
29889,
739,
947,
451,
3394,
304,
25010,
2794,
310,
12969,
310,
263,
17268,
29892,
1584,
565,
896,
526,
9904,
297,
278,
1002,
1082,
310,
7602,
2463,
29889,
13,
20335,
29889,
5792,
275,
750,
1063,
7602,
18186,
2999,
3064,
310,
26072,
29880,
653,
1090,
25327,
4307,
29892,
607,
10664,
443,
10653,
1319,
6251,
964,
376,
1384,
5214,
29892,
3829,
29892,
518,
272,
29962,
2982,
29892,
4094,
470,
4799,
19716,
1213,
5792,
275,
29892,
269,
3466,
1015,
29889,
472,
29871,
29945,
29889,
1152,
319,
4174,
29909,
11976,
29892,
373,
278,
916,
1361,
29892,
278,
9245,
750,
4934,
2440,
8859,
393,
263,
7602,
2463,
871,
29115,
408,
376,
3074,
29880,
653,
29908,
565,
372,
9701,
443,
10653,
1319,
6251,
964,
263,
5214,
470,
916,
3829,
29889,
450,
1139,
4550,
3897,
3692,
697,
1033,
1106,
472,
278,
2407,
310,
3237,
29889,
5792,
275,
29915,
29879,
7602,
2463,
304,
1074,
3692,
540,
750,
1063,
7602,
18186,
310,
16679,
964,
263,
5214,
470,
916,
3829,
29892,
373,
278,
697,
1361,
29892,
607,
723,
4021,
1598,
408,
319,
4174,
29909,
376,
3074,
29880,
653,
613,
470,
16679,
964,
263,
2982,
29892,
4094,
470,
4799,
19716,
29892,
607,
723,
451,
577,
4021,
1598,
29889,
25327,
1206,
4307,
1754,
2821,
393,
1438,
892,
13586,
8671,
2794,
310,
844,
5367,
263,
2323,
17268,
29892,
322,
393,
263,
432,
2857,
1033,
7602,
919,
4856,
310,
26072,
29880,
653,
1728,
8661,
292,
373,
3692,
278,
822,
5818,
750,
26072,
839,
263,
5214,
470,
263,
19716,
29889,
450,
5874,
18365,
2360,
16561,
29892,
3138,
29892,
304,
27754,
2729,
373,
10701,
515,
3237,
29889,
5792,
275,
29915,
29879,
7536,
22161,
4251,
393,
540,
750,
297,
2114,
1063,
7602,
18186,
310,
26072,
4675,
5281,
263,
3699,
322,
451,
263,
19716,
29889,
13,
797,
263,
10608,
3971,
491,
17181,
476,
18939,
322,
8772,
491,
14546,
17181,
27253,
322,
3387,
1575,
23166,
29892,
5569,
29892,
322,
317,
327,
290,
388,
272,
29892,
278,
9245,
4934,
393,
445,
471,
451,
3635,
790,
1821,
29889,
26748,
29879,
373,
607,
263,
432,
2857,
817,
451,
367,
443,
11576,
681,
526,
451,
3161,
310,
263,
17268,
29892,
322,
577,
278,
25327,
26072,
29880,
653,
1002,
1082,
472,
2228,
471,
451,
19781,
8572,
1821,
29901,
372,
2825,
871,
697,
17268,
29892,
451,
1784,
29889,
7311,
278,
11608,
936,
2948,
8569,
267,
373,
825,
278,
822,
5818,
471,
7602,
18186,
310,
2599,
29892,
322,
451,
825,
540,
470,
1183,
1122,
505,
2869,
2309,
29892,
697,
2609,
5839,
1549,
278,
2407,
310,
263,
7536,
1206,
322,
1580,
5987,
11211,
3692,
278,
432,
2857,
1795,
505,
15502,
373,
1554,
393,
278,
4307,
1258,
451,
1996,
372,
304,
8661,
373,
297,
1797,
304,
7602,
919,
29889,
13,
14084,
625,
5826,
7598,
29892,
8772,
491,
17181,
402,
1144,
3074,
29892,
16317,
14927,
29892,
1852,
26420,
393,
565,
278,
2407,
17845,
393,
10757,
6969,
7602,
2463,
310,
263,
822,
5818,
871,
12359,
29884,
424,
304,
697,
1002,
329,
706,
1734,
470,
16549,
29892,
372,
881,
451,
4383,
3692,
393,
1734,
470,
16549,
471,
1840,
287,
385,
1543,
470,
263,
2794,
29889,
17181,
838,
2049,
29892,
5007,
871,
363,
3654,
29892,
9401,
278,
9245,
29915,
29879,
319,
4174,
29909,
1206,
4307,
304,
263,
9923,
713,
6114,
1058,
750,
731,
714,
363,
1771,
1558,
1379,
322,
9698,
701,
297,
20173,
15203,
29892,
8764,
28752,
29892,
491,
1494,
902,
402,
7024,
2086,
443,
12470,
11687,
29889,
4831,
4746,
278,
13638,
310,
376,
3149,
2222,
11595,
1608,
613,
540,
13752,
267,
1891,
263,
3309,
29891,
5644,
29874,
784,
417,
339,
29891,
297,
607,
263,
822,
5818,
20186,
304,
26072,
4675,
5281,
263,
3699,
472,
376,
29896,
29900,
4241,
624,
1213,
297,
278,
3700,
310,
3309,
29891,
1139,
292,
515,
278,
16833,
11211,
3692,
445,
3211,
1795,
2755,
263,
343,
5860,
29892,
3699,
13006,
29892,
1020,
3955,
29892,
470,
12033,
29889,
313,
1576,
13752,
300,
936,
822,
5818,
29915,
29879,
2933,
304,
445,
1833,
2346,
471,
1497,
304,
367,
29892,
376,
3782,
29892,
372,
29915,
29879,
1754,
310,
28645,
29889,
306,
885,
2390,
287,
590,
17905,
29872,
373,
278,
28645,
23157,
7753,
297,
445,
1206,
29892,
540,
301,
1166,
287,
29892,
14704,
29874,
29962,
29879,
278,
9245,
18553,
2712,
29892,
5642,
310,
445,
723,
367,
3307,
1213,
13,
18743,
3211,
292,
777,
310,
278,
2411,
5795,
310,
445,
10608,
363,
5198,
16783,
11976,
29892,
306,
674,
19957,
304,
4443,
393,
17181,
838,
2049,
29915,
29879,
3309,
29891,
13752,
300,
936,
784,
417,
339,
29891,
29892,
3165,
20657,
2466,
372,
1122,
367,
29892,
19492,
267,
592,
408,
451,
2289,
20382,
670,
2980,
29892,
322,
6060,
1584,
408,
8062,
8333,
372,
29889,
512,
278,
1855,
3186,
29892,
263,
16833,
723,
4359,
2360,
748,
1549,
1316,
263,
3309,
29891,
10679,
310,
278,
5469,
310,
278,
5188,
4637,
26072,
839,
565,
393,
1258,
451,
6602,
278,
17268,
310,
607,
278,
822,
5818,
471,
304,
367,
7602,
18186,
470,
278,
6035,
18310,
363,
607,
278,
822,
5818,
471,
560,
335,
1821,
29889,
960,
278,
822,
5818,
750,
2869,
9391,
964,
263,
1559,
14089,
287,
1492,
5377,
278,
7171,
482,
310,
278,
3699,
472,
29871,
29896,
29900,
4241,
7103,
304,
1886,
284,
6909,
322,
432,
809,
295,
719,
29892,
3265,
1135,
16679,
964,
263,
1559,
14089,
287,
2768,
278,
7171,
482,
322,
278,
7171,
482,
3528,
304,
1886,
284,
278,
1021,
6909,
322,
432,
809,
295,
719,
29892,
697,
723,
451,
2149,
1075,
304,
1369,
385,
2980,
411,
278,
16833,
29892,
297,
278,
3030,
310,
263
] | 1,024 |
CommonCrawl
| 0.712624 | 1.474927 | 0.427801 | 1.144461 |
law which made the penalty for the two versions of the crime exactly the same. Rather, if asked whether he broke into a "structure" at 10 Main Street, there is a good chance that such a hypothetical defendant would simply say "yes", and that would be the end of it. To quote from Justice Kagan's majority opinion:
"At trial, and still more at plea hearings, a defendant may have no incentive to contest what does not matter under the law; to the contrary, he "may have good reason not to"—or even be precluded from doing so by the court. . . . When that is true, a prosecutor's or judge's mistake as to means, reflected in the record, is likely to go uncorrected. See ibid. Such inaccuracies should not come back to haunt the defendant many years down the road by triggering a lengthy mandatory sentence."
Justice Alito's hypothetical colloquy, which I admit was quite funny, draws its humor partly from its unrealistic nature.
And just as the sorts of uncorrected inaccuracies to which Justice Kagan refers "should not come back to haunt the defendant many years down the road by triggering a lengthy mandatory sentence", as Mathis makes clear, they also should not come back to haunt the defendant by triggering deportation. In footnote 3 of Mathis, Justice Kagan specifically notes a scenario in which the Mathis rule will apply to immigration cases:
To see the point most clearly, consider an example arising in the immigration context: A defendant charged under a statute that criminalizes "intentionally, knowingly, or recklessly" assaulting another—as exists in many States, see, e.g., Tex. Penal Code Ann. §22.01(a)(1) (West Cum. Supp. 2015)—has no apparent reason to dispute a prosecutor's statement that he committed the crime intentionally (as opposed to recklessly) if those mental states are interchangeable means of satisfying a single mens rea element. But such a statement, if treated as reliable, could make a huge difference in a deportation proceeding years in the future, because an intentional assault (unlike a reckless one) qualifies as a "crime involving moral turpitude," and so requires removal from the country.
Under Mathis, if recklessness and intentional assault are indeed interchangeable means of satisfying the same mens rea requirement under a particular statute, then for immigration purposes the statute cannot be divided between them.
That is, even someone who seemingly pled guilty to intentional conduct, under such an indivisible statute, should be considered as if he or she had only pled guilty to reckless conduct, because, as Moncrieffe explained, "we must presume that the conviction "rested upon [nothing] more than the least of th[e] acts" criminalized," 133 S. Ct. at 1684. As Mathis's footnote 3 explains, this makes more sense than might at first glance appear, because someone convicted under such a statute would have had no reason to dispute the allegation that he or she had acted intentionally rather than recklessly—unless perhaps he or she had consulted immigration counsel prior to entering a plea. Allowing the proverbial hairs of a state statute's text to be split, adversely to the noncitizen, beyond the point where the distinction makes any difference under state law, would penalize those who did not have immigration consequences in mind at the time of their plea or trial. Certainly, for a variety of reasons, all noncitizens charged with a crime should consult with a competent immigration attorney before pleading to any charge or otherwise proceeding with their criminal case, but the law should not unnecessarily and unfairly penalize those who fail to heed this advice.
The distinction between recklessness and intentional conduct is not the only context in which this means/elements distinction may have relevance for immigration law. For example, attorneys whose clients have been convicted of possession of a controlled substance, under state laws covering at least some substances not federally controlled, should explore whether the identity of the substance is an element or a means under the relevant state law—whether, in order to obtain a conviction, the state is required to prove which controlled substance a defendant possessed. If the identity of the substance is a means and not an element, then the conviction may, under Mathis,
|
[
4307,
607,
1754,
278,
27368,
363,
278,
1023,
6910,
310,
278,
17268,
3721,
278,
1021,
29889,
390,
1624,
29892,
565,
4433,
3692,
540,
14455,
964,
263,
376,
23905,
29908,
472,
29871,
29896,
29900,
4241,
7103,
29892,
727,
338,
263,
1781,
8825,
393,
1316,
263,
13752,
300,
936,
822,
5818,
723,
3763,
1827,
376,
3582,
613,
322,
393,
723,
367,
278,
1095,
310,
372,
29889,
1763,
14978,
515,
17181,
476,
18939,
29915,
29879,
13638,
9426,
29901,
13,
29908,
4178,
14260,
29892,
322,
1603,
901,
472,
5644,
29874,
8293,
886,
29892,
263,
822,
5818,
1122,
505,
694,
297,
1760,
573,
304,
17793,
825,
947,
451,
4383,
1090,
278,
4307,
29936,
304,
278,
21138,
29892,
540,
376,
13029,
505,
1781,
2769,
451,
304,
29908,
30003,
272,
1584,
367,
758,
13347,
515,
2599,
577,
491,
278,
8973,
29889,
869,
869,
869,
1932,
393,
338,
1565,
29892,
263,
410,
3471,
3406,
29915,
29879,
470,
16833,
29915,
29879,
10171,
408,
304,
2794,
29892,
25312,
297,
278,
2407,
29892,
338,
5517,
304,
748,
443,
15728,
287,
29889,
2823,
474,
23883,
29889,
10506,
297,
5753,
2002,
2478,
881,
451,
2041,
1250,
304,
447,
1657,
278,
822,
5818,
1784,
2440,
1623,
278,
6520,
491,
7135,
292,
263,
3309,
29891,
9619,
7606,
10541,
1213,
13,
14084,
625,
838,
2049,
29915,
29879,
13752,
300,
936,
784,
417,
339,
29891,
29892,
607,
306,
20000,
471,
3755,
2090,
1460,
29892,
4216,
29879,
967,
27448,
22669,
515,
967,
443,
6370,
4695,
5469,
29889,
13,
2855,
925,
408,
278,
23551,
310,
443,
15728,
287,
297,
5753,
2002,
2478,
304,
607,
17181,
476,
18939,
14637,
376,
9344,
451,
2041,
1250,
304,
447,
1657,
278,
822,
5818,
1784,
2440,
1623,
278,
6520,
491,
7135,
292,
263,
3309,
29891,
9619,
7606,
10541,
613,
408,
5792,
275,
3732,
2821,
29892,
896,
884,
881,
451,
2041,
1250,
304,
447,
1657,
278,
822,
5818,
491,
7135,
292,
21455,
362,
29889,
512,
3661,
6812,
29871,
29941,
310,
5792,
275,
29892,
17181,
476,
18939,
10816,
11486,
263,
10483,
297,
607,
278,
5792,
275,
5751,
674,
3394,
304,
5198,
16783,
4251,
29901,
13,
1762,
1074,
278,
1298,
1556,
9436,
29892,
2050,
385,
1342,
564,
5921,
297,
278,
5198,
16783,
3030,
29901,
319,
822,
5818,
20139,
1090,
263,
1002,
1082,
393,
22161,
7093,
376,
524,
2509,
635,
29892,
1073,
11687,
29892,
470,
25527,
23769,
29908,
29159,
292,
1790,
30003,
294,
4864,
297,
1784,
3900,
29892,
1074,
29892,
321,
29889,
29887,
1696,
8490,
29889,
7363,
284,
5920,
8081,
29889,
16683,
29906,
29906,
29889,
29900,
29896,
29898,
29874,
5033,
29896,
29897,
313,
16128,
315,
398,
29889,
9179,
29889,
29871,
29906,
29900,
29896,
29945,
29897,
30003,
5349,
694,
20295,
2769,
304,
28447,
263,
410,
3471,
3406,
29915,
29879,
3229,
393,
540,
19355,
278,
17268,
16392,
635,
313,
294,
15869,
304,
25527,
23769,
29897,
565,
1906,
19119,
5922,
526,
1006,
3167,
519,
2794,
310,
24064,
263,
2323,
18664,
337,
29874,
1543,
29889,
1205,
1316,
263,
3229,
29892,
565,
14914,
408,
23279,
29892,
1033,
1207,
263,
12176,
4328,
297,
263,
21455,
362,
8469,
292,
2440,
297,
278,
5434,
29892,
1363,
385,
7609,
1848,
29159,
313,
348,
4561,
263,
25527,
2222,
697,
29897,
4021,
11057,
408,
263,
376,
7283,
603,
21677,
14731,
7013,
29886,
4279,
1699,
322,
577,
6858,
28744,
515,
278,
4234,
29889,
13,
29177,
5792,
275,
29892,
565,
25527,
2222,
2264,
322,
7609,
1848,
29159,
526,
6200,
1006,
3167,
519,
2794,
310,
24064,
278,
1021,
18664,
337,
29874,
11809,
1090,
263,
3153,
1002,
1082,
29892,
769,
363,
5198,
16783,
11976,
278,
1002,
1082,
2609,
367,
13931,
1546,
963,
29889,
13,
7058,
338,
29892,
1584,
4856,
1058,
2833,
11687,
282,
839,
27719,
304,
7609,
1848,
7512,
29892,
1090,
1316,
385,
1399,
440,
275,
1821,
1002,
1082,
29892,
881,
367,
5545,
408,
565,
540,
470,
1183,
750,
871,
282,
839,
27719,
304,
25527,
2222,
7512,
29892,
1363,
29892,
408,
2598,
29883,
2546,
17615,
10824,
29892,
376,
705,
1818,
2225,
2017,
393,
278,
7602,
2463,
376,
29878,
2868,
2501,
518,
28450,
29962,
901,
1135,
278,
3203,
310,
266,
29961,
29872,
29962,
14741,
29908,
22161,
1891,
1699,
29871,
29896,
29941,
29941,
317,
29889,
315,
29873,
29889,
472,
29871,
29896,
29953,
29947,
29946,
29889,
1094,
5792,
275,
29915,
29879,
3661,
6812,
29871,
29941,
18568,
29892,
445,
3732,
901,
4060,
1135,
1795,
472,
937,
21798,
2615,
29892,
1363,
4856,
7602,
18186,
1090,
1316,
263,
1002,
1082,
723,
505,
750,
694,
2769,
304,
28447,
278,
16831,
362,
393,
540,
470,
1183,
750,
27320,
16392,
635,
3265,
1135,
25527,
23769,
30003,
28952,
6060,
540,
470,
1183,
750,
8799,
287,
5198,
16783,
2613,
2838,
7536,
304,
18055,
263,
5644,
29874,
29889,
29408,
292,
278,
410,
18248,
616,
11315,
29879,
310,
263,
2106,
1002,
1082,
29915,
29879,
1426,
304,
367,
6219,
29892,
19901,
873,
304,
278,
1661,
20752,
19642,
29892,
8724,
278,
1298,
988,
278,
21578,
3732,
738,
4328,
1090,
2106,
4307,
29892,
723,
6584,
284,
675,
1906,
1058,
1258,
451,
505,
5198,
16783,
27721,
297,
3458,
472,
278,
931,
310,
1009,
5644,
29874,
470,
14260,
29889,
315,
13946,
368,
29892,
363,
263,
12875,
310,
9590,
29892,
599,
1661,
20752,
466,
575,
20139,
411,
263,
17268,
881,
8799,
411,
263,
5100,
296,
5198,
16783,
1098,
25252,
1434,
5644,
9382,
304,
738,
8323,
470,
6467,
8469,
292,
411,
1009,
22161,
1206,
29892,
541,
278,
4307,
881,
451,
17128,
6275,
322,
29395,
1466,
368,
6584,
284,
675,
1906,
1058,
4418,
304,
540,
287,
445,
9848,
29889,
13,
1576,
21578,
1546,
25527,
2222,
2264,
322,
7609,
1848,
7512,
338,
451,
278,
871,
3030,
297,
607,
445,
2794,
29914,
17664,
21578,
1122,
505,
29527,
749,
363,
5198,
16783,
4307,
29889,
1152,
1342,
29892,
1098,
272,
484,
952,
5069,
13154,
505,
1063,
7602,
18186,
310,
17715,
310,
263,
20704,
5960,
749,
29892,
1090,
2106,
14243,
21653,
472,
3203,
777,
5960,
2925,
451,
12067,
635,
20704,
29892,
881,
26987,
3692,
278,
10110,
310,
278,
5960,
749,
338,
385,
1543,
470,
263,
2794,
1090,
278,
8018,
2106,
4307,
30003,
1332,
1979,
29892,
297,
1797,
304,
4017,
263,
7602,
2463,
29892,
278,
2106,
338,
3734,
304,
6356,
607,
20704,
5960,
749,
263,
822,
5818,
25622,
29889,
960,
278,
10110,
310,
278,
5960,
749,
338,
263,
2794,
322,
451,
385,
1543,
29892,
769,
278,
7602,
2463,
1122,
29892,
1090,
5792,
275,
29892
] | 1,024 |
CommonCrawl
| 0.593073 | 1.438589 | 0.210821 | 0.802944 |
fall within the protection of the Supreme Court's decision last year in Mellouli v. Lynch, which required a controlled substance conviction to relate to a federally controlled substance in order to cause adverse immigration consequences. (The Third Circuit's 2013 en banc decision in Rojas v. Attorney General rejected what it called the "formal categorical approach" in this controlled-substance context, but it is not clear that this aspect of Rojas can survive the combination of Mellouli and Mathis in states where the identity of the controlled substance is a means and not an element under state law, although we will have to wait and see how the case law develops to be sure.) There will be other areas, as well, where a statute which lists multiple ways of committing a crime is actually indivisible under state law, and so a noncitizen is entitled under Moncrieffe and Mathis to the assumption that he or she committed the crime in whichever way is least harmful for immigration purposes.
Mathis is therefore good news for a significant number of immigrants, and their attorneys, even though this small piece of good news may pale in comparison to the disappointment of U.S. v. Texas and the continued injunction against DAPA and DACA+, which are bad news for substantially more immigrants. With respect to the latter, we can hope for, and fight for, the possibility that the November elections may bring more good news.
All Checks but No Balances — The Systemic Failure to Protect EB-5 Investors
by Angelo Paparelli, Past ABIL President
Nation of Immigrators
Several widely-publicized actions by the Securities Exchange Commission (SEC), and the inevitable litigation that has piled on in consequence, have pressure-tested the EB-5 ecosystem and found it defective. Simply stated, the system too likely and too often fails. Sadly, many EB-5 investors, after writing half-million-dollar or greater checks, ultimately have learned that no balances remain in their capital accounts and no green cards land or stay in their own and their family's wallets.
Stripped of cash and cards, an unacceptable number of EB-5 investors find out that they have been fleeced. This calamity is not necessarily attributable to the ever-present moral hazard of failing to thoroughly investigate a proposed investment, and imprudently placing at-risk funds into an unsound business opportunity. Rather, the flaw lies in structural and process failures of misfeasant government officials and market participants to take care that EB-5 investors are adequately protected from errant or unscrupulous promoters, developers, regional-center principals and migration agents, and from sundry Ponzi schemers, garden-variety fraudsters and sophisticated con artists.
No EB-5 stakeholders and the ecosystem itself cannot continue to thrive if too many investors are defrauded of money and the promised permanent residency. Clearly, a healthy, sustainable EB-5 ecosystem cannot endure without meaningful investor protections and reliable safeguards that promote real job creation.
Culprit number one — thoughtless lawmakers. The successive Congresses that crafted and amended the EB-5 program surely must have known that some degree of fraud, waste, and abuse are the inevitable byproducts of all government programs. Yet, lawmakers never included investor protections in the enabling EB-5 legislation. Even more appalling, our legislators should have foreseen that foreign investors, many of whom lack rudimentary English fluency or business savvy, would at once be dazzled by the allure of a green card and likely unaware of the risk that the U.S. government's own EB-5 program would be unsafe.
The Immigration Act of 1990, the law that ushered in the EB-5 program, and follow-on amendments, never authorized federal immigration administrators to debar from program participation blatantly out-of-compliance regional centers, developers, hucksters and their co-conspirators, or to protect investors from the harsh consequences of clearly foreseeable fraud and financial crimes. On the contrary, Congress knew from the outset that some deals will likely go south because scoundrels might hoodwink invest
|
[
6416,
2629,
278,
13047,
310,
278,
22569,
9245,
29915,
29879,
10608,
1833,
1629,
297,
341,
514,
283,
492,
325,
29889,
18989,
305,
29892,
607,
3734,
263,
20704,
5960,
749,
7602,
2463,
304,
29279,
304,
263,
12067,
635,
20704,
5960,
749,
297,
1797,
304,
4556,
594,
3901,
5198,
16783,
27721,
29889,
313,
1576,
18008,
12594,
3121,
29915,
29879,
29871,
29906,
29900,
29896,
29941,
427,
289,
4564,
10608,
297,
1528,
14196,
325,
29889,
6212,
25252,
4593,
22225,
825,
372,
2000,
278,
376,
689,
284,
11608,
936,
2948,
29908,
297,
445,
20704,
29899,
22492,
749,
3030,
29892,
541,
372,
338,
451,
2821,
393,
445,
9565,
310,
1528,
14196,
508,
10503,
573,
278,
10296,
310,
341,
514,
283,
492,
322,
5792,
275,
297,
5922,
988,
278,
10110,
310,
278,
20704,
5960,
749,
338,
263,
2794,
322,
451,
385,
1543,
1090,
2106,
4307,
29892,
5998,
591,
674,
505,
304,
4480,
322,
1074,
920,
278,
1206,
4307,
2693,
29879,
304,
367,
1854,
1846,
1670,
674,
367,
916,
10161,
29892,
408,
1532,
29892,
988,
263,
1002,
1082,
607,
8857,
2999,
5837,
310,
844,
5367,
263,
17268,
338,
2869,
1399,
440,
275,
1821,
1090,
2106,
4307,
29892,
322,
577,
263,
1661,
20752,
19642,
338,
23437,
1090,
2598,
29883,
2546,
17615,
322,
5792,
275,
304,
278,
11833,
393,
540,
470,
1183,
19355,
278,
17268,
297,
377,
4070,
369,
982,
338,
3203,
10311,
1319,
363,
5198,
16783,
11976,
29889,
13,
11309,
275,
338,
5480,
1781,
9763,
363,
263,
7282,
1353,
310,
5198,
4481,
1934,
29892,
322,
1009,
1098,
272,
484,
952,
29892,
1584,
2466,
445,
2319,
8424,
310,
1781,
9763,
1122,
15509,
297,
10230,
304,
278,
23451,
358,
310,
501,
29889,
29903,
29889,
325,
29889,
10319,
322,
278,
7572,
10899,
651,
2750,
360,
3301,
29909,
322,
360,
2477,
29909,
29974,
29892,
607,
526,
4319,
9763,
363,
23228,
368,
901,
5198,
4481,
1934,
29889,
2973,
3390,
304,
278,
7480,
29892,
591,
508,
4966,
363,
29892,
322,
8589,
363,
29892,
278,
13331,
393,
278,
3979,
20209,
1122,
6963,
901,
1781,
9763,
29889,
13,
3596,
5399,
29879,
541,
1939,
7392,
2925,
813,
450,
2184,
293,
29098,
545,
304,
14409,
312,
382,
29933,
29899,
29945,
512,
10147,
943,
13,
1609,
3218,
7078,
8579,
17243,
492,
29892,
19793,
17571,
6227,
7178,
13,
29940,
362,
310,
1954,
29885,
335,
3605,
943,
13,
29903,
1310,
284,
17644,
29899,
3597,
1891,
8820,
491,
278,
5356,
332,
1907,
24004,
11444,
313,
1660,
29907,
511,
322,
278,
297,
5750,
8270,
11872,
335,
362,
393,
756,
282,
2356,
373,
297,
17004,
29892,
505,
12959,
29899,
1688,
287,
278,
382,
29933,
29899,
29945,
321,
3944,
973,
322,
1476,
372,
23503,
573,
29889,
3439,
17632,
8703,
29892,
278,
1788,
2086,
5517,
322,
2086,
4049,
8465,
29889,
21772,
368,
29892,
1784,
382,
29933,
29899,
29945,
13258,
943,
29892,
1156,
5007,
4203,
29899,
19958,
291,
29899,
29881,
26810,
470,
7621,
12747,
29892,
18973,
505,
10972,
393,
694,
6411,
2925,
3933,
297,
1009,
7483,
15303,
322,
694,
7933,
15889,
2982,
470,
7952,
297,
1009,
1914,
322,
1009,
3942,
29915,
29879,
17042,
10376,
29889,
13,
855,
374,
2986,
310,
274,
1161,
322,
15889,
29892,
385,
443,
16044,
519,
1353,
310,
382,
29933,
29899,
29945,
13258,
943,
1284,
714,
393,
896,
505,
1063,
9115,
687,
287,
29889,
910,
1208,
314,
537,
338,
451,
12695,
1098,
1091,
9246,
304,
278,
3926,
29899,
6338,
14731,
447,
29920,
538,
310,
17581,
304,
26606,
23033,
263,
7972,
13258,
358,
29892,
322,
16921,
566,
2705,
24421,
472,
29899,
3780,
29895,
29199,
964,
385,
9644,
618,
5381,
15130,
29889,
390,
1624,
29892,
278,
285,
10653,
12185,
297,
2281,
3631,
322,
1889,
4418,
1973,
310,
3984,
1725,
13186,
5874,
24921,
322,
9999,
27138,
304,
2125,
2562,
393,
382,
29933,
29899,
29945,
13258,
943,
526,
19967,
339,
2486,
6364,
515,
4589,
424,
470,
443,
1557,
17827,
352,
681,
2504,
327,
414,
29892,
18777,
29892,
14014,
29899,
5064,
3420,
1338,
322,
20332,
19518,
29892,
322,
515,
480,
299,
719,
26317,
2526,
1364,
331,
414,
29892,
16423,
29899,
5927,
3305,
5227,
566,
23080,
322,
269,
3021,
4695,
630,
378,
17906,
29889,
13,
3782,
382,
29933,
29899,
29945,
380,
1296,
8948,
414,
322,
278,
321,
3944,
973,
3528,
2609,
6773,
304,
266,
4401,
565,
2086,
1784,
13258,
943,
526,
822,
336,
566,
287,
310,
6909,
322,
278,
22399,
17667,
620,
3615,
1270,
29889,
17732,
368,
29892,
263,
9045,
29891,
29892,
15075,
475,
519,
382,
29933,
29899,
29945,
321,
3944,
973,
2609,
1095,
545,
1728,
6593,
1319,
13258,
272,
3279,
1953,
322,
23279,
9437,
24024,
3163,
393,
27391,
1855,
4982,
11265,
29889,
13,
29907,
352,
19428,
1353,
697,
813,
2714,
2222,
4307,
29885,
21079,
29889,
450,
2551,
573,
11559,
267,
393,
25554,
287,
322,
626,
2760,
278,
382,
29933,
29899,
29945,
1824,
18880,
1818,
505,
2998,
393,
777,
7426,
310,
5227,
566,
29892,
19863,
29892,
322,
633,
1509,
526,
278,
297,
5750,
8270,
491,
14456,
310,
599,
5874,
11104,
29889,
15175,
29892,
4307,
29885,
21079,
2360,
5134,
13258,
272,
3279,
1953,
297,
278,
427,
17961,
382,
29933,
29899,
29945,
13332,
362,
29889,
7753,
901,
623,
27855,
29892,
1749,
13332,
4097,
881,
505,
363,
968,
264,
393,
9117,
13258,
943,
29892,
1784,
310,
6029,
10225,
29051,
2073,
653,
4223,
20501,
3819,
470,
5381,
4048,
13308,
29892,
723,
472,
2748,
367,
270,
7511,
839,
491,
278,
599,
545,
310,
263,
7933,
5881,
322,
5517,
1185,
2519,
310,
278,
12045,
393,
278,
501,
29889,
29903,
29889,
5874,
29915,
29879,
1914,
382,
29933,
29899,
29945,
1824,
723,
367,
25110,
29889,
13,
1576,
1954,
29885,
16783,
3185,
310,
29871,
29896,
29929,
29929,
29900,
29892,
278,
4307,
393,
502,
2276,
287,
297,
278,
382,
29933,
29899,
29945,
1824,
29892,
322,
1101,
29899,
265,
626,
355,
1860,
29892,
2360,
4148,
1891,
17097,
5198,
16783,
6343,
4097,
304,
316,
1646,
515,
1824,
27577,
1999,
271,
10835,
714,
29899,
974,
29899,
2388,
13036,
14014,
1644,
414,
29892,
18777,
29892,
298,
2707,
23080,
322,
1009,
1302,
29899,
535,
1028,
381,
4097,
29892,
470,
304,
12566,
13258,
943,
515,
278,
4023,
845,
27721,
310,
9436,
363,
968,
29872,
519,
5227,
566,
322,
18161,
2181,
1355,
29889,
1551,
278,
21138,
29892,
11559,
6363,
515,
278,
714,
842,
393,
777,
316,
1338,
674,
5517,
748,
7062,
1363,
885,
618,
2674,
29879,
1795,
298,
2092,
29893,
682,
13258
] | 1,024 |
CommonCrawl
| 0.723492 | 1.288046 | 0.836656 | 0.572736 |
ors, or because other investments involve business plans with little likelihood of actual success.
Nonetheless, from inception, the law has provided that conditional residency would end and EB-5 investors brought before immigration judges in deportation proceedings if investments were not sustained — something that's hard to do if one has been swindled — or the requisite jobs were not created.
Congress should also have known that securities law enforcers and immigration administrators would probably not quickly uncover the shenanigans hiding in some unregistered investments whose job-creating worthiness — by legislative design — would only be determined some two years hence.
Regrettably, Congress never provided for a real-time, on-the-scene watchdog to protect investors' interests and make sure that promised actions supposedly leading to job creation for U.S. workers would actually happen.
Culprit number two — unaware and insensitive bureaucrats. As the SEC has acknowledged, it arrived several years late to the EB-5 program, having been oblivious for too many years to the Commission's established statutory duty to protect investors in all securities transactions, including EB-5 deals. While SEC enforcement of EB-5 investments has fortunately intensified in the last few years and months, the Commission most often will likely play only an ex post facto enforcement role in punishing wrongdoers and trying to make investors whole once a deal has already failed or investors have been hurt, unless a whistleblower or grieving investor surfaces sooner.
So too have immigration administrators been derelict. Regulations of the legacy agency, the Immigration and Naturalization Service (INS), and the current Department of Homeland Security immigration components — U.S. Immigration & Customs Enforcement (ICE) and U.S. Citizenship and Immigration Services (USCIS) — offer defrauded investors no relief or even the possibility that discretion might be favorably exercised. To be sure, USCIS has tasked its Fraud Detection and National Security Directorate to make unannounced site visits to projects and regional centers and called for investor interviews when the issue of removing conditions on residency are to be ultimately adjudicated. These actions, however, have yet to occur in great numbers, may not be efficacious, or may just be too little too late.
Culprit number three — EB-5 market participants. The construction, lending and escrow industries — sectors especially active in EB-5 projects — have long been accustomed to the salutary, sentinel effect of meaningful oversight. Money doesn't move from one contracting party to another unless predetermined and mutually agreed conditions are satisfied. The ever-present risk that funds or assets will be stolen, lost or wasted, or that desired outcomes might not be achieved, can be eliminated or mitigated if a real-time watchdog monitors contract compliance and fulfillment. The same safeguard could readily be applied for the protection of EB-5 investors and the assurance of job creation — but until now EB-5 market players seemed not to care. Instead, conflicts of interests continue to abound in the EB-5 industry and wrongdoers still go undetected until it's too late.
What's to be done?
All three culprits can expiate their sins if they act together or at least separately to introduce a new participant in the EB-5 ecosystem — the independent fiduciary — a person or firm that would represent the interests of investors in trying to assure the ultimate return of principal, the maximization of profits (once job creation at the requisite level has been confirmed by USCIS), and the ultimate prize — unconditional permanent residence.
An independent fiduciary would act much like a bank officer who decides whether agreed conditions have been satisfied before releasing progress payments of loan proceeds. The investors' independent fiduciary could also perform:
an auditing function to confirm that required business and accounting records are maintained, and only permissible expenditures are paid,
corporate secretary/treasurer functions to make timely disclosures to investors of financial activities, monitor fund administration and job creation/hiring, give notice of the time, date and methodology for investors to vote on corporate matters, and report any significant developments or material changes to the business plan,
a
|
[
943,
29892,
470,
1363,
916,
13258,
1860,
25135,
5381,
13900,
411,
2217,
4188,
22342,
310,
3935,
2551,
29889,
13,
12283,
621,
6393,
29892,
515,
297,
1441,
29892,
278,
4307,
756,
4944,
393,
15047,
620,
3615,
1270,
723,
1095,
322,
382,
29933,
29899,
29945,
13258,
943,
6296,
1434,
5198,
16783,
6577,
2710,
297,
21455,
362,
8469,
886,
565,
13258,
1860,
892,
451,
15075,
7114,
813,
1554,
393,
29915,
29879,
2898,
304,
437,
565,
697,
756,
1063,
2381,
513,
839,
813,
470,
278,
5054,
275,
568,
17643,
892,
451,
2825,
29889,
13,
29907,
549,
1253,
881,
884,
505,
2998,
393,
409,
2764,
1907,
4307,
24555,
22543,
322,
5198,
16783,
6343,
4097,
723,
3117,
451,
9098,
443,
11911,
278,
528,
264,
273,
335,
550,
25508,
297,
777,
443,
9573,
287,
13258,
1860,
5069,
4982,
29899,
1037,
1218,
7088,
3335,
813,
491,
13332,
1230,
2874,
813,
723,
871,
367,
10087,
777,
1023,
2440,
8151,
29889,
13,
4597,
13158,
2197,
29892,
11559,
2360,
4944,
363,
263,
1855,
29899,
2230,
29892,
373,
29899,
1552,
29899,
24645,
6505,
26169,
304,
12566,
13258,
943,
29915,
20017,
322,
1207,
1854,
393,
22399,
8820,
7424,
368,
8236,
304,
4982,
11265,
363,
501,
29889,
29903,
29889,
17162,
723,
2869,
3799,
29889,
13,
29907,
352,
19428,
1353,
1023,
813,
1185,
2519,
322,
1663,
575,
3321,
289,
13411,
7283,
1446,
29889,
1094,
278,
3725,
29907,
756,
24084,
3192,
29892,
372,
11977,
3196,
2440,
5683,
304,
278,
382,
29933,
29899,
29945,
1824,
29892,
2534,
1063,
704,
17843,
2738,
363,
2086,
1784,
2440,
304,
278,
11444,
29915,
29879,
7841,
1002,
329,
706,
13360,
304,
12566,
13258,
943,
297,
599,
409,
2764,
1907,
22160,
29892,
3704,
382,
29933,
29899,
29945,
316,
1338,
29889,
5806,
3725,
29907,
24555,
13561,
310,
382,
29933,
29899,
29945,
13258,
1860,
756,
5162,
348,
2486,
12838,
2164,
297,
278,
1833,
2846,
2440,
322,
7378,
29892,
278,
11444,
1556,
4049,
674,
5517,
1708,
871,
385,
429,
1400,
2114,
29877,
24555,
13561,
6297,
297,
6035,
14424,
2743,
1867,
414,
322,
1811,
304,
1207,
13258,
943,
3353,
2748,
263,
5376,
756,
2307,
5229,
470,
13258,
943,
505,
1063,
21682,
29892,
6521,
263,
377,
391,
19982,
13609,
470,
867,
15387,
13258,
272,
28001,
29548,
29889,
13,
6295,
2086,
505,
5198,
16783,
6343,
4097,
1063,
589,
295,
919,
29889,
2169,
8250,
310,
278,
25000,
946,
3819,
29892,
278,
1954,
29885,
16783,
322,
18385,
2133,
6692,
313,
1177,
29903,
511,
322,
278,
1857,
10317,
310,
15089,
14045,
14223,
5198,
16783,
7117,
813,
501,
29889,
29903,
29889,
1954,
29885,
16783,
669,
8701,
29879,
1174,
1454,
13561,
313,
12107,
29897,
322,
501,
29889,
29903,
29889,
21353,
466,
575,
4034,
322,
1954,
29885,
16783,
15538,
313,
3308,
29907,
3235,
29897,
813,
5957,
822,
336,
566,
287,
13258,
943,
694,
18892,
470,
1584,
278,
13331,
393,
766,
4838,
291,
1795,
367,
7853,
2197,
11382,
3368,
29889,
1763,
367,
1854,
29892,
3148,
29907,
3235,
756,
3414,
287,
967,
7347,
566,
360,
2650,
428,
322,
3086,
14223,
15944,
403,
304,
1207,
443,
812,
20979,
3268,
1998,
1169,
304,
9279,
322,
14014,
1644,
414,
322,
2000,
363,
13258,
272,
1006,
7406,
746,
278,
2228,
310,
11077,
5855,
373,
620,
3615,
1270,
526,
304,
367,
18973,
12109,
566,
293,
630,
29889,
4525,
8820,
29892,
3138,
29892,
505,
3447,
304,
6403,
297,
2107,
3694,
29892,
1122,
451,
367,
1801,
983,
8802,
29892,
470,
1122,
925,
367,
2086,
2217,
2086,
5683,
29889,
13,
29907,
352,
19428,
1353,
2211,
813,
382,
29933,
29899,
29945,
9999,
27138,
29889,
450,
7632,
29892,
301,
2548,
322,
3966,
798,
6397,
2722,
813,
409,
14359,
7148,
6136,
297,
382,
29933,
29899,
29945,
9279,
813,
505,
1472,
1063,
1035,
1796,
287,
304,
278,
4497,
329,
653,
29892,
2665,
262,
295,
2779,
310,
6593,
1319,
288,
874,
523,
29889,
341,
4992,
1838,
29915,
29873,
4337,
515,
697,
8078,
292,
6263,
304,
1790,
6521,
4450,
300,
837,
1312,
322,
5478,
1474,
15502,
5855,
526,
15787,
29889,
450,
3926,
29899,
6338,
12045,
393,
29199,
470,
21608,
674,
367,
380,
18975,
29892,
5714,
470,
471,
9446,
29892,
470,
393,
7429,
714,
26807,
1795,
451,
367,
14363,
29892,
508,
367,
10397,
630,
470,
1380,
335,
630,
565,
263,
1855,
29899,
2230,
6505,
26169,
1601,
17259,
8078,
752,
13036,
322,
6095,
5589,
358,
29889,
450,
1021,
9437,
24024,
538,
1033,
28520,
367,
7436,
363,
278,
13047,
310,
382,
29933,
29899,
29945,
13258,
943,
322,
278,
1223,
18541,
310,
4982,
11265,
813,
541,
2745,
1286,
382,
29933,
29899,
29945,
9999,
10769,
6140,
451,
304,
2562,
29889,
8669,
29892,
28792,
310,
20017,
6773,
304,
633,
618,
297,
278,
382,
29933,
29899,
29945,
13661,
322,
2743,
1867,
414,
1603,
748,
563,
300,
26458,
2745,
372,
29915,
29879,
2086,
5683,
29889,
13,
5618,
29915,
29879,
304,
367,
2309,
29973,
13,
3596,
2211,
13949,
558,
1169,
508,
1518,
29347,
1009,
269,
1144,
565,
896,
1044,
4208,
470,
472,
3203,
16949,
304,
14944,
263,
716,
5221,
424,
297,
278,
382,
29933,
29899,
29945,
321,
3944,
973,
813,
278,
7417,
25947,
29884,
455,
653,
813,
263,
2022,
470,
9226,
393,
723,
2755,
278,
20017,
310,
13258,
943,
297,
1811,
304,
1223,
545,
278,
8494,
6490,
736,
310,
5882,
29892,
278,
5256,
2133,
310,
2600,
1169,
313,
10646,
4982,
11265,
472,
278,
5054,
275,
568,
3233,
756,
1063,
16725,
491,
3148,
29907,
3235,
511,
322,
278,
8494,
6490,
26836,
813,
443,
1116,
3245,
17667,
25488,
29889,
13,
2744,
7417,
25947,
29884,
455,
653,
723,
1044,
1568,
763,
263,
9124,
12139,
1058,
1602,
2247,
3692,
15502,
5855,
505,
1063,
15787,
1434,
337,
280,
5832,
6728,
5146,
1860,
310,
24806,
8469,
29879,
29889,
450,
13258,
943,
29915,
7417,
25947,
29884,
455,
653,
1033,
884,
2189,
29901,
13,
273,
12990,
11407,
740,
304,
9659,
393,
3734,
5381,
322,
3633,
292,
6475,
526,
19949,
29892,
322,
871,
3635,
790,
1821,
1518,
355,
277,
1973,
526,
12530,
29892,
13,
2616,
1971,
403,
28274,
29914,
2484,
294,
9945,
3168,
304,
1207,
5335,
873,
766,
11291,
1973,
304,
13258,
943,
310,
18161,
14188,
29892,
11819,
5220,
17517,
322,
4982,
11265,
29914,
29882,
8491,
29892,
2367,
8369,
310,
278,
931,
29892,
2635,
322,
1158,
3002,
363,
13258,
943,
304,
11719,
373,
17266,
403,
13750,
29892,
322,
3461,
738,
7282,
2693,
1860,
470,
5518,
3620,
304,
278,
5381,
3814,
29892,
13,
29874
] | 1,024 |
CommonCrawl
| 0.766965 | 1.267281 | 0.829483 | 0.782706 |
watchdog function to confirm that insurance underwriter conditions are fulfilled and required permits and licenses are maintained; and
an education and information function to explain to investors the role and activities of the independent fiduciary, and formally confirm to government officials how EB-5 program requirements are met when and as they are due.
Other loss mitigation safeguards could also be established. These include (a) the expanded availability and acquisition of EB-5 insurance to pay a form of indemnity (the return of the investment) if investor petitions are not approved or conditions on residence are not removed, (b) the addition of other traditional forms of insurance protection (e.g., fidelity bonds, and coverage for errors and omissions, business interruption, directors and officers liability and general liability), and (c) the use of standby letters of credit.
Without eliminating moral hazard, Congress could provide in the forthcoming integrity measures that a regional center's or project developer's engagement of an independent fiduciary, purchase of particular types of insurance, and/or arrangement for a standby letter of credit would serve as a good-faith prohibition against the denial or loss of lawful permanent resident status in cases where the SEC, ICE or USCIS determines that EB-5 investors have been defrauded.
USCIS could add similar protections in its (reportedly) soon-to-be-published EB-5 regulations. The agency could also specify by regulation or policy memorandum circumstances where the exercise of prosecutorial discretion should be exercised and of the full range of its parole authority and employment authorization are granted to defrauded but otherwise admissible EB-5 investors.
Market players should also have "skin in the game" of protecting EB-5 investors. They should require the engagement of independent fiduciaries and procurement of appropriate insurance coverages or letters of credit that reduce but do not eliminate all investment risk so that USCIS's at-risk capital requirements are nonetheless satisfied. Market participants should also recognize that risk reduction will add costs but likely generate overriding cost savings that will redound to the benefit of primary lenders, project developers and regional centers while also attracting migration agents, investment advisors and the EB-5 investors themselves. Ultimately and ideally, only deals with fiduciary and insurance protections will be marketable.
Checks and balances — the brainchildren of the Founding Fathers — established our constitutional form of government that has endured for centuries. One of these features is the protection of minorities against majority power. So too will a new system of checks and balances protect vulnerable EB-5 investors likely reduce the prospect that investors will write very sizable checks but yield no balances in their capital accounts or place green cards in their wallets.
Trump and the Snake
Donald Trump is fond of reading the lyrics from Al Wilson's 1968 R&B hit song "The Snake" in his campaign rallies. While this is a catchy tune, Trump has now corrupted the song by associating it with his opposition to Muslims. He first called for a ban on Muslims entering the United States, including Syrian refugees, and recently modified it by calling for a suspension of immigration from areas of the world when there is a proven history of terrorism against the United States or its allies. Trump most recently said that the United States should consider more racial profiling, in response to a question about whether he supported greater law enforcement scrutiny of Muslim Americans after the Orlando mass shooting. If all of these proposals were implemented, it would impede the ability of millions of temporary visa holders and immigrants to legitimately enter the United States.
This video depicting Trump's reading of The Snake in his rally in Greensboro, NC on June 14, 2016 is too chilling to watch, as the reading is interspersed with the ejection of a protestor amidst frenzied chants of "USA… USA". Although the lyrics are inspired by Aesop's fable of the Farmer and the Viper, the lyrics appear very sinister when Trump associates them with his war on Muslims. The lyrics revolve around a tender hearted woman who rescues a half frozen snake. After the snake is rescued, he bites the woman, and when she is dying, the snake tells her that she
|
[
6505,
26169,
740,
304,
9659,
393,
1663,
18541,
1090,
13236,
5855,
526,
6095,
26940,
322,
3734,
3635,
1169,
322,
7794,
11259,
526,
19949,
29936,
322,
13,
273,
9793,
322,
2472,
740,
304,
5649,
304,
13258,
943,
278,
6297,
322,
14188,
310,
278,
7417,
25947,
29884,
455,
653,
29892,
322,
28269,
9659,
304,
5874,
24921,
920,
382,
29933,
29899,
29945,
1824,
11780,
526,
1539,
746,
322,
408,
896,
526,
2861,
29889,
13,
16107,
6410,
1380,
335,
362,
9437,
24024,
3163,
1033,
884,
367,
7841,
29889,
4525,
3160,
313,
29874,
29897,
278,
17832,
20847,
3097,
322,
1274,
23493,
310,
382,
29933,
29899,
29945,
1663,
18541,
304,
5146,
263,
883,
310,
297,
2310,
29876,
537,
313,
1552,
736,
310,
278,
13258,
358,
29897,
565,
13258,
272,
5697,
2187,
526,
451,
23454,
470,
5855,
373,
25488,
526,
451,
6206,
29892,
313,
29890,
29897,
278,
6124,
310,
916,
13807,
7190,
310,
1663,
18541,
13047,
313,
29872,
29889,
29887,
1696,
285,
10652,
537,
289,
13788,
29892,
322,
23746,
363,
4436,
322,
2703,
6847,
29892,
5381,
1006,
18953,
29892,
1513,
943,
322,
13049,
619,
3097,
322,
2498,
619,
3097,
511,
322,
313,
29883,
29897,
278,
671,
310,
2317,
1609,
8721,
310,
16200,
29889,
13,
3047,
449,
10397,
1218,
14731,
447,
29920,
538,
29892,
11559,
1033,
3867,
297,
278,
11483,
11506,
28410,
15366,
393,
263,
14014,
4818,
29915,
29879,
470,
2060,
13897,
29915,
29879,
3033,
5049,
310,
385,
7417,
25947,
29884,
455,
653,
29892,
20590,
310,
3153,
4072,
310,
1663,
18541,
29892,
322,
29914,
272,
24628,
363,
263,
2317,
1609,
5497,
310,
16200,
723,
9080,
408,
263,
1781,
29899,
5444,
389,
21460,
654,
2750,
278,
972,
616,
470,
6410,
310,
4307,
1319,
17667,
20201,
4660,
297,
4251,
988,
278,
3725,
29907,
29892,
306,
4741,
470,
3148,
29907,
3235,
3683,
1475,
393,
382,
29933,
29899,
29945,
13258,
943,
505,
1063,
822,
336,
566,
287,
29889,
13,
3308,
29907,
3235,
1033,
788,
2788,
3279,
1953,
297,
967,
313,
12276,
23244,
29897,
4720,
29899,
517,
29899,
915,
29899,
5467,
3726,
382,
29933,
29899,
29945,
1072,
8250,
29889,
450,
946,
3819,
1033,
884,
6084,
491,
1072,
2785,
470,
8898,
26959,
392,
398,
14209,
988,
278,
15058,
310,
410,
3471,
6072,
766,
4838,
291,
881,
367,
11382,
3368,
322,
310,
278,
2989,
3464,
310,
967,
610,
1772,
14329,
322,
5703,
358,
28733,
526,
16896,
304,
822,
336,
566,
287,
541,
6467,
7336,
790,
1821,
382,
29933,
29899,
29945,
13258,
943,
29889,
13,
9802,
300,
10769,
881,
884,
505,
376,
808,
262,
297,
278,
3748,
29908,
310,
12566,
292,
382,
29933,
29899,
29945,
13258,
943,
29889,
2688,
881,
1996,
278,
3033,
5049,
310,
7417,
25947,
29884,
455,
4314,
322,
9580,
545,
358,
310,
8210,
1663,
18541,
4612,
1179,
470,
8721,
310,
16200,
393,
10032,
541,
437,
451,
27399,
599,
13258,
358,
12045,
577,
393,
3148,
29907,
3235,
29915,
29879,
472,
29899,
3780,
29895,
7483,
11780,
526,
1661,
621,
6393,
15787,
29889,
28794,
27138,
881,
884,
18720,
393,
12045,
20376,
674,
788,
21544,
541,
5517,
5706,
20831,
292,
3438,
4048,
886,
393,
674,
2654,
618,
304,
278,
14169,
310,
7601,
301,
21043,
29892,
2060,
18777,
322,
14014,
1644,
414,
1550,
884,
13978,
292,
20332,
19518,
29892,
13258,
358,
25228,
943,
322,
278,
382,
29933,
29899,
29945,
13258,
943,
6053,
29889,
18514,
15084,
322,
1957,
635,
29892,
871,
316,
1338,
411,
25947,
29884,
455,
653,
322,
1663,
18541,
3279,
1953,
674,
367,
9999,
519,
29889,
13,
5596,
29879,
322,
6411,
2925,
813,
278,
17294,
11991,
310,
278,
7460,
292,
383,
19467,
813,
7841,
1749,
16772,
284,
883,
310,
5874,
393,
756,
1095,
2955,
363,
21726,
29889,
3118,
310,
1438,
5680,
338,
278,
13047,
310,
9461,
1907,
2750,
13638,
3081,
29889,
1105,
2086,
674,
263,
716,
1788,
310,
12747,
322,
6411,
2925,
12566,
23180,
519,
382,
29933,
29899,
29945,
13258,
943,
5517,
10032,
278,
27289,
393,
13258,
943,
674,
2436,
1407,
269,
13902,
12747,
541,
7709,
694,
6411,
2925,
297,
1009,
7483,
15303,
470,
2058,
7933,
15889,
297,
1009,
17042,
10376,
29889,
13,
2308,
3427,
322,
278,
317,
21040,
13,
28080,
27504,
338,
6299,
310,
5183,
278,
26627,
1199,
515,
838,
13015,
29915,
29879,
29871,
29896,
29929,
29953,
29947,
390,
29987,
29933,
7124,
4823,
376,
1576,
317,
21040,
29908,
297,
670,
11531,
364,
284,
3687,
29889,
5806,
445,
338,
263,
4380,
29891,
260,
1540,
29892,
27504,
756,
1286,
1034,
14214,
278,
4823,
491,
4067,
1218,
372,
411,
670,
19626,
304,
23772,
29879,
29889,
940,
937,
2000,
363,
263,
9892,
373,
23772,
29879,
18055,
278,
3303,
3900,
29892,
3704,
8713,
6392,
25447,
267,
29892,
322,
10325,
9120,
372,
491,
5432,
363,
263,
8872,
2673,
310,
5198,
16783,
515,
10161,
310,
278,
3186,
746,
727,
338,
263,
16413,
4955,
310,
15115,
1608,
2750,
278,
3303,
3900,
470,
967,
394,
3687,
29889,
27504,
1556,
10325,
1497,
393,
278,
3303,
3900,
881,
2050,
901,
1153,
1455,
20077,
292,
29892,
297,
2933,
304,
263,
1139,
1048,
3692,
540,
6969,
7621,
4307,
24555,
13561,
14387,
329,
4901,
310,
23772,
23035,
1156,
278,
1394,
28514,
4158,
27904,
29889,
960,
599,
310,
1438,
9551,
1338,
892,
8762,
29892,
372,
723,
2411,
2742,
278,
11509,
310,
14746,
310,
13201,
1998,
29874,
4808,
414,
322,
5198,
4481,
1934,
304,
25204,
15084,
3896,
278,
3303,
3900,
29889,
13,
4013,
4863,
1401,
919,
292,
27504,
29915,
29879,
5183,
310,
450,
317,
21040,
297,
670,
364,
635,
297,
4122,
575,
4089,
29877,
29892,
25166,
373,
5306,
29871,
29896,
29946,
29892,
29871,
29906,
29900,
29896,
29953,
338,
2086,
521,
8873,
304,
6505,
29892,
408,
278,
5183,
338,
1006,
1028,
414,
287,
411,
278,
321,
6929,
310,
263,
10021,
272,
28655,
303,
285,
1267,
29920,
1000,
521,
1934,
310,
376,
27019,
30098,
8278,
1642,
8512,
278,
26627,
1199,
526,
20603,
491,
319,
267,
459,
29915,
29879,
285,
519,
310,
278,
8413,
1050,
322,
278,
10630,
546,
29892,
278,
26627,
1199,
2615,
1407,
4457,
1531,
746,
27504,
4067,
1078,
963,
411,
670,
1370,
373,
23772,
29879,
29889,
450,
26627,
1199,
13819,
345,
2820,
263,
22707,
5192,
287,
6114,
1058,
620,
29883,
1041,
263,
4203,
14671,
2256,
269,
21040,
29889,
2860,
278,
269,
21040,
338,
620,
4979,
287,
29892,
540,
2586,
267,
278,
6114,
29892,
322,
746,
1183,
338,
27116,
29892,
278,
269,
21040,
10603,
902,
393,
1183
] | 1,024 |
CommonCrawl
| 0.443635 | 0.919475 | 0.357866 | 0.549969 |
knew very well that she took in a poisonous snake. One view regarding the moral of this fable is to teach the lesson not to expect a reward from the wicked. Another view is that the rescuer realizes that it is his own fault for pitying a scoundrel. Trump first associated these lyrics with Syrian refugees, fully realizing that almost all the refugees have genuinely escaped harm in Syria, and many have been desperate enough to even die, including children, while trying to reach safer shores.
Read the lyrics yourself to see how they have been twisted to suit Trump's agenda:
On her way to work one morning
Down the path alongside the lake
A tender hearted woman saw a poor half frozen snake
His pretty colored skin had been all frosted with the dew
"Oh well," she cried, "I'll take you in and I'll take care of you"
"Take me in oh tender woman
Take me in, for heaven's sake
Take me in oh tender woman," sighed the snake
She wrapped him up all cozy in a curvature of silk
And then laid him by the fireside with some honey and some milk
Now she hurried home from work that night as soon as she arrived
She found that pretty snake she'd taking in had been revived
"Take me in, oh tender woman
Now she clutched him to her bosom, "You're so beautiful, " she cried
"But if I hadn't brought you in by now you might have died"
Now she stroked his pretty skin and then she kissed and held him tight
But instead of saying thanks, that snake gave her a vicious bite
"I saved you," cried that woman
"And you've bit me even, why?
You know your bite is poisonous and now I'm going to die"
"Oh shut up, silly woman," said the reptile with a grin
"You knew damn well I was a snake before you took me in
Trump has even more shamelessly exploited these lyrics after the massacre of innocent LGBT party goers in an Orlando night club by Omar Mateen, who was discovered to be a Muslim and born in the United States. The snake, according to Trump, represents the Muslim immigrant who was let into the country, and who now viciously bites the people who let him in. Even though Mateen was a US citizen by virtue of his birth in this country, Trump falsely asserted in one of his speeches that he "was born in Afghan, of Afghan parents, who immigrated to the United States." Trump went on to add that the "only reason the killer was in America in the first place was because we allowed his family to come here." While there was profiling of Muslim immigrants following the terrorist attacks on September 11, 2001, Trump's proposals would far exceed the profiling policies that were put into place following 9/11.
Following 9/11, the Bush administration through Attorney General Ashcroft tweaked the rules to make it easier to detain immigrants. The expanded regulation, which took effect on September 20, 2001, authorized the then INS to hold any non-citizen in custody for 48 hours or an unspecified "additional reasonable time" before charging the person with an offense. In the post 9/11 sweep, immigrants from mainly Muslim countries were detained and deported in secret. Although they were detained because of immigration violations, it was under the pretext of investigating them for suspected links to terrorism. In the end, the 1000+ immigrants who were detained and deported in secret were not charged or convicted of terrorism.
The Bush Administration then implemented Special Registration, which applied to males from 26 countries, 25 of which had significant Islamic populations. Dutifully, 85,000 people lined up to register, thinking that they should cooperate with the government. 13,000 men who were found to have immigration violations, many of whom may have been on the path to getting green cards, were placed in deportation proceedings. Not a single terrorist was discovered under the Special Registration program, which proved to be a colossal waste of tax payer money and was disbanded.
Trump now wishes to take these discredited policies even further. Although there was profiling since 9/
|
[
6363,
1407,
1532,
393,
1183,
3614,
297,
263,
27908,
681,
269,
21040,
29889,
3118,
1776,
11211,
278,
14731,
310,
445,
285,
519,
338,
304,
6860,
278,
3109,
265,
451,
304,
2149,
263,
20751,
515,
278,
281,
17840,
29889,
7280,
1776,
338,
393,
278,
620,
29883,
2853,
1855,
7093,
393,
372,
338,
670,
1914,
12570,
363,
28875,
292,
263,
885,
618,
2674,
29889,
27504,
937,
6942,
1438,
26627,
1199,
411,
8713,
6392,
25447,
267,
29892,
8072,
1855,
5281,
393,
4359,
599,
278,
25447,
267,
505,
29120,
262,
873,
19824,
10311,
297,
8713,
2849,
29892,
322,
1784,
505,
1063,
553,
21194,
3307,
304,
1584,
762,
29892,
3704,
4344,
29892,
1550,
1811,
304,
6159,
872,
571,
528,
2361,
29889,
13,
6359,
278,
26627,
1199,
7535,
304,
1074,
920,
896,
505,
1063,
3252,
12652,
304,
14726,
27504,
29915,
29879,
946,
8395,
29901,
13,
2951,
902,
982,
304,
664,
697,
7250,
13,
6767,
278,
2224,
19963,
278,
19437,
13,
29909,
22707,
5192,
287,
6114,
4446,
263,
6460,
4203,
14671,
2256,
269,
21040,
13,
29950,
275,
5051,
28684,
19309,
750,
1063,
599,
14671,
16470,
411,
278,
316,
29893,
13,
29908,
9048,
1532,
1699,
1183,
10680,
29892,
376,
29902,
29915,
645,
2125,
366,
297,
322,
306,
29915,
645,
2125,
2562,
310,
366,
29908,
13,
29908,
26772,
592,
297,
9360,
22707,
6114,
13,
26772,
592,
297,
29892,
363,
18356,
29915,
29879,
16563,
13,
26772,
592,
297,
9360,
22707,
6114,
1699,
269,
25398,
278,
269,
21040,
13,
13468,
21021,
1075,
701,
599,
1302,
1537,
297,
263,
27686,
1535,
310,
4047,
29895,
13,
2855,
769,
12341,
1075,
491,
278,
24237,
680,
411,
777,
298,
4992,
322,
777,
27274,
13,
10454,
1183,
29117,
3271,
515,
664,
393,
4646,
408,
4720,
408,
1183,
11977,
13,
13468,
1476,
393,
5051,
269,
21040,
1183,
29915,
29881,
5622,
297,
750,
1063,
6664,
2347,
13,
29908,
26772,
592,
297,
29892,
9360,
22707,
6114,
13,
10454,
1183,
1067,
10519,
287,
1075,
304,
902,
13601,
290,
29892,
376,
3492,
29915,
276,
577,
9560,
29892,
376,
1183,
10680,
13,
29908,
6246,
565,
306,
27222,
29915,
29873,
6296,
366,
297,
491,
1286,
366,
1795,
505,
6423,
29908,
13,
10454,
1183,
380,
16475,
287,
670,
5051,
19309,
322,
769,
1183,
20057,
287,
322,
4934,
1075,
19932,
13,
6246,
2012,
310,
5934,
3969,
29892,
393,
269,
21040,
4846,
902,
263,
325,
14803,
289,
568,
13,
29908,
29902,
7160,
366,
1699,
10680,
393,
6114,
13,
29908,
2855,
366,
29915,
345,
2586,
592,
1584,
29892,
2020,
29973,
13,
3492,
1073,
596,
289,
568,
338,
27908,
681,
322,
1286,
306,
29915,
29885,
2675,
304,
762,
29908,
13,
29908,
9048,
12522,
701,
29892,
24866,
6114,
1699,
1497,
278,
337,
415,
488,
411,
263,
867,
262,
13,
29908,
3492,
6363,
5625,
29876,
1532,
306,
471,
263,
269,
21040,
1434,
366,
3614,
592,
297,
13,
2308,
3427,
756,
1584,
901,
528,
314,
6393,
368,
16035,
1573,
1438,
26627,
1199,
1156,
278,
4158,
21160,
310,
21458,
296,
365,
7210,
29911,
6263,
748,
414,
297,
385,
1394,
28514,
4646,
4402,
491,
438,
3034,
29566,
264,
29892,
1058,
471,
10943,
304,
367,
263,
23772,
322,
6345,
297,
278,
3303,
3900,
29889,
450,
269,
21040,
29892,
5034,
304,
27504,
29892,
11524,
278,
23772,
5198,
4481,
424,
1058,
471,
1235,
964,
278,
4234,
29892,
322,
1058,
1286,
325,
1654,
5794,
2586,
267,
278,
2305,
1058,
1235,
1075,
297,
29889,
7753,
2466,
29566,
264,
471,
263,
3148,
14497,
264,
491,
27935,
310,
670,
12060,
297,
445,
4234,
29892,
27504,
285,
1338,
873,
4974,
287,
297,
697,
310,
670,
12032,
267,
393,
540,
376,
11102,
6345,
297,
27135,
29892,
310,
27135,
11825,
29892,
1058,
5198,
4481,
630,
304,
278,
3303,
3900,
1213,
27504,
3512,
373,
304,
788,
393,
278,
376,
6194,
2769,
278,
413,
5495,
471,
297,
6813,
297,
278,
937,
2058,
471,
1363,
591,
6068,
670,
3942,
304,
2041,
1244,
1213,
5806,
727,
471,
20077,
292,
310,
23772,
5198,
4481,
1934,
1494,
278,
15115,
391,
16661,
373,
3839,
29871,
29896,
29896,
29892,
29871,
29906,
29900,
29900,
29896,
29892,
27504,
29915,
29879,
9551,
1338,
723,
2215,
13461,
278,
20077,
292,
24833,
393,
892,
1925,
964,
2058,
1494,
29871,
29929,
29914,
29896,
29896,
29889,
13,
29943,
2952,
292,
29871,
29929,
29914,
29896,
29896,
29892,
278,
24715,
17517,
1549,
6212,
25252,
4593,
12835,
24077,
615,
7780,
12535,
278,
6865,
304,
1207,
372,
6775,
304,
1439,
475,
5198,
4481,
1934,
29889,
450,
17832,
1072,
2785,
29892,
607,
3614,
2779,
373,
3839,
29871,
29906,
29900,
29892,
29871,
29906,
29900,
29900,
29896,
29892,
4148,
1891,
278,
769,
2672,
29903,
304,
4808,
738,
1661,
29899,
20752,
19642,
297,
25387,
1486,
363,
29871,
29946,
29947,
6199,
470,
385,
443,
6550,
2164,
376,
1202,
3245,
15590,
931,
29908,
1434,
9151,
292,
278,
2022,
411,
385,
1283,
1947,
29889,
512,
278,
1400,
29871,
29929,
29914,
29896,
29896,
7901,
1022,
29892,
5198,
4481,
1934,
515,
14364,
23772,
10916,
892,
1439,
7114,
322,
21455,
287,
297,
7035,
29889,
8512,
896,
892,
1439,
7114,
1363,
310,
5198,
16783,
5537,
800,
29892,
372,
471,
1090,
278,
758,
726,
310,
7405,
1218,
963,
363,
2858,
6021,
2988,
304,
15115,
1608,
29889,
512,
278,
1095,
29892,
278,
29871,
29896,
29900,
29900,
29900,
29974,
5198,
4481,
1934,
1058,
892,
1439,
7114,
322,
21455,
287,
297,
7035,
892,
451,
20139,
470,
7602,
18186,
310,
15115,
1608,
29889,
13,
1576,
24715,
23303,
769,
8762,
12630,
2169,
8306,
29892,
607,
7436,
304,
25269,
515,
29871,
29906,
29953,
10916,
29892,
29871,
29906,
29945,
310,
607,
750,
7282,
16427,
293,
23093,
29889,
360,
329,
6845,
368,
29892,
29871,
29947,
29945,
29892,
29900,
29900,
29900,
2305,
301,
1312,
701,
304,
6036,
29892,
7291,
393,
896,
881,
1302,
3372,
403,
411,
278,
5874,
29889,
29871,
29896,
29941,
29892,
29900,
29900,
29900,
1757,
1058,
892,
1476,
304,
505,
5198,
16783,
5537,
800,
29892,
1784,
310,
6029,
1122,
505,
1063,
373,
278,
2224,
304,
2805,
7933,
15889,
29892,
892,
7180,
297,
21455,
362,
8469,
886,
29889,
2216,
263,
2323,
15115,
391,
471,
10943,
1090,
278,
12630,
2169,
8306,
1824,
29892,
607,
11827,
304,
367,
263,
784,
2209,
284,
19863,
310,
8818,
282,
2747,
6909,
322,
471,
766,
4980,
287,
29889,
13,
2308,
3427,
1286,
28688,
304,
2125,
1438,
766,
11944,
1573,
24833,
1584,
4340,
29889,
8512,
727,
471,
20077,
292,
1951,
29871,
29929,
29914
] | 1,024 |
CommonCrawl
| 0.070039 | 0.341961 | 0.387454 | -0.459877 |
11, and every application for an immigration benefit since those attacks is viewed through the prism of national security, immigration did not stop. The basic architecture of our immigration system remained intact, and eligible applicants have been admitted while undergoing more extensive security checks. If Trump's proposals are implemented, there will be a complete ban on immigration from countries where there is a proven history of terrorism against the United States. Just as finding out who is a Muslim would be unclear, it is equally unclear whether this ban would include people from countries such as Syria or Pakistan, or whether it would also involve certain European countries such as France, the United Kingdom and Belgium. Would it also include countries like India or The Philippines, which sends one of the largest numbers of immigrants to the United States? The ban would cover visitors, students and people from these countries, which have all inspired terrorist attacks on its soil, who are legitimately immigrating, including spouses of US citizens. To blame immigrants for the Orlando killings goes beyond the pale, which was perpetrated by a mentally unstable American citizen who may have been inspired by terrorism but also by hate against LGBTs. And where does this stop? Trump said that if the parents were not allowed into the country, this massacre would not have happened. But what about the countless gun deaths caused by other mentally unstable US citizens? Is Trump blaming these killers' ancestors who may have at some point in time come from another country? Trump is inappropriately casting doubt on an entire religion of over 1.2 billion adherents worldwide who are essentially peaceful.
While Trump's rhetoric is frightening enough, there is ample authority in the law that would allow him to implement his proposed ban if he became President. Section 212(f) of the Immigration and Nationality Act provides in part, as follows:
(f) Suspension of entry or imposition of restrictions by President – Whenever the President finds that the entry of any aliens or of any class of aliens into the United States would be detrimental to the interests of the United States, he may by proclamation, and for such period as he shall deem necessary, suspend the entry of all aliens or ay class of aliens as immigrants or nonimmigrants, or impose on the entry of aliens any restrictions he may deem to be appropriate.
Apart from Congress putting a check on the President's authority under INA 212(f), and possibly the courts, the only likely limitations on the exercise of this authority is with respect to lawful permanent residents who have taken brief trips abroad and would be assimilated to the status of a continuously-present resident under Delgadillo v. Carmichael, Kwong Hai Chew v. Colding and Landon v. Plasencia. Even they would be at some risk of being denied readmission, and would probably be better advised not to travel outside the US under a hypothetical President Trump.
The good news is that despite playing to irrational fear and reciting the lyrics of The Snake, Trump's poll numbers have slipped. The conventional wisdom used to be that a Republican presidential candidate who was forceful on security issues would gain an advantage prior to an election. It appears that the attack in Orlando has not helped Trump, and fear mongering may have lost its appeal. This could well change if there was another attack orchestrated by a foreign terrorist organization rather than by an unstable US citizen, but so far Trump's war on Muslims does not seem to be helping him. After all the senseless racial profiling following 9/11, it should become pretty obvious to the American people that profiling a whole community for the acts of one person is not a good law enforcement tactic. It would only alienate the community whose members are well integrated into the American fabric and contributing to the country, and who would also be willing to cooperate with law enforcement. It is also most un-American to profile a whole community as a substitute for individualized guilt, which goes against the principles upon which this nation was founded and has set an example for scores of countries around the world.
If Trump continues to slip, it is hoped that The Snake again be viewed as a cool R&B song in the soul music genre rather than a hate anthem against Muslims.
Can a STEM OPT Student Be Employed At A Third Party Client Site?
by Cora-Ann Pestaina, Partner with ABIL member, Cyrus D. Mehta
The most frequently asked question
|
[
29896,
29896,
29892,
322,
1432,
2280,
363,
385,
5198,
16783,
14169,
1951,
1906,
16661,
338,
24774,
1549,
278,
544,
1608,
310,
4797,
6993,
29892,
5198,
16783,
1258,
451,
5040,
29889,
450,
6996,
11258,
310,
1749,
5198,
16783,
1788,
9488,
938,
627,
29892,
322,
560,
335,
1821,
15576,
1934,
505,
1063,
20186,
1550,
1090,
17696,
901,
20607,
6993,
12747,
29889,
960,
27504,
29915,
29879,
9551,
1338,
526,
8762,
29892,
727,
674,
367,
263,
4866,
9892,
373,
5198,
16783,
515,
10916,
988,
727,
338,
263,
16413,
4955,
310,
15115,
1608,
2750,
278,
3303,
3900,
29889,
3387,
408,
9138,
714,
1058,
338,
263,
23772,
723,
367,
20871,
29892,
372,
338,
18018,
20871,
3692,
445,
9892,
723,
3160,
2305,
515,
10916,
1316,
408,
8713,
2849,
470,
21215,
29892,
470,
3692,
372,
723,
884,
25135,
3058,
7824,
10916,
1316,
408,
3444,
29892,
278,
3303,
12626,
322,
9923,
1974,
29889,
10878,
372,
884,
3160,
10916,
763,
7513,
470,
450,
26260,
29892,
607,
16003,
697,
310,
278,
10150,
3694,
310,
5198,
4481,
1934,
304,
278,
3303,
3900,
29973,
450,
9892,
723,
4612,
26824,
29892,
8041,
322,
2305,
515,
1438,
10916,
29892,
607,
505,
599,
20603,
15115,
391,
16661,
373,
967,
22473,
29892,
1058,
526,
25204,
15084,
5198,
4481,
1218,
29892,
3704,
805,
23676,
310,
3148,
18363,
29889,
1763,
1999,
420,
5198,
4481,
1934,
363,
278,
1394,
28514,
12088,
886,
5771,
8724,
278,
15509,
29892,
607,
471,
25722,
29878,
630,
491,
263,
6042,
635,
443,
13844,
3082,
14497,
264,
1058,
1122,
505,
1063,
20603,
491,
15115,
1608,
541,
884,
491,
26277,
2750,
365,
7210,
29911,
29879,
29889,
1126,
988,
947,
445,
5040,
29973,
27504,
1497,
393,
565,
278,
11825,
892,
451,
6068,
964,
278,
4234,
29892,
445,
4158,
21160,
723,
451,
505,
9559,
29889,
1205,
825,
1048,
278,
2302,
2222,
13736,
4892,
29879,
8581,
491,
916,
6042,
635,
443,
13844,
3148,
18363,
29973,
1317,
27504,
1999,
11500,
1438,
12088,
414,
29915,
19525,
943,
1058,
1122,
505,
472,
777,
1298,
297,
931,
2041,
515,
1790,
4234,
29973,
27504,
338,
297,
932,
6649,
2486,
23013,
7404,
373,
385,
4152,
13433,
310,
975,
29871,
29896,
29889,
29906,
24464,
594,
2276,
1237,
3186,
8157,
1058,
526,
13674,
10776,
1319,
29889,
13,
8809,
488,
27504,
29915,
29879,
364,
9188,
272,
293,
338,
22739,
8333,
3307,
29892,
727,
338,
626,
552,
14329,
297,
278,
4307,
393,
723,
2758,
1075,
304,
2334,
670,
7972,
9892,
565,
540,
3897,
7178,
29889,
9779,
29871,
29906,
29896,
29906,
29898,
29888,
29897,
310,
278,
1954,
29885,
16783,
322,
3086,
537,
3185,
8128,
297,
760,
29892,
408,
4477,
29901,
13,
29898,
29888,
29897,
9511,
29886,
2673,
310,
6251,
470,
527,
3283,
310,
25091,
491,
7178,
785,
1932,
1310,
278,
7178,
14061,
393,
278,
6251,
310,
738,
394,
11689,
470,
310,
738,
770,
310,
394,
11689,
964,
278,
3303,
3900,
723,
367,
1439,
29878,
2073,
284,
304,
278,
20017,
310,
278,
3303,
3900,
29892,
540,
1122,
491,
410,
15719,
362,
29892,
322,
363,
1316,
3785,
408,
540,
4091,
316,
331,
5181,
29892,
8872,
355,
278,
6251,
310,
599,
394,
11689,
470,
10156,
770,
310,
394,
11689,
408,
5198,
4481,
1934,
470,
1661,
6727,
4481,
1934,
29892,
470,
2411,
852,
373,
278,
6251,
310,
394,
11689,
738,
25091,
540,
1122,
316,
331,
304,
367,
8210,
29889,
13,
29909,
1595,
515,
11559,
10594,
263,
1423,
373,
278,
7178,
29915,
29879,
14329,
1090,
2672,
29909,
29871,
29906,
29896,
29906,
29898,
29888,
511,
322,
10075,
278,
28033,
29892,
278,
871,
5517,
27028,
373,
278,
15058,
310,
445,
14329,
338,
411,
3390,
304,
4307,
1319,
17667,
24060,
1058,
505,
4586,
11473,
3367,
567,
28177,
322,
723,
367,
24443,
309,
630,
304,
278,
4660,
310,
263,
3133,
5794,
29899,
6338,
20201,
1090,
5556,
29887,
328,
9093,
325,
29889,
1704,
29885,
436,
4271,
29892,
476,
29893,
549,
379,
1794,
6561,
29893,
325,
29889,
26731,
292,
322,
3172,
265,
325,
29889,
1858,
294,
5760,
29889,
7753,
896,
723,
367,
472,
777,
12045,
310,
1641,
17935,
1303,
6737,
29892,
322,
723,
3117,
367,
2253,
594,
11292,
451,
304,
9850,
5377,
278,
3148,
1090,
263,
13752,
300,
936,
7178,
27504,
29889,
13,
1576,
1781,
9763,
338,
393,
15020,
8743,
304,
3805,
29878,
1288,
8866,
322,
1162,
11407,
278,
26627,
1199,
310,
450,
317,
21040,
29892,
27504,
29915,
29879,
21180,
3694,
505,
20662,
2986,
29889,
450,
28557,
29714,
1304,
304,
367,
393,
263,
21178,
6673,
616,
14020,
1058,
471,
4889,
1319,
373,
6993,
5626,
723,
11581,
385,
10631,
7536,
304,
385,
8271,
29889,
739,
5692,
393,
278,
5337,
297,
1394,
28514,
756,
451,
9213,
27504,
29892,
322,
8866,
286,
549,
3241,
1122,
505,
5714,
967,
25530,
29889,
910,
1033,
1532,
1735,
565,
727,
471,
1790,
5337,
22624,
16444,
630,
491,
263,
9117,
15115,
391,
13013,
3265,
1135,
491,
385,
443,
13844,
3148,
14497,
264,
29892,
541,
577,
2215,
27504,
29915,
29879,
1370,
373,
23772,
29879,
947,
451,
2833,
304,
367,
19912,
1075,
29889,
2860,
599,
278,
4771,
6393,
1153,
1455,
20077,
292,
1494,
29871,
29929,
29914,
29896,
29896,
29892,
372,
881,
4953,
5051,
6924,
304,
278,
3082,
2305,
393,
20077,
292,
263,
3353,
7881,
363,
278,
14741,
310,
697,
2022,
338,
451,
263,
1781,
4307,
24555,
13561,
260,
17911,
29889,
739,
723,
871,
394,
819,
403,
278,
7881,
5069,
5144,
526,
1532,
23387,
964,
278,
3082,
18187,
322,
17737,
17068,
304,
278,
4234,
29892,
322,
1058,
723,
884,
367,
17762,
304,
1302,
3372,
403,
411,
4307,
24555,
13561,
29889,
739,
338,
884,
1556,
443,
29899,
14689,
304,
8722,
263,
3353,
7881,
408,
263,
23764,
363,
5375,
1891,
1410,
2782,
29892,
607,
5771,
2750,
278,
18671,
2501,
607,
445,
5233,
471,
11091,
322,
756,
731,
385,
1342,
363,
19435,
310,
10916,
2820,
278,
3186,
29889,
13,
3644,
27504,
18172,
304,
269,
3466,
29892,
372,
338,
24936,
393,
450,
317,
21040,
1449,
367,
24774,
408,
263,
12528,
390,
29987,
29933,
4823,
297,
278,
10752,
4696,
16151,
3265,
1135,
263,
26277,
24612,
331,
2750,
23772,
29879,
29889,
13,
6028,
263,
317,
4330,
29924,
6418,
29911,
15740,
1522,
7361,
2376,
287,
2180,
319,
18008,
9173,
12477,
10781,
29973,
13,
1609,
315,
2207,
29899,
2744,
29876,
349,
342,
475,
29874,
29892,
3455,
1089,
411,
17571,
6227,
4509,
29892,
315,
29646,
360,
29889,
2191,
400,
29874,
13,
1576,
1556,
13672,
4433,
1139
] | 1,024 |
CommonCrawl
| 0.455182 | 0.761145 | 0.287706 | 0.572736 |
in response to my recent blog entitled, "A Closer Look At The Form I-983 – Training Plan for STEM OPT Students", is whether a STEM OPT student can be employed at a third party client site or at multiple client sites. I would argue that the answer to this question ought to be YES! Since the new rule only took effect on May 10, 2016, there isn't yet any strong anecdotal evidence on whether Designated Student Officers (DSO) will approve Forms I-983 which set forth training to take place at client sites. However, there isn't anything in the governing regulations that expressly forbids this type of employment.
This is a big issue for many students and employers because under the standard 12-month OPT program, the employer may employ the student in a regular job, even at third party sites, as long as the employment is related to their major area of study in the US. However, in order for the student to obtain a 24-month STEM OPT extension, the employer and student, through the submission of the Form I-983 to the DSO, must demonstrate that the student will be employed only as a trainee. The Form I-983 specifically requires the employer to attest that the student will "receive on-site supervision and training by experienced and knowledgeable staff" and that the employer "has sufficient resources and personnel to provide the training and is prepared to implement the program." Department of Homeland Security (DHS) has made it clear in the preamble to the new regulations that the STEM OPT extension is not apt for certain types of employment arrangements which include multiple employer arrangements, sole proprietorships, employment through "temp" agencies, employment through consulting firm arrangements that provide labor for hire, and other relationships that do not constitute a bona fide employer-employee relationship. Students cannot qualify for STEM OPT extensions unless they will be bona fide employees of the employer signing the Form I-983, and the employer that signs the Form I-983 must be the same entity that employs the student and provides the practical training experience.
None of the above listed requirements prevent the employment of a STEM OPT student at a third-party client site. I would argue that the issues surrounding such employment are similar to the issues surrounding the employment of an H-1B beneficiary at a third party client site. In the case of the H-1B, the employer must also establish that a valid employer-employee relationship will exist with the H-1B beneficiary throughout the requested H-1B validity period. By now, most H-1B employers are used to the USCIS requirements published in its memo entitled, "Determining Employer-Employee for Adjudication of H-1B Petitions, Including Third-Party Site Placements" ("the Neufeld Memo"). Under the Neufeld Memo, in considering whether or not there is a valid "employer-employee relationship," USCIS must determine if the employer exercises a sufficient level of "control" over the prospective H-1B employee. To demonstrate control, the employer can submit various evidence including a copy of its employment agreement with the prospective employee; copies of its contractual agreement(s) with the end client where the employee will work; a letter from the end client describing its relationship with the employer and the prospective employee; sample staff evaluation forms to demonstrate how the employee will be evaluated; a clear description of how employee supervision will be conducted; a list of the various benefits provided to the employee by the employer; and so on.
I would argue that similarly, in the case of the STEM OPT student, the employer should be able to satisfactorily demonstrate its control over the student despite placement of the student at an end client site. Under the final rule, the Form I-983 must, among other things: (1) Identify the goals for the STEM practical training opportunity, including specific knowledge, skills, or techniques that will be imparted to the student; (2) explain how those goals will be achieved through the work-based learning opportunity with the employer; (3) describe a performance evaluation process; and (4) describe methods of oversight and supervision. Admittedly, having the student work at a client site makes for a more difficult case. However, if the employer already has employees at that site who can implement the employer's training program by providing the
|
[
297,
2933,
304,
590,
7786,
12618,
23437,
29892,
376,
29909,
2233,
22969,
7419,
2180,
450,
3812,
306,
29899,
29929,
29947,
29941,
785,
26101,
8402,
363,
317,
4330,
29924,
6418,
29911,
5088,
1237,
613,
338,
3692,
263,
317,
4330,
29924,
6418,
29911,
8368,
508,
367,
15723,
472,
263,
4654,
6263,
3132,
3268,
470,
472,
2999,
3132,
11840,
29889,
306,
723,
27754,
393,
278,
1234,
304,
445,
1139,
12722,
304,
367,
22483,
29991,
4001,
278,
716,
5751,
871,
3614,
2779,
373,
2610,
29871,
29896,
29900,
29892,
29871,
29906,
29900,
29896,
29953,
29892,
727,
3508,
29915,
29873,
3447,
738,
4549,
385,
687,
6333,
284,
10757,
373,
3692,
12037,
630,
15740,
20339,
414,
313,
29928,
6156,
29897,
674,
2134,
345,
3812,
29879,
306,
29899,
29929,
29947,
29941,
607,
731,
11483,
6694,
304,
2125,
2058,
472,
3132,
11840,
29889,
2398,
29892,
727,
3508,
29915,
29873,
3099,
297,
278,
14765,
1076,
1072,
8250,
393,
4653,
368,
19752,
4841,
445,
1134,
310,
5703,
358,
29889,
13,
4013,
338,
263,
4802,
2228,
363,
1784,
8041,
322,
5703,
414,
1363,
1090,
278,
3918,
29871,
29896,
29906,
29899,
10874,
6418,
29911,
1824,
29892,
278,
5703,
261,
1122,
5703,
278,
8368,
297,
263,
4943,
4982,
29892,
1584,
472,
4654,
6263,
11840,
29892,
408,
1472,
408,
278,
5703,
358,
338,
4475,
304,
1009,
4655,
4038,
310,
6559,
297,
278,
3148,
29889,
2398,
29892,
297,
1797,
363,
278,
8368,
304,
4017,
263,
29871,
29906,
29946,
29899,
10874,
317,
4330,
29924,
6418,
29911,
6081,
29892,
278,
5703,
261,
322,
8368,
29892,
1549,
278,
29240,
310,
278,
3812,
306,
29899,
29929,
29947,
29941,
304,
278,
360,
6156,
29892,
1818,
22222,
393,
278,
8368,
674,
367,
15723,
871,
408,
263,
1020,
457,
29872,
29889,
450,
3812,
306,
29899,
29929,
29947,
29941,
10816,
6858,
278,
5703,
261,
304,
1098,
342,
393,
278,
8368,
674,
376,
13556,
573,
373,
29899,
2746,
2428,
4924,
322,
6694,
491,
18860,
322,
7134,
519,
13925,
29908,
322,
393,
278,
5703,
261,
376,
5349,
8002,
7788,
322,
24127,
304,
3867,
278,
6694,
322,
338,
13240,
304,
2334,
278,
1824,
1213,
10317,
310,
15089,
14045,
14223,
313,
29928,
14851,
29897,
756,
1754,
372,
2821,
297,
278,
758,
314,
569,
304,
278,
716,
1072,
8250,
393,
278,
317,
4330,
29924,
6418,
29911,
6081,
338,
451,
10882,
363,
3058,
4072,
310,
5703,
358,
15196,
4110,
607,
3160,
2999,
5703,
261,
15196,
4110,
29892,
14419,
24440,
272,
9981,
29892,
5703,
358,
1549,
376,
7382,
29908,
946,
15942,
29892,
5703,
358,
1549,
8799,
292,
9226,
15196,
4110,
393,
3867,
10212,
363,
298,
533,
29892,
322,
916,
21702,
393,
437,
451,
1040,
12356,
263,
289,
2681,
285,
680,
5703,
261,
29899,
26143,
9443,
29889,
5088,
1237,
2609,
4021,
1598,
363,
317,
4330,
29924,
6418,
29911,
17752,
6521,
896,
674,
367,
289,
2681,
285,
680,
22873,
310,
278,
5703,
261,
26188,
278,
3812,
306,
29899,
29929,
29947,
29941,
29892,
322,
278,
5703,
261,
393,
18906,
278,
3812,
306,
29899,
29929,
29947,
29941,
1818,
367,
278,
1021,
7855,
393,
3710,
417,
952,
278,
8368,
322,
8128,
278,
15031,
6694,
7271,
29889,
13,
8516,
310,
278,
2038,
9904,
11780,
5557,
278,
5703,
358,
310,
263,
317,
4330,
29924,
6418,
29911,
8368,
472,
263,
4654,
29899,
22633,
3132,
3268,
29889,
306,
723,
27754,
393,
278,
5626,
18830,
1316,
5703,
358,
526,
2788,
304,
278,
5626,
18830,
278,
5703,
358,
310,
385,
379,
29899,
29896,
29933,
7795,
1654,
653,
472,
263,
4654,
6263,
3132,
3268,
29889,
512,
278,
1206,
310,
278,
379,
29899,
29896,
29933,
29892,
278,
5703,
261,
1818,
884,
10127,
393,
263,
2854,
5703,
261,
29899,
26143,
9443,
674,
1863,
411,
278,
379,
29899,
29896,
29933,
7795,
1654,
653,
10106,
278,
13877,
379,
29899,
29896,
29933,
2854,
537,
3785,
29889,
2648,
1286,
29892,
1556,
379,
29899,
29896,
29933,
5703,
414,
526,
1304,
304,
278,
3148,
29907,
3235,
11780,
6369,
297,
967,
2626,
29877,
23437,
29892,
376,
6362,
837,
2827,
7361,
2376,
261,
29899,
19461,
363,
2087,
17675,
293,
362,
310,
379,
29899,
29896,
29933,
5879,
2187,
29892,
512,
22368,
18008,
29899,
7439,
29891,
10781,
349,
4620,
4110,
29908,
4852,
1552,
2448,
1137,
2495,
8133,
29877,
2564,
7634,
278,
2448,
1137,
2495,
8133,
29877,
29892,
297,
13858,
3692,
470,
451,
727,
338,
263,
2854,
376,
3451,
2376,
261,
29899,
26143,
9443,
1699,
3148,
29907,
3235,
1818,
8161,
565,
278,
5703,
261,
24472,
3476,
267,
263,
8002,
3233,
310,
376,
6451,
29908,
975,
278,
10791,
12645,
379,
29899,
29896,
29933,
19001,
29889,
1763,
22222,
2761,
29892,
278,
5703,
261,
508,
9752,
5164,
10757,
3704,
263,
3509,
310,
967,
5703,
358,
17327,
411,
278,
10791,
12645,
19001,
29936,
14591,
310,
967,
8078,
950,
17327,
29898,
29879,
29897,
411,
278,
1095,
3132,
988,
278,
19001,
674,
664,
29936,
263,
5497,
515,
278,
1095,
3132,
20766,
967,
9443,
411,
278,
5703,
261,
322,
278,
10791,
12645,
19001,
29936,
4559,
13925,
17983,
7190,
304,
22222,
920,
278,
19001,
674,
367,
19030,
29936,
263,
2821,
6139,
310,
920,
19001,
2428,
4924,
674,
367,
18043,
29936,
263,
1051,
310,
278,
5164,
23633,
4944,
304,
278,
19001,
491,
278,
5703,
261,
29936,
322,
577,
373,
29889,
13,
29902,
723,
27754,
393,
22829,
29892,
297,
278,
1206,
310,
278,
317,
4330,
29924,
6418,
29911,
8368,
29892,
278,
5703,
261,
881,
367,
2221,
304,
5119,
7168,
2354,
22222,
967,
2761,
975,
278,
8368,
15020,
2174,
13561,
310,
278,
8368,
472,
385,
1095,
3132,
3268,
29889,
7634,
278,
2186,
5751,
29892,
278,
3812,
306,
29899,
29929,
29947,
29941,
1818,
29892,
4249,
916,
2712,
29901,
313,
29896,
29897,
13355,
1598,
278,
14433,
363,
278,
317,
4330,
29924,
15031,
6694,
15130,
29892,
3704,
2702,
7134,
29892,
25078,
29892,
470,
13698,
393,
674,
367,
527,
1595,
287,
304,
278,
8368,
29936,
313,
29906,
29897,
5649,
920,
1906,
14433,
674,
367,
14363,
1549,
278,
664,
29899,
6707,
6509,
15130,
411,
278,
5703,
261,
29936,
313,
29941,
29897,
8453,
263,
4180,
17983,
1889,
29936,
322,
313,
29946,
29897,
8453,
3519,
310,
288,
874,
523,
322,
2428,
4924,
29889,
2087,
29885,
4430,
368,
29892,
2534,
278,
8368,
664,
472,
263,
3132,
3268,
3732,
363,
263,
901,
5189,
1206,
29889,
2398,
29892,
565,
278,
5703,
261,
2307,
756,
22873,
472,
393,
3268,
1058,
508,
2334,
278,
5703,
261,
29915,
29879,
6694,
1824,
491,
13138,
278
] | 1,024 |
CommonCrawl
| 0.156305 | 1.163459 | 0.020292 | 1.144461 |
training, on-site supervision and evaluation of the student, then the Form I-983 ought to be approvable. The employer may face a more insurmountable hurdle in a case where the student would be its only employee stationed at the client site. In such a case it would be very difficult to argue that the employer will provide a structured and guided work-based learning experience for the student, although a case could still potentially be made for a bona fide training program if the employer has ready access at the site to supervise the trainee.
With regard to multiple worksites, in the preamble to the regulations, DHS made it clear that the Form I-983 may incorporate provisions for project, position, or department rotations that directly relate to the STEM student's field of study, provided there will be appropriate supervision during each rotation and the employer otherwise meets all relevant requirements. Similarly, changes in client site locations can be well documented and explained upon submission of the Form I-983. New and previously unforeseen changes can always be addressed through the preparation and submission of a modified Training Plan to the DSO.
The fact that the Form I-983 must be submitted to the DSO and not to DHS is significant because with filings submitted to DHS, there is usually a filing fee and the potential for costly (time and money) rejections by an inaccessible, unseen and unknown officer. A DSO is an individual who is typically more accessible. Should the DSO not approve the initial Form I-983, there should, hopefully, be more of an opportunity for the employer and student to understand the Training Plan's defects and to provide additional information in a new submission.
The new STEM OPT rule would allow talented students who have graduated from US universities in vital STEM fields to remain for an additional 24 months. As a result, the rule must encompass all kinds of modern work arrangements, including working at third party sites. Otherwise, entire industries, including IT, management consulting or accounting, would be deprived of engaging talented foreign students. Foreign students can also benefit by receiving training in industries whose business model relies on assigning employees to third party client sites. It is industries that rely on assigning workers to third party sites that give American businesses a competitive edge by providing them with much needed flexibility. They should not be left out from the new rule!
Were the DOJ Lawyers Really Unethical in Texas v. USA?
Judge Hanen's order dated May 19, 2016 reprimanding thousands of Department of Justice lawyers for unethical conduct is astounding because it does not even appear that their conduct was unethical.
Much has already been written about Judge Hanen's strange order. Professor Orin Kerr questions whether the judge can even impose ethics classes on hundreds of DOJ lawyers who are not remotely connected to the case. Professor Shobha Sivaprasad Wadhia is justifiably concerned that the order, in addition to reprimanding DOJ attorneys, also threatens to 'out' the names of more than hundred thousand recipients of the Deferred Action for Childhood Arrival (DACA) program who were granted 3 year extensions instead of 2 year extensions. Professor Stephen Legomsky does not even think the DOJ lawyers did anything wrong.
I completely agree. Let's look at Rule 3.3 of the American Bar Association Model rules of Professional Conduct and the corresponding Texas Disciplinary Rules of Professional conduct, which Judge Hanen used, along with a fair sprinkling of dialogs from popular films, for finding that the DOJ lawyers were not truthful to the court. One of the cardinal ethical cannons is that a lawyer has a duty of candor to a tribunal. ABA Model Rule 3.3 provides in relevant part:
a) A lawyer shall not knowingly:
1) Make a false statement of fact or law to a tribunal or fail to correct a false statement of fact or law to a tribunal or fail to correct a false statement of material fact or law previously made to the tribunal by the lawyer.
The relevant potions of the Texas version of Rule 3.3 are similar:
1) Make a false statement of material fact or law to a tribunal
2) Fail to disclose a fact to as tribunal when disclosure is necessary to avoid assisting a criminal or fraudulent act;
5) offer or use evidence that the lawyer knows to
|
[
6694,
29892,
373,
29899,
2746,
2428,
4924,
322,
17983,
310,
278,
8368,
29892,
769,
278,
3812,
306,
29899,
29929,
29947,
29941,
12722,
304,
367,
2134,
29894,
519,
29889,
450,
5703,
261,
1122,
3700,
263,
901,
1663,
332,
16476,
519,
12166,
29881,
280,
297,
263,
1206,
988,
278,
8368,
723,
367,
967,
871,
19001,
5073,
287,
472,
278,
3132,
3268,
29889,
512,
1316,
263,
1206,
372,
723,
367,
1407,
5189,
304,
27754,
393,
278,
5703,
261,
674,
3867,
263,
2281,
2955,
322,
1410,
2618,
664,
29899,
6707,
6509,
7271,
363,
278,
8368,
29892,
5998,
263,
1206,
1033,
1603,
19998,
367,
1754,
363,
263,
289,
2681,
285,
680,
6694,
1824,
565,
278,
5703,
261,
756,
7960,
2130,
472,
278,
3268,
304,
2428,
29894,
895,
278,
1020,
457,
29872,
29889,
13,
3047,
4880,
304,
2999,
1736,
3246,
29892,
297,
278,
758,
314,
569,
304,
278,
1072,
8250,
29892,
360,
14851,
1754,
372,
2821,
393,
278,
3812,
306,
29899,
29929,
29947,
29941,
1122,
11039,
403,
1326,
12112,
363,
2060,
29892,
2602,
29892,
470,
14311,
5731,
800,
393,
4153,
29279,
304,
278,
317,
4330,
29924,
8368,
29915,
29879,
1746,
310,
6559,
29892,
4944,
727,
674,
367,
8210,
2428,
4924,
2645,
1269,
13733,
322,
278,
5703,
261,
6467,
28103,
599,
8018,
11780,
29889,
20175,
29892,
3620,
297,
3132,
3268,
14354,
508,
367,
1532,
23531,
322,
10824,
2501,
29240,
310,
278,
3812,
306,
29899,
29929,
29947,
29941,
29889,
1570,
322,
9251,
443,
1454,
968,
264,
3620,
508,
2337,
367,
20976,
1549,
278,
10223,
362,
322,
29240,
310,
263,
9120,
26101,
8402,
304,
278,
360,
6156,
29889,
13,
1576,
2114,
393,
278,
3812,
306,
29899,
29929,
29947,
29941,
1818,
367,
18397,
304,
278,
360,
6156,
322,
451,
304,
360,
14851,
338,
7282,
1363,
411,
977,
886,
18397,
304,
360,
14851,
29892,
727,
338,
5491,
263,
977,
292,
27684,
322,
278,
7037,
363,
3438,
368,
313,
2230,
322,
6909,
29897,
337,
24247,
491,
385,
297,
5943,
1821,
29892,
443,
28026,
322,
9815,
12139,
29889,
319,
360,
6156,
338,
385,
5375,
1058,
338,
12234,
901,
15579,
29889,
10575,
278,
360,
6156,
451,
2134,
345,
278,
2847,
3812,
306,
29899,
29929,
29947,
29941,
29892,
727,
881,
29892,
27581,
29892,
367,
901,
310,
385,
15130,
363,
278,
5703,
261,
322,
8368,
304,
2274,
278,
26101,
8402,
29915,
29879,
23503,
29879,
322,
304,
3867,
5684,
2472,
297,
263,
716,
29240,
29889,
13,
1576,
716,
317,
4330,
29924,
6418,
29911,
5751,
723,
2758,
5969,
14927,
8041,
1058,
505,
23588,
515,
3148,
4946,
1907,
297,
27131,
317,
4330,
29924,
4235,
304,
3933,
363,
385,
5684,
29871,
29906,
29946,
7378,
29889,
1094,
263,
1121,
29892,
278,
5751,
1818,
427,
2388,
465,
599,
17690,
310,
5400,
664,
15196,
4110,
29892,
3704,
1985,
472,
4654,
6263,
11840,
29889,
13466,
29892,
4152,
6397,
2722,
29892,
3704,
13315,
29892,
10643,
8799,
292,
470,
3633,
292,
29892,
723,
367,
1401,
1150,
287,
310,
3033,
6751,
5969,
14927,
9117,
8041,
29889,
19358,
8041,
508,
884,
14169,
491,
13442,
6694,
297,
6397,
2722,
5069,
5381,
1904,
337,
3687,
373,
23188,
22873,
304,
4654,
6263,
3132,
11840,
29889,
739,
338,
6397,
2722,
393,
19104,
373,
23188,
17162,
304,
4654,
6263,
11840,
393,
2367,
3082,
5381,
267,
263,
5100,
3321,
7636,
491,
13138,
963,
411,
1568,
4312,
8525,
4127,
29889,
2688,
881,
451,
367,
2175,
714,
515,
278,
716,
5751,
29991,
13,
29956,
406,
278,
11662,
29967,
7927,
29891,
414,
830,
635,
853,
621,
936,
297,
10319,
325,
29889,
8278,
29973,
13,
29967,
566,
479,
7169,
264,
29915,
29879,
1797,
29797,
2610,
29871,
29896,
29929,
29892,
29871,
29906,
29900,
29896,
29953,
2062,
326,
392,
292,
17202,
310,
10317,
310,
17181,
4307,
29891,
414,
363,
443,
621,
936,
7512,
338,
8717,
12449,
1363,
372,
947,
451,
1584,
2615,
393,
1009,
7512,
471,
443,
621,
936,
29889,
13,
29924,
987,
756,
2307,
1063,
3971,
1048,
26817,
7169,
264,
29915,
29879,
8515,
1797,
29889,
11386,
1394,
262,
476,
3127,
5155,
3692,
278,
16833,
508,
1584,
2411,
852,
11314,
1199,
4413,
373,
21006,
310,
11662,
29967,
4307,
29891,
414,
1058,
526,
451,
1083,
327,
873,
6631,
304,
278,
1206,
29889,
11386,
1383,
711,
2350,
317,
440,
481,
3417,
328,
399,
328,
29882,
423,
338,
925,
6832,
2197,
15041,
393,
278,
1797,
29892,
297,
6124,
304,
2062,
326,
392,
292,
11662,
29967,
1098,
272,
484,
952,
29892,
884,
28469,
575,
304,
525,
449,
29915,
278,
2983,
310,
901,
1135,
6893,
10405,
23957,
10070,
310,
278,
897,
14373,
9123,
363,
10517,
6614,
826,
15081,
313,
29928,
2477,
29909,
29897,
1824,
1058,
892,
16896,
29871,
29941,
1629,
17752,
2012,
310,
29871,
29906,
1629,
17752,
29889,
11386,
14317,
5682,
290,
7912,
947,
451,
1584,
1348,
278,
11662,
29967,
4307,
29891,
414,
1258,
3099,
2743,
29889,
13,
29902,
6446,
8661,
29889,
2803,
29915,
29879,
1106,
472,
27308,
29871,
29941,
29889,
29941,
310,
278,
3082,
2261,
7993,
8125,
6865,
310,
20031,
1281,
2199,
322,
278,
6590,
10319,
3295,
13326,
3821,
390,
2540,
310,
20031,
7512,
29892,
607,
26817,
7169,
264,
1304,
29892,
3412,
411,
263,
6534,
7689,
682,
1847,
310,
7928,
29879,
515,
5972,
12298,
29892,
363,
9138,
393,
278,
11662,
29967,
4307,
29891,
414,
892,
451,
8760,
1319,
304,
278,
8973,
29889,
3118,
310,
278,
21008,
11314,
936,
508,
29876,
787,
338,
393,
263,
25008,
756,
263,
13360,
310,
23794,
272,
304,
263,
9434,
17507,
29889,
319,
5688,
8125,
27308,
29871,
29941,
29889,
29941,
8128,
297,
8018,
760,
29901,
13,
29874,
29897,
319,
25008,
4091,
451,
1073,
11687,
29901,
13,
29896,
29897,
8561,
263,
2089,
3229,
310,
2114,
470,
4307,
304,
263,
9434,
17507,
470,
4418,
304,
1959,
263,
2089,
3229,
310,
2114,
470,
4307,
304,
263,
9434,
17507,
470,
4418,
304,
1959,
263,
2089,
3229,
310,
5518,
2114,
470,
4307,
9251,
1754,
304,
278,
9434,
17507,
491,
278,
25008,
29889,
13,
1576,
8018,
3104,
1080,
310,
278,
10319,
1873,
310,
27308,
29871,
29941,
29889,
29941,
526,
2788,
29901,
13,
29896,
29897,
8561,
263,
2089,
3229,
310,
5518,
2114,
470,
4307,
304,
263,
9434,
17507,
13,
29906,
29897,
29098,
304,
2313,
2226,
263,
2114,
304,
408,
9434,
17507,
746,
766,
25071,
338,
5181,
304,
4772,
6985,
292,
263,
22161,
470,
5227,
566,
352,
296,
1044,
29936,
13,
29945,
29897,
5957,
470,
671,
10757,
393,
278,
25008,
9906,
304
] | 1,024 |
CommonCrawl
| 0.209967 | 0.839012 | 0.34621 | 0.937021 |
be false.
In order for a lawyer to violate Rule 3.3, he or she must have knowingly made a false statement to the tribunal. Was there such a knowing violation of Rule 3.3 here?
On June 15, 2012, the Obama administration announced DACA that allowed young people who came to the United States prior to the age of 16 and had lived continuously since June 15, 2007, and were not in a lawful status, to be granted deferred action. On November 20, 2014, DHS Secretary Jeh Johnson issued a memo expanding DACA by changing the eligibility criteria to cover those who had come to the United States prior to January 1, 2010 instead of June 15, 2007 and by removing the maximum age limit of 31 ("Johnson Memo"). The Johnson Memo also lengthened the deferred action time from two to three years. The Johnson Memo further granted deferred action to parents of US citizens or resident children, known as the Deferred Action for Parent Accountability (DAPA), if they had arrived into the United States on or before January 1, 2010.
A group of states challenged the Johnson Memo in Texas v. USA by filing in a court in Brownsville, TX, where Judge Hanen sat who had already expressed strong views against the Obama administration on immigration. Judge Hanen granted a preliminary injunction on February 16, 2015 blocking DAPA and expanded DACA. Much has already been written to rebut the conclusions in this flawed decision, and the further flaw in the Fifth Circuit's affirmation of Judge Hanen's preliminary injunction. The preliminary injunction order did not expressly block the original DACA 2012 program. Qualified applicants thus continued to apply for DACA 2012 benefits. Under the terms of the Johnson Memo, qualified applicants under DACA 2012 started receiving grants of deferred action for 3 years instead of 2 years as of November 24, 2014.
Prior to the preliminary injunction of February 16, 2015, in conversations between Judge Hanen and DOJ attorneys, the DOJ attorneys indicated to the court that USCIS had not taken any actions pursuant to the Johnson Memo. Although actions had been taken since November 24, 2014 to grant three year deferred action periods rather than two years, those stemmed from the DACA 2012 program. They were also well publicized. The expanded DACA, which brought forward the entry date from June 15, 2007 to January 1, 2010, was to take effect on February 18, 2015. Thus, when DOJ attorneys denied that the government had not taken any actions regarding expanded DACA, it was well conceivable that issuing three year deferred action periods instead of two years were actions stemming from the DACA 2012 program and had nothing to do with the expanded DACA program, which had not gone into effect.
After the preliminary injunction was issued, which applied to "expansions (including any and all changes)" to DACA 2012, the DOJ filed an Advisory indicating that out of an abundance of caution it was informing the court that it had granted three year periods of deferred action under the original DACA 2012 guidelines in the event of any misunderstanding.
Given this lack of clarity, as well as the fact that DACA 2012 was never the subject of the lawsuit, could the DOJ attorneys have knowingly made a false statement to be sanctioned under Rule 3.3? This Ethics Committee of the American Immigration Lawyers Association first questioned whether this was so in 2015, but it has become even more important to assert whether there was a Rule 3.3 violation Judge Hanen's order.ABA Rule 1.0(f) defines the terms "knowingly," "known" or "knows" as "actual knowledge of the fact in question." Rule 1.0(f) goes on to state that a "person's knowledge may be inferred from circumstances." When the DOJ attorneys were giving an assurance to
|
[
367,
2089,
29889,
13,
797,
1797,
363,
263,
25008,
304,
5537,
403,
27308,
29871,
29941,
29889,
29941,
29892,
540,
470,
1183,
1818,
505,
1073,
11687,
1754,
263,
2089,
3229,
304,
278,
9434,
17507,
29889,
12547,
727,
1316,
263,
13797,
5537,
362,
310,
27308,
29871,
29941,
29889,
29941,
1244,
29973,
13,
2951,
5306,
29871,
29896,
29945,
29892,
29871,
29906,
29900,
29896,
29906,
29892,
278,
4250,
3304,
17517,
9326,
360,
2477,
29909,
393,
6068,
4123,
2305,
1058,
2996,
304,
278,
3303,
3900,
7536,
304,
278,
5046,
310,
29871,
29896,
29953,
322,
750,
10600,
3133,
5794,
1951,
5306,
29871,
29896,
29945,
29892,
29871,
29906,
29900,
29900,
29955,
29892,
322,
892,
451,
297,
263,
4307,
1319,
4660,
29892,
304,
367,
16896,
316,
14373,
3158,
29889,
1551,
3979,
29871,
29906,
29900,
29892,
29871,
29906,
29900,
29896,
29946,
29892,
360,
14851,
17719,
2581,
29882,
11717,
16610,
263,
2626,
29877,
7985,
292,
360,
2477,
29909,
491,
6480,
278,
560,
335,
4127,
16614,
304,
4612,
1906,
1058,
750,
2041,
304,
278,
3303,
3900,
7536,
304,
5490,
29871,
29896,
29892,
29871,
29906,
29900,
29896,
29900,
2012,
310,
5306,
29871,
29896,
29945,
29892,
29871,
29906,
29900,
29900,
29955,
322,
491,
11077,
278,
7472,
5046,
4046,
310,
29871,
29941,
29896,
4852,
11639,
1100,
8133,
29877,
2564,
450,
11717,
8133,
29877,
884,
3309,
6419,
278,
316,
14373,
3158,
931,
515,
1023,
304,
2211,
2440,
29889,
450,
11717,
8133,
29877,
4340,
16896,
316,
14373,
3158,
304,
11825,
310,
3148,
18363,
470,
20201,
4344,
29892,
2998,
408,
278,
897,
14373,
9123,
363,
22280,
16535,
3097,
313,
29928,
3301,
29909,
511,
565,
896,
750,
11977,
964,
278,
3303,
3900,
373,
470,
1434,
5490,
29871,
29896,
29892,
29871,
29906,
29900,
29896,
29900,
29889,
13,
29909,
2318,
310,
5922,
18066,
287,
278,
11717,
8133,
29877,
297,
10319,
325,
29889,
8278,
491,
977,
292,
297,
263,
8973,
297,
22415,
1983,
4909,
29892,
323,
29990,
29892,
988,
26817,
7169,
264,
3290,
1058,
750,
2307,
13384,
4549,
8386,
2750,
278,
4250,
3304,
17517,
373,
5198,
16783,
29889,
26817,
7169,
264,
16896,
263,
758,
2576,
3821,
10899,
651,
373,
6339,
29871,
29896,
29953,
29892,
29871,
29906,
29900,
29896,
29945,
23473,
360,
3301,
29909,
322,
17832,
360,
2477,
29909,
29889,
18927,
756,
2307,
1063,
3971,
304,
337,
4187,
278,
21204,
1080,
297,
445,
17422,
8734,
10608,
29892,
322,
278,
4340,
285,
10653,
297,
278,
29008,
386,
12594,
3121,
29915,
29879,
2756,
3568,
362,
310,
26817,
7169,
264,
29915,
29879,
758,
2576,
3821,
10899,
651,
29889,
450,
758,
2576,
3821,
10899,
651,
1797,
1258,
451,
4653,
368,
2908,
278,
2441,
360,
2477,
29909,
29871,
29906,
29900,
29896,
29906,
1824,
29889,
15146,
2164,
15576,
1934,
4550,
7572,
304,
3394,
363,
360,
2477,
29909,
29871,
29906,
29900,
29896,
29906,
23633,
29889,
7634,
278,
4958,
310,
278,
11717,
8133,
29877,
29892,
18698,
15576,
1934,
1090,
360,
2477,
29909,
29871,
29906,
29900,
29896,
29906,
4687,
13442,
867,
1934,
310,
316,
14373,
3158,
363,
29871,
29941,
2440,
2012,
310,
29871,
29906,
2440,
408,
310,
3979,
29871,
29906,
29946,
29892,
29871,
29906,
29900,
29896,
29946,
29889,
13,
29925,
13479,
304,
278,
758,
2576,
3821,
10899,
651,
310,
6339,
29871,
29896,
29953,
29892,
29871,
29906,
29900,
29896,
29945,
29892,
297,
9678,
800,
1546,
26817,
7169,
264,
322,
11662,
29967,
1098,
272,
484,
952,
29892,
278,
11662,
29967,
1098,
272,
484,
952,
18694,
304,
278,
8973,
393,
3148,
29907,
3235,
750,
451,
4586,
738,
8820,
12359,
29884,
424,
304,
278,
11717,
8133,
29877,
29889,
8512,
8820,
750,
1063,
4586,
1951,
3979,
29871,
29906,
29946,
29892,
29871,
29906,
29900,
29896,
29946,
304,
16690,
2211,
1629,
316,
14373,
3158,
23704,
3265,
1135,
1023,
2440,
29892,
1906,
20805,
2168,
515,
278,
360,
2477,
29909,
29871,
29906,
29900,
29896,
29906,
1824,
29889,
2688,
892,
884,
1532,
970,
1891,
29889,
450,
17832,
360,
2477,
29909,
29892,
607,
6296,
6375,
278,
6251,
2635,
515,
5306,
29871,
29896,
29945,
29892,
29871,
29906,
29900,
29900,
29955,
304,
5490,
29871,
29896,
29892,
29871,
29906,
29900,
29896,
29900,
29892,
471,
304,
2125,
2779,
373,
6339,
29871,
29896,
29947,
29892,
29871,
29906,
29900,
29896,
29945,
29889,
6549,
29892,
746,
11662,
29967,
1098,
272,
484,
952,
17935,
393,
278,
5874,
750,
451,
4586,
738,
8820,
11211,
17832,
360,
2477,
29909,
29892,
372,
471,
1532,
10628,
440,
519,
393,
17759,
292,
2211,
1629,
316,
14373,
3158,
23704,
2012,
310,
1023,
2440,
892,
8820,
20805,
4056,
515,
278,
360,
2477,
29909,
29871,
29906,
29900,
29896,
29906,
1824,
322,
750,
3078,
304,
437,
411,
278,
17832,
360,
2477,
29909,
1824,
29892,
607,
750,
451,
7695,
964,
2779,
29889,
13,
13555,
278,
758,
2576,
3821,
10899,
651,
471,
16610,
29892,
607,
7436,
304,
376,
4548,
550,
1080,
313,
18271,
738,
322,
599,
3620,
5513,
304,
360,
2477,
29909,
29871,
29906,
29900,
29896,
29906,
29892,
278,
11662,
29967,
934,
29881,
385,
2087,
1730,
706,
23941,
393,
714,
310,
385,
18666,
749,
310,
5777,
918,
372,
471,
1871,
292,
278,
8973,
393,
372,
750,
16896,
2211,
1629,
23704,
310,
316,
14373,
3158,
1090,
278,
2441,
360,
2477,
29909,
29871,
29906,
29900,
29896,
29906,
1410,
10652,
1475,
297,
278,
1741,
310,
738,
19818,
11235,
29889,
13,
29954,
5428,
445,
10225,
310,
7542,
537,
29892,
408,
1532,
408,
278,
2114,
393,
360,
2477,
29909,
29871,
29906,
29900,
29896,
29906,
471,
2360,
278,
4967,
310,
278,
4307,
29658,
29892,
1033,
278,
11662,
29967,
1098,
272,
484,
952,
505,
1073,
11687,
1754,
263,
2089,
3229,
304,
367,
9753,
428,
287,
1090,
27308,
29871,
29941,
29889,
29941,
29973,
910,
13772,
1199,
12930,
310,
278,
3082,
1954,
29885,
16783,
7927,
29891,
414,
7993,
937,
1139,
287,
3692,
445,
471,
577,
297,
29871,
29906,
29900,
29896,
29945,
29892,
541,
372,
756,
4953,
1584,
901,
4100,
304,
4974,
3692,
727,
471,
263,
27308,
29871,
29941,
29889,
29941,
5537,
362,
26817,
7169,
264,
29915,
29879,
1797,
29889,
2882,
29909,
27308,
29871,
29896,
29889,
29900,
29898,
29888,
29897,
17645,
278,
4958,
376,
28385,
11687,
1699,
376,
5203,
29908,
470,
376,
3959,
1242,
29908,
408,
376,
19304,
7134,
310,
278,
2114,
297,
1139,
1213,
27308,
29871,
29896,
29889,
29900,
29898,
29888,
29897,
5771,
373,
304,
2106,
393,
263,
376,
10532,
29915,
29879,
7134,
1122,
367,
10115,
1127,
515,
14209,
1213,
1932,
278,
11662,
29967,
1098,
272,
484,
952,
892,
6820,
385,
1223,
18541,
304
] | 1,024 |
CommonCrawl
| -0.214234 | 0.935049 | 0.333657 | 0.33494 |
from savu.plugins.plugin import Plugin
"""
.. module:: plugins_test
:platform: Unix
:synopsis: unittest test classes for plugins
.. moduleauthor:: Mark Basham <scientificsoftware@diamond.ac.uk>
"""
import unittest
from savu.plugins import utils as pu
from savu.test import test_utils as tu
from savu.plugins.cpu_plugin import CpuPlugin
base_class_name = "savu.plugins.plugin"
class PluginTest(unittest.TestCase):
def setUp(self):
self.plugin_name = base_class_name
def test_get_plugin(self):
try :
plugin = pu.load_plugin(self.plugin_name)
self.assertIsNotNone(plugin)
except ImportError as e:
print("Failed to run plugin test as libraries not available (%s), passing test" % (e))
pass
def test_process(self):
try:
plugin = pu.load_plugin(self.plugin_name)
if self.plugin_name == base_class_name:
self.assertRaises(NotImplementedError, plugin.process,
"test", "test", 1, 1)
return
# load appropriate data
data = tu.get_appropriate_input_data(plugin)
self.assertGreater(len(data), 0, "Cannot find appropriate test data")
# generate somewhere for the data to go
output = tu.get_appropriate_output_data(plugin, data)
self.assertGreater(len(output), 0,
"Cannot create appropriate output data")
plugin.set_parameters(None)
for i in range(len(data)):
plugin.run_process(data[i], output[i], ["CPU0"], 0)
print("Output from plugin under test ( %s ) is in %s" %
(plugin.name, output[i].backing_file.filename))
data[i].complete()
output[i].complete()
except ImportError as e:
print("Failed to run plugin test as libraries not available (%s), passing test" % (e))
pass
class CpuPluginWrapper(Plugin, CpuPlugin):
def __init__(self):
super(CpuPluginWrapper, self).__init__()
self.data = None
self.output = None
self.processes = None
self.process_number = None
def process(self, data, output, processes, process):
self.data = data
self.output = output
self.processes = processes
self.process_number = process
class CpuPluginTest(unittest.TestCase):
def setUp(self):
self.plugin = None
def test_run_process(self):
self.plugin = CpuPluginWrapper()
self.plugin.run_process("data", "out", ["CPU0"], 0)
self.assertEqual(self.plugin.processes, ["CPU0"])
self.assertEqual(self.plugin.process_number, 0)
self.plugin = CpuPluginWrapper()
self.plugin.run_process("data", "out",
["CPU0", "CPU1", "CPU2", "CPU3"], 0)
self.assertEqual(self.plugin.processes,
["CPU0", "CPU1", "CPU2", "CPU3"])
self.assertEqual(self.plugin.process_number, 0)
self.plugin = CpuPluginWrapper()
self.plugin.run_process("data", "out",
["CPU0", "CPU1", "CPU2", "CPU3"], 1)
self.assertEqual(self.plugin.processes,
["CPU0", "CPU1", "CPU2", "CPU3"])
self.assertEqual(self.plugin.process_number, 1)
self.plugin = CpuPluginWrapper()
|
[
515,
4048,
29884,
29889,
12800,
29889,
8582,
1053,
1858,
3851,
13,
13,
13,
15945,
29908,
13,
636,
3883,
1057,
18224,
29918,
1688,
13,
259,
584,
12120,
29901,
26663,
13,
259,
584,
19274,
15368,
29901,
443,
27958,
1243,
4413,
363,
18224,
13,
13,
636,
3883,
8921,
1057,
4485,
29677,
314,
529,
29879,
15566,
928,
20415,
29992,
29881,
2829,
898,
29889,
562,
29889,
2679,
29958,
13,
13,
15945,
29908,
13,
13,
5215,
443,
27958,
13,
13,
3166,
4048,
29884,
29889,
12800,
1053,
3667,
29879,
408,
2653,
13,
3166,
4048,
29884,
29889,
1688,
1053,
1243,
29918,
13239,
408,
5291,
13,
3166,
4048,
29884,
29889,
12800,
29889,
21970,
29918,
8582,
1053,
315,
3746,
16288,
13,
13,
3188,
29918,
1990,
29918,
978,
353,
376,
29879,
485,
29884,
29889,
12800,
29889,
8582,
29908,
13,
13,
13,
1990,
1858,
3851,
3057,
29898,
348,
27958,
29889,
3057,
8259,
1125,
13,
13,
1678,
822,
731,
3373,
29898,
1311,
1125,
13,
4706,
1583,
29889,
8582,
29918,
978,
353,
2967,
29918,
1990,
29918,
978,
13,
13,
1678,
822,
1243,
29918,
657,
29918,
8582,
29898,
1311,
1125,
13,
4706,
1018,
584,
13,
9651,
7079,
353,
2653,
29889,
1359,
29918,
8582,
29898,
1311,
29889,
8582,
29918,
978,
29897,
13,
9651,
1583,
29889,
9294,
3624,
3664,
8516,
29898,
8582,
29897,
13,
4706,
5174,
16032,
2392,
408,
321,
29901,
13,
9651,
1596,
703,
17776,
304,
1065,
7079,
1243,
408,
9562,
451,
3625,
313,
29995,
29879,
511,
6819,
1243,
29908,
1273,
313,
29872,
876,
13,
9651,
1209,
13,
13,
1678,
822,
1243,
29918,
5014,
29898,
1311,
1125,
13,
4706,
1018,
29901,
13,
9651,
7079,
353,
2653,
29889,
1359,
29918,
8582,
29898,
1311,
29889,
8582,
29918,
978,
29897,
13,
9651,
565,
1583,
29889,
8582,
29918,
978,
1275,
2967,
29918,
1990,
29918,
978,
29901,
13,
18884,
1583,
29889,
9294,
29934,
1759,
267,
29898,
3664,
1888,
2037,
287,
2392,
29892,
7079,
29889,
5014,
29892,
13,
462,
462,
29871,
376,
1688,
613,
376,
1688,
613,
29871,
29896,
29892,
29871,
29896,
29897,
13,
18884,
736,
13,
9651,
396,
2254,
8210,
848,
13,
9651,
848,
353,
5291,
29889,
657,
29918,
932,
6649,
403,
29918,
2080,
29918,
1272,
29898,
8582,
29897,
13,
9651,
1583,
29889,
9294,
25120,
1008,
29898,
2435,
29898,
1272,
511,
29871,
29900,
29892,
376,
29089,
1284,
8210,
1243,
848,
1159,
13,
13,
9651,
396,
5706,
9051,
363,
278,
848,
304,
748,
13,
9651,
1962,
353,
5291,
29889,
657,
29918,
932,
6649,
403,
29918,
4905,
29918,
1272,
29898,
8582,
29892,
848,
29897,
13,
9651,
1583,
29889,
9294,
25120,
1008,
29898,
2435,
29898,
4905,
511,
29871,
29900,
29892,
13,
462,
1669,
376,
29089,
1653,
8210,
1962,
848,
1159,
13,
13,
9651,
7079,
29889,
842,
29918,
16744,
29898,
8516,
29897,
13,
13,
9651,
363,
474,
297,
3464,
29898,
2435,
29898,
1272,
22164,
13,
18884,
7079,
29889,
3389,
29918,
5014,
29898,
1272,
29961,
29875,
1402,
1962,
29961,
29875,
1402,
6796,
6271,
29965,
29900,
12436,
29871,
29900,
29897,
13,
18884,
1596,
703,
6466,
515,
7079,
1090,
1243,
313,
1273,
29879,
1723,
338,
297,
1273,
29879,
29908,
1273,
13,
462,
418,
313,
8582,
29889,
978,
29892,
1962,
29961,
29875,
1822,
1627,
292,
29918,
1445,
29889,
9507,
876,
13,
13,
18884,
848,
29961,
29875,
1822,
8835,
580,
13,
18884,
1962,
29961,
29875,
1822,
8835,
580,
13,
13,
4706,
5174,
16032,
2392,
408,
321,
29901,
13,
9651,
1596,
703,
17776,
304,
1065,
7079,
1243,
408,
9562,
451,
3625,
313,
29995,
29879,
511,
6819,
1243,
29908,
1273,
313,
29872,
876,
13,
9651,
1209,
13,
13,
13,
1990,
315,
3746,
16288,
15646,
29898,
16288,
29892,
315,
3746,
16288,
1125,
13,
13,
1678,
822,
4770,
2344,
12035,
1311,
1125,
13,
4706,
2428,
29898,
29907,
3746,
16288,
15646,
29892,
1583,
467,
1649,
2344,
1649,
580,
13,
4706,
1583,
29889,
1272,
353,
6213,
13,
4706,
1583,
29889,
4905,
353,
6213,
13,
4706,
1583,
29889,
5014,
267,
353,
6213,
13,
4706,
1583,
29889,
5014,
29918,
4537,
353,
6213,
13,
13,
1678,
822,
1889,
29898,
1311,
29892,
848,
29892,
1962,
29892,
10174,
29892,
1889,
1125,
13,
4706,
1583,
29889,
1272,
353,
848,
13,
4706,
1583,
29889,
4905,
353,
1962,
13,
4706,
1583,
29889,
5014,
267,
353,
10174,
13,
4706,
1583,
29889,
5014,
29918,
4537,
353,
1889,
13,
13,
13,
1990,
315,
3746,
16288,
3057,
29898,
348,
27958,
29889,
3057,
8259,
1125,
13,
13,
1678,
822,
731,
3373,
29898,
1311,
1125,
13,
4706,
1583,
29889,
8582,
353,
6213,
13,
13,
1678,
822,
1243,
29918,
3389,
29918,
5014,
29898,
1311,
1125,
13,
4706,
1583,
29889,
8582,
353,
315,
3746,
16288,
15646,
580,
13,
4706,
1583,
29889,
8582,
29889,
3389,
29918,
5014,
703,
1272,
613,
376,
449,
613,
6796,
6271,
29965,
29900,
12436,
29871,
29900,
29897,
13,
4706,
1583,
29889,
9294,
9843,
29898,
1311,
29889,
8582,
29889,
5014,
267,
29892,
6796,
6271,
29965,
29900,
20068,
13,
4706,
1583,
29889,
9294,
9843,
29898,
1311,
29889,
8582,
29889,
5014,
29918,
4537,
29892,
29871,
29900,
29897,
13,
13,
4706,
1583,
29889,
8582,
353,
315,
3746,
16288,
15646,
580,
13,
4706,
1583,
29889,
8582,
29889,
3389,
29918,
5014,
703,
1272,
613,
376,
449,
613,
13,
462,
18884,
6796,
6271,
29965,
29900,
613,
376,
6271,
29965,
29896,
613,
376,
6271,
29965,
29906,
613,
376,
6271,
29965,
29941,
12436,
29871,
29900,
29897,
13,
4706,
1583,
29889,
9294,
9843,
29898,
1311,
29889,
8582,
29889,
5014,
267,
29892,
13,
462,
308,
6796,
6271,
29965,
29900,
613,
376,
6271,
29965,
29896,
613,
376,
6271,
29965,
29906,
613,
376,
6271,
29965,
29941,
20068,
13,
4706,
1583,
29889,
9294,
9843,
29898,
1311,
29889,
8582,
29889,
5014,
29918,
4537,
29892,
29871,
29900,
29897,
13,
13,
4706,
1583,
29889,
8582,
353,
315,
3746,
16288,
15646,
580,
13,
4706,
1583,
29889,
8582,
29889,
3389,
29918,
5014,
703,
1272,
613,
376,
449,
613,
13,
462,
18884,
6796,
6271,
29965,
29900,
613,
376,
6271,
29965,
29896,
613,
376,
6271,
29965,
29906,
613,
376,
6271,
29965,
29941,
12436,
29871,
29896,
29897,
13,
4706,
1583,
29889,
9294,
9843,
29898,
1311,
29889,
8582,
29889,
5014,
267,
29892,
13,
462,
308,
6796,
6271,
29965,
29900,
613,
376,
6271,
29965,
29896,
613,
376,
6271,
29965,
29906,
613,
376,
6271,
29965,
29941,
20068,
13,
4706,
1583,
29889,
9294,
9843,
29898,
1311,
29889,
8582,
29889,
5014,
29918,
4537,
29892,
29871,
29896,
29897,
13,
13,
4706,
1583,
29889,
8582,
353,
315,
3746,
16288,
15646,
580,
13
] | 1,024 |
Github
| -1.477328 | 0.242031 | -0.786211 | 0.08481 |
Begin your exploration of Fog a Dox by Bruce Pascoe by exploring the cover. Allow an open response and if prompting is necessary, draw attention to the artwork on the cover, the techniques used in the background and the colour of the fox. Encourage some discussion around the title of the book in relation to the image of the fox. Then go to the back cover where there is another image of the fox and read the blurb. Several themes for the book are introduced here. Discuss the purpose of the blurb and how well these three sentences fulfil that purpose in terms of enticing the reader. Students might like to discuss this after checking some blurbs on a few other texts in the class library.
Provide some time for students to work in groups to do some research with the purpose of reporting back to the group.
One group of students can research Bruce Pascoe beginning with the short bio in the back of the book.
One group can find out about the other books authored by Bruce Pascoe.
Students can read the book independently, however, as this is a fast moving engaging tale, if time allows it is recommended the book is read aloud in one or two sessions (maximum three). The book is written in third person, is in past tense and is sometimes 'poetic'. The narrative is gripping making it a perfect read aloud experience for older students.
If you choose to read aloud but have limited time, try to read to page 49 in the first reading. For the next reading to page 58 and then the remainder of the book. Do not ask questions or quiz the students. Read, close the book and move to the next lesson. Allow students to take in the narrative and personally reflect on what they have heard before coming together again the next week. It is likely you will overhear students talking about the book, what they have heard and what they are thinking about in their own time.
There are many aspects of this book that individual students might connect to personally across the themes of friendship, loneliness, prejudice, Indigenous perspective and nature. In groups of four or five invite students to talk about their personal reaction to this book and about anything that resonated with them. Conclude by inviting students to share their thoughts with the whole class.
Then in the same groups ask students to give a personal response to the following prompts.
'Albert cuts down trees, not because he doesn't like trees or even wants to, it is his job'.
Albert knew some words from his grandfather's mother's Maap language. He thought a few words were better than none.
Maria's mother wrung her hands and Maria knew that the wringing would 'squeeze tears' from her mother's eyes.
Dave wanted the company of a living creature, someone he could talk to.
'But people can't abide anything different and many can't relax until the difference is destroyed.' (p. 45).
Ask students to write a personal reaction to the text. This should not be a retelling but a response to the whole text or an event or character in the text and personal reflections coming from that. Given the wide-ranging possibilities, this task will be assessed on the connections being made, depth of understanding and clarity of the writing. While this is personal writing, ensure students understand it will be read and assessed by the teacher.
Ask if any students are willing to share the personal reflections prepared for the assessment task. If so select three or four to be read to the whole class to reinforce how each reader (listener) has a personal response.
Provide copies of the text and in small groups ask students to discuss the following quotes and statements from the text. Groups can use popplet to organise their ideas.
Albert is worried that Fog will be disliked because he is a fox. Engage in a discussion about stereotypes and prejudice.
This seemingly simple narrative provides opportunities for a range of responses that might be quite personal. The characters are diverse and each is quite complex, alone, and their paths cross throughout the text. Begin by having small groups explore each character by reading the text and extracting key vocabulary, quotes and events that the author uses to create these characters before doing a group character description. Ask students to explain how the author's language choices influence their personal response to that character.
Conclude by having a short whole class discussion so that each small group discusses the character they explored.
Working independently, ask students to explore the character of Dave in a similar way to the last activity. Remind them that the
|
[
14893,
596,
3902,
12418,
310,
383,
468,
263,
1938,
29916,
491,
18885,
10043,
1111,
29872,
491,
3902,
8253,
278,
4612,
29889,
29408,
385,
1722,
2933,
322,
565,
9508,
292,
338,
5181,
29892,
4216,
8570,
304,
278,
1616,
1287,
373,
278,
4612,
29892,
278,
13698,
1304,
297,
278,
3239,
322,
278,
12384,
310,
278,
1701,
29916,
29889,
11346,
283,
6617,
777,
10679,
2820,
278,
3611,
310,
278,
3143,
297,
8220,
304,
278,
1967,
310,
278,
1701,
29916,
29889,
1987,
748,
304,
278,
1250,
4612,
988,
727,
338,
1790,
1967,
310,
278,
1701,
29916,
322,
1303,
278,
1999,
9265,
29889,
21882,
963,
267,
363,
278,
3143,
526,
9129,
1244,
29889,
8565,
1558,
278,
6437,
310,
278,
1999,
9265,
322,
920,
1532,
1438,
2211,
25260,
6095,
1777,
393,
6437,
297,
4958,
310,
875,
18499,
278,
9591,
29889,
5088,
1237,
1795,
763,
304,
5353,
445,
1156,
8454,
777,
1999,
332,
5824,
373,
263,
2846,
916,
26442,
297,
278,
770,
3489,
29889,
13,
1184,
29894,
680,
777,
931,
363,
8041,
304,
664,
297,
6471,
304,
437,
777,
5925,
411,
278,
6437,
310,
23415,
1250,
304,
278,
2318,
29889,
13,
6716,
2318,
310,
8041,
508,
5925,
18885,
10043,
1111,
29872,
6763,
411,
278,
3273,
17799,
297,
278,
1250,
310,
278,
3143,
29889,
13,
6716,
2318,
508,
1284,
714,
1048,
278,
916,
8277,
4148,
287,
491,
18885,
10043,
1111,
29872,
29889,
13,
855,
566,
1237,
508,
1303,
278,
3143,
25499,
29892,
3138,
29892,
408,
445,
338,
263,
5172,
8401,
3033,
6751,
17694,
29892,
565,
931,
6511,
372,
338,
13622,
278,
3143,
338,
1303,
394,
2736,
297,
697,
470,
1023,
21396,
313,
27525,
398,
2211,
467,
450,
3143,
338,
3971,
297,
4654,
2022,
29892,
338,
297,
4940,
260,
1947,
322,
338,
6041,
525,
1129,
7492,
4286,
450,
15474,
1230,
338,
330,
374,
3262,
3907,
372,
263,
4922,
1303,
394,
2736,
7271,
363,
9642,
8041,
29889,
13,
3644,
366,
6755,
304,
1303,
394,
2736,
541,
505,
9078,
931,
29892,
1018,
304,
1303,
304,
1813,
29871,
29946,
29929,
297,
278,
937,
5183,
29889,
1152,
278,
2446,
5183,
304,
1813,
29871,
29945,
29947,
322,
769,
278,
21162,
310,
278,
3143,
29889,
1938,
451,
2244,
5155,
470,
439,
466,
278,
8041,
29889,
7523,
29892,
3802,
278,
3143,
322,
4337,
304,
278,
2446,
3109,
265,
29889,
29408,
8041,
304,
2125,
297,
278,
15474,
1230,
322,
22345,
9432,
373,
825,
896,
505,
6091,
1434,
6421,
4208,
1449,
278,
2446,
4723,
29889,
739,
338,
5517,
366,
674,
975,
354,
279,
8041,
9963,
1048,
278,
3143,
29892,
825,
896,
505,
6091,
322,
825,
896,
526,
7291,
1048,
297,
1009,
1914,
931,
29889,
13,
8439,
526,
1784,
21420,
310,
445,
3143,
393,
5375,
8041,
1795,
4511,
304,
22345,
4822,
278,
963,
267,
310,
27994,
29892,
301,
11064,
3335,
29892,
758,
17675,
625,
29892,
1894,
2101,
681,
18520,
322,
5469,
29889,
512,
6471,
310,
3023,
470,
5320,
2437,
568,
8041,
304,
5193,
1048,
1009,
7333,
19848,
304,
445,
3143,
322,
1048,
3099,
393,
27396,
630,
411,
963,
29889,
1281,
2325,
491,
2437,
11407,
8041,
304,
6232,
1009,
13133,
411,
278,
3353,
770,
29889,
13,
11760,
297,
278,
1021,
6471,
2244,
8041,
304,
2367,
263,
7333,
2933,
304,
278,
1494,
9508,
29879,
29889,
13,
29915,
2499,
2151,
5700,
29879,
1623,
10697,
29892,
451,
1363,
540,
1838,
29915,
29873,
763,
10697,
470,
1584,
10753,
304,
29892,
372,
338,
670,
4982,
4286,
13,
2499,
2151,
6363,
777,
3838,
515,
670,
4595,
22212,
29915,
29879,
5637,
29915,
29879,
3219,
481,
4086,
29889,
940,
2714,
263,
2846,
3838,
892,
2253,
1135,
5642,
29889,
13,
29924,
4568,
29915,
29879,
5637,
2358,
686,
902,
6567,
322,
6217,
6363,
393,
278,
2358,
292,
292,
723,
525,
29879,
802,
29872,
911,
20190,
29915,
515,
902,
5637,
29915,
29879,
5076,
29889,
13,
29928,
1351,
5131,
278,
5001,
310,
263,
8471,
23940,
29892,
4856,
540,
1033,
5193,
304,
29889,
13,
29915,
6246,
2305,
508,
29915,
29873,
633,
680,
3099,
1422,
322,
1784,
508,
29915,
29873,
26681,
2745,
278,
4328,
338,
14416,
6169,
313,
29886,
29889,
29871,
29946,
29945,
467,
13,
29909,
808,
8041,
304,
2436,
263,
7333,
19848,
304,
278,
1426,
29889,
910,
881,
451,
367,
263,
3240,
7807,
541,
263,
2933,
304,
278,
3353,
1426,
470,
385,
1741,
470,
2931,
297,
278,
1426,
322,
7333,
2143,
5942,
6421,
515,
393,
29889,
11221,
278,
9377,
29899,
29878,
9776,
24496,
29892,
445,
3414,
674,
367,
1223,
11517,
373,
278,
12368,
1641,
1754,
29892,
10809,
310,
8004,
322,
7542,
537,
310,
278,
5007,
29889,
5806,
445,
338,
7333,
5007,
29892,
9801,
8041,
2274,
372,
674,
367,
1303,
322,
1223,
11517,
491,
278,
15703,
29889,
13,
29909,
808,
565,
738,
8041,
526,
17762,
304,
6232,
278,
7333,
2143,
5942,
13240,
363,
278,
24809,
358,
3414,
29889,
960,
577,
1831,
2211,
470,
3023,
304,
367,
1303,
304,
278,
3353,
770,
304,
15561,
10118,
920,
1269,
9591,
313,
25894,
29897,
756,
263,
7333,
2933,
29889,
13,
1184,
29894,
680,
14591,
310,
278,
1426,
322,
297,
2319,
6471,
2244,
8041,
304,
5353,
278,
1494,
11839,
322,
9506,
515,
278,
1426,
29889,
1632,
4410,
508,
671,
772,
407,
1026,
304,
2894,
895,
1009,
7014,
29889,
13,
2499,
2151,
338,
6365,
1255,
393,
383,
468,
674,
367,
766,
5081,
287,
1363,
540,
338,
263,
1701,
29916,
29889,
2201,
482,
297,
263,
10679,
1048,
269,
12358,
327,
7384,
322,
758,
17675,
625,
29889,
13,
4013,
2833,
11687,
2560,
15474,
1230,
8128,
28602,
1907,
363,
263,
3464,
310,
20890,
393,
1795,
367,
3755,
7333,
29889,
450,
4890,
526,
16984,
322,
1269,
338,
3755,
4280,
29892,
7432,
29892,
322,
1009,
10898,
4891,
10106,
278,
1426,
29889,
14893,
491,
2534,
2319,
6471,
26987,
1269,
2931,
491,
5183,
278,
1426,
322,
6597,
292,
1820,
7931,
370,
352,
653,
29892,
11839,
322,
4959,
393,
278,
4148,
3913,
304,
1653,
1438,
4890,
1434,
2599,
263,
2318,
2931,
6139,
29889,
26579,
8041,
304,
5649,
920,
278,
4148,
29915,
29879,
4086,
19995,
9949,
1009,
7333,
2933,
304,
393,
2931,
29889,
13,
1168,
2325,
491,
2534,
263,
3273,
3353,
770,
10679,
577,
393,
1269,
2319,
2318,
5353,
267,
278,
2931,
896,
3902,
4395,
29889,
13,
5531,
292,
25499,
29892,
2244,
8041,
304,
26987,
278,
2931,
310,
17841,
297,
263,
2788,
982,
304,
278,
1833,
6354,
29889,
5240,
513,
963,
393,
278
] | 1,024 |
C4
| 0.818589 | -0.022717 | -0.098061 | 2.252493 |
reader first meets Dave when Albert recalled him getting one of Brim's pups and then later when he visited bringing a collar for Fog. Finally Dave was key in the rescue of Albert.
Students independently collect information from the text before writing a paragraph in their own words describing and summing up the character Dave.
Assess the planning sheet (above) and the final paragraph. Look for connections the students make to the author's craft and how the words, actions, events and relationships create the character.
Fog a Dox is a narrative. Check your students are familiar with the structure of a narrative and revise if necessary. This novel is organised into seven chapters. Draw the students' attention to the small pencil drawing and the title at the beginning of each chapter.
Explain the illustration and the connection to the content of the chapter.
Explain the chapter heading and the connection to the content of the chapter.
Each group shares their findings with the whole class. Finally ask each small group: Are there any illustrations or chapter headings you would change and why?
There are two main stories within this text that come together; Albert's story and Maria's story. These intersect towards the end at the bush hospital. Discuss how the author crafted this moment and the effect on the reader.
While Fog A Dox is a narrative, it contains some factual information. Have students examine how the author incorporates factual information.
Use page 25 where some information is given about the chough bird and page 103 as an example where Albert gives some information about the stingless bush bee. Share opinions and findings with the whole class.
Finally ask students to comment on the overall text structure used by Bruce Pascoe: his organisation of chapters, stories within the text and the snippets of factual information.
This text is rich in vocabulary and figurative language used so effectively that often a few words can transport the reader through this text.
In three groups have students engage in word searches. Ask all groups to note the page numbers of the words they find in preparation for the next activity.
Group 1: Search for words that might be more commonly used in the bush or considered slang. Use an audio dictionary or App to learn how to pronounce the words, e.g. Cambridge Dictionary.
Group 2: Search for poetic devices such as similes, personification and examples of imagery.
Group 3: Search for words that are unfamiliar.
Use an audio dictionary or App to learn how to pronounce the words, e.g. Cambridge Dictionary.
As the groups search for words, they can briefly discuss the words and the context in which they are found as they record them.
Do a whole class share, allowing each group to select some examples from their lists that demonstrate how vocabulary choices and figurative language can express shades of meaning. Then back in their groups, have students work through their lists recording the meaning for the word or phrase. First step is to return to the text using the recorded page numbers to check the context where the word of phrase has been used.
To conclude, remind the students that Albert learned some words from the Maap language from his grandfather. Conclude by asking students to work in small groups to make a list of Maap words occurring in the text.
In this 8-minute video, Louise Alexander introduces Uncle Ernie Grant's Framework for an Indigenous Perspective. This framework has six elements: Land, Language, Culture, Time, Place and Relationships.
2. Ask students to review the text and the lists again and identify words and phrases that make links to the six elements of an Indigenous Perspective. For example, aspects of Indigenous culture can be found throughout the text, particularly with the references to the characters' respect and love of the land and nature. Have student find these references and think about how these references occur.
Note the use of 'we', 'our people' and the connection to nature.
Friendship and love are strong themes in this book. These themes are presented in a number of ways between the various characters, sometimes in unexpected ways.
As a whole class brainstorm the characters connected by the theme of friendship or love.
This may include the list below and other examples found in the text.
Albert and his love for the bush and country.
Albert and the reciprocal love between him and Brim.
Brim and her love for her pups.
Friendship between Albert and Dave.
Together look at the brainstormed lists to map the friendship/love connections using the text. Use the IWB or mind mapping software.
For
|
[
9591,
937,
28103,
17841,
746,
10537,
337,
13998,
1075,
2805,
697,
310,
1771,
326,
29915,
29879,
2653,
567,
322,
769,
2678,
746,
540,
16669,
20794,
263,
784,
4675,
363,
383,
468,
29889,
9788,
17841,
471,
1820,
297,
278,
26429,
310,
10537,
29889,
13,
855,
566,
1237,
25499,
6314,
2472,
515,
278,
1426,
1434,
5007,
263,
14880,
297,
1009,
1914,
3838,
20766,
322,
2533,
4056,
701,
278,
2931,
17841,
29889,
13,
7900,
404,
278,
18987,
9869,
313,
27215,
29897,
322,
278,
2186,
14880,
29889,
7419,
363,
12368,
278,
8041,
1207,
304,
278,
4148,
29915,
29879,
25554,
322,
920,
278,
3838,
29892,
8820,
29892,
4959,
322,
21702,
1653,
278,
2931,
29889,
13,
29943,
468,
263,
1938,
29916,
338,
263,
15474,
1230,
29889,
5399,
596,
8041,
526,
9985,
411,
278,
3829,
310,
263,
15474,
1230,
322,
6664,
895,
565,
5181,
29889,
910,
9554,
338,
2894,
3368,
964,
9881,
10708,
2153,
29889,
18492,
278,
8041,
29915,
8570,
304,
278,
2319,
282,
3977,
309,
11580,
322,
278,
3611,
472,
278,
6763,
310,
1269,
16385,
29889,
13,
9544,
7420,
278,
8632,
362,
322,
278,
3957,
304,
278,
2793,
310,
278,
16385,
29889,
13,
9544,
7420,
278,
16385,
28435,
322,
278,
3957,
304,
278,
2793,
310,
278,
16385,
29889,
13,
9760,
2318,
29358,
1009,
1284,
886,
411,
278,
3353,
770,
29889,
9788,
2244,
1269,
2319,
2318,
29901,
4683,
727,
738,
8632,
800,
470,
16385,
2343,
886,
366,
723,
1735,
322,
2020,
29973,
13,
8439,
526,
1023,
1667,
15874,
2629,
445,
1426,
393,
2041,
4208,
29936,
10537,
29915,
29879,
5828,
322,
6217,
29915,
29879,
5828,
29889,
4525,
25869,
7113,
278,
1095,
472,
278,
27089,
13457,
29889,
8565,
1558,
920,
278,
4148,
25554,
287,
445,
3256,
322,
278,
2779,
373,
278,
9591,
29889,
13,
8809,
488,
383,
468,
319,
1938,
29916,
338,
263,
15474,
1230,
29892,
372,
3743,
777,
2114,
950,
2472,
29889,
6975,
8041,
25917,
920,
278,
4148,
11039,
1078,
2114,
950,
2472,
29889,
13,
11403,
1813,
29871,
29906,
29945,
988,
777,
2472,
338,
2183,
1048,
278,
521,
820,
11199,
322,
1813,
29871,
29896,
29900,
29941,
408,
385,
1342,
988,
10537,
4076,
777,
2472,
1048,
278,
380,
292,
2222,
27089,
367,
29872,
29889,
26849,
26971,
322,
1284,
886,
411,
278,
3353,
770,
29889,
13,
12881,
635,
2244,
8041,
304,
3440,
373,
278,
12463,
1426,
3829,
1304,
491,
18885,
10043,
1111,
29872,
29901,
670,
24788,
310,
10708,
2153,
29892,
15874,
2629,
278,
1426,
322,
278,
9830,
27421,
310,
2114,
950,
2472,
29889,
13,
4013,
1426,
338,
8261,
297,
7931,
370,
352,
653,
322,
28477,
1230,
4086,
1304,
577,
17583,
393,
4049,
263,
2846,
3838,
508,
8608,
278,
9591,
1549,
445,
1426,
29889,
13,
797,
2211,
6471,
505,
8041,
3033,
482,
297,
1734,
29645,
29889,
26579,
599,
6471,
304,
4443,
278,
1813,
3694,
310,
278,
3838,
896,
1284,
297,
10223,
362,
363,
278,
2446,
6354,
29889,
13,
4782,
29871,
29896,
29901,
11856,
363,
3838,
393,
1795,
367,
901,
15574,
1304,
297,
278,
27089,
470,
5545,
2243,
574,
29889,
4803,
385,
10348,
8600,
470,
2401,
304,
5110,
920,
304,
11504,
21543,
278,
3838,
29892,
321,
29889,
29887,
29889,
12585,
13343,
29889,
13,
4782,
29871,
29906,
29901,
11856,
363,
772,
7492,
9224,
1316,
408,
1027,
5475,
29892,
2022,
2450,
322,
6455,
310,
6382,
708,
29889,
13,
4782,
29871,
29941,
29901,
11856,
363,
3838,
393,
526,
443,
8302,
4447,
29889,
13,
11403,
385,
10348,
8600,
470,
2401,
304,
5110,
920,
304,
11504,
21543,
278,
3838,
29892,
321,
29889,
29887,
29889,
12585,
13343,
29889,
13,
2887,
278,
6471,
2740,
363,
3838,
29892,
896,
508,
23359,
5353,
278,
3838,
322,
278,
3030,
297,
607,
896,
526,
1476,
408,
896,
2407,
963,
29889,
13,
6132,
263,
3353,
770,
6232,
29892,
14372,
1269,
2318,
304,
1831,
777,
6455,
515,
1009,
8857,
393,
22222,
920,
7931,
370,
352,
653,
19995,
322,
28477,
1230,
4086,
508,
4653,
528,
3076,
310,
6593,
29889,
1987,
1250,
297,
1009,
6471,
29892,
505,
8041,
664,
1549,
1009,
8857,
16867,
278,
6593,
363,
278,
1734,
470,
16549,
29889,
3824,
4331,
338,
304,
736,
304,
278,
1426,
773,
278,
10478,
1813,
3694,
304,
1423,
278,
3030,
988,
278,
1734,
310,
16549,
756,
1063,
1304,
29889,
13,
1762,
17668,
29892,
1083,
513,
278,
8041,
393,
10537,
10972,
777,
3838,
515,
278,
3219,
481,
4086,
515,
670,
4595,
22212,
29889,
1281,
2325,
491,
6721,
8041,
304,
664,
297,
2319,
6471,
304,
1207,
263,
1051,
310,
3219,
481,
3838,
13920,
292,
297,
278,
1426,
29889,
13,
797,
445,
29871,
29947,
29899,
1195,
1082,
4863,
29892,
24417,
9428,
4547,
778,
29108,
1425,
2786,
18102,
29915,
29879,
16657,
363,
385,
1894,
2101,
681,
9034,
12645,
29889,
910,
6890,
756,
4832,
3161,
29901,
3172,
29892,
17088,
29892,
14062,
29892,
5974,
29892,
15484,
322,
6376,
800,
14587,
29889,
13,
29906,
29889,
26579,
8041,
304,
9076,
278,
1426,
322,
278,
8857,
1449,
322,
12439,
3838,
322,
12216,
2129,
393,
1207,
2988,
304,
278,
4832,
3161,
310,
385,
1894,
2101,
681,
9034,
12645,
29889,
1152,
1342,
29892,
21420,
310,
1894,
2101,
681,
9257,
508,
367,
1476,
10106,
278,
1426,
29892,
10734,
411,
278,
9282,
304,
278,
4890,
29915,
3390,
322,
5360,
310,
278,
2982,
322,
5469,
29889,
6975,
8368,
1284,
1438,
9282,
322,
1348,
1048,
920,
1438,
9282,
6403,
29889,
13,
9842,
278,
671,
310,
525,
705,
742,
525,
473,
2305,
29915,
322,
278,
3957,
304,
5469,
29889,
13,
27034,
355,
3527,
322,
5360,
526,
4549,
963,
267,
297,
445,
3143,
29889,
4525,
963,
267,
526,
9132,
297,
263,
1353,
310,
5837,
1546,
278,
5164,
4890,
29892,
6041,
297,
15668,
5837,
29889,
13,
2887,
263,
3353,
770,
4105,
2611,
555,
278,
4890,
6631,
491,
278,
10929,
310,
27994,
470,
5360,
29889,
13,
4013,
1122,
3160,
278,
1051,
2400,
322,
916,
6455,
1476,
297,
278,
1426,
29889,
13,
2499,
2151,
322,
670,
5360,
363,
278,
27089,
322,
4234,
29889,
13,
2499,
2151,
322,
278,
9522,
771,
1052,
5360,
1546,
1075,
322,
1771,
326,
29889,
13,
29933,
5632,
322,
902,
5360,
363,
902,
2653,
567,
29889,
13,
27034,
355,
3527,
1546,
10537,
322,
17841,
29889,
13,
29911,
12966,
1106,
472,
278,
4105,
2611,
555,
287,
8857,
304,
2910,
278,
27994,
29914,
417,
345,
12368,
773,
278,
1426,
29889,
4803,
278,
306,
29956,
29933,
470,
3458,
10417,
7047,
29889,
13,
2831
] | 1,024 |
C4
| 0.367557 | 0.117444 | 0.553327 | 3.031657 |
Microeconomics of collaboration and network configuration
Robert D. Weaver
Agricultural Economics, Sociology and Education
Purpose – The past decade has witnessed a trend toward unbundling of enterprises that were once highly integrated in vertical forms or horizontally as conglomerates. The economic forces behind these changes simultaneously enabled collaborative relationships that replaced command-control coordination. While such change has been widespread, the food industry serves as an example of an industry where such strategies have been incompletely pursued. This paper aims to provide a microeconomic explanation of three bases for the emergence of collaboration and network formation: transaction costs, interdependence in value creation processes, and shared resources. Design/methodology/approach – Within a setting of the food industry, a microeconomic theory of firm level choice of transactions is presented and extended to consider market level equilibrium in where persistent relationships are defined to compose an integrated economic network. Findings – The paper presents a framework for identification of the optimal pattern of relationships across firms to compose networks that could enhance the competitive advantage and the economic performance of the food sector. Originality/value – While performance of the food system has been the target of extensive public policy, that policy has typically viewed food enterprises as autonomous units operating through competitive markets. This paper provides a framework that highlights an economic rationale for collaboration across food enterprises that offers enhanced performance while maintaining essential aspects of competition.
British Food Journal
Weaver, R. D. (2009). Microeconomics of collaboration and network configuration. British Food Journal, 111(8), 746-761. https://doi.org/10.1108/00070700910980892
Weaver, Robert D. / Microeconomics of collaboration and network configuration. In: British Food Journal. 2009 ; Vol. 111, No. 8. pp. 746-761.
@article{759b00c7d31b4b389d518052987c6d71,
title = "Microeconomics of collaboration and network configuration",
abstract = "Purpose – The past decade has witnessed a trend toward unbundling of enterprises that were once highly integrated in vertical forms or horizontally as conglomerates. The economic forces behind these changes simultaneously enabled collaborative relationships that replaced command-control coordination. While such change has been widespread, the food industry serves as an example of an industry where such strategies have been incompletely pursued. This paper aims to provide a microeconomic explanation of three bases for the emergence of collaboration and network formation: transaction costs, interdependence in value creation processes, and shared resources. Design/methodology/approach – Within a setting of the food industry, a microeconomic theory of firm level choice of transactions is presented and extended to consider market level equilibrium in where persistent relationships are defined to compose an integrated economic network. Findings – The paper presents a framework for identification of the optimal pattern of relationships across firms to compose networks that could enhance the competitive advantage and the economic performance of the food sector. Originality/value – While performance of the food system has been the target of extensive public policy, that policy has typically viewed food enterprises as autonomous units operating through competitive markets. This paper provides a framework that highlights an economic rationale for collaboration across food enterprises that offers enhanced performance while maintaining essential aspects of competition.",
author = "Weaver, {Robert D.}",
journal = "British Food Journal",
publisher = "Emerald Group Publishing Ltd.",
Weaver, RD 2009, 'Microeconomics of collaboration and network configuration', British Food Journal, vol. 111, no. 8, pp. 746-761. https://doi.org/10.1108/00070700910980892
Microeconomics of collaboration and network configuration. / Weaver, Robert D.
In: British Food Journal, Vol. 111, No. 8, 08.08.2009, p. 746-761.
T1 - Microeconomics of collaboration and network configuration
AU - Weaver, Robert D.
N2 - Purpose – The past decade has witnessed
|
[
20140,
29872,
4599,
1199,
310,
24771,
322,
3564,
5285,
13,
9588,
2151,
360,
29889,
1334,
12483,
13,
29909,
629,
3953,
3631,
12884,
1199,
29892,
5329,
3002,
322,
13151,
13,
29925,
332,
4220,
785,
450,
4940,
316,
6332,
756,
16277,
287,
263,
534,
355,
11183,
443,
16293,
1847,
310,
3896,
558,
4637,
393,
892,
2748,
10712,
23387,
297,
11408,
7190,
470,
4029,
6753,
635,
408,
378,
3820,
12392,
1078,
29889,
450,
17407,
8249,
5742,
1438,
3620,
21699,
9615,
11465,
1230,
21702,
393,
8611,
1899,
29899,
6451,
29311,
3381,
29889,
5806,
1316,
1735,
756,
1063,
281,
2247,
29886,
949,
29892,
278,
9687,
13661,
19700,
408,
385,
1342,
310,
385,
13661,
988,
1316,
16650,
583,
505,
1063,
297,
5729,
29873,
873,
12359,
6742,
29889,
910,
5650,
263,
9893,
304,
3867,
263,
9200,
29872,
4599,
293,
8252,
310,
2211,
22561,
363,
278,
11176,
10238,
310,
24771,
322,
3564,
12409,
29901,
10804,
21544,
29892,
1006,
2716,
355,
663,
297,
995,
11265,
10174,
29892,
322,
7258,
7788,
29889,
12037,
29914,
5696,
3002,
29914,
9961,
496,
785,
23732,
263,
4444,
310,
278,
9687,
13661,
29892,
263,
9200,
29872,
4599,
293,
6368,
310,
9226,
3233,
7348,
310,
22160,
338,
9132,
322,
10410,
304,
2050,
9999,
3233,
26440,
297,
988,
28152,
21702,
526,
3342,
304,
27435,
385,
23387,
17407,
3564,
29889,
10987,
886,
785,
450,
5650,
22981,
263,
6890,
363,
29769,
310,
278,
14413,
4766,
310,
21702,
4822,
13734,
1516,
304,
27435,
14379,
393,
1033,
26371,
749,
278,
5100,
3321,
10631,
322,
278,
17407,
4180,
310,
278,
9687,
17535,
29889,
8533,
537,
29914,
1767,
785,
5806,
4180,
310,
278,
9687,
1788,
756,
1063,
278,
3646,
310,
20607,
970,
8898,
29892,
393,
8898,
756,
12234,
24774,
9687,
3896,
558,
4637,
408,
28273,
681,
10340,
13598,
1549,
5100,
3321,
2791,
1691,
29889,
910,
5650,
8128,
263,
6890,
393,
12141,
29879,
385,
17407,
364,
11464,
363,
24771,
4822,
9687,
3896,
558,
4637,
393,
16688,
427,
29308,
4180,
1550,
7344,
292,
18853,
21420,
310,
13888,
29889,
13,
29933,
768,
728,
25453,
8237,
13,
4806,
12483,
29892,
390,
29889,
360,
29889,
313,
29906,
29900,
29900,
29929,
467,
20140,
29872,
4599,
1199,
310,
24771,
322,
3564,
5285,
29889,
4908,
25453,
8237,
29892,
29871,
29896,
29896,
29896,
29898,
29947,
511,
29871,
29955,
29946,
29953,
29899,
29955,
29953,
29896,
29889,
2045,
597,
1867,
29875,
29889,
990,
29914,
29896,
29900,
29889,
29896,
29896,
29900,
29947,
29914,
29900,
29900,
29900,
29955,
29900,
29955,
29900,
29900,
29929,
29896,
29900,
29929,
29947,
29900,
29947,
29929,
29906,
13,
4806,
12483,
29892,
4755,
360,
29889,
847,
20140,
29872,
4599,
1199,
310,
24771,
322,
3564,
5285,
29889,
512,
29901,
4908,
25453,
8237,
29889,
29871,
29906,
29900,
29900,
29929,
2056,
3684,
29889,
29871,
29896,
29896,
29896,
29892,
1939,
29889,
29871,
29947,
29889,
6499,
29889,
29871,
29955,
29946,
29953,
29899,
29955,
29953,
29896,
29889,
13,
29992,
7914,
29912,
29955,
29945,
29929,
29890,
29900,
29900,
29883,
29955,
29881,
29941,
29896,
29890,
29946,
29890,
29941,
29947,
29929,
29881,
29945,
29896,
29947,
29900,
29945,
29906,
29929,
29947,
29955,
29883,
29953,
29881,
29955,
29896,
29892,
13,
3257,
353,
376,
29924,
2357,
29872,
4599,
1199,
310,
24771,
322,
3564,
5285,
613,
13,
16595,
353,
376,
29925,
332,
4220,
785,
450,
4940,
316,
6332,
756,
16277,
287,
263,
534,
355,
11183,
443,
16293,
1847,
310,
3896,
558,
4637,
393,
892,
2748,
10712,
23387,
297,
11408,
7190,
470,
4029,
6753,
635,
408,
378,
3820,
12392,
1078,
29889,
450,
17407,
8249,
5742,
1438,
3620,
21699,
9615,
11465,
1230,
21702,
393,
8611,
1899,
29899,
6451,
29311,
3381,
29889,
5806,
1316,
1735,
756,
1063,
281,
2247,
29886,
949,
29892,
278,
9687,
13661,
19700,
408,
385,
1342,
310,
385,
13661,
988,
1316,
16650,
583,
505,
1063,
297,
5729,
29873,
873,
12359,
6742,
29889,
910,
5650,
263,
9893,
304,
3867,
263,
9200,
29872,
4599,
293,
8252,
310,
2211,
22561,
363,
278,
11176,
10238,
310,
24771,
322,
3564,
12409,
29901,
10804,
21544,
29892,
1006,
2716,
355,
663,
297,
995,
11265,
10174,
29892,
322,
7258,
7788,
29889,
12037,
29914,
5696,
3002,
29914,
9961,
496,
785,
23732,
263,
4444,
310,
278,
9687,
13661,
29892,
263,
9200,
29872,
4599,
293,
6368,
310,
9226,
3233,
7348,
310,
22160,
338,
9132,
322,
10410,
304,
2050,
9999,
3233,
26440,
297,
988,
28152,
21702,
526,
3342,
304,
27435,
385,
23387,
17407,
3564,
29889,
10987,
886,
785,
450,
5650,
22981,
263,
6890,
363,
29769,
310,
278,
14413,
4766,
310,
21702,
4822,
13734,
1516,
304,
27435,
14379,
393,
1033,
26371,
749,
278,
5100,
3321,
10631,
322,
278,
17407,
4180,
310,
278,
9687,
17535,
29889,
8533,
537,
29914,
1767,
785,
5806,
4180,
310,
278,
9687,
1788,
756,
1063,
278,
3646,
310,
20607,
970,
8898,
29892,
393,
8898,
756,
12234,
24774,
9687,
3896,
558,
4637,
408,
28273,
681,
10340,
13598,
1549,
5100,
3321,
2791,
1691,
29889,
910,
5650,
8128,
263,
6890,
393,
12141,
29879,
385,
17407,
364,
11464,
363,
24771,
4822,
9687,
3896,
558,
4637,
393,
16688,
427,
29308,
4180,
1550,
7344,
292,
18853,
21420,
310,
13888,
19602,
13,
8921,
353,
376,
4806,
12483,
29892,
426,
9588,
2151,
360,
5003,
613,
13,
29926,
4659,
353,
376,
29933,
768,
728,
25453,
8237,
613,
13,
23679,
261,
353,
376,
29923,
1050,
2741,
6431,
19088,
19806,
19602,
13,
4806,
12483,
29892,
390,
29928,
29871,
29906,
29900,
29900,
29929,
29892,
525,
29924,
2357,
29872,
4599,
1199,
310,
24771,
322,
3564,
5285,
742,
4908,
25453,
8237,
29892,
1700,
29889,
29871,
29896,
29896,
29896,
29892,
694,
29889,
29871,
29947,
29892,
6499,
29889,
29871,
29955,
29946,
29953,
29899,
29955,
29953,
29896,
29889,
2045,
597,
1867,
29875,
29889,
990,
29914,
29896,
29900,
29889,
29896,
29896,
29900,
29947,
29914,
29900,
29900,
29900,
29955,
29900,
29955,
29900,
29900,
29929,
29896,
29900,
29929,
29947,
29900,
29947,
29929,
29906,
13,
29924,
2357,
29872,
4599,
1199,
310,
24771,
322,
3564,
5285,
29889,
847,
1334,
12483,
29892,
4755,
360,
29889,
13,
797,
29901,
4908,
25453,
8237,
29892,
3684,
29889,
29871,
29896,
29896,
29896,
29892,
1939,
29889,
29871,
29947,
29892,
29871,
29900,
29947,
29889,
29900,
29947,
29889,
29906,
29900,
29900,
29929,
29892,
282,
29889,
29871,
29955,
29946,
29953,
29899,
29955,
29953,
29896,
29889,
13,
29911,
29896,
448,
20140,
29872,
4599,
1199,
310,
24771,
322,
3564,
5285,
13,
25951,
448,
1334,
12483,
29892,
4755,
360,
29889,
13,
29940,
29906,
448,
15247,
4220,
785,
450,
4940,
316,
6332,
756,
16277,
287
] | 1,024 |
CommonCrawl
| 0.542807 | 1.474927 | 0.608917 | 1.761721 |
Every week, we post seven to ten news stories from Israel with a suggested prayer focus and scripture for each one, guiding readers how to pray for Israel's most urgent needs. This Prayer Update is also sent to over 18,000 subscribers every Friday by e-mail. Please contact us at intl.office@bridgesforpeace.com if you would like to receive this Prayer Update by e-mail.
Bennett, Biden Discuss "Steps to Block" Iran Nuke Program
Israel, Bahrain Ink Historic Defense Cooperation Agreement
Israel Launches US $70 Million to Help Propel More Arabs into Tech Industry
US Hopes to Seal Israeli–Lebanese Maritime Agreement to Unlock Natural Gas for Lebanon
Anti-Semitism Soars to Record High in UK
Erdoğan: Turkey, Israel to Discuss Energy Cooperation, Transporting Gas to Europe
Rocket Sirens Wail as Syrian Antiaircraft Missile Explodes over Northern Israel
African Union Divided over Israel's "Observer Status"
UAE Textbooks Teach Tolerance as Palestinian Curriculum Calls for Violence, Anti-Jewish Incitement
by Joshua Spurlock
Israeli and American flags fly side by side.
Monday, 7 February 2022 | Israeli Prime Minister Naftali Bennett on Sunday discussed "steps to block the Iranian nuclear program" in a call with United States President Joe Biden, just hours after Bennett indicated that his country retains the right to attack Iran "with or without" a renewed nuclear deal between Iran and the major world powers.
The Israeli leader's conversation with Biden on Sunday evening covered a range of topics, including "regional challenges, especially the growing Iranian aggression," according to a summary of the call published by Israel.
That "growing Iranian aggression" was certainly on Bennett's mind earlier in the day during Israel's weekly cabinet meeting, when the Prime Minister made it clear that a nuclear deal will make things worse, not better.
"Our position is well-known and clear: An agreement—according to the apparent terms—will damage the ability to deal with the nuclear program," said Bennett in comments released by his office. "Whoever thinks that an agreement will increase stability—is mistaken. It will temporarily delay enrichment, but all of us in the region will pay a heavy, disproportionate price for it."
Bennett later issued a veiled threat to attack Iran as needed, regardless of the outcome of the nuclear negotiations. "We are currently closing gaps and building up Israel's military strength for years and even decades to come. Israel will maintain freedom of action in any case, with or without an agreement," said Bennett.
The implied warning follows comments last week in which Bennett spelled out some of the ways his country is already working to "weaken" Iran—including "cyber, overt and covert operations"—and as news reports continue to indicate Israel is preparing to launch a military strike on Iran's nuclear program if necessary.
In the speech last Tuesday, also published by his office, Bennett laid out the dangerous impact of sanctions relief for Iran as part of a nuclear agreement.
"If an agreement is signed, and the flow of dollars is renewed, we know that their aggressive behavior will only intensify. We in Israel are ready. We will continue to stand against them in every way," said Bennett.
"No agreement will prevent us from protecting the citizens of Israel. The Israeli strategy remains the same in the case of an agreement, which in any case only buys a very short amount of time until the sunset clause, or in the event that there is no agreement. In both cases, our campaign continues."
The strong words from Bennett come as the US said they had reached the "final stretch" of talks with Iran. In comments on background last Monday, an unnamed US senior State Department official told reporters that there are only "a handful of weeks left to get a deal" before Iran's nuclear advances make the old terms of the preexisting deal obsolete.
That deal, known as the Joint Comprehensive Plan of Action (JCPOA), was signed in 2015, but the US withdrew from the accord in 2018, after a failed attempt to strengthen the restrictions on Iran. In response, Iran walked away from their own obligations under the JCPOA and then some, making disturb
|
[
7569,
4723,
29892,
591,
1400,
9881,
304,
3006,
9763,
15874,
515,
11996,
411,
263,
7829,
27402,
8569,
322,
2471,
545,
363,
1269,
697,
29892,
1410,
4821,
22176,
920,
304,
12475,
363,
11996,
29915,
29879,
1556,
5065,
5362,
4225,
29889,
910,
349,
764,
261,
10318,
338,
884,
2665,
304,
975,
29871,
29896,
29947,
29892,
29900,
29900,
29900,
21696,
2596,
1432,
28728,
491,
321,
29899,
2549,
29889,
3529,
6958,
502,
472,
938,
29880,
29889,
20205,
29992,
19515,
2710,
1454,
412,
815,
29889,
510,
565,
366,
723,
763,
304,
7150,
445,
349,
764,
261,
10318,
491,
321,
29899,
2549,
29889,
13,
20841,
20764,
29892,
350,
3615,
8565,
1558,
376,
7789,
567,
304,
15658,
29908,
14883,
12487,
446,
7835,
13,
3624,
9058,
29892,
15174,
6038,
512,
29895,
17939,
5282,
1947,
3189,
16453,
4059,
276,
882,
13,
3624,
9058,
997,
3322,
267,
3148,
395,
29955,
29900,
8649,
291,
304,
22305,
1019,
13111,
5853,
10387,
29879,
964,
1920,
305,
12157,
719,
13,
3308,
20727,
267,
304,
922,
284,
22895,
5037,
29994,
29931,
774,
273,
968,
1085,
21998,
4059,
276,
882,
304,
853,
908,
18385,
19141,
363,
9388,
19930,
13,
13448,
29875,
29899,
29903,
21976,
1608,
1105,
1503,
304,
14164,
5057,
297,
10261,
13,
2110,
1867,
30200,
273,
29901,
26459,
29892,
11996,
304,
8565,
1558,
24836,
3189,
16453,
29892,
15710,
292,
19141,
304,
4092,
13,
29934,
9749,
6290,
575,
399,
737,
408,
8713,
6392,
5459,
423,
12664,
4750,
488,
1222,
572,
2631,
975,
14299,
11996,
13,
29909,
1341,
2185,
7761,
4910,
2618,
975,
11996,
29915,
29879,
376,
28066,
16034,
29908,
13,
29965,
16036,
3992,
12733,
1920,
496,
16977,
261,
749,
408,
22053,
262,
713,
315,
1038,
12906,
398,
315,
4293,
363,
10630,
324,
663,
29892,
18473,
29899,
29967,
809,
728,
9266,
17334,
13,
1609,
22838,
3357,
1706,
332,
908,
13,
3624,
336,
5037,
322,
3082,
13449,
11340,
2625,
491,
2625,
29889,
13,
29924,
898,
388,
29892,
29871,
29955,
6339,
29871,
29906,
29900,
29906,
29906,
891,
22895,
5037,
15512,
7668,
405,
2051,
2606,
4111,
20764,
373,
16340,
15648,
376,
24530,
304,
2908,
278,
14883,
713,
20346,
1824,
29908,
297,
263,
1246,
411,
3303,
3900,
7178,
11131,
350,
3615,
29892,
925,
6199,
1156,
4111,
20764,
18694,
393,
670,
4234,
11551,
29879,
278,
1492,
304,
5337,
14883,
376,
2541,
470,
1728,
29908,
263,
23011,
287,
20346,
5376,
1546,
14883,
322,
278,
4655,
3186,
10801,
29889,
13,
1576,
22895,
5037,
11822,
29915,
29879,
14983,
411,
350,
3615,
373,
16340,
11005,
10664,
263,
3464,
310,
23820,
29892,
3704,
376,
1727,
1848,
18066,
267,
29892,
7148,
278,
15678,
14883,
713,
946,
11476,
1699,
5034,
304,
263,
15837,
310,
278,
1246,
6369,
491,
11996,
29889,
13,
7058,
376,
29887,
798,
292,
14883,
713,
946,
11476,
29908,
471,
8959,
373,
4111,
20764,
29915,
29879,
3458,
8859,
297,
278,
2462,
2645,
11996,
29915,
29879,
4723,
368,
28966,
11781,
29892,
746,
278,
15512,
7668,
1754,
372,
2821,
393,
263,
20346,
5376,
674,
1207,
2712,
15029,
29892,
451,
2253,
29889,
13,
29908,
29949,
332,
2602,
338,
1532,
29899,
5203,
322,
2821,
29901,
530,
17327,
30003,
5753,
3278,
304,
278,
20295,
4958,
30003,
14043,
18658,
278,
11509,
304,
5376,
411,
278,
20346,
1824,
1699,
1497,
4111,
20764,
297,
6589,
5492,
491,
670,
8034,
29889,
376,
22110,
1310,
22405,
393,
385,
17327,
674,
7910,
25806,
30003,
275,
25257,
29889,
739,
674,
5382,
6275,
9055,
427,
4018,
358,
29892,
541,
599,
310,
502,
297,
278,
5120,
674,
5146,
263,
9416,
29892,
766,
771,
637,
291,
403,
8666,
363,
372,
1213,
13,
20841,
20764,
2678,
16610,
263,
2453,
2356,
28469,
304,
5337,
14883,
408,
4312,
29892,
17126,
310,
278,
21957,
310,
278,
20346,
27214,
800,
29889,
376,
4806,
526,
5279,
14382,
330,
2547,
322,
5214,
701,
11996,
29915,
29879,
9121,
9324,
363,
2440,
322,
1584,
1602,
3076,
304,
2041,
29889,
11996,
674,
7344,
16082,
310,
3158,
297,
738,
1206,
29892,
411,
470,
1728,
385,
17327,
1699,
1497,
4111,
20764,
29889,
13,
1576,
2411,
2957,
9177,
4477,
6589,
1833,
4723,
297,
607,
4111,
20764,
805,
14356,
714,
777,
310,
278,
5837,
670,
4234,
338,
2307,
1985,
304,
376,
705,
9424,
29908,
14883,
30003,
18271,
376,
1270,
495,
29892,
975,
29873,
322,
4612,
29873,
6931,
29908,
30003,
392,
408,
9763,
13676,
6773,
304,
12266,
11996,
338,
10223,
292,
304,
6826,
263,
9121,
21283,
373,
14883,
29915,
29879,
20346,
1824,
565,
5181,
29889,
13,
797,
278,
12032,
1833,
323,
1041,
3250,
29892,
884,
6369,
491,
670,
8034,
29892,
4111,
20764,
12341,
714,
278,
18215,
10879,
310,
9753,
1953,
18892,
363,
14883,
408,
760,
310,
263,
20346,
17327,
29889,
13,
29908,
3644,
385,
17327,
338,
8794,
29892,
322,
278,
4972,
310,
17208,
338,
23011,
287,
29892,
591,
1073,
393,
1009,
946,
3663,
573,
6030,
674,
871,
12838,
1598,
29889,
1334,
297,
11996,
526,
7960,
29889,
1334,
674,
6773,
304,
2317,
2750,
963,
297,
1432,
982,
1699,
1497,
4111,
20764,
29889,
13,
29908,
3782,
17327,
674,
5557,
502,
515,
12566,
292,
278,
18363,
310,
11996,
29889,
450,
22895,
5037,
13705,
9242,
278,
1021,
297,
278,
1206,
310,
385,
17327,
29892,
607,
297,
738,
1206,
871,
1321,
952,
263,
1407,
3273,
5253,
310,
931,
2745,
278,
6575,
842,
11845,
29892,
470,
297,
278,
1741,
393,
727,
338,
694,
17327,
29889,
512,
1716,
4251,
29892,
1749,
11531,
18172,
1213,
13,
1576,
4549,
3838,
515,
4111,
20764,
2041,
408,
278,
3148,
1497,
896,
750,
7450,
278,
376,
8394,
16116,
29908,
310,
5969,
2039,
411,
14883,
29889,
512,
6589,
373,
3239,
1833,
27822,
29892,
385,
443,
17514,
3148,
16336,
4306,
10317,
6221,
5429,
1634,
272,
2153,
393,
727,
526,
871,
376,
29874,
1361,
1319,
310,
11405,
2175,
304,
679,
263,
5376,
29908,
1434,
14883,
29915,
29879,
20346,
3061,
2925,
1207,
278,
2030,
4958,
310,
278,
758,
735,
15423,
5376,
704,
2170,
371,
29889,
13,
7058,
5376,
29892,
2998,
408,
278,
435,
2461,
422,
1457,
29882,
6270,
8402,
310,
9123,
313,
29967,
6271,
29949,
29909,
511,
471,
8794,
297,
29871,
29906,
29900,
29896,
29945,
29892,
541,
278,
3148,
411,
29881,
3973,
515,
278,
15017,
297,
29871,
29906,
29900,
29896,
29947,
29892,
1156,
263,
5229,
4218,
304,
9324,
264,
278,
25091,
373,
14883,
29889,
512,
2933,
29892,
14883,
17096,
3448,
515,
1009,
1914,
10788,
800,
1090,
278,
435,
6271,
29949,
29909,
322,
769,
777,
29892,
3907,
29543
] | 1,024 |
CommonCrawl
| 0.489825 | 0.428912 | 0.634022 | 0.136354 |
ing progress towards enriching enough uranium to serve as nuclear fuel for a weapon.
The US and other major world powers have been trying to resurrect the JCPOA for around 10 months without success, as talks continue in Vienna. The Senior US State Department official, whose comments were published by the State Department, claimed last week that "seniormost former Israeli officials" have called the US withdrawal from the JCPOA a "terrible mistake."
By contrast, the current leadership in Israel certainly does not believe that returning to the JCPOA would be better—and the current Iranian actions only confirm this. "The greatest threat against the State of Israel is Iran," said Bennett in the cabinet meeting on Sunday. "…In recent weeks, precisely during the negotiations, Iran is increasing its aggression and repeatedly using terrorism in the region, as you all have seen. This is how you conduct negotiations, Tehran-style."
Despite the obvious disagreement between Bennett and Biden on how to best handle Iran, in the call on Sunday, Bennett thanked Biden for "his steadfast support of Israel as well as the support of his entire administration, especially with regards to American assistance towards the Iron Dome." He also invited Biden and his wife to visit Israel.
His future outlook for Iran was not so rosy. In the cabinet meeting, Bennett warned, "Every knowledgeable investor knows that investing in the Iranian regime, in the Iranian economy, is an unwise investment in both the long and intermediate terms."
Source: (This article was originally published by the Mideast Update on January 6, 2022. Time-related language has been modified to reflect our publication today.)
Photo Credit: U.S. Embassy Jerusalem/Flickr.com
Photo License: Flickr
Pray that the Lord will bless Israel's leaders with great wisdom as they chart a course of action regarding Iran. Cry out to the Lord for a solution that will stop Iranian nuclear ambitions once and for all while also bringing an end to their support of terrorism in the region.
For the LORD gives wisdom; from His mouth come knowledge and understanding.
- Proverbs 2:6
by Liad Osmo ~ Ynetnews
Bahrain skyline
Friday, 4 February 2022 | Defense Minister Benny Gantz met on Thursday with King Hamad Bin Isa al Khalifa of Bahrain in his palace during [Gantz's] first official visit to the kingdom, where he signed a defense cooperation agreement.
Gantz also met with Crown Prince and Prime Minister Salman Bin Hamad al Khalifa as well as the commander of the Bahrain armed forces and Defense Minister Abdulla bin Hasan Al Nuaimi, who co-signed the historic Memorandum of Understanding (MOU) between the defense establishments of the two countries.
The MOU comes 18 months after the Abraham Accords formalizing Israel's relations with Bahrain and the UAE [United Arab Emirates] and will promote intelligence sharing, joint military drills and closer ties between the defense industries in both nations, and was the pinnacle of the visit, along with the successful meeting with the king and the Bahraini leaders.
Sources close to Gantz expressed their optimism that the visit will help advance a close and intimate relationship between Israel and Bahrain.
Gantz, who was received with honors, thanked the king for his hospitality and his support of broadening bilateral ties and the economic and civilian agreements signed thus far, solidifying the defense cooperation.
"The strategic cooperation that we are bringing to a new high point today with the signing of this agreement and with the important meeting with the king is the continuation of the historic Abraham Accords and of the developing relationship between our nations and people," Gantz said at the ceremony.
"Only a year after the signing of the accords, we are already here signing a significant security agreement, which will allow robust cooperation and strengthen the security of both countries and of the entire region," the defense minister said.
Speaking at a joint Israeli–Emirati–Bahraini panel during an annual conference of the Institute for National Security Studies last month, Bahrain's Foreign Minister Abdullatif ben Rashid Al Zayani said the country saw eye to eye on many aspects of the Iranian expansion in the region.
"A common regional position on these issues will allow us to have greater influence over the United States," he said, stressing that the issues were of high importance for
|
[
292,
6728,
7113,
427,
4018,
292,
3307,
318,
661,
1974,
304,
9080,
408,
20346,
26413,
363,
263,
28639,
29889,
13,
1576,
3148,
322,
916,
4655,
3186,
10801,
505,
1063,
1811,
304,
620,
332,
1621,
278,
435,
6271,
29949,
29909,
363,
2820,
29871,
29896,
29900,
7378,
1728,
2551,
29892,
408,
5969,
2039,
6773,
297,
25309,
29889,
450,
24260,
3148,
4306,
10317,
6221,
29892,
5069,
6589,
892,
6369,
491,
278,
4306,
10317,
29892,
17049,
1833,
4723,
393,
376,
4881,
29875,
555,
520,
4642,
22895,
5037,
24921,
29908,
505,
2000,
278,
3148,
28679,
284,
515,
278,
435,
6271,
29949,
29909,
263,
376,
357,
11710,
10171,
1213,
13,
2059,
12814,
29892,
278,
1857,
26001,
297,
11996,
8959,
947,
451,
4658,
393,
7863,
304,
278,
435,
6271,
29949,
29909,
723,
367,
2253,
30003,
392,
278,
1857,
14883,
713,
8820,
871,
9659,
445,
29889,
376,
1576,
14176,
28469,
2750,
278,
4306,
310,
11996,
338,
14883,
1699,
1497,
4111,
20764,
297,
278,
28966,
11781,
373,
16340,
29889,
376,
30098,
797,
7786,
11405,
29892,
17503,
2645,
278,
27214,
800,
29892,
14883,
338,
10231,
967,
946,
11476,
322,
28424,
773,
15115,
1608,
297,
278,
5120,
29892,
408,
366,
599,
505,
3595,
29889,
910,
338,
920,
366,
7512,
27214,
800,
29892,
1920,
29882,
661,
29899,
3293,
1213,
13,
4002,
29886,
568,
278,
6924,
22941,
276,
882,
1546,
4111,
20764,
322,
350,
3615,
373,
920,
304,
1900,
4386,
14883,
29892,
297,
278,
1246,
373,
16340,
29892,
4111,
20764,
6452,
287,
350,
3615,
363,
376,
22880,
28325,
11255,
2304,
310,
11996,
408,
1532,
408,
278,
2304,
310,
670,
4152,
17517,
29892,
7148,
411,
21778,
304,
3082,
18872,
7113,
278,
20492,
360,
608,
1213,
940,
884,
23610,
350,
3615,
322,
670,
6532,
304,
6493,
11996,
29889,
13,
29950,
275,
5434,
714,
6914,
363,
14883,
471,
451,
577,
14652,
29891,
29889,
512,
278,
28966,
11781,
29892,
4111,
20764,
1370,
9571,
29892,
376,
26526,
7134,
519,
13258,
272,
9906,
393,
13258,
292,
297,
278,
14883,
713,
22384,
29892,
297,
278,
14883,
713,
26504,
29892,
338,
385,
443,
3538,
13258,
358,
297,
1716,
278,
1472,
322,
19697,
4958,
1213,
13,
4435,
29901,
313,
4013,
4274,
471,
10437,
6369,
491,
278,
341,
680,
579,
10318,
373,
5490,
29871,
29953,
29892,
29871,
29906,
29900,
29906,
29906,
29889,
5974,
29899,
12817,
4086,
756,
1063,
9120,
304,
9432,
1749,
17745,
9826,
1846,
13,
25971,
24596,
277,
29901,
501,
29889,
29903,
29889,
28735,
465,
29891,
23204,
29914,
29943,
1406,
29878,
29889,
510,
13,
25971,
19245,
29901,
383,
1406,
29878,
13,
29925,
764,
393,
278,
6171,
674,
17065,
11996,
29915,
29879,
20251,
411,
2107,
29714,
408,
896,
8727,
263,
3236,
310,
3158,
11211,
14883,
29889,
22121,
714,
304,
278,
6171,
363,
263,
1650,
393,
674,
5040,
14883,
713,
20346,
3181,
2187,
2748,
322,
363,
599,
1550,
884,
20794,
385,
1095,
304,
1009,
2304,
310,
15115,
1608,
297,
278,
5120,
29889,
13,
2831,
278,
365,
25593,
4076,
29714,
29936,
515,
3600,
13394,
2041,
7134,
322,
8004,
29889,
13,
29899,
1019,
369,
5824,
29871,
29906,
29901,
29953,
13,
1609,
2718,
328,
438,
3844,
29877,
3695,
612,
1212,
15753,
13,
29933,
801,
6038,
14744,
1220,
13,
29943,
2429,
388,
29892,
29871,
29946,
6339,
29871,
29906,
29900,
29906,
29906,
891,
5282,
1947,
7668,
4111,
1460,
402,
424,
29920,
1539,
373,
498,
1295,
3250,
411,
4088,
7904,
328,
27662,
19899,
394,
476,
4077,
26056,
310,
15174,
6038,
297,
670,
24369,
2645,
518,
29954,
424,
29920,
29915,
29879,
29962,
937,
6221,
6493,
304,
278,
20748,
29892,
988,
540,
8794,
263,
26406,
1302,
16453,
17327,
29889,
13,
29954,
424,
29920,
884,
1539,
411,
25306,
10787,
322,
15512,
7668,
3956,
1171,
27662,
7904,
328,
394,
476,
4077,
26056,
408,
1532,
408,
278,
22079,
310,
278,
15174,
6038,
23926,
8249,
322,
5282,
1947,
7668,
17860,
21797,
9016,
11699,
273,
838,
405,
3357,
10233,
29892,
1058,
1302,
29899,
7433,
278,
22879,
8133,
272,
392,
398,
310,
7634,
11235,
313,
6720,
29965,
29897,
1546,
278,
26406,
10127,
1860,
310,
278,
1023,
10916,
29889,
13,
1576,
16999,
29965,
5304,
29871,
29896,
29947,
7378,
1156,
278,
24763,
4831,
4339,
11595,
5281,
11996,
29915,
29879,
5302,
411,
15174,
6038,
322,
278,
501,
16036,
518,
2525,
1573,
10387,
2812,
381,
1078,
29962,
322,
674,
27391,
21082,
19383,
29892,
14002,
9121,
4192,
6090,
322,
17649,
260,
583,
1546,
278,
26406,
6397,
2722,
297,
1716,
19079,
29892,
322,
471,
278,
282,
2559,
6436,
310,
278,
6493,
29892,
3412,
411,
278,
9150,
11781,
411,
278,
6989,
322,
278,
15174,
336,
2172,
20251,
29889,
13,
29903,
2863,
3802,
304,
402,
424,
29920,
13384,
1009,
5994,
1608,
393,
278,
6493,
674,
1371,
6564,
263,
3802,
322,
938,
6490,
9443,
1546,
11996,
322,
15174,
6038,
29889,
13,
29954,
424,
29920,
29892,
1058,
471,
4520,
411,
4207,
943,
29892,
6452,
287,
278,
6989,
363,
670,
13457,
537,
322,
670,
2304,
310,
2545,
4858,
292,
13181,
1008,
284,
260,
583,
322,
278,
17407,
322,
7631,
713,
8571,
4110,
8794,
4550,
2215,
29892,
7773,
9215,
278,
26406,
1302,
16453,
29889,
13,
29908,
1576,
16650,
293,
1302,
16453,
393,
591,
526,
20794,
304,
263,
716,
1880,
1298,
9826,
411,
278,
26188,
310,
445,
17327,
322,
411,
278,
4100,
11781,
411,
278,
6989,
338,
278,
3133,
362,
310,
278,
22879,
24763,
4831,
4339,
322,
310,
278,
14338,
9443,
1546,
1749,
19079,
322,
2305,
1699,
402,
424,
29920,
1497,
472,
278,
25672,
29889,
13,
29908,
11730,
263,
1629,
1156,
278,
26188,
310,
278,
1035,
4339,
29892,
591,
526,
2307,
1244,
26188,
263,
7282,
6993,
17327,
29892,
607,
674,
2758,
16424,
1302,
16453,
322,
9324,
264,
278,
6993,
310,
1716,
10916,
322,
310,
278,
4152,
5120,
1699,
278,
26406,
11050,
1497,
29889,
13,
10649,
5086,
472,
263,
14002,
22895,
5037,
29994,
6026,
381,
2219,
29994,
29933,
801,
336,
2172,
9451,
2645,
385,
17568,
21362,
310,
278,
8907,
363,
3086,
14223,
16972,
1833,
4098,
29892,
15174,
6038,
29915,
29879,
19358,
7668,
17860,
913,
25572,
3856,
390,
1161,
333,
838,
796,
388,
3270,
1497,
278,
4234,
4446,
10977,
304,
10977,
373,
1784,
21420,
310,
278,
14883,
713,
13184,
297,
278,
5120,
29889,
13,
29908,
29909,
3619,
14014,
2602,
373,
1438,
5626,
674,
2758,
502,
304,
505,
7621,
9949,
975,
278,
3303,
3900,
1699,
540,
1497,
29892,
22884,
292,
393,
278,
5626,
892,
310,
1880,
13500,
363
] | 1,024 |
CommonCrawl
| 0.810438 | 0.369214 | 0.714717 | -0.111562 |
regional stability.
"Any future agreement with Iran must reflect the new reality in the region and be acceptable to all states in the region," he added.
Source: (This article was originally published by Ynetnews on February 3, 2022. Time-related language has been modified to reflect our publication today. See original article at this link.)
Photo License: B.alotaby/Commons.wikimedia.org
Photo License: wikimedia.org
Praise the Lord for the signing of the MOU between the defense establishments of Israel and Bahrain. Pray that this growing relationship will bring stability to the region and will encourage the US to take the new Middle East reality into consideration in any negotiations or interactions with Iran.
I will give peace in the land, and you shall lie down, and none will make you afraid; I will rid the land of evil beasts, and the sword will not go through your land.
- Leviticus 26:6
by JNS
Israel is making every effort to include more Arabs in the high-tech industry.
Tuesday, 8 February 2022 | Israel launched a [US] $70 million plan to promote the Arab sector in the high-tech industry.
"We see a change in Arab employment in high-tech in the past years, but it's not enough," Karina Rubinstein, director of business development for the Startup Division of the Israel Innovation Authority, told i24News.
"[Arab Israelis] have to compete with other workers, and they sometimes lack the knowledge of how to pass an interview or how to have these 'soft skills,'" she said.
To that end, innovation centers will be established in Israeli Arab communities to foster communication and consultation, including "investors clubs" to encourage funding for Arab start-ups, said Rubinstein.
Arab Israelis make up about 20% of the population, though only 3.5% of workers in high-tech.
Source: (This article was originally published by the Jewish News Syndicate on February 7, 2022. Time-related language has been modified to reflect our publication today. See original article at this link.)
Photo Credit: Pixabay/JNS.org
Praise the Lord for this innovative program to help Arab Israelis, and pray that it will be successful. Also pray that this type of genuine concern for the Arab sector will help many to recognize that Israel is not the enemy or the purveyor of hatred in the region. Ask the Lord to reveal Himself to many through Israel's actions of care and compassion.
The LORD has made bare His holy arm in the eyes of all the nations; and all the ends of the earth shall see the salvation of our God.
- Isaiah 52:10
by Yaakov Lappin ~ JNS
Israeli navy boats are seen from Rosh Hanikra on the Israeli and Lebanese border in northern Israel.
Wednesday, 9 February 2022 | In recent days, Lebanese President Michel Aoun told a UN envoy that his country is ready to return to indirect negotiations with Israel over the maritime border between the two countries.
Days later, the US envoy to the maritime negotiations, Amos Hochstein, began conducting shuttle diplomacy, involving meetings with Israel's Energy Minister Karine Elharrar and scheduled meetings with Lebanese officials in Beirut.
The developments represent an American push to seal a border agreement as part of Washington's overall desire to get additional gas to Lebanon, Maj. (res.) Tal Beeri, head of the research department at the Alma Center, a defense research center in northern Israel, told JNS.
"The Americans are pushing strongly for an agreement," he said.
Last year, regional Arab states and the United States hammered out the details of a plan designed to rescue Lebanon from its severe energy crisis by transferring Egyptian natural gas to Lebanon via pipelines that run from the Sinai Peninsula into Jordan and through Syria to feed Lebanon's Deir Ammar power plant, which has a 450-megawatt capacity. In addition, according to the plan, Jordan will link its electrical grid to Lebanon's via Syria to increase the power supply.
Aoun's announcement comes after Lebanon once again found itself "on the bench" on the issue of underwater gas reserves as other regional actors finalized maritime borders and secured their access to such resources, said Be
|
[
14014,
25806,
29889,
13,
29908,
10773,
5434,
17327,
411,
14883,
1818,
9432,
278,
716,
16832,
297,
278,
5120,
322,
367,
22691,
304,
599,
5922,
297,
278,
5120,
1699,
540,
2715,
29889,
13,
4435,
29901,
313,
4013,
4274,
471,
10437,
6369,
491,
612,
1212,
15753,
373,
6339,
29871,
29941,
29892,
29871,
29906,
29900,
29906,
29906,
29889,
5974,
29899,
12817,
4086,
756,
1063,
9120,
304,
9432,
1749,
17745,
9826,
29889,
2823,
2441,
4274,
472,
445,
1544,
1846,
13,
25971,
19245,
29901,
350,
29889,
284,
327,
10798,
29914,
5261,
787,
29889,
2851,
3393,
29889,
990,
13,
25971,
19245,
29901,
281,
638,
3393,
29889,
990,
13,
29925,
22692,
278,
6171,
363,
278,
26188,
310,
278,
16999,
29965,
1546,
278,
26406,
10127,
1860,
310,
11996,
322,
15174,
6038,
29889,
349,
764,
393,
445,
15678,
9443,
674,
6963,
25806,
304,
278,
5120,
322,
674,
13731,
6617,
278,
3148,
304,
2125,
278,
716,
14253,
6932,
16832,
964,
19220,
297,
738,
27214,
800,
470,
22060,
411,
14883,
29889,
13,
29902,
674,
2367,
10776,
297,
278,
2982,
29892,
322,
366,
4091,
3804,
1623,
29892,
322,
5642,
674,
1207,
366,
13421,
29936,
306,
674,
8177,
278,
2982,
310,
16126,
367,
19416,
29892,
322,
278,
22378,
674,
451,
748,
1549,
596,
2982,
29889,
13,
29899,
20708,
277,
12473,
29871,
29906,
29953,
29901,
29953,
13,
1609,
435,
3059,
13,
3624,
9058,
338,
3907,
1432,
7225,
304,
3160,
901,
10387,
29879,
297,
278,
1880,
29899,
11345,
13661,
29889,
13,
29911,
1041,
3250,
29892,
29871,
29947,
6339,
29871,
29906,
29900,
29906,
29906,
891,
11996,
15241,
263,
518,
3308,
29962,
395,
29955,
29900,
7284,
3814,
304,
27391,
278,
10387,
17535,
297,
278,
1880,
29899,
11345,
13661,
29889,
13,
29908,
4806,
1074,
263,
1735,
297,
10387,
5703,
358,
297,
1880,
29899,
11345,
297,
278,
4940,
2440,
29892,
541,
372,
29915,
29879,
451,
3307,
1699,
3467,
1099,
14985,
262,
5465,
29892,
8881,
310,
5381,
5849,
363,
278,
7370,
786,
7946,
310,
278,
11996,
512,
13715,
362,
13361,
537,
29892,
5429,
474,
29906,
29946,
29328,
29889,
13,
29908,
29961,
29909,
4201,
11996,
275,
29962,
505,
304,
752,
2650,
411,
916,
17162,
29892,
322,
896,
6041,
10225,
278,
7134,
310,
920,
304,
1209,
385,
15593,
470,
920,
304,
505,
1438,
525,
2695,
25078,
5501,
29908,
1183,
1497,
29889,
13,
1762,
393,
1095,
29892,
24233,
362,
1644,
414,
674,
367,
7841,
297,
22895,
5037,
10387,
23507,
304,
9926,
261,
12084,
322,
8799,
362,
29892,
3704,
376,
262,
10147,
943,
17651,
29908,
304,
13731,
6617,
5220,
292,
363,
10387,
1369,
29899,
14340,
29892,
1497,
14985,
262,
5465,
29889,
13,
29909,
4201,
11996,
275,
1207,
701,
1048,
29871,
29906,
29900,
29995,
310,
278,
4665,
29892,
2466,
871,
29871,
29941,
29889,
29945,
29995,
310,
17162,
297,
1880,
29899,
11345,
29889,
13,
4435,
29901,
313,
4013,
4274,
471,
10437,
6369,
491,
278,
16728,
10130,
8713,
299,
9593,
373,
6339,
29871,
29955,
29892,
29871,
29906,
29900,
29906,
29906,
29889,
5974,
29899,
12817,
4086,
756,
1063,
9120,
304,
9432,
1749,
17745,
9826,
29889,
2823,
2441,
4274,
472,
445,
1544,
1846,
13,
25971,
24596,
277,
29901,
349,
861,
370,
388,
29914,
29967,
3059,
29889,
990,
13,
29925,
22692,
278,
6171,
363,
445,
24233,
1230,
1824,
304,
1371,
10387,
11996,
275,
29892,
322,
12475,
393,
372,
674,
367,
9150,
29889,
3115,
12475,
393,
445,
1134,
310,
29120,
457,
5932,
363,
278,
10387,
17535,
674,
1371,
1784,
304,
18720,
393,
11996,
338,
451,
278,
11103,
470,
278,
3708,
6950,
272,
310,
3056,
1127,
297,
278,
5120,
29889,
26579,
278,
6171,
304,
10320,
284,
18136,
1311,
304,
1784,
1549,
11996,
29915,
29879,
8820,
310,
2562,
322,
752,
465,
291,
29889,
13,
1576,
365,
25593,
756,
1754,
16079,
3600,
26630,
5075,
297,
278,
5076,
310,
599,
278,
19079,
29936,
322,
599,
278,
10614,
310,
278,
8437,
4091,
1074,
278,
24754,
362,
310,
1749,
4177,
29889,
13,
29899,
19899,
21071,
29871,
29945,
29906,
29901,
29896,
29900,
13,
1609,
25887,
27695,
365,
932,
262,
3695,
435,
3059,
13,
3624,
336,
5037,
302,
5301,
25462,
526,
3595,
515,
5678,
29882,
7169,
638,
336,
373,
278,
22895,
5037,
322,
9388,
273,
968,
5139,
297,
14622,
11996,
29889,
13,
29956,
287,
4515,
3250,
29892,
29871,
29929,
6339,
29871,
29906,
29900,
29906,
29906,
891,
512,
7786,
3841,
29892,
9388,
273,
968,
7178,
12255,
319,
1309,
5429,
263,
8291,
427,
18644,
393,
670,
4234,
338,
7960,
304,
736,
304,
26377,
27214,
800,
411,
11996,
975,
278,
1766,
21998,
5139,
1546,
278,
1023,
10916,
29889,
13,
25991,
2678,
29892,
278,
3148,
427,
18644,
304,
278,
1766,
21998,
27214,
800,
29892,
1913,
359,
10955,
5465,
29892,
4689,
7512,
292,
528,
4774,
280,
13487,
4135,
29892,
21677,
5870,
886,
411,
11996,
29915,
29879,
24836,
7668,
3467,
457,
1260,
29882,
2749,
279,
322,
21467,
5870,
886,
411,
9388,
273,
968,
24921,
297,
1522,
381,
329,
29889,
13,
1576,
2693,
1860,
2755,
385,
3082,
5503,
304,
409,
284,
263,
5139,
17327,
408,
760,
310,
7660,
29915,
29879,
12463,
13521,
304,
679,
5684,
10489,
304,
9388,
19930,
29892,
6973,
29889,
313,
690,
1846,
10288,
1522,
13613,
29892,
2343,
310,
278,
5925,
14311,
472,
278,
838,
655,
7817,
29892,
263,
26406,
5925,
4818,
297,
14622,
11996,
29892,
5429,
435,
3059,
29889,
13,
29908,
1576,
23035,
526,
27556,
13818,
363,
385,
17327,
1699,
540,
1497,
29889,
13,
8897,
1629,
29892,
14014,
10387,
5922,
322,
278,
3303,
3900,
16366,
1050,
287,
714,
278,
4902,
310,
263,
3814,
8688,
304,
26429,
9388,
19930,
515,
967,
22261,
5864,
24161,
491,
6782,
5393,
12892,
713,
5613,
10489,
304,
9388,
19930,
3025,
8450,
24210,
393,
1065,
515,
278,
317,
1099,
29875,
7363,
1144,
2497,
964,
18284,
322,
1549,
8713,
2849,
304,
8343,
9388,
19930,
29915,
29879,
897,
381,
1913,
3034,
3081,
8024,
29892,
607,
756,
263,
29871,
29946,
29945,
29900,
29899,
29885,
387,
1450,
1131,
13284,
29889,
512,
6124,
29892,
5034,
304,
278,
3814,
29892,
18284,
674,
1544,
967,
3546,
16888,
6856,
304,
9388,
19930,
29915,
29879,
3025,
8713,
2849,
304,
7910,
278,
3081,
11421,
29889,
13,
29909,
1309,
29915,
29879,
7475,
13561,
5304,
1156,
9388,
19930,
2748,
1449,
1476,
3528,
376,
265,
278,
3856,
305,
29908,
373,
278,
2228,
310,
1090,
13405,
10489,
620,
20098,
408,
916,
14014,
29701,
2186,
1891,
1766,
21998,
28199,
322,
26130,
1009,
2130,
304,
1316,
7788,
29892,
1497,
1522
] | 1,024 |
CommonCrawl
| 0.409671 | -0.250478 | -0.088647 | 0.022515 |
eri, who served for 20 years in the Israel Defense Forces [IDF] Military Intelligence Directorate.
"Lebanon has been out of the economic waters game for a long time. The Syrians have 'taken a bite' from gas reserves to the north—and the maritime border between Syria and Lebanon has never been finalized," noted Beeri.
Instead, Lebanon demarcated its maritime borders independently in 2011, causing a rare complaint from Syria in 2014 to the United Nations over the issue.
According to the current situation, Syria's Block 1 zone overlaps with Lebanon's Block 2—meaning that Syria has taken 750 square kilometers [290 sq. mi.] of maritime territory and began searching for gas using Russian company Capital Oil.
Since then, "no one in the Lebanese government has dared confront the Syrians," said Beeri. "The Syrians have conducted gas searches, and no one in Lebanon, including Hezbollah, has said a word. The only exception is the protest over the matter by the head of the [Christian Lebanese movement] Lebanese Forces, Samir Gaegae, in 2021," he stated.
Syria and Russia are not the only winners from this situation; Hezbollah is likely to receive "gas coupons" from discoveries in the area from [Syrian President Bashar al-Assad's] regime in the event of a discovery of a gas field.
'Not a Green Light to Normalization with Israel'
Meanwhile, Lebanon has found itself shut out from other regional arrangements as well due to its paralysis. In 2010, Israel and Cyprus reached a former maritime border agreement, giving Cyprus access to the Aphrodite gas reserve.
"Lebanon did not want to take part in this due to Israel's involvement; hence, it stayed outside of the talks," said Beeri. Now, Beirut finds itself able to complete for only one maritime area—the sea zone to its south, in which the Karish gas field exists.
Beeri noted that the country "decided that in order to avoid losing again, it had to make its stance extreme."
That approach has resulted in Lebanon demanding a southern maritime border in 2021, which is far south of the line it presented in 2010. The line "just happens" to run halfway through the known Karish gas-field zone—meaning that if the demand is expected, Lebanon can hire companies to simply arrive and begin drilling, skipping over the lengthy search stage, pointed out Beeri.
Hezbollah, for its part, has allowed the Lebanese government to take part in indirect talks, primarily for economic reasons.
"It is clear that some of the gas profits will 'drain' into Hezbollah's accounts," said Beeri. "In addition, politically, it does not want to be remembered as the side that blocked Lebanon from reaching gas. Hence, suddenly, it strikes 'moderate' stances and allows negotiations after Lebanon has faced years of economic crisis that is directly influenced by Hezbollah's activities."
Although Hezbollah chief Hassan Nasrallah routinely accuses Israel of stealing Lebanese resources, including gas, the Lebanese terror organization has left the issue under the portfolio of the Lebanese government.
Its chief concern, Beeri said, is to make it known that it is not a side to the negotiations with Israel and to market itself as the defender of Lebanon's borders.
"Hezbollah always stresses that the negotiations are part of a national Lebanese requirement, but are not a green light to normalization with Israel," said Beeri. "Hezbollah is telling itself a story that this is purely a military-technical issue, not a normalization affair. That there are no political ties between Israel and Lebanon.
"When Lebanon's former Foreign Minister, Jubran Bassil, the head of the Free Patriotic Movement and the nephew of President Aoun, offered in April 2021 to speed-up talks by investing together with Israel in a third-party company to search for gas, he was strongly reprimanded by Hezbollah," noted Beeri. "Hezbollah wants to dance at two weddings—to say that it is present on this issue, but also that it is absent."
|
[
13613,
29892,
1058,
6766,
363,
29871,
29906,
29900,
2440,
297,
278,
11996,
5282,
1947,
25571,
518,
1367,
29943,
29962,
20080,
3159,
28286,
15944,
403,
29889,
13,
29908,
29931,
774,
19930,
756,
1063,
714,
310,
278,
17407,
19922,
3748,
363,
263,
1472,
931,
29889,
450,
8713,
374,
550,
505,
525,
29873,
9424,
263,
289,
568,
29915,
515,
10489,
620,
20098,
304,
278,
6641,
30003,
392,
278,
1766,
21998,
5139,
1546,
8713,
2849,
322,
9388,
19930,
756,
2360,
1063,
2186,
1891,
1699,
11682,
1522,
13613,
29889,
13,
3379,
1479,
29892,
9388,
19930,
1261,
5666,
630,
967,
1766,
21998,
28199,
25499,
297,
29871,
29906,
29900,
29896,
29896,
29892,
10805,
263,
10812,
15313,
524,
515,
8713,
2849,
297,
29871,
29906,
29900,
29896,
29946,
304,
278,
3303,
18269,
975,
278,
2228,
29889,
13,
7504,
3278,
304,
278,
1857,
6434,
29892,
8713,
2849,
29915,
29879,
15658,
29871,
29896,
10640,
975,
14128,
411,
9388,
19930,
29915,
29879,
15658,
29871,
29906,
30003,
12676,
292,
393,
8713,
2849,
756,
4586,
29871,
29955,
29945,
29900,
6862,
20052,
2699,
518,
29906,
29929,
29900,
18074,
29889,
3737,
5586,
310,
1766,
21998,
20123,
322,
4689,
11975,
363,
10489,
773,
10637,
5001,
25343,
438,
309,
29889,
13,
23036,
769,
29892,
376,
1217,
697,
297,
278,
9388,
273,
968,
5874,
756,
270,
1965,
21751,
278,
8713,
374,
550,
1699,
1497,
1522,
13613,
29889,
376,
1576,
8713,
374,
550,
505,
18043,
10489,
29645,
29892,
322,
694,
697,
297,
9388,
19930,
29892,
3704,
940,
29920,
2095,
8083,
29892,
756,
1497,
263,
1734,
29889,
450,
871,
3682,
338,
278,
10021,
975,
278,
4383,
491,
278,
2343,
310,
278,
518,
18687,
713,
9388,
273,
968,
10298,
29962,
9388,
273,
968,
25571,
29892,
3685,
381,
10415,
2442,
29872,
29892,
297,
29871,
29906,
29900,
29906,
29896,
1699,
540,
8703,
29889,
13,
29903,
29891,
2849,
322,
12710,
526,
451,
278,
871,
281,
16697,
515,
445,
6434,
29936,
940,
29920,
2095,
8083,
338,
5517,
304,
7150,
376,
25496,
17103,
787,
29908,
515,
6523,
583,
297,
278,
4038,
515,
518,
29903,
29891,
6392,
7178,
29677,
279,
394,
29899,
7900,
328,
29915,
29879,
29962,
22384,
297,
278,
1741,
310,
263,
20699,
310,
263,
10489,
1746,
29889,
13,
29915,
3664,
263,
7646,
12790,
304,
21981,
2133,
411,
11996,
29915,
13,
6816,
273,
8000,
29892,
9388,
19930,
756,
1476,
3528,
12522,
714,
515,
916,
14014,
15196,
4110,
408,
1532,
2861,
304,
967,
610,
284,
4848,
29889,
512,
29871,
29906,
29900,
29896,
29900,
29892,
11996,
322,
8045,
558,
375,
7450,
263,
4642,
1766,
21998,
5139,
17327,
29892,
6820,
8045,
558,
375,
2130,
304,
278,
319,
561,
5964,
568,
10489,
23986,
29889,
13,
29908,
29931,
774,
19930,
1258,
451,
864,
304,
2125,
760,
297,
445,
2861,
304,
11996,
29915,
29879,
5297,
29841,
29936,
8151,
29892,
372,
27661,
5377,
310,
278,
5969,
2039,
1699,
1497,
1522,
13613,
29889,
2567,
29892,
1522,
381,
329,
14061,
3528,
2221,
304,
4866,
363,
871,
697,
1766,
21998,
4038,
30003,
1552,
7205,
10640,
304,
967,
7062,
29892,
297,
607,
278,
3467,
728,
10489,
1746,
4864,
29889,
13,
3629,
13613,
11682,
393,
278,
4234,
376,
7099,
2618,
393,
297,
1797,
304,
4772,
19035,
1449,
29892,
372,
750,
304,
1207,
967,
380,
749,
18677,
1213,
13,
7058,
2948,
756,
20601,
297,
9388,
19930,
9667,
292,
263,
14841,
1766,
21998,
5139,
297,
29871,
29906,
29900,
29906,
29896,
29892,
607,
338,
2215,
7062,
310,
278,
1196,
372,
9132,
297,
29871,
29906,
29900,
29896,
29900,
29889,
450,
1196,
376,
5143,
5930,
29908,
304,
1065,
4203,
1582,
1549,
278,
2998,
3467,
728,
10489,
29899,
2671,
10640,
30003,
12676,
292,
393,
565,
278,
9667,
338,
3806,
29892,
9388,
19930,
508,
298,
533,
14582,
304,
3763,
18331,
322,
3380,
4192,
8873,
29892,
14993,
3262,
975,
278,
3309,
29891,
2740,
7408,
29892,
11520,
714,
1522,
13613,
29889,
13,
3868,
29920,
2095,
8083,
29892,
363,
967,
760,
29892,
756,
6068,
278,
9388,
273,
968,
5874,
304,
2125,
760,
297,
26377,
5969,
2039,
29892,
19434,
363,
17407,
9590,
29889,
13,
29908,
3112,
338,
2821,
393,
777,
310,
278,
10489,
2600,
1169,
674,
525,
29881,
6038,
29915,
964,
940,
29920,
2095,
8083,
29915,
29879,
15303,
1699,
1497,
1522,
13613,
29889,
376,
797,
6124,
29892,
2832,
1711,
29892,
372,
947,
451,
864,
304,
367,
21832,
408,
278,
2625,
393,
24370,
9388,
19930,
515,
20888,
10489,
29889,
10133,
29892,
11584,
29892,
372,
19492,
267,
525,
1545,
261,
403,
29915,
380,
2925,
322,
6511,
27214,
800,
1156,
9388,
19930,
756,
20050,
2440,
310,
17407,
24161,
393,
338,
4153,
28482,
491,
940,
29920,
2095,
8083,
29915,
29879,
14188,
1213,
13,
2499,
3592,
940,
29920,
2095,
8083,
9087,
27558,
273,
22318,
1705,
8083,
6745,
262,
873,
1035,
6394,
11996,
310,
1886,
12818,
9388,
273,
968,
7788,
29892,
3704,
10489,
29892,
278,
9388,
273,
968,
15115,
13013,
756,
2175,
278,
2228,
1090,
278,
2011,
25648,
310,
278,
9388,
273,
968,
5874,
29889,
13,
29902,
1372,
9087,
5932,
29892,
1522,
13613,
1497,
29892,
338,
304,
1207,
372,
2998,
393,
372,
338,
451,
263,
2625,
304,
278,
27214,
800,
411,
11996,
322,
304,
9999,
3528,
408,
278,
822,
1581,
310,
9388,
19930,
29915,
29879,
28199,
29889,
13,
29908,
3868,
29920,
2095,
8083,
2337,
851,
15322,
393,
278,
27214,
800,
526,
760,
310,
263,
4797,
9388,
273,
968,
11809,
29892,
541,
526,
451,
263,
7933,
3578,
304,
4226,
2133,
411,
11996,
1699,
1497,
1522,
13613,
29889,
376,
3868,
29920,
2095,
8083,
338,
14509,
3528,
263,
5828,
393,
445,
338,
24837,
263,
9121,
29899,
21695,
936,
2228,
29892,
451,
263,
4226,
2133,
26195,
29889,
2193,
727,
526,
694,
8604,
260,
583,
1546,
11996,
322,
9388,
19930,
29889,
13,
29908,
10401,
9388,
19930,
29915,
29879,
4642,
19358,
7668,
29892,
435,
431,
661,
17328,
309,
29892,
278,
2343,
310,
278,
12362,
23168,
13574,
14104,
882,
322,
278,
23446,
13636,
310,
7178,
319,
1309,
29892,
12520,
297,
3786,
29871,
29906,
29900,
29906,
29896,
304,
6210,
29899,
786,
5969,
2039,
491,
13258,
292,
4208,
411,
11996,
297,
263,
4654,
29899,
22633,
5001,
304,
2740,
363,
10489,
29892,
540,
471,
13818,
2062,
326,
392,
287,
491,
940,
29920,
2095,
8083,
1699,
11682,
1522,
13613,
29889,
376,
3868,
29920,
2095,
8083,
10753,
304,
17948,
472,
1023,
14837,
29881,
886,
30003,
517,
1827,
393,
372,
338,
2198,
373,
445,
2228,
29892,
541,
884,
393,
372,
338,
29207,
1213
] | 1,024 |
CommonCrawl
| 0.508844 | 0.85199 | 1.102053 | -0.182395 |
Photo Credit: Jamal Awad/Flash90/JNS.org
Pray that Israel will be successful in negotiating its maritime border with Lebanon, which may help lessen the economic crisis that has plagued that nation for years. Pray also that Lebanon will accept Israel's other offers of assistance, bringing relief to the beleaguered country and weakening the stranglehold that keeps Lebanon in a state of chaos through Iranian proxy Hezbollah.
The LORD will give strength to His people; the LORD will bless His people with peace.
- Psalm 29:11
by Ilse Strauss
Anti-Semitism reached record highs in the UK in 2021.
Thursday, 10 February 2022 | Anti-Semitism in Britain soared to a record high in 2021, Community Security Trust (CST) revealed in a report published yesterday.
According to the UK-based anti-Semitism watchdog, British Jews and authorities reported 2,255 anti-Semitic incidents in the UK during the 12-month period, the highest number in recent history anywhere in Europe and the highest number in a single calendar year since the CST started recording anti-Jewish hate occurrences nearly 40 years ago.
The 2,255 incidents mark a 34% increase from the 1,684 recorded in 2020 and a 24% spike from the previous record of 1,813 offences in 2019.
"These record levels of anti-Jewish racism, reported by our Jewish community to CST and police, show how difficult last year was for Jews across Britain. These hatreds boil away, taking any excuse to publicly burst out against Jews," Mark Gardner, CST chief executive.
The CST ascribes the record escalation to the "huge rise in anti-Jewish hate and extremism during and following the escalation in violence in Israel and Palestine last year."
May 2021, the month in which Operation Guardian of the Walls—the 11-day conflict between Israel and Gaza-based terror groups flared up, saw a record of 661 anti-Semitic incidents in the UK, while 210 events were logged in June.
"It's no exaggeration to say that the landscape of anti-Semitism in the UK in 2021 is largely defined by responses to the conflict in Israel and Palestine, as indeed is the unprecedented annual figure," the report states.
Nearly 200 of the 2,255 incidents last year were violent, three of which involved grievous bodily harm or endangering a life. There were also 502 occurrences where far-right or Nazi themes were used and 90 incidents of celebrating the Holocaust.
Source: (Bridges for Peace, February 10, 2022)
Photo Credit: Beny Shlevich/Commons.wikimedia.org
Photo License: Wikimedia
Cry out to the Lord for His protection to surround the Jews of the UK as anti-Semitism continues to soar to record numbers. Pray that the government of the UK will take a strong stand against this heinous racism, criminalizing such actions and punishing all perpetrators. Pray also that many Jews in the UK will choose to make aliyah, taking refuge in the land of their forefathers.
You are my hiding place; You shall preserve me from trouble; You shall surround me with songs of deliverance.
- Psalm 32:7
Turkish President Recep Tayyip Erdoğan
Tuesday, 8 February 2022 | Turkish President Recep Tayyip Erdoğan said a discussion over energy cooperation with Israel would take place in March and that on the table for discussion is a plan to work together to transport natural gas to Europe.
"We can use Israeli natural gas in our country, and beyond using it, we can also engage in a joint effort on its passage to Europe," said Erdoğan on January 4, reported Reuters.
"Now, God willing, these issues will be on our agenda with Mr. Herzog during their visit to Turkey," he said, as reported by Turkish media.
Israeli President Isaac Herzog plans to visit Turkey next month, Erdoğan said on Thursday.
|
[
13,
25971,
24596,
277,
29901,
16131,
284,
319,
13829,
29914,
8754,
1161,
29929,
29900,
29914,
29967,
3059,
29889,
990,
13,
29925,
764,
393,
11996,
674,
367,
9150,
297,
27214,
1218,
967,
1766,
21998,
5139,
411,
9388,
19930,
29892,
607,
1122,
1371,
3109,
264,
278,
17407,
24161,
393,
756,
715,
351,
6742,
393,
5233,
363,
2440,
29889,
349,
764,
884,
393,
9388,
19930,
674,
3544,
11996,
29915,
29879,
916,
16688,
310,
18872,
29892,
20794,
18892,
304,
278,
367,
280,
351,
2853,
287,
4234,
322,
8062,
8333,
278,
851,
2521,
8948,
393,
14874,
9388,
19930,
297,
263,
2106,
310,
10329,
359,
1549,
14883,
713,
10166,
940,
29920,
2095,
8083,
29889,
13,
1576,
365,
25593,
674,
2367,
9324,
304,
3600,
2305,
29936,
278,
365,
25593,
674,
17065,
3600,
2305,
411,
10776,
29889,
13,
29899,
10144,
17120,
29871,
29906,
29929,
29901,
29896,
29896,
13,
1609,
1720,
344,
7357,
1558,
13,
13448,
29875,
29899,
29903,
21976,
1608,
7450,
2407,
1880,
29879,
297,
278,
10261,
297,
29871,
29906,
29900,
29906,
29896,
29889,
13,
1349,
1295,
3250,
29892,
29871,
29896,
29900,
6339,
29871,
29906,
29900,
29906,
29906,
891,
18473,
29899,
29903,
21976,
1608,
297,
14933,
577,
1965,
304,
263,
2407,
1880,
297,
29871,
29906,
29900,
29906,
29896,
29892,
19184,
14223,
20692,
313,
29907,
1254,
29897,
17845,
297,
263,
3461,
6369,
22600,
29889,
13,
7504,
3278,
304,
278,
10261,
29899,
6707,
9418,
29899,
29903,
21976,
1608,
6505,
26169,
29892,
4908,
22057,
322,
21142,
8967,
29871,
29906,
29892,
29906,
29945,
29945,
9418,
29899,
29903,
21976,
293,
5528,
16719,
297,
278,
10261,
2645,
278,
29871,
29896,
29906,
29899,
10874,
3785,
29892,
278,
9939,
1353,
297,
7786,
4955,
12214,
297,
4092,
322,
278,
9939,
1353,
297,
263,
2323,
17684,
1629,
1951,
278,
315,
1254,
4687,
16867,
9418,
29899,
29967,
809,
728,
26277,
13920,
2063,
8886,
29871,
29946,
29900,
2440,
8020,
29889,
13,
1576,
29871,
29906,
29892,
29906,
29945,
29945,
5528,
16719,
2791,
263,
29871,
29941,
29946,
29995,
7910,
515,
278,
29871,
29896,
29892,
29953,
29947,
29946,
10478,
297,
29871,
29906,
29900,
29906,
29900,
322,
263,
29871,
29906,
29946,
29995,
805,
9345,
515,
278,
3517,
2407,
310,
29871,
29896,
29892,
29947,
29896,
29941,
1283,
2063,
297,
29871,
29906,
29900,
29896,
29929,
29889,
13,
29908,
1349,
968,
2407,
11174,
310,
9418,
29899,
29967,
809,
728,
11021,
1608,
29892,
8967,
491,
1749,
16728,
7881,
304,
315,
1254,
322,
10974,
29892,
1510,
920,
5189,
1833,
1629,
471,
363,
22057,
4822,
14933,
29889,
4525,
3056,
1127,
29879,
1045,
309,
3448,
29892,
5622,
738,
5566,
1509,
304,
970,
368,
20887,
714,
2750,
22057,
1699,
4485,
15971,
1089,
29892,
315,
1254,
9087,
22760,
29889,
13,
1576,
315,
1254,
408,
699,
5707,
278,
2407,
831,
1052,
362,
304,
278,
376,
29882,
5710,
14451,
297,
9418,
29899,
29967,
809,
728,
26277,
322,
9413,
1608,
2645,
322,
1494,
278,
831,
1052,
362,
297,
21448,
297,
11996,
322,
22053,
457,
1833,
1629,
1213,
13,
12703,
29871,
29906,
29900,
29906,
29896,
29892,
278,
4098,
297,
607,
20462,
29429,
310,
278,
399,
4293,
30003,
1552,
29871,
29896,
29896,
29899,
3250,
14529,
1546,
11996,
322,
15853,
29874,
29899,
6707,
15115,
6471,
17422,
1127,
701,
29892,
4446,
263,
2407,
310,
29871,
29953,
29953,
29896,
9418,
29899,
29903,
21976,
293,
5528,
16719,
297,
278,
10261,
29892,
1550,
29871,
29906,
29896,
29900,
4959,
892,
13817,
297,
5306,
29889,
13,
29908,
3112,
29915,
29879,
694,
429,
9921,
362,
304,
1827,
393,
278,
24400,
310,
9418,
29899,
29903,
21976,
1608,
297,
278,
10261,
297,
29871,
29906,
29900,
29906,
29896,
338,
18425,
3342,
491,
20890,
304,
278,
14529,
297,
11996,
322,
22053,
457,
29892,
408,
6200,
338,
278,
443,
1457,
1133,
14927,
17568,
4377,
1699,
278,
3461,
5922,
29889,
13,
29940,
799,
368,
29871,
29906,
29900,
29900,
310,
278,
29871,
29906,
29892,
29906,
29945,
29945,
5528,
16719,
1833,
1629,
892,
24252,
29892,
2211,
310,
607,
9701,
867,
10384,
681,
13004,
2354,
10311,
470,
1095,
574,
3241,
263,
2834,
29889,
1670,
892,
884,
29871,
29945,
29900,
29906,
13920,
2063,
988,
2215,
29899,
1266,
470,
11409,
29875,
963,
267,
892,
1304,
322,
29871,
29929,
29900,
5528,
16719,
310,
10894,
1218,
278,
4168,
6400,
504,
29889,
13,
4435,
29901,
313,
29933,
2429,
2710,
363,
24819,
29892,
6339,
29871,
29896,
29900,
29892,
29871,
29906,
29900,
29906,
29906,
29897,
13,
25971,
24596,
277,
29901,
4111,
29891,
1383,
2608,
436,
29914,
5261,
787,
29889,
2851,
3393,
29889,
990,
13,
25971,
19245,
29901,
7494,
13,
29907,
719,
714,
304,
278,
6171,
363,
3600,
13047,
304,
8388,
618,
278,
22057,
310,
278,
10261,
408,
9418,
29899,
29903,
21976,
1608,
18172,
304,
577,
279,
304,
2407,
3694,
29889,
349,
764,
393,
278,
5874,
310,
278,
10261,
674,
2125,
263,
4549,
2317,
2750,
445,
540,
262,
681,
11021,
1608,
29892,
22161,
5281,
1316,
8820,
322,
6035,
14424,
599,
25722,
3605,
943,
29889,
349,
764,
884,
393,
1784,
22057,
297,
278,
10261,
674,
6755,
304,
1207,
14430,
29891,
801,
29892,
5622,
25447,
297,
278,
2982,
310,
1009,
363,
1389,
19467,
29889,
13,
3492,
526,
590,
25508,
2058,
29936,
887,
4091,
19905,
592,
515,
7458,
29936,
887,
4091,
8388,
618,
592,
411,
12516,
310,
12021,
749,
29889,
13,
29899,
10144,
17120,
29871,
29941,
29906,
29901,
29955,
13,
29911,
29641,
728,
7178,
830,
13300,
323,
388,
29891,
666,
1425,
1867,
30200,
273,
13,
29911,
1041,
3250,
29892,
29871,
29947,
6339,
29871,
29906,
29900,
29906,
29906,
891,
24682,
7178,
830,
13300,
323,
388,
29891,
666,
1425,
1867,
30200,
273,
1497,
263,
10679,
975,
5864,
1302,
16453,
411,
11996,
723,
2125,
2058,
297,
4779,
322,
393,
373,
278,
1591,
363,
10679,
338,
263,
3814,
304,
664,
4208,
304,
8608,
5613,
10489,
304,
4092,
29889,
13,
29908,
4806,
508,
671,
22895,
5037,
5613,
10489,
297,
1749,
4234,
29892,
322,
8724,
773,
372,
29892,
591,
508,
884,
3033,
482,
297,
263,
14002,
7225,
373,
967,
13382,
304,
4092,
1699,
1497,
1425,
1867,
30200,
273,
373,
5490,
29871,
29946,
29892,
8967,
830,
329,
414,
29889,
13,
29908,
10454,
29892,
4177,
17762,
29892,
1438,
5626,
674,
367,
373,
1749,
946,
8395,
411,
3237,
29889,
23804,
2645,
1009,
6493,
304,
26459,
1699,
540,
1497,
29892,
408,
8967,
491,
24682,
5745,
29889,
13,
3624,
336,
5037,
7178,
28156,
23804,
13900,
304,
6493,
26459,
2446,
4098,
29892,
1425,
1867,
30200,
273,
1497,
373,
498,
1295,
3250,
29889
] | 1,024 |
CommonCrawl
| 0.496617 | 0.10836 | 0.696785 | -0.034404 |
Tense relations have existed in recent years between the countries as Erdoğan has repeatedly put Israel in the crosshairs on a number of issues, as well as hosted members of the Hamas terror organization that rules the Gaza Strip.
Photo Credit: Turkish Presidency/Twitter/JNS.org
Pray for the relationship between Israel and Turkey, which has been strained in recent years. Pray that energy cooperation will be a first step in restoring diplomatic and cooperative ties that actually began in 1948.
And the LORD, He is the One who goes before you. He will be with you, He will not leave you nor forsake you; do not fear nor be dismayed.
- Deuteronomy 31:8
by Kate Norman
The Syrian missile triggered sirens in the Arab Israeli city of Umm al-Fahm.
Wednesday, 9 February 2022 | An antiaircraft missile was fired from Syria into northern Israel in the pre-dawn hours of Wednesday morning, prompting a responsive strike from the Israeli military.
The Syrian missile triggered sirens in the Arab Israeli city of Umm al-Fahm and part of northern Samaria. The missile exploded midair, the Israel Defense Forces (IDF) reported, so no interception was necessary.
In response, the IDF said it struck "surface-to-air missile targets in Syria, including radar and antiaircraft batteries."
"The IDF will continue to protect the skies of the State of Israel and its security," the Israeli military said on its website this morning.
The Syrian antiaircraft missile was reportedly launched just after an alleged Israeli air strike near Damascus.
The IDF launched an air strike from the direction of neighboring Beirut just before 1 a.m. and fired another round of surface-to-surface missiles from the Golan Heights around 1:10 a.m., killing one Syrian soldier, wounding five others and causing material damage, state-affiliated Syrian Arab News Agency (SANA) reported.
The target of the alleged Israeli strikes remains unknown, but they may have been targeting sites of the Syrian regime as well as Iranian militias, the Jerusalem Post reported.
Israel did not comment on its involvement in the early morning strikes in keeping with its longstanding policy. The Jewish state has vowed, however, to prevent Iran from entrenching itself in the region and has acknowledged that it has conducted some hundreds of air strikes in Syria over the past decade, often targeting Iranian weapons shipments to its proxies in the region, particularly the Lebanese-based Hezbollah terror organization.
Last week another air strike outside of Damascus was attributed to Israel. The target there too was ambiguous, as Ynetnews reported it was a Hezbollah warehouse, while the Jerusalem Post, citing Syrian opposition media, said it was Syrian regime sites.
That strike took place just a week after a joint Russian–Syrian flyby along the Israeli–Syrian border.
The Jerusalem Post also pointed out that independent flight trackers have logged multiple cargo flights shipping weapons from Iran to Syria over the past few weeks.
Source: (Bridges for Peace, February 9, 2022)
Source: Moataz1997/Commons.wikimedia.org
Cry out to the Lord that the IDF will, in fact, continue to protect the skies of the State of Israel and will be successful in keeping Iran from becoming further entrenched in Syria. Pray for the safety of the citizens of northern Israel as the enemy sits on their doorstep.
"Fear not, for I am with you; be not dismayed, for I am your God. I will strengthen you, yes, I will help you, I will uphold you with My righteous right hand."
by David Isaac ~ JNS
The heads of state in the Africa Union in 2016
Thursday, 10 February 2022 | The African Union pushed off debate on a vote over Israel's observer status at its meeting on Sunday. Instead, it set up a committee to examine the matter, which will report its findings at next year's summit. Reports say the issue has proved divisive in the 55-member bloc.
The African Union granted Israel observer status in July. However, opposition to the decision only gained
|
[
13,
29911,
1947,
5302,
505,
22856,
297,
7786,
2440,
1546,
278,
10916,
408,
1425,
1867,
30200,
273,
756,
28424,
1925,
11996,
297,
278,
274,
1883,
845,
7121,
373,
263,
1353,
310,
5626,
29892,
408,
1532,
408,
17791,
5144,
310,
278,
7904,
294,
15115,
13013,
393,
6865,
278,
15853,
29874,
624,
6472,
29889,
13,
25971,
24596,
277,
29901,
24682,
4360,
3615,
1270,
29914,
27418,
5171,
29914,
29967,
3059,
29889,
990,
13,
29925,
764,
363,
278,
9443,
1546,
11996,
322,
26459,
29892,
607,
756,
1063,
5312,
1312,
297,
7786,
2440,
29889,
349,
764,
393,
5864,
1302,
16453,
674,
367,
263,
937,
4331,
297,
1791,
8253,
13487,
2454,
322,
1302,
3372,
1230,
260,
583,
393,
2869,
4689,
297,
29871,
29896,
29929,
29946,
29947,
29889,
13,
2855,
278,
365,
25593,
29892,
940,
338,
278,
3118,
1058,
5771,
1434,
366,
29889,
940,
674,
367,
411,
366,
29892,
940,
674,
451,
5967,
366,
3643,
363,
29879,
1296,
366,
29936,
437,
451,
8866,
3643,
367,
766,
13029,
287,
29889,
13,
29899,
897,
12811,
21926,
29871,
29941,
29896,
29901,
29947,
13,
1609,
23738,
21296,
13,
1576,
8713,
6392,
3052,
488,
19799,
8889,
575,
297,
278,
10387,
22895,
5037,
4272,
310,
501,
4317,
394,
29899,
29943,
801,
29885,
29889,
13,
29956,
287,
4515,
3250,
29892,
29871,
29929,
6339,
29871,
29906,
29900,
29906,
29906,
891,
530,
3677,
423,
12664,
3052,
488,
471,
17285,
515,
8713,
2849,
964,
14622,
11996,
297,
278,
758,
29899,
25516,
6199,
310,
15050,
4515,
3250,
7250,
29892,
9508,
292,
263,
5544,
573,
21283,
515,
278,
22895,
5037,
9121,
29889,
13,
1576,
8713,
6392,
3052,
488,
19799,
8889,
575,
297,
278,
10387,
22895,
5037,
4272,
310,
501,
4317,
394,
29899,
29943,
801,
29885,
322,
760,
310,
14622,
3685,
4568,
29889,
450,
3052,
488,
3902,
6797,
286,
1458,
381,
29892,
278,
11996,
5282,
1947,
25571,
313,
1367,
29943,
29897,
8967,
29892,
577,
694,
1006,
1441,
471,
5181,
29889,
13,
797,
2933,
29892,
278,
3553,
29943,
1497,
372,
15469,
376,
7610,
2161,
29899,
517,
29899,
1466,
3052,
488,
22525,
297,
8713,
2849,
29892,
3704,
2971,
279,
322,
3677,
423,
12664,
10193,
583,
1213,
13,
29908,
1576,
3553,
29943,
674,
6773,
304,
12566,
278,
2071,
583,
310,
278,
4306,
310,
11996,
322,
967,
6993,
1699,
278,
22895,
5037,
9121,
1497,
373,
967,
4700,
445,
7250,
29889,
13,
1576,
8713,
6392,
3677,
423,
12664,
3052,
488,
471,
8967,
368,
15241,
925,
1156,
385,
16831,
287,
22895,
5037,
4799,
21283,
2978,
9865,
6151,
375,
29889,
13,
1576,
3553,
29943,
15241,
385,
4799,
21283,
515,
278,
5305,
310,
12307,
292,
1522,
381,
329,
925,
1434,
29871,
29896,
263,
29889,
29885,
29889,
322,
17285,
1790,
4513,
310,
7101,
29899,
517,
29899,
7610,
2161,
3052,
5475,
515,
278,
20268,
273,
940,
5861,
2820,
29871,
29896,
29901,
29896,
29900,
263,
29889,
29885,
1696,
23393,
697,
8713,
6392,
20767,
29892,
281,
12449,
5320,
4045,
322,
10805,
5518,
18658,
29892,
2106,
29899,
3470,
2638,
630,
8713,
6392,
10387,
10130,
29353,
313,
29903,
2190,
29909,
29897,
8967,
29889,
13,
1576,
3646,
310,
278,
16831,
287,
22895,
5037,
19492,
267,
9242,
9815,
29892,
541,
896,
1122,
505,
1063,
3646,
292,
11840,
310,
278,
8713,
6392,
22384,
408,
1532,
408,
14883,
713,
5549,
3173,
29892,
278,
23204,
4918,
8967,
29889,
13,
3624,
9058,
1258,
451,
3440,
373,
967,
5297,
29841,
297,
278,
4688,
7250,
19492,
267,
297,
12515,
411,
967,
1472,
11235,
8898,
29889,
450,
16728,
2106,
756,
325,
20937,
29892,
3138,
29892,
304,
5557,
14883,
515,
875,
4615,
292,
3528,
297,
278,
5120,
322,
756,
24084,
3192,
393,
372,
756,
18043,
777,
21006,
310,
4799,
19492,
267,
297,
8713,
2849,
975,
278,
4940,
316,
6332,
29892,
4049,
3646,
292,
14883,
713,
25340,
7751,
1860,
304,
967,
410,
29916,
583,
297,
278,
5120,
29892,
10734,
278,
9388,
273,
968,
29899,
6707,
940,
29920,
2095,
8083,
15115,
13013,
29889,
13,
8897,
4723,
1790,
4799,
21283,
5377,
310,
9865,
6151,
375,
471,
29393,
304,
11996,
29889,
450,
3646,
727,
2086,
471,
22363,
681,
29892,
408,
612,
1212,
15753,
8967,
372,
471,
263,
940,
29920,
2095,
8083,
281,
598,
8697,
29892,
1550,
278,
23204,
4918,
29892,
7537,
292,
8713,
6392,
19626,
5745,
29892,
1497,
372,
471,
8713,
6392,
22384,
11840,
29889,
13,
7058,
21283,
3614,
2058,
925,
263,
4723,
1156,
263,
14002,
10637,
29994,
29903,
29891,
6392,
11340,
1609,
3412,
278,
22895,
5037,
29994,
29903,
29891,
6392,
5139,
29889,
13,
1576,
23204,
4918,
884,
11520,
714,
393,
7417,
16286,
5702,
414,
505,
13817,
2999,
17040,
1652,
5861,
528,
17347,
25340,
515,
14883,
304,
8713,
2849,
975,
278,
4940,
2846,
11405,
29889,
13,
4435,
29901,
313,
29933,
2429,
2710,
363,
24819,
29892,
6339,
29871,
29929,
29892,
29871,
29906,
29900,
29906,
29906,
29897,
13,
4435,
29901,
4546,
532,
29920,
29896,
29929,
29929,
29955,
29914,
5261,
787,
29889,
2851,
3393,
29889,
990,
13,
29907,
719,
714,
304,
278,
6171,
393,
278,
3553,
29943,
674,
29892,
297,
2114,
29892,
6773,
304,
12566,
278,
2071,
583,
310,
278,
4306,
310,
11996,
322,
674,
367,
9150,
297,
12515,
14883,
515,
14171,
4340,
875,
4615,
287,
297,
8713,
2849,
29889,
349,
764,
363,
278,
15332,
310,
278,
18363,
310,
14622,
11996,
408,
278,
11103,
269,
1169,
373,
1009,
3050,
10568,
29889,
13,
29908,
29943,
799,
451,
29892,
363,
306,
626,
411,
366,
29936,
367,
451,
766,
13029,
287,
29892,
363,
306,
626,
596,
4177,
29889,
306,
674,
9324,
264,
366,
29892,
4874,
29892,
306,
674,
1371,
366,
29892,
306,
674,
318,
561,
1025,
366,
411,
1619,
364,
1141,
371,
681,
1492,
1361,
1213,
13,
1609,
4699,
28156,
3695,
435,
3059,
13,
1576,
15883,
310,
2106,
297,
278,
10557,
7761,
297,
29871,
29906,
29900,
29896,
29953,
13,
1349,
1295,
3250,
29892,
29871,
29896,
29900,
6339,
29871,
29906,
29900,
29906,
29906,
891,
450,
11715,
7761,
18760,
1283,
27836,
373,
263,
11719,
975,
11996,
29915,
29879,
22944,
4660,
472,
967,
11781,
373,
16340,
29889,
8669,
29892,
372,
731,
701,
263,
21118,
304,
25917,
278,
4383,
29892,
607,
674,
3461,
967,
1284,
886,
472,
2446,
1629,
29915,
29879,
2533,
2415,
29889,
830,
4011,
1827,
278,
2228,
756,
11827,
8572,
573,
297,
278,
29871,
29945,
29945,
29899,
14242,
289,
2029,
29889,
13,
1576,
11715,
7761,
16896,
11996,
22944,
4660,
297,
5468,
29889,
2398,
29892,
19626,
304,
278,
10608,
871,
17515
] | 1,024 |
CommonCrawl
| 0.222194 | 0.173249 | 0.556914 | -0.401298 |
momentum recently, coming to a head at the day-two summit this weekend in Addis Ababa, which also celebrated the 20th anniversary of the member organization's founding.
Although observer status in the African Union is not unusual and many non-African countries and groups hold observer status ("Palestine" being one of them), offering it to Israel created a stir—so much so that despite more pressing problems, the chairperson of the African Union Commission, Moussa Faki Mahamat, felt compelled to mention the issue in his opening address and put it on the agenda.
"South Africa and Algeria are the dominant countries trying to change this decision. South Africa, a very important member of the African Union, has since the 1990s taken a very hardline, anti-Israel stance," Irit Back, head of the Inter-University Program of African Studies at Tel Aviv University, told JNS.
She said the two countries paved the way for Palestinian [Authority] Prime Minister Mohammed Shtayyeh to address the summit on Saturday. He called for the African Union to reverse its decision, saying "Israel should never be rewarded for its violation and for the apartheid regime it does impose on the Palestinian people."
Israel has been trying to win back observer status for 20 years, ever since it lost it when the Organization of African Unity was disbanded in 2002 to make way for its replacement organization, the African Union. Libyan leader Moammar Gaddafi was behind Israel's original ouster, said Back, "and since then, we've been trying to regain our status."
Back described observer status as "mainly symbolic," a "stamp of approval." Nevertheless, she noted that it comes with practical economic and diplomatic benefits. For instance, as an observer at the African Union, Israel has a greater ability to influence the bloc to change its generally anti-Israel voting record at the United Nations.
"So this can be a turning point. I'm not sure it will happen even if Israel is an observer, but it's better to be inside than outside," she said.
'An Instrument in the Service of Peace'
Efraim Inbar, president of the Jerusalem Institute for Strategy and Security (JISS), expressed reservations about Israeli efforts to join the African Union. "Africa is not as strategically important as other places," he said. "The Horn of Africa is important for us. Morocco is important for us."
Inbar says that if Israel wants to fight efforts to strip it of its newfound status, that's fine as long as it doesn't mean expending political capital. Israel should focus more on bilateral relations and less on seeking influence in multilateral organizations, he said.
Israel's Foreign Ministry sees it differently. In a statement on Sunday, it said that "Israel attaches great importance to expanding the dialogue and cooperation with the African Union in line with changes in the Middle East and views it as an important expression of our shared activities for the continent's next generation."
Israel's history with Africa goes back to the early years of the state when the country's founding father and first prime minister, David Ben-Gurion, saw friendship with African countries as a way to break Israel's diplomatic isolation. However, with the Yom Kippur War in 1973, Arab countries successfully pressured most African countries to sever ties with Israel.
That breakdown still casts a shadow over Israel–Africa relations. "It was a traumatic event from Israel's side, and it still affects Israel's attitude towards Africa. There is this kind of suspicion that it could happen again," said Back.
She said a major milestone was Sudan joining the Abraham Accords. "Sudan is really important in this context because it's part of the Arab League. It sometimes identified itself as an Arab country as well, not just an African country," she noted.
Back said it was surprising when Israel gained observer status over the summer, and it's equally surprising now that it might be taken away.
In his opening speech, Mahamat gave no indication of which side will eventually win. He both reaffirmed the group's commitment to "the cause of the Palestinian people" and defended the decision to give Israel observer status, calling it "an instrument in the service of peace."
Whatever happens, Back emphasized that Israel should invest more effort in forging ties with African nations and that the Foreign Ministry isn't doing enough in this regard.
|
[
19399,
10325,
29892,
6421,
304,
263,
2343,
472,
278,
2462,
29899,
10184,
2533,
2415,
445,
4723,
355,
297,
3462,
275,
1976,
5363,
29892,
607,
884,
26301,
278,
29871,
29906,
29900,
386,
6957,
27547,
310,
278,
4509,
13013,
29915,
29879,
1476,
292,
29889,
13,
2499,
3592,
22944,
4660,
297,
278,
11715,
7761,
338,
451,
22910,
322,
1784,
1661,
29899,
29909,
1341,
2185,
10916,
322,
6471,
4808,
22944,
4660,
4852,
18210,
342,
457,
29908,
1641,
697,
310,
963,
511,
27032,
372,
304,
11996,
2825,
263,
23546,
30003,
578,
1568,
577,
393,
15020,
901,
24795,
4828,
29892,
278,
11774,
10532,
310,
278,
11715,
7761,
11444,
29892,
341,
681,
4977,
383,
9940,
10082,
314,
271,
29892,
7091,
752,
14356,
304,
3585,
278,
2228,
297,
670,
8718,
3211,
322,
1925,
372,
373,
278,
946,
8395,
29889,
13,
29908,
29903,
2438,
10557,
322,
838,
914,
423,
526,
278,
28526,
10916,
1811,
304,
1735,
445,
10608,
29889,
4275,
10557,
29892,
263,
1407,
4100,
4509,
310,
278,
11715,
7761,
29892,
756,
1951,
278,
29871,
29896,
29929,
29929,
29900,
29879,
4586,
263,
1407,
2898,
1220,
29892,
9418,
29899,
3624,
9058,
380,
749,
1699,
306,
768,
7437,
29892,
2343,
310,
278,
4124,
29899,
11574,
537,
7835,
310,
11715,
16972,
472,
18815,
7740,
440,
3014,
29892,
5429,
435,
3059,
29889,
13,
13468,
1497,
278,
1023,
10916,
282,
10511,
278,
982,
363,
22053,
262,
713,
518,
13720,
537,
29962,
15512,
7668,
12929,
28913,
317,
400,
388,
4099,
29882,
304,
3211,
278,
2533,
2415,
373,
24211,
29889,
940,
2000,
363,
278,
11715,
7761,
304,
11837,
967,
10608,
29892,
5934,
376,
3624,
9058,
881,
2360,
367,
20751,
287,
363,
967,
5537,
362,
322,
363,
278,
12435,
9722,
22384,
372,
947,
2411,
852,
373,
278,
22053,
262,
713,
2305,
1213,
13,
3624,
9058,
756,
1063,
1811,
304,
5401,
1250,
22944,
4660,
363,
29871,
29906,
29900,
2440,
29892,
3926,
1951,
372,
5714,
372,
746,
278,
9205,
2133,
310,
11715,
20872,
471,
766,
4980,
287,
297,
29871,
29906,
29900,
29900,
29906,
304,
1207,
982,
363,
967,
16920,
13013,
29892,
278,
11715,
7761,
29889,
8153,
10094,
11822,
4546,
314,
3034,
402,
1202,
2142,
29875,
471,
5742,
11996,
29915,
29879,
2441,
2123,
2475,
29892,
1497,
7437,
29892,
376,
392,
1951,
769,
29892,
591,
29915,
345,
1063,
1811,
304,
1072,
475,
1749,
4660,
1213,
13,
5841,
5439,
22944,
4660,
408,
376,
3396,
368,
5829,
293,
1699,
263,
376,
303,
1160,
310,
2134,
791,
1213,
25678,
29892,
1183,
11682,
393,
372,
5304,
411,
15031,
17407,
322,
13487,
2454,
23633,
29889,
1152,
2777,
29892,
408,
385,
22944,
472,
278,
11715,
7761,
29892,
11996,
756,
263,
7621,
11509,
304,
9949,
278,
289,
2029,
304,
1735,
967,
6892,
9418,
29899,
3624,
9058,
28931,
2407,
472,
278,
3303,
18269,
29889,
13,
29908,
6295,
445,
508,
367,
263,
14712,
1298,
29889,
306,
29915,
29885,
451,
1854,
372,
674,
3799,
1584,
565,
11996,
338,
385,
22944,
29892,
541,
372,
29915,
29879,
2253,
304,
367,
2768,
1135,
5377,
1699,
1183,
1497,
29889,
13,
29915,
2744,
2799,
15461,
297,
278,
6692,
310,
24819,
29915,
13,
29923,
20910,
326,
512,
1646,
29892,
6673,
310,
278,
23204,
8907,
363,
3767,
8963,
322,
14223,
313,
29967,
29902,
1799,
511,
13384,
620,
6972,
800,
1048,
22895,
5037,
14231,
304,
5988,
278,
11715,
7761,
29889,
376,
29909,
18961,
338,
451,
408,
16650,
1711,
4100,
408,
916,
7600,
1699,
540,
1497,
29889,
376,
1576,
19674,
310,
10557,
338,
4100,
363,
502,
29889,
3879,
542,
1111,
338,
4100,
363,
502,
1213,
13,
797,
1646,
4083,
393,
565,
11996,
10753,
304,
8589,
14231,
304,
17820,
372,
310,
967,
716,
11940,
4660,
29892,
393,
29915,
29879,
2691,
408,
1472,
408,
372,
1838,
29915,
29873,
2099,
1518,
2548,
8604,
7483,
29889,
11996,
881,
8569,
901,
373,
13181,
1008,
284,
5302,
322,
3109,
373,
25738,
9949,
297,
1773,
309,
1008,
284,
25700,
29892,
540,
1497,
29889,
13,
3624,
9058,
29915,
29879,
19358,
20886,
18553,
372,
17587,
29889,
512,
263,
3229,
373,
16340,
29892,
372,
1497,
393,
376,
3624,
9058,
10641,
267,
2107,
13500,
304,
7985,
292,
278,
7928,
434,
322,
1302,
16453,
411,
278,
11715,
7761,
297,
1196,
411,
3620,
297,
278,
14253,
6932,
322,
8386,
372,
408,
385,
4100,
4603,
310,
1749,
7258,
14188,
363,
278,
25523,
29915,
29879,
2446,
12623,
1213,
13,
3624,
9058,
29915,
29879,
4955,
411,
10557,
5771,
1250,
304,
278,
4688,
2440,
310,
278,
2106,
746,
278,
4234,
29915,
29879,
1476,
292,
4783,
322,
937,
6019,
11050,
29892,
4699,
4111,
29899,
29954,
332,
291,
29892,
4446,
27994,
411,
11715,
10916,
408,
263,
982,
304,
2867,
11996,
29915,
29879,
13487,
2454,
11695,
362,
29889,
2398,
29892,
411,
278,
612,
290,
476,
8377,
332,
3362,
297,
29871,
29896,
29929,
29955,
29941,
29892,
10387,
10916,
8472,
3965,
2955,
1556,
11715,
10916,
304,
2775,
260,
583,
411,
11996,
29889,
13,
7058,
2867,
3204,
1603,
4320,
29879,
263,
15504,
975,
11996,
29994,
29909,
18961,
5302,
29889,
376,
3112,
471,
263,
1020,
398,
2454,
1741,
515,
11996,
29915,
29879,
2625,
29892,
322,
372,
1603,
6602,
29879,
11996,
29915,
29879,
26309,
7113,
10557,
29889,
1670,
338,
445,
2924,
310,
8872,
15353,
393,
372,
1033,
3799,
1449,
1699,
1497,
7437,
29889,
13,
13468,
1497,
263,
4655,
2316,
27744,
471,
8383,
273,
22960,
278,
24763,
4831,
4339,
29889,
376,
29903,
566,
273,
338,
2289,
4100,
297,
445,
3030,
1363,
372,
29915,
29879,
760,
310,
278,
10387,
5165,
29889,
739,
6041,
15659,
3528,
408,
385,
10387,
4234,
408,
1532,
29892,
451,
925,
385,
11715,
4234,
1699,
1183,
11682,
29889,
13,
5841,
1497,
372,
471,
26800,
746,
11996,
17515,
22944,
4660,
975,
278,
11801,
29892,
322,
372,
29915,
29879,
18018,
26800,
1286,
393,
372,
1795,
367,
4586,
3448,
29889,
13,
797,
670,
8718,
12032,
29892,
10082,
314,
271,
4846,
694,
4221,
362,
310,
607,
2625,
674,
10201,
5401,
29889,
940,
1716,
337,
3470,
381,
2168,
278,
2318,
29915,
29879,
9063,
358,
304,
376,
1552,
4556,
310,
278,
22053,
262,
713,
2305,
29908,
322,
822,
2760,
278,
10608,
304,
2367,
11996,
22944,
4660,
29892,
5432,
372,
376,
273,
11395,
297,
278,
2669,
310,
10776,
1213,
13,
8809,
5564,
5930,
29892,
7437,
19310,
1891,
393,
11996,
881,
13258,
901,
7225,
297,
363,
3460,
260,
583,
411,
11715,
19079,
322,
393,
278,
19358,
20886,
3508,
29915,
29873,
2599,
3307,
297,
445,
4880,
29889
] | 1,024 |
CommonCrawl
| 0.89195 | 0.665109 | 0.843829 | 0.555028 |
"Israel actually needs to be more active and increase the importance of Africa in its foreign policy," she said.
Inbar, however, maintained that Israel should not put too much store in the African Union, saying "Israel is doing quite well internationally, and we have some excellent relations with African countries."
Photo Credit: Paul Kagame/Flickr/JNS.org
Pray that South Africa will be unsuccessful in influencing the African Union with its anti-Semitic views, especially since Israel has established relations with so many African nations in the last few years. Pray that Israel will maintain its observer status in the union, successfully leveraging that relationship to change the bloc's anti-Israel voting record at the United Nations.
Pronounce them guilty, O God! Let them fall by their own counsels; cast them out in the multitude of their transgressions.
- Psalm 5:10a
by Israel Kasnett ~ JNS
United Arab Emirates Ministry of Education
Monday, 7 February 2022 | The Institute for Monitoring Peace and Tolerance in School Education (IMPACT-se) has uncovered thousands of pages of new teaching material produced by the Palestinian Authority [PA] that directly calls for violence and promotes anti-Semitism, even after promising European Union donors it would implement changes.
IMPACT-se found that rather than publishing revised textbooks as promised, the PA reprinted last years' criticized textbooks for use in the current school year. In parallel, it produced thousands of pages of new material, roughly equivalent in size to all the textbooks in the curriculum. The new material contains content that has been determined as worse than current or previous Palestinian textbooks, with a greater number of lessons that directly incite violence and propagate overt anti-Semitism.
For example, the material demands that students die as martyrs to liberate the Al-Aqsa Mosque and explains that those who die as martyrs killing infidels (Christians and Jews) will receive grace and be greatly rewarded.
IMPACT-se CEO Marcus Sheff said the PA "seems to have gone to a great deal of effort to hoodwink its donors. Faced with a clear call by the EU for them to create new textbooks free of hate and anti-Semitism, the PA simply reprinted the old ones, then produced thousands of pages of new teaching material with content worse than the textbooks themselves."
IMPACT-se presented its findings to representatives from the EU Commission, as well as to Parliament members in Brussels, who had no knowledge that the old textbooks were still being used in Palestinian and UNRWA [United Nations Relief and Works Agency] schools, or that a set of new, hate-filled materials had been produced in 2021.
In contrast to students in the PA, those in the United Arab Emirates related to the school curriculum have been updated to teach values of peace and tolerance. In what can be viewed as a sure sign of change in the region, many examples of anti-Semitism or incitement have been removed from the curriculum.
IMPACT-se also released a report on January 20 examining 220 Arab-language textbooks in grades 1–12 from the UAE's national curriculum, printed between 2016 and 2021.
Still Some Room for Improvement
Niclas Herbst, a German politician of the Christian Democratic Union and a member of the European Parliament who is familiar with the issue, told JNS the improvements in the UAE curriculum "are a step in the right direction" as they "promote tolerance and trustful interfaith relations."
Notably, much of the anti-Israel material has been moderated. Passages that previously demonized Israel, presented anti-Semitic conspiracies and blamed "the Zionist enemy" for seeking to exterminate the Palestinian people have been removed. Passages focusing on tolerance towards Jews are widespread throughout the textbooks. Especially noteworthy is the removal of a passage that presented the Palestinian issue as "the basis of conflicts in the Middle East."
IMPACT-se CEO Marcus Sheff told JNS, "Emirati textbooks are reflective of the assessment made by Crown Prince Mohamed bin Zayed over a decade ago that the West is a potential ally and not a colonial threat; that radicalism is categorically wrong and self-defe
|
[
376,
3624,
9058,
2869,
4225,
304,
367,
901,
6136,
322,
7910,
278,
13500,
310,
10557,
297,
967,
9117,
8898,
1699,
1183,
1497,
29889,
13,
797,
1646,
29892,
3138,
29892,
19949,
393,
11996,
881,
451,
1925,
2086,
1568,
3787,
297,
278,
11715,
7761,
29892,
5934,
376,
3624,
9058,
338,
2599,
3755,
1532,
2836,
362,
635,
29892,
322,
591,
505,
777,
15129,
5302,
411,
11715,
10916,
1213,
13,
25971,
24596,
277,
29901,
3739,
476,
351,
420,
29914,
29943,
1406,
29878,
29914,
29967,
3059,
29889,
990,
13,
29925,
764,
393,
4275,
10557,
674,
367,
443,
8698,
1319,
297,
13787,
3277,
278,
11715,
7761,
411,
967,
9418,
29899,
29903,
21976,
293,
8386,
29892,
7148,
1951,
11996,
756,
7841,
5302,
411,
577,
1784,
11715,
19079,
297,
278,
1833,
2846,
2440,
29889,
349,
764,
393,
11996,
674,
7344,
967,
22944,
4660,
297,
278,
9833,
29892,
8472,
26610,
6751,
393,
9443,
304,
1735,
278,
289,
2029,
29915,
29879,
9418,
29899,
3624,
9058,
28931,
2407,
472,
278,
3303,
18269,
29889,
13,
29925,
1617,
21543,
963,
27719,
29892,
438,
4177,
29991,
2803,
963,
6416,
491,
1009,
1914,
2613,
29879,
1379,
29936,
4320,
963,
714,
297,
278,
1773,
4279,
310,
1009,
1301,
3663,
1080,
29889,
13,
29899,
10144,
17120,
29871,
29945,
29901,
29896,
29900,
29874,
13,
1609,
11996,
19157,
20764,
3695,
435,
3059,
13,
2525,
1573,
10387,
2812,
381,
1078,
20886,
310,
13151,
13,
29924,
898,
388,
29892,
29871,
29955,
6339,
29871,
29906,
29900,
29906,
29906,
891,
450,
8907,
363,
2598,
2105,
292,
24819,
322,
16977,
261,
749,
297,
4523,
13151,
313,
29902,
3580,
17923,
29899,
344,
29897,
756,
443,
11911,
287,
17202,
310,
6515,
310,
716,
18819,
5518,
7371,
491,
278,
22053,
262,
713,
13361,
537,
518,
7228,
29962,
393,
4153,
5717,
363,
21448,
322,
2504,
4769,
9418,
29899,
29903,
21976,
1608,
29892,
1584,
1156,
2504,
5921,
7824,
7761,
1016,
943,
372,
723,
2334,
3620,
29889,
13,
29902,
3580,
17923,
29899,
344,
1476,
393,
3265,
1135,
27256,
337,
11292,
1426,
12733,
408,
22399,
29892,
278,
17687,
2062,
524,
287,
1833,
2440,
29915,
11164,
1891,
1426,
12733,
363,
671,
297,
278,
1857,
3762,
1629,
29889,
512,
8943,
29892,
372,
7371,
17202,
310,
6515,
310,
716,
5518,
29892,
20928,
7126,
297,
2159,
304,
599,
278,
1426,
12733,
297,
278,
16256,
12906,
398,
29889,
450,
716,
5518,
3743,
2793,
393,
756,
1063,
10087,
408,
15029,
1135,
1857,
470,
3517,
22053,
262,
713,
1426,
12733,
29892,
411,
263,
7621,
1353,
310,
3109,
787,
393,
4153,
297,
2036,
21448,
322,
13089,
403,
975,
29873,
9418,
29899,
29903,
21976,
1608,
29889,
13,
2831,
1342,
29892,
278,
5518,
1261,
4167,
393,
8041,
762,
408,
14436,
29891,
2288,
304,
7866,
403,
278,
838,
29899,
29909,
29939,
4977,
6630,
802,
322,
18568,
393,
1906,
1058,
762,
408,
14436,
29891,
2288,
23393,
3041,
333,
1379,
313,
18687,
5834,
322,
22057,
29897,
674,
7150,
17659,
322,
367,
11180,
20751,
287,
29889,
13,
29902,
3580,
17923,
29899,
344,
14645,
29949,
27390,
2296,
600,
1497,
278,
17687,
376,
344,
1567,
304,
505,
7695,
304,
263,
2107,
5376,
310,
7225,
304,
298,
2092,
29893,
682,
967,
1016,
943,
29889,
14184,
287,
411,
263,
2821,
1246,
491,
278,
19007,
363,
963,
304,
1653,
716,
1426,
12733,
3889,
310,
26277,
322,
9418,
29899,
29903,
21976,
1608,
29892,
278,
17687,
3763,
2062,
524,
287,
278,
2030,
6743,
29892,
769,
7371,
17202,
310,
6515,
310,
716,
18819,
5518,
411,
2793,
15029,
1135,
278,
1426,
12733,
6053,
1213,
13,
29902,
3580,
17923,
29899,
344,
9132,
967,
1284,
886,
304,
2755,
5056,
515,
278,
19007,
11444,
29892,
408,
1532,
408,
304,
13212,
5144,
297,
1771,
1558,
1379,
29892,
1058,
750,
694,
7134,
393,
278,
2030,
1426,
12733,
892,
1603,
1641,
1304,
297,
22053,
262,
713,
322,
8291,
29934,
12982,
518,
2525,
1573,
18269,
6376,
2575,
322,
13976,
29353,
29962,
12462,
29892,
470,
393,
263,
731,
310,
716,
29892,
26277,
29899,
26940,
17279,
750,
1063,
7371,
297,
29871,
29906,
29900,
29906,
29896,
29889,
13,
797,
12814,
304,
8041,
297,
278,
17687,
29892,
1906,
297,
278,
3303,
10387,
2812,
381,
1078,
4475,
304,
278,
3762,
16256,
12906,
398,
505,
1063,
4784,
304,
6860,
1819,
310,
10776,
322,
20341,
749,
29889,
512,
825,
508,
367,
24774,
408,
263,
1854,
1804,
310,
1735,
297,
278,
5120,
29892,
1784,
6455,
310,
9418,
29899,
29903,
21976,
1608,
470,
5528,
17334,
505,
1063,
6206,
515,
278,
16256,
12906,
398,
29889,
13,
29902,
3580,
17923,
29899,
344,
884,
5492,
263,
3461,
373,
5490,
29871,
29906,
29900,
4392,
2827,
29871,
29906,
29906,
29900,
10387,
29899,
11675,
1426,
12733,
297,
867,
3076,
29871,
29896,
29994,
29896,
29906,
515,
278,
501,
16036,
29915,
29879,
4797,
16256,
12906,
398,
29892,
13350,
1546,
29871,
29906,
29900,
29896,
29953,
322,
29871,
29906,
29900,
29906,
29896,
29889,
13,
855,
453,
3834,
25114,
363,
1954,
16123,
882,
13,
29940,
293,
3333,
2439,
7891,
29892,
263,
5332,
14099,
310,
278,
6111,
19083,
7761,
322,
263,
4509,
310,
278,
7824,
13212,
1058,
338,
9985,
411,
278,
2228,
29892,
5429,
435,
3059,
278,
28473,
297,
278,
501,
16036,
16256,
12906,
398,
376,
598,
263,
4331,
297,
278,
1492,
5305,
29908,
408,
896,
376,
14032,
866,
20341,
749,
322,
9311,
1319,
1006,
5444,
389,
5302,
1213,
13,
3664,
2197,
29892,
1568,
310,
278,
9418,
29899,
3624,
9058,
5518,
756,
1063,
17768,
630,
29889,
6978,
1179,
393,
9251,
8033,
1891,
11996,
29892,
9132,
9418,
29899,
29903,
21976,
293,
21588,
3055,
2478,
322,
1999,
2795,
376,
1552,
796,
291,
391,
11103,
29908,
363,
25738,
304,
429,
18821,
403,
278,
22053,
262,
713,
2305,
505,
1063,
6206,
29889,
6978,
1179,
12789,
4746,
373,
20341,
749,
7113,
22057,
526,
281,
2247,
29886,
949,
10106,
278,
1426,
12733,
29889,
3423,
25009,
451,
809,
2072,
29891,
338,
278,
28744,
310,
263,
13382,
393,
9132,
278,
22053,
262,
713,
2228,
408,
376,
1552,
8405,
310,
28792,
297,
278,
14253,
6932,
1213,
13,
29902,
3580,
17923,
29899,
344,
14645,
29949,
27390,
2296,
600,
5429,
435,
3059,
29892,
376,
6026,
381,
2219,
1426,
12733,
526,
9432,
573,
310,
278,
24809,
358,
1754,
491,
25306,
10787,
12929,
2795,
9016,
796,
388,
287,
975,
263,
316,
6332,
8020,
393,
278,
3122,
338,
263,
7037,
394,
368,
322,
451,
263,
25539,
28469,
29936,
393,
24818,
1608,
338,
11608,
1711,
2743,
322,
1583,
29899,
311,
1725
] | 1,024 |
CommonCrawl
| 0.48439 | 0.659918 | 0.958596 | 0.714403 |
While you were sleeping: Putin's former bodyguard won't debate, prefers to duel
Maybe the opposition leader would have been more amenable to a dance off. As well, we check in on Hurricane Willa.
Men transport a boat on a truck in Teacapan, Mexico, Oct. 22, 2018, before the arrival of Hurricane Willa. ALFREDO ESTRELLA / AFP/Getty Images
Here are a few things you might have missed while you were sleeping.
It's a pretty wide stopgap: The Canadian Coast Guard says three "interim" icebreakers that were recently purchased without a competition will be used for the next 15 to 20 years. Coast Guard officials revealed the timeframe in interviews with The Canadian Press while playing down concerns about the state of their aging fleet — and the challenges in building replacements. The government in August agreed to sole-sourcing the purchase of three used icebreakers from Davie Shipbuilding for $610 million, saying a stop-gap was needed until replacements could be built. The deal represented a win for the Quebec-based shipyard, which had been lobbying hard for additional federal work, and should ease pressure on the Coast Guard's icebreaking fleet.
HAPPY BIRTHDAY CCGS GRIFFON! #OTD 26 SEPT 1969 CCGS GRIFFON (Hull#664) was launched September 26, 1969 by Davie Shipbuilding Ltd. Lauzon, Quebec for the Canadian Coast Guard 49 years old today. @chantierdavie @CoastGuardCAN @DFO_Central @CoastGuardShips @MarineDelivers #CCGProud pic.twitter.com/hvVBlJguSQ
— Joanne Crack (@PrescottAnchor) September 26, 2018
63 days till Christmas: Small businesses called for a speedy end to rotating walkouts in four cities by Canada Post employees, citing concern that a prolonged strike could chew into their profits ahead of the busy holiday shopping season. The Canadian Federation of Independent Business also urged the post office to rein in spending to reduce costs and warned postal workers that continued job action could hurt them and their employer. The Canadian Union of Postal Workers (CUPW), which represents 50,000 postal employees, launched rotating demonstrations in Victoria, Edmonton, Windsor, Ont., and Halifax on Monday to pressure Canada Post into accepting contract changes. The union said almost 9,000 workers in the Greater Toronto Area would walk off the job Tuesday at 12:01 a.m. "For us to reach any agreements, Canada Post has to talk about the changing nature of postal work and the issues that stem from the growth of parcels," said the CUPW national president.
Willa's landfall: Authorities rushed to evacuate low-lying areas and set up shelters as an "extremely dangerous" Hurricane Willa with winds of 230 kilometres per hour headed toward a Tuesday afternoon landfall along a stretch of Mexico's Pacific coast dotted with high-rise resorts, surfing beaches and fishing villages. Farther south, meanwhile, Mexican officials there had been 12 deaths related to heavy rains from Tropical Storm Vicente. Willa briefly reached Category 5 strength, then weakened a bit to Category 4. But the U.S. National Hurricane Center warned that it still was likely to bring "life-threatening storm surge, wind and rainfall" to parts of west-central and southwestern Mexico. The hurricane was expected to first pass over or near the Islas Marias, which includes a nature preserve and a federal prison.
No debate, thanks: President Vladimir Putin's former bodyguard has rejected an opposition leader's invitation for a television debate after challenging him to a duel. General Viktor Zolotov, chief of Russia's National Guard and Putin's longtime bodyguard, recorded a rare video address to opposition leader Alexei Navalny last month, challenging him to a duel over his investigation into corruption at the National Guard. Navalny, who was released Sunday after spending 50 days in jail for an unsanctioned protest, put out a video Thursday, reiterating his claims and inviting Zolotov to
|
[
5806,
366,
892,
8709,
292,
29901,
12065,
262,
29915,
29879,
4642,
3573,
17728,
2113,
29915,
29873,
27836,
29892,
758,
25534,
304,
868,
295,
13,
22762,
278,
19626,
11822,
723,
505,
1063,
901,
626,
12007,
304,
263,
17948,
1283,
29889,
1094,
1532,
29892,
591,
1423,
297,
373,
379,
1038,
26655,
399,
2911,
29889,
13,
28154,
8608,
263,
13006,
373,
263,
534,
2707,
297,
1920,
562,
21419,
29892,
12568,
29892,
4756,
29889,
29871,
29906,
29906,
29892,
29871,
29906,
29900,
29896,
29947,
29892,
1434,
278,
18517,
310,
379,
1038,
26655,
399,
2911,
29889,
14445,
29943,
1525,
3970,
382,
1254,
1525,
2208,
29909,
847,
23844,
29925,
29914,
2577,
1017,
1954,
1179,
13,
10605,
526,
263,
2846,
2712,
366,
1795,
505,
13726,
1550,
366,
892,
8709,
292,
29889,
13,
3112,
29915,
29879,
263,
5051,
9377,
5040,
29887,
481,
29901,
450,
11443,
17700,
13211,
4083,
2211,
376,
1639,
326,
29908,
14890,
8690,
414,
393,
892,
10325,
20848,
1728,
263,
13888,
674,
367,
1304,
363,
278,
2446,
29871,
29896,
29945,
304,
29871,
29906,
29900,
2440,
29889,
17700,
13211,
24921,
17845,
278,
931,
2557,
297,
1006,
7406,
411,
450,
11443,
5254,
1550,
8743,
1623,
21838,
1048,
278,
2106,
310,
1009,
946,
292,
22338,
813,
322,
278,
18066,
267,
297,
5214,
1634,
4620,
4110,
29889,
450,
5874,
297,
3111,
15502,
304,
14419,
29899,
29879,
473,
3277,
278,
20590,
310,
2211,
1304,
14890,
8690,
414,
515,
3589,
347,
1383,
666,
25237,
363,
395,
29953,
29896,
29900,
7284,
29892,
5934,
263,
5040,
29899,
29887,
481,
471,
4312,
2745,
1634,
4620,
4110,
1033,
367,
4240,
29889,
450,
5376,
9875,
263,
5401,
363,
278,
27605,
29899,
6707,
7751,
19852,
29892,
607,
750,
1063,
658,
1327,
5414,
2898,
363,
5684,
17097,
664,
29892,
322,
881,
16326,
12959,
373,
278,
17700,
13211,
29915,
29879,
14890,
1030,
5086,
22338,
29889,
13,
29950,
3301,
20055,
350,
8193,
4690,
28658,
19178,
10749,
402,
3960,
4198,
1164,
29991,
396,
2891,
29928,
29871,
29906,
29953,
3725,
7982,
29871,
29896,
29929,
29953,
29929,
19178,
10749,
402,
3960,
4198,
1164,
313,
29950,
913,
29937,
29953,
29953,
29946,
29897,
471,
15241,
3839,
29871,
29906,
29953,
29892,
29871,
29896,
29929,
29953,
29929,
491,
3589,
347,
1383,
666,
25237,
19806,
29889,
25092,
6626,
29892,
27605,
363,
278,
11443,
17700,
13211,
29871,
29946,
29929,
2440,
2030,
9826,
29889,
732,
13775,
631,
29881,
485,
347,
732,
7967,
579,
9485,
538,
29907,
2190,
732,
4037,
29949,
29918,
23369,
1705,
732,
7967,
579,
9485,
538,
2713,
4512,
732,
7083,
457,
13157,
1536,
396,
4174,
29954,
4040,
2736,
11942,
29889,
24946,
29889,
510,
29914,
29882,
29894,
29963,
10358,
29967,
2543,
29903,
29984,
13,
30003,
3650,
11276,
14279,
384,
20164,
13504,
29883,
1501,
24458,
29897,
3839,
29871,
29906,
29953,
29892,
29871,
29906,
29900,
29896,
29947,
13,
29953,
29941,
3841,
3428,
17661,
29901,
18285,
5381,
267,
2000,
363,
263,
6210,
29891,
1095,
304,
5731,
1218,
6686,
17718,
297,
3023,
14368,
491,
7400,
4918,
22873,
29892,
7537,
292,
5932,
393,
263,
27044,
287,
21283,
1033,
923,
29893,
964,
1009,
2600,
1169,
14432,
310,
278,
19587,
8753,
22394,
17394,
3262,
4259,
29889,
450,
11443,
20438,
310,
25266,
15197,
884,
5065,
3192,
278,
1400,
8034,
304,
15561,
297,
805,
2548,
304,
10032,
21544,
322,
1370,
9571,
29767,
17162,
393,
7572,
4982,
3158,
1033,
21682,
963,
322,
1009,
5703,
261,
29889,
450,
11443,
7761,
310,
4918,
284,
5244,
414,
313,
29907,
4897,
29956,
511,
607,
11524,
29871,
29945,
29900,
29892,
29900,
29900,
29900,
29767,
22873,
29892,
15241,
5731,
1218,
9004,
800,
297,
14212,
29892,
2155,
14132,
265,
29892,
399,
12772,
272,
29892,
18265,
1696,
322,
8142,
361,
1165,
373,
27822,
304,
12959,
7400,
4918,
964,
25967,
8078,
3620,
29889,
450,
9833,
1497,
4359,
29871,
29929,
29892,
29900,
29900,
29900,
17162,
297,
278,
4122,
1008,
17612,
18320,
723,
6686,
1283,
278,
4982,
323,
1041,
3250,
472,
29871,
29896,
29906,
29901,
29900,
29896,
263,
29889,
29885,
29889,
376,
2831,
502,
304,
6159,
738,
8571,
4110,
29892,
7400,
4918,
756,
304,
5193,
1048,
278,
6480,
5469,
310,
29767,
664,
322,
278,
5626,
393,
20805,
515,
278,
14321,
310,
20067,
1379,
1699,
1497,
278,
315,
4897,
29956,
4797,
6673,
29889,
13,
29956,
2911,
29915,
29879,
2982,
11950,
29901,
13361,
1907,
364,
15392,
304,
3415,
22061,
403,
4482,
29899,
5890,
10161,
322,
731,
701,
528,
295,
2153,
408,
385,
376,
1062,
1745,
873,
18215,
29908,
379,
1038,
26655,
399,
2911,
411,
8805,
29879,
310,
29871,
29906,
29941,
29900,
24016,
639,
7234,
2343,
287,
11183,
263,
323,
1041,
3250,
17724,
2982,
11950,
3412,
263,
16116,
310,
12568,
29915,
29879,
14328,
12180,
270,
15048,
411,
1880,
29899,
29878,
895,
25362,
29879,
29892,
1190,
29888,
292,
367,
14520,
322,
9427,
292,
18481,
29889,
8413,
721,
7062,
29892,
2099,
8000,
29892,
24777,
24921,
727,
750,
1063,
29871,
29896,
29906,
4892,
29879,
4475,
304,
9416,
1153,
1144,
515,
21601,
936,
24444,
13423,
2016,
29889,
399,
2911,
23359,
7450,
17943,
29871,
29945,
9324,
29892,
769,
8062,
6419,
263,
2586,
304,
17943,
29871,
29946,
29889,
1205,
278,
501,
29889,
29903,
29889,
3086,
379,
1038,
26655,
7817,
1370,
9571,
393,
372,
1603,
471,
5517,
304,
6963,
376,
19264,
29899,
386,
276,
2579,
292,
14280,
1190,
479,
29892,
8805,
322,
1153,
7192,
497,
29908,
304,
5633,
310,
5833,
29899,
25171,
322,
7062,
22741,
12568,
29889,
450,
298,
1038,
26655,
471,
3806,
304,
937,
1209,
975,
470,
2978,
278,
1317,
3333,
1085,
3173,
29892,
607,
7805,
263,
5469,
19905,
322,
263,
17097,
8475,
29889,
13,
3782,
27836,
29892,
3969,
29901,
7178,
27880,
12065,
262,
29915,
29879,
4642,
3573,
17728,
756,
22225,
385,
19626,
11822,
29915,
29879,
2437,
7018,
363,
263,
11456,
27836,
1156,
18066,
292,
1075,
304,
263,
868,
295,
29889,
4593,
18503,
7345,
796,
324,
327,
586,
29892,
9087,
310,
12710,
29915,
29879,
3086,
13211,
322,
12065,
262,
29915,
29879,
1472,
2230,
3573,
17728,
29892,
10478,
263,
10812,
4863,
3211,
304,
19626,
11822,
4827,
10096,
21590,
1460,
1833,
4098,
29892,
18066,
292,
1075,
304,
263,
868,
295,
975,
670,
22522,
964,
1034,
18953,
472,
278,
3086,
13211,
29889,
21590,
1460,
29892,
1058,
471,
5492,
16340,
1156,
805,
2548,
29871,
29945,
29900,
3841,
297,
432,
737,
363,
385,
9644,
273,
428,
287,
10021,
29892,
1925,
714,
263,
4863,
498,
1295,
3250,
29892,
337,
1524,
1218,
670,
16726,
322,
2437,
11407,
796,
324,
327,
586,
304
] | 1,024 |
CommonCrawl
| -0.501903 | -1.128267 | -0.09044 | -1.152555 |
January in Four Weeks
Here's the thing: I have this list of "BIG GOALS FOR THE MONTH" that are large, recurring things I would like to accomplish once a month: things like reading a book, completing a large home improvement project, and writing a new blog post. If you're a longtime reader, you know how well that goal has been going (hint: BADLY). I personally benefit from reading many blogs, instructions, and musings on the internet. I believe to my core that we must be creators and consumers; yet I incessantly consume, consume, consume the hearts and stories of others, but I struggle to share my own. It's big scary.
So here I am, on January 31st, with my "BIG GOALS FOR THE MONTH" list that is entirely checked off except for this one last blogging task. Usually I have at least two goals that are unchecked by the end of the month; this allows me to shrug it off and say "I never actually aim to finish them all, just want to try and finish as many as I can." I make excuses like "my home improvement project this month was so big. Of course I can't write a blog." Eventually so much time goes on between posts that it feels like I have too much to say, but when I start typing it feels like I don't actually have anything important to contribute to internet humanity. I pass it off until next month. But I have to be able to check it off this time. I mean, it's January. I cannot start the year off by leaving one box unchecked out of sheer laziness.
As a result, you get this unplanned, rambling, likely unedited post.
In defense of my procrastination: it was a full month.
Week One: Art of the State 2019 Install
"Homecoming" by Lara Whitley at Art of the State 2019; Arvada Center for the Arts and Humanities
I spent the first week of January working as a Gallery Assistant at the Arvada Center for the Arts and Humanities, hanging Art of the State 2019. It's a gig that I came to by a sort of cloud-parting magic early last year. I was on a behind-the-scenes tour of the Arvada Center with my Leadership Arvada group; while the tour centered around the theater spaces, we ended standing in the main gallery in the midst of one of the most compelling exhibition concepts I have ever seen. I could write pages about it, about why I thought it was pure genius, about how edifying it was to see as an artist.
I straggled behind the rest of my group to talk to the curator, Collin, and asked if I could help out in the galleries to learn how this aspect of the local art world worked (this makes me sound a lot more bold and awesome than it actually was; I nervously babbled about how I knew how to sort of do lots of stuff and had a pretty open schedule and could kind of build things). Although I've worked in museums and with smaller community art spaces, I knew nearly nothing about the business of being an artist and the role that regional art centers played in the larger art scene. This seemed like somewhere I could learn. Thankfully, my random-hat-full-of-half-skills and flexible schedule fit with what the galleries needed, and I was hired as a Gallery Assistant to help out seasonally during show turnovers and installs.
It is my favorite. I get to do so much cool stuff; exhibition graphics, packaging art, transporting art to and from collectors, hanging work (so much hanging), and aiding with installation work. We also move a lot of white cubes.
One of my favorite works in Art of the State 2019 is Homecoming by Lara Whitley; it is a house made of suspended glass bottles which Lara found on treks with her dog through a home-adjacent meadow. As we worked with Lara to unpack and install Homecoming, she told us beauitful and heartshaking stories of how the work came into existence, the history and meaning the bottles held for her, and ways that she interacted with the work during its first exhibition near her home.
Week Two: I ran the very long Disney Half-Marathon.
After running the Bolder Boulder last year, Jake realized his upcoming Disney vacation happened to coincide with Disney Marathon Weekend. He asked me
|
[
5490,
297,
12458,
1334,
14541,
13,
10605,
29915,
29879,
278,
2655,
29901,
306,
505,
445,
1051,
310,
376,
29933,
6259,
21947,
1964,
29903,
15842,
6093,
341,
1164,
4690,
29908,
393,
526,
2919,
29892,
1162,
1038,
292,
2712,
306,
723,
763,
304,
12709,
2748,
263,
4098,
29901,
2712,
763,
5183,
263,
3143,
29892,
1614,
1259,
263,
2919,
3271,
20414,
2060,
29892,
322,
5007,
263,
716,
12618,
1400,
29889,
960,
366,
29915,
276,
263,
1472,
2230,
9591,
29892,
366,
1073,
920,
1532,
393,
7306,
756,
1063,
2675,
313,
29882,
524,
29901,
350,
3035,
16786,
467,
29871,
30166,
29902,
22345,
14169,
515,
5183,
1784,
12618,
29879,
29892,
11994,
29892,
322,
2301,
886,
373,
278,
8986,
29889,
306,
4658,
304,
590,
7136,
393,
591,
1818,
367,
907,
4097,
322,
11233,
414,
29936,
3447,
306,
297,
985,
10835,
29151,
29892,
29151,
29892,
29151,
278,
26490,
322,
15874,
310,
4045,
29892,
541,
306,
21117,
304,
6232,
590,
1914,
29889,
739,
29915,
29879,
4802,
885,
653,
29889,
13,
6295,
1244,
306,
626,
29892,
373,
5490,
29871,
29941,
29896,
303,
29892,
411,
590,
376,
29933,
6259,
21947,
1964,
29903,
15842,
6093,
341,
1164,
4690,
29908,
1051,
393,
338,
9186,
7120,
1283,
5174,
363,
445,
697,
1833,
12618,
3460,
3414,
29889,
26991,
306,
505,
472,
3203,
1023,
14433,
393,
526,
443,
11238,
491,
278,
1095,
310,
278,
4098,
29936,
445,
6511,
592,
304,
528,
11124,
372,
1283,
322,
1827,
376,
29902,
2360,
2869,
12242,
304,
8341,
963,
599,
29892,
925,
864,
304,
1018,
322,
8341,
408,
1784,
408,
306,
508,
1213,
306,
1207,
5566,
6394,
763,
376,
1357,
3271,
20414,
2060,
445,
4098,
471,
577,
4802,
29889,
4587,
3236,
306,
508,
29915,
29873,
2436,
263,
12618,
1213,
6864,
1474,
577,
1568,
931,
5771,
373,
1546,
11803,
393,
372,
23880,
763,
306,
505,
2086,
1568,
304,
1827,
29892,
541,
746,
306,
1369,
19229,
372,
23880,
763,
306,
1016,
29915,
29873,
2869,
505,
3099,
4100,
304,
29126,
304,
8986,
5199,
537,
29889,
306,
1209,
372,
1283,
2745,
2446,
4098,
29889,
1205,
306,
505,
304,
367,
2221,
304,
1423,
372,
1283,
445,
931,
29889,
306,
2099,
29892,
372,
29915,
29879,
5490,
29889,
306,
2609,
1369,
278,
1629,
1283,
491,
10124,
697,
3800,
443,
11238,
714,
310,
1183,
261,
425,
29920,
3335,
29889,
13,
2887,
263,
1121,
29892,
366,
679,
445,
443,
572,
11310,
29892,
364,
1117,
1847,
29892,
5517,
443,
287,
1573,
1400,
29889,
13,
797,
26406,
310,
590,
410,
7283,
579,
3381,
29901,
372,
471,
263,
2989,
4098,
29889,
13,
22606,
3118,
29901,
3012,
310,
278,
4306,
29871,
29906,
29900,
29896,
29929,
16052,
13,
29908,
11184,
11506,
29908,
491,
365,
2518,
806,
1740,
29891,
472,
3012,
310,
278,
4306,
29871,
29906,
29900,
29896,
29929,
29936,
826,
29894,
1114,
7817,
363,
278,
11401,
322,
12968,
1907,
13,
29902,
10398,
278,
937,
4723,
310,
5490,
1985,
408,
263,
15230,
4007,
22137,
472,
278,
826,
29894,
1114,
7817,
363,
278,
11401,
322,
12968,
1907,
29892,
298,
9776,
3012,
310,
278,
4306,
29871,
29906,
29900,
29896,
29929,
30166,
29889,
739,
29915,
29879,
263,
19340,
393,
306,
2996,
304,
491,
263,
2656,
310,
9570,
29899,
1595,
292,
15709,
4688,
1833,
1629,
29889,
306,
471,
373,
263,
5742,
29899,
1552,
29899,
1557,
25487,
6282,
310,
278,
826,
29894,
1114,
7817,
411,
590,
951,
328,
10475,
826,
29894,
1114,
2318,
29936,
1550,
278,
6282,
24764,
2820,
278,
278,
1008,
8162,
29892,
591,
9698,
13407,
297,
278,
1667,
23363,
297,
278,
29214,
310,
697,
310,
278,
1556,
752,
7807,
29130,
22001,
306,
505,
3926,
3595,
29889,
306,
1033,
2436,
6515,
1048,
372,
29892,
1048,
2020,
306,
2714,
372,
471,
8296,
26858,
29892,
1048,
920,
1226,
9215,
372,
471,
304,
1074,
408,
385,
7664,
29889,
13,
29902,
380,
1431,
29887,
839,
5742,
278,
1791,
310,
590,
2318,
304,
5193,
304,
278,
3151,
1061,
29892,
1530,
1915,
29892,
322,
4433,
565,
306,
1033,
1371,
714,
297,
278,
11798,
6358,
304,
5110,
920,
445,
9565,
310,
278,
1887,
1616,
3186,
3796,
313,
1366,
3732,
592,
6047,
263,
3287,
901,
14288,
322,
29663,
1135,
372,
2869,
471,
29936,
306,
23547,
5794,
289,
370,
27225,
1048,
920,
306,
6363,
920,
304,
2656,
310,
437,
14568,
310,
6433,
322,
750,
263,
5051,
1722,
20410,
322,
1033,
2924,
310,
2048,
2712,
467,
8512,
306,
29915,
345,
3796,
297,
19133,
29879,
322,
411,
7968,
7881,
1616,
8162,
29892,
306,
6363,
8886,
3078,
1048,
278,
5381,
310,
1641,
385,
7664,
322,
278,
6297,
393,
14014,
1616,
1644,
414,
5318,
297,
278,
7200,
1616,
9088,
29889,
910,
6140,
763,
9051,
306,
1033,
5110,
29889,
3374,
3730,
29892,
590,
4036,
29899,
2455,
29899,
8159,
29899,
974,
29899,
24498,
29899,
808,
6090,
322,
25706,
20410,
6216,
411,
825,
278,
11798,
6358,
4312,
29892,
322,
306,
471,
298,
2859,
408,
263,
15230,
4007,
22137,
304,
1371,
714,
4259,
635,
2645,
1510,
24562,
874,
322,
2601,
29879,
29889,
13,
3112,
338,
590,
25448,
29889,
306,
679,
304,
437,
577,
1568,
12528,
6433,
29936,
29130,
18533,
29892,
4870,
6751,
1616,
29892,
8608,
292,
1616,
304,
322,
515,
6314,
943,
29892,
298,
9776,
664,
313,
578,
1568,
298,
9776,
511,
322,
263,
4821,
411,
11161,
664,
29889,
1334,
884,
4337,
263,
3287,
310,
4796,
13630,
267,
29889,
13,
30166,
6716,
310,
590,
25448,
1736,
297,
3012,
310,
278,
4306,
29871,
29906,
29900,
29896,
29929,
30166,
338,
8778,
11506,
30166,
491,
365,
2518,
806,
1740,
29891,
29936,
372,
338,
263,
3699,
1754,
310,
8872,
2760,
12917,
18046,
793,
607,
365,
2518,
1476,
373,
2578,
2039,
411,
902,
11203,
1549,
263,
3271,
29899,
26859,
16648,
592,
6986,
29889,
1094,
591,
3796,
411,
365,
2518,
304,
443,
4058,
322,
2601,
8778,
11506,
29892,
1183,
5429,
502,
367,
585,
277,
1319,
322,
5192,
845,
5086,
15874,
310,
920,
278,
664,
2996,
964,
10379,
29892,
278,
4955,
322,
6593,
278,
18046,
793,
4934,
363,
902,
29892,
322,
5837,
393,
1183,
16254,
287,
411,
278,
664,
2645,
967,
937,
29130,
2978,
902,
3271,
29889,
13,
22606,
7803,
29901,
306,
6350,
278,
1407,
1472,
17944,
28144,
29899,
7083,
25206,
29889,
13,
13555,
2734,
278,
350,
3194,
350,
483,
261,
1833,
1629,
29892,
435,
1296,
16387,
670,
701,
11506,
17944,
11757,
362,
9559,
304,
1302,
2173,
311,
411,
17944,
1085,
25206,
15511,
355,
29889,
940,
4433,
592
] | 1,024 |
CommonCrawl
| 1.285924 | -0.937006 | -0.398874 | 0.346324 |
if I wanted to make the trip to Florida and run the half marathon together, and it sounded like a great excuse to return to humidity and see a few old friends. We started training in the summer and ran together nearly every single Saturday for six months, slowly building our distances.
I enjoyed running along Denver's amazing trails and parks with Jake and rewarding ourselves with smoothies and thrift store trips afterwards. Then we started going seven miles at a time, and I did not like seven miles. I did not like eight miles either. You can guess how I felt about all the miles past that. The best thing about training was that it gave Jake and I a routine of hanging out every week; that's hard when you're adult humans who have responsibilities and jobs and Netflix.
We finished our training and flew to Florida the day before the race. It was there that we found out that although the race started at 5:30am, we had to be at the front of our hotel ready to get on a bus at 3:30am. EW. Despite the early wake-up and a nervous night of sleep, we made it to the starting line, met up with two more vacationing pals, and ran the very long race.
Me, unhappy to be awake at 3:00am for a race that doesn't start until 5:30am
Me and Jake, unhappy to be on a bus at 3:30am on our way to the race that starts at 5:30am
Me and Jake, really happy to be done with the very long race that started too early in the morning
Propelled by the insane amounts of oxygen in Florida, we ran the entire race with no walking breaks (which was our goal) and didn't feel like we were going to die afterwards! I didn't even hate running until mile ten.
Week Three: Besties Forever
A Florida "hike" with these bits of my organs
Linden on a swing at the Hostel in the Forest
After the race, general merriment, and lots of my favorite Florida foods, Linden and I drove up to Georgia to go to the Hostel in the Forest. The Hostel stays with me in a sunset-magic kind of way and has me looking at my things, my food, and my people in a more beautifully warm light.
Week Four: New Basement Floors
Our cat Nemo has been very reliable at peeing on the carpet everywhere I have lived with him. Our pup, Luna, would swiftly follow the unwritten rule where one animal goes, the other follows. Since the basement is the only area of our home that has carpet, it was disgusting. Rented shampooers and a friend's fancy Rainbow vacuum hid the surface issues, but I hated being downstairs. It just felt gross all the time.
In late November, after several months of no animal-carpet wars, Nemo went at it again. THIS IS MY LIMIT. NO MORE. I immediately rage-measured the basement, rage-ordered laminate flooring (thankfully there was some sort of Black Friday/Cyber Monday/American Buying Holiday deal), and rage-told Dave that I would be rage-ripping out all of the carpet as soon as I got back from Florida. And I did it with such rage.
GOOD RIDDANCE, GROSS CARPET
#GOODBYEFOREVER #SORRYNOTSORRY
I also did it rather awkwardly. Our basement is full of stuff; stuff that I did not want to carry up the stairs. I piled everything into one half of the room, ripped up the carpet from the other side, moved all the stuff to the exposed floor, and ripped out the rest of the carpet. I repeated the process for cleaning the floor, then for laying the underlayment and the laminate. I needed all that rage to fuel the bonkers method of moving heavy furniture so many times. Thankfully, college had taught me many strategies for rearranging entire apartments of furniture as a single weakling human, so I was well-trained for the task.
The new basement floors and new basement arrangement meant that we had enough room for a real-size coffee table, and we decided it was time to retire my beloved white bookshelf--purchased by my Dad as my first "grown-up furn
|
[
565,
306,
5131,
304,
1207,
278,
17487,
304,
13813,
322,
1065,
278,
4203,
1766,
25206,
4208,
29892,
322,
372,
6047,
287,
763,
263,
2107,
5566,
1509,
304,
736,
304,
3165,
333,
537,
322,
1074,
263,
2846,
2030,
7875,
29889,
1334,
4687,
6694,
297,
278,
11801,
322,
6350,
4208,
8886,
1432,
2323,
24211,
363,
4832,
7378,
29892,
14205,
5214,
1749,
24610,
29889,
13,
29902,
27849,
2734,
3412,
3384,
369,
29915,
29879,
21863,
292,
1020,
2719,
322,
610,
2039,
411,
435,
1296,
322,
20751,
292,
20278,
411,
10597,
583,
322,
1468,
2027,
3787,
3367,
567,
12335,
29889,
1987,
591,
4687,
2675,
9881,
7800,
472,
263,
931,
29892,
322,
306,
1258,
451,
763,
9881,
7800,
29889,
306,
1258,
451,
763,
9475,
7800,
2845,
29889,
887,
508,
4140,
920,
306,
7091,
1048,
599,
278,
7800,
4940,
393,
29889,
450,
1900,
2655,
1048,
6694,
471,
393,
372,
4846,
435,
1296,
322,
306,
263,
26529,
310,
298,
9776,
714,
1432,
4723,
29936,
393,
29915,
29879,
2898,
746,
366,
29915,
276,
16157,
25618,
1058,
505,
5544,
747,
9770,
322,
17643,
322,
12670,
20157,
29916,
29889,
13,
4806,
7743,
1749,
6694,
322,
9115,
29893,
304,
13813,
278,
2462,
1434,
278,
8175,
29889,
739,
471,
727,
393,
591,
1476,
714,
393,
5998,
278,
8175,
4687,
472,
29871,
29945,
29901,
29941,
29900,
314,
29892,
591,
750,
304,
367,
472,
278,
4565,
310,
1749,
16730,
7960,
304,
679,
373,
263,
3593,
472,
29871,
29941,
29901,
29941,
29900,
314,
29889,
382,
29956,
29889,
19454,
278,
4688,
281,
1296,
29899,
786,
322,
263,
23547,
681,
4646,
310,
8709,
29892,
591,
1754,
372,
304,
278,
6257,
1196,
29892,
1539,
701,
411,
1023,
901,
11757,
362,
292,
282,
1338,
29892,
322,
6350,
278,
1407,
1472,
8175,
29889,
13,
6816,
29892,
28859,
14862,
304,
367,
3773,
1296,
472,
29871,
29941,
29901,
29900,
29900,
314,
363,
263,
8175,
393,
1838,
29915,
29873,
1369,
2745,
29871,
29945,
29901,
29941,
29900,
314,
13,
6816,
322,
435,
1296,
29892,
28859,
14862,
304,
367,
373,
263,
3593,
472,
29871,
29941,
29901,
29941,
29900,
314,
373,
1749,
982,
304,
278,
8175,
393,
8665,
472,
29871,
29945,
29901,
29941,
29900,
314,
13,
6816,
322,
435,
1296,
29892,
2289,
9796,
304,
367,
2309,
411,
278,
1407,
1472,
8175,
393,
4687,
2086,
4688,
297,
278,
7250,
13,
1184,
13111,
839,
491,
278,
1663,
1662,
26999,
310,
288,
28596,
297,
13813,
29892,
591,
6350,
278,
4152,
8175,
411,
694,
22049,
16706,
313,
4716,
471,
1749,
7306,
29897,
322,
3282,
29915,
29873,
4459,
763,
591,
892,
2675,
304,
762,
12335,
29991,
306,
3282,
29915,
29873,
1584,
26277,
2734,
2745,
17967,
3006,
29889,
13,
22606,
12753,
29901,
6407,
583,
28297,
369,
13,
29909,
13813,
376,
2918,
446,
29908,
411,
1438,
9978,
310,
590,
1638,
550,
13,
29931,
10060,
373,
263,
24500,
472,
278,
16956,
295,
297,
278,
17300,
13,
13555,
278,
8175,
29892,
2498,
2778,
29878,
2073,
29892,
322,
14568,
310,
590,
25448,
13813,
9687,
29879,
29892,
365,
10060,
322,
306,
26603,
701,
304,
16762,
304,
748,
304,
278,
16956,
295,
297,
278,
17300,
29889,
450,
16956,
295,
27111,
411,
592,
297,
263,
6575,
842,
29899,
11082,
293,
2924,
310,
982,
322,
756,
592,
3063,
472,
590,
2712,
29892,
590,
9687,
29892,
322,
590,
2305,
297,
263,
901,
9560,
368,
14294,
3578,
29889,
13,
22606,
12458,
29901,
1570,
4886,
882,
26043,
943,
13,
29949,
332,
6635,
405,
15810,
756,
1063,
1407,
23279,
472,
1236,
22430,
373,
278,
1559,
10963,
16978,
306,
505,
10600,
411,
1075,
29889,
8680,
23449,
29892,
365,
4347,
29892,
723,
12086,
368,
1101,
278,
443,
17625,
5751,
988,
697,
13019,
5771,
29892,
278,
916,
4477,
29889,
29871,
30166,
23036,
278,
2362,
882,
338,
278,
871,
4038,
310,
1749,
3271,
393,
756,
1559,
10963,
29892,
372,
471,
766,
29887,
504,
292,
29889,
390,
14927,
528,
314,
1129,
29877,
414,
322,
263,
5121,
29915,
29879,
19231,
21431,
17729,
11757,
29884,
398,
20552,
278,
7101,
5626,
29892,
541,
306,
298,
630,
1641,
1623,
303,
7121,
29889,
739,
925,
7091,
22683,
599,
278,
931,
29889,
13,
797,
5683,
3979,
29892,
1156,
3196,
7378,
310,
694,
13019,
29899,
4287,
10963,
29129,
29892,
405,
15810,
3512,
472,
372,
1449,
29889,
3446,
3235,
8519,
19519,
27848,
29889,
11698,
16999,
1525,
29889,
306,
7389,
1153,
479,
29899,
1004,
294,
2955,
278,
2362,
882,
29892,
1153,
479,
29899,
21693,
301,
9103,
403,
5685,
8253,
313,
386,
804,
3730,
727,
471,
777,
2656,
310,
6054,
28728,
29914,
29733,
495,
27822,
29914,
14689,
5373,
5414,
4168,
22394,
5376,
511,
322,
1153,
479,
29899,
29873,
1025,
17841,
393,
306,
723,
367,
1153,
479,
29899,
374,
3262,
714,
599,
310,
278,
1559,
10963,
408,
4720,
408,
306,
2355,
1250,
515,
13813,
29889,
1126,
306,
1258,
372,
411,
29871,
30166,
14565,
1153,
479,
29889,
13,
17080,
13668,
390,
1367,
29928,
23219,
29892,
402,
1672,
1799,
315,
1718,
29925,
2544,
13,
29937,
17080,
29949,
4051,
29979,
29923,
5800,
1525,
5348,
396,
29903,
1955,
13207,
12256,
29903,
1955,
13207,
13,
29902,
884,
1258,
372,
3265,
13689,
1328,
368,
29889,
8680,
2362,
882,
338,
2989,
310,
6433,
29936,
6433,
393,
306,
1258,
451,
864,
304,
8677,
701,
278,
380,
7121,
29889,
306,
282,
2356,
4129,
964,
697,
4203,
310,
278,
5716,
29892,
10107,
2986,
701,
278,
1559,
10963,
515,
278,
916,
2625,
29892,
6153,
599,
278,
6433,
304,
278,
19884,
11904,
29892,
322,
10107,
2986,
714,
278,
1791,
310,
278,
1559,
10963,
29889,
306,
10324,
278,
1889,
363,
5941,
292,
278,
11904,
29892,
769,
363,
6568,
292,
278,
1090,
8387,
358,
322,
278,
301,
9103,
403,
29889,
306,
4312,
599,
393,
1153,
479,
304,
26413,
278,
10814,
29895,
414,
1158,
310,
8401,
9416,
15252,
17252,
577,
1784,
3064,
29889,
3374,
3730,
29892,
12755,
750,
16187,
592,
1784,
16650,
583,
363,
337,
2749,
9776,
4152,
12435,
1860,
310,
15252,
17252,
408,
263,
2323,
8062,
1847,
5199,
29892,
577,
306,
471,
1532,
29899,
3018,
1312,
363,
278,
3414,
29889,
13,
1576,
716,
2362,
882,
5685,
943,
322,
716,
2362,
882,
24628,
6839,
393,
591,
750,
3307,
5716,
363,
263,
1855,
29899,
2311,
26935,
1591,
29892,
322,
591,
8459,
372,
471,
931,
304,
3240,
533,
590,
1339,
8238,
4796,
3143,
845,
761,
489,
29886,
2458,
1463,
491,
590,
360,
328,
408,
590,
937,
376,
29887,
4708,
29899,
786,
15252
] | 1,024 |
CommonCrawl
| 1.101163 | -1.277675 | -0.711792 | -0.259552 |
The options for Syrian female activists in Lebanon: Fear, imprisonment or departure
Suffering in silence
Why are Syrian female activists now leaving Lebanon? What are the obstacles, hardships and threats they are facing? And what are the options for those who choose to stay?
Nehmeh al-Alwani
Syrian journalist living in Lebanon.
This article is published as part of the "Syria In Depth" project, in collaboration and partnership with International Media Support, the Guardian Foundation and SyriaUntold.
BEIRUT: "For the sake of tarnishing my reputation and that of other Syrian women, they accused me of prostitution simply because I participated in the protests supporting the rights of refugees in Lebanon," said Dima Bsaiss, a 31-year-old Syrian woman who had to leave Lebanon in search of any safe haven due to the harassment.
After obtaining a protection visa, Dima arrived in France with her child.
And yet she hasn't forgotten the dangers closing in on her back in Lebanon, dangers that emerged just because she was a woman.
Hoping for change, Dima had participated in the peaceful protests in the streets of Syria in 2011. But by 2012, Homs was already a wreck, and Dima was being summoned for interrogation several times by the notorious security branches over "accusations" that she was participating in public protests.
Shortly afterwards, Dima he and her child left Syria.
Because of my political activism, I face direct threats.
Carrying her dreams for political change with her, from Syria to Lebanon's Tripoli, Dima there continued her activism and advocacy.
She faced security risks from the Syrian regime and its supporters there, Dima says, while displacement also took its toll on the family, on their health and psychological well-being.
"My finances and health were not the only matters at stake. My child Ward, who was only 11 at the time, was also threatened with abduction," she explains.
Dima worked in schools for Syrian refugees in Tripoli and hoped to contribute to building a better future for them, as she saw in those kids her own child. But the "lamentable situation" at school prompted her to expose corrupt practices, involving the teachers and curricula, through the media.
She gave a journalist several pieces of evidence—including videos of the teaching staff beating students violently as well as documents revealing the administration's manipulation of salaries by paying partial amounts and stealing the rest. The documents led to the school being shut down, the students moved to better schools.
"My quest against corruption bore fruit," Dima says. "But my hopes to change reality did not fade away: I took up social work and work on rights, attending conferences and meetings about Syrian affairs—especially those concerning women and children. My circle of contacts widened, and I formed a communication network with journalists and rights activists."
"But because of my political activism, I face direct threats," she tells SyriaUntold.
Fabricated photos of her were posted on the Internet intended to blackmail her, Dima says, on top of threats that she received by phone and social media. Other threats included offers of marriage and dating propositions.
A friend told me out of the blue that there were inappropriate photos of me in one of the Syrian groups. I could not contain my anger, and when I signed in to check, I found out that a person was trying to provoke me by altering photos from my personal accounts and putting them on indecent images to force me to meet him."
The threats negatively affected Dima's work and financial situation because she could not accept certain jobs—like those that required transportation in Lebanon or crossing military checkpoints. She started to seriously think about moving to another country, somewhere that would guarantee her rights and offer her family protection.
The Challenges of Education For Syrian Children in Lebanon
Irene Tuzi Sonia Grieco |
Without serious investment in their education, Syrian children will miss out on education not only as a result of the war but also as a consequence of life in exile.
'Change your address'
Dima turned to the UN to help her seek asylum in a European country, but they only advised her to change her address. She contacted other organizations to put pressure on the UN, and
|
[
450,
3987,
363,
8713,
6392,
12944,
5039,
2879,
297,
9388,
19930,
29901,
383,
799,
29892,
29535,
358,
470,
25619,
13,
29903,
3043,
292,
297,
15142,
13,
11008,
526,
8713,
6392,
12944,
5039,
2879,
1286,
10124,
9388,
19930,
29973,
1724,
526,
278,
14979,
23435,
29892,
2898,
9981,
322,
12455,
1446,
896,
526,
14870,
29973,
1126,
825,
526,
278,
3987,
363,
1906,
1058,
6755,
304,
7952,
29973,
13,
8139,
29882,
1004,
29882,
394,
29899,
2499,
29893,
3270,
13,
29903,
29891,
6392,
24452,
8471,
297,
9388,
19930,
29889,
13,
4013,
4274,
338,
6369,
408,
760,
310,
278,
376,
29903,
29891,
2849,
512,
10034,
386,
29908,
2060,
29892,
297,
24771,
322,
22056,
4034,
411,
4623,
8213,
18601,
29892,
278,
29429,
10606,
322,
8713,
2849,
29965,
593,
1025,
29889,
13,
15349,
8193,
2692,
29901,
376,
2831,
278,
16563,
310,
260,
2753,
14424,
590,
19821,
322,
393,
310,
916,
8713,
6392,
5866,
29892,
896,
28886,
592,
310,
16810,
5008,
3763,
1363,
306,
25223,
297,
278,
10021,
29879,
20382,
278,
10462,
310,
25447,
267,
297,
9388,
19930,
1699,
1497,
360,
2946,
350,
4977,
790,
29892,
263,
29871,
29941,
29896,
29899,
6360,
29899,
1025,
8713,
6392,
6114,
1058,
750,
304,
5967,
9388,
19930,
297,
2740,
310,
738,
9109,
7359,
2861,
304,
278,
4023,
465,
358,
29889,
13,
13555,
4017,
292,
263,
13047,
1998,
29874,
29892,
360,
2946,
11977,
297,
3444,
411,
902,
2278,
29889,
13,
2855,
3447,
1183,
22602,
29915,
29873,
20898,
278,
270,
13873,
14382,
297,
373,
902,
1250,
297,
9388,
19930,
29892,
270,
13873,
393,
11176,
3192,
925,
1363,
1183,
471,
263,
6114,
29889,
13,
29950,
459,
292,
363,
1735,
29892,
360,
2946,
750,
25223,
297,
278,
10776,
1319,
10021,
29879,
297,
278,
19756,
310,
8713,
2849,
297,
29871,
29906,
29900,
29896,
29896,
29889,
1205,
491,
29871,
29906,
29900,
29896,
29906,
29892,
379,
4835,
471,
2307,
263,
281,
18217,
29892,
322,
360,
2946,
471,
1641,
2533,
3712,
287,
363,
1006,
9102,
362,
3196,
3064,
491,
278,
451,
23308,
6993,
14202,
975,
376,
5753,
375,
800,
29908,
393,
1183,
471,
5221,
1218,
297,
970,
10021,
29879,
29889,
13,
21322,
368,
12335,
29892,
360,
2946,
540,
322,
902,
2278,
2175,
8713,
2849,
29889,
13,
29933,
5658,
310,
590,
8604,
5039,
1608,
29892,
306,
3700,
1513,
12455,
1446,
29889,
13,
8179,
719,
292,
902,
12561,
29879,
363,
8604,
1735,
411,
902,
29892,
515,
8713,
2849,
304,
9388,
19930,
29915,
29879,
8602,
3733,
29875,
29892,
360,
2946,
727,
7572,
902,
5039,
1608,
322,
22545,
4135,
29889,
13,
13468,
20050,
6993,
5161,
2039,
515,
278,
8713,
6392,
22384,
322,
967,
1462,
272,
2153,
727,
29892,
360,
2946,
4083,
29892,
1550,
12272,
9552,
884,
3614,
967,
304,
645,
373,
278,
3942,
29892,
373,
1009,
9045,
322,
11643,
5996,
1532,
29899,
915,
292,
29889,
13,
29908,
3421,
1436,
2925,
322,
9045,
892,
451,
278,
871,
13750,
472,
380,
1296,
29889,
1619,
2278,
21910,
29892,
1058,
471,
871,
29871,
29896,
29896,
472,
278,
931,
29892,
471,
884,
29513,
411,
633,
700,
428,
1699,
1183,
18568,
29889,
13,
29928,
2946,
3796,
297,
12462,
363,
8713,
6392,
25447,
267,
297,
8602,
3733,
29875,
322,
24936,
304,
29126,
304,
5214,
263,
2253,
5434,
363,
963,
29892,
408,
1183,
4446,
297,
1906,
413,
4841,
902,
1914,
2278,
29889,
1205,
278,
376,
12598,
519,
6434,
29908,
472,
3762,
9508,
287,
902,
304,
24396,
1034,
6685,
23274,
29892,
21677,
278,
27335,
322,
16256,
293,
2497,
29892,
1549,
278,
5745,
29889,
13,
13468,
4846,
263,
24452,
3196,
12785,
310,
10757,
30003,
18271,
19707,
310,
278,
18819,
13925,
367,
1218,
8041,
5537,
2705,
408,
1532,
408,
10701,
10320,
12818,
278,
17517,
29915,
29879,
11525,
2785,
310,
4497,
4314,
491,
5146,
292,
7687,
26999,
322,
1886,
12818,
278,
1791,
29889,
450,
10701,
5331,
304,
278,
3762,
1641,
12522,
1623,
29892,
278,
8041,
6153,
304,
2253,
12462,
29889,
13,
29908,
3421,
21126,
2750,
1034,
18953,
23888,
15774,
1699,
360,
2946,
4083,
29889,
376,
6246,
590,
26926,
304,
1735,
16832,
1258,
451,
28747,
3448,
29901,
306,
3614,
701,
5264,
664,
322,
664,
373,
10462,
29892,
1098,
2548,
378,
10662,
322,
5870,
886,
1048,
8713,
6392,
19469,
30003,
267,
25009,
1906,
19813,
5866,
322,
4344,
29889,
1619,
8607,
310,
25957,
281,
3615,
287,
29892,
322,
306,
8429,
263,
12084,
3564,
411,
8955,
2879,
322,
10462,
5039,
2879,
1213,
13,
29908,
6246,
1363,
310,
590,
8604,
5039,
1608,
29892,
306,
3700,
1513,
12455,
1446,
1699,
1183,
10603,
8713,
2849,
29965,
593,
1025,
29889,
13,
29943,
370,
2200,
630,
20612,
310,
902,
892,
8059,
373,
278,
4685,
9146,
304,
4628,
2549,
902,
29892,
360,
2946,
4083,
29892,
373,
2246,
310,
12455,
1446,
393,
1183,
4520,
491,
9008,
322,
5264,
5745,
29889,
5901,
12455,
1446,
5134,
16688,
310,
13718,
322,
270,
1218,
9551,
2187,
29889,
13,
29909,
5121,
5429,
592,
714,
310,
278,
7254,
393,
727,
892,
297,
932,
6649,
403,
20612,
310,
592,
297,
697,
310,
278,
8713,
6392,
6471,
29889,
306,
1033,
451,
1712,
590,
27343,
29892,
322,
746,
306,
8794,
297,
304,
1423,
29892,
306,
1476,
714,
393,
263,
2022,
471,
1811,
304,
1326,
6946,
592,
491,
10551,
292,
20612,
515,
590,
7333,
15303,
322,
10594,
963,
373,
5704,
1760,
4558,
304,
4889,
592,
304,
5870,
1075,
1213,
13,
1576,
12455,
1446,
3480,
6703,
15201,
360,
2946,
29915,
29879,
664,
322,
18161,
6434,
1363,
1183,
1033,
451,
3544,
3058,
17643,
30003,
4561,
1906,
393,
3734,
8608,
362,
297,
9388,
19930,
470,
24067,
9121,
1423,
9748,
29889,
2296,
4687,
304,
25798,
1348,
1048,
8401,
304,
1790,
4234,
29892,
9051,
393,
723,
18818,
902,
10462,
322,
5957,
902,
3942,
13047,
29889,
13,
1576,
678,
16047,
267,
310,
13151,
1152,
8713,
6392,
20986,
297,
9388,
19930,
13,
29902,
28593,
12603,
2526,
5791,
423,
1632,
347,
1111,
891,
13,
3047,
449,
10676,
13258,
358,
297,
1009,
9793,
29892,
8713,
6392,
4344,
674,
3052,
714,
373,
9793,
451,
871,
408,
263,
1121,
310,
278,
1370,
541,
884,
408,
263,
17004,
310,
2834,
297,
429,
488,
29889,
13,
29915,
7277,
596,
3211,
29915,
13,
29928,
2946,
6077,
304,
278,
8291,
304,
1371,
902,
16508,
408,
2904,
398,
297,
263,
7824,
4234,
29892,
541,
896,
871,
594,
11292,
902,
304,
1735,
902,
3211,
29889,
2296,
6958,
287,
916,
25700,
304,
1925,
12959,
373,
278,
8291,
29892,
322
] | 1,024 |
CommonCrawl
| 1.285924 | 0.631367 | 1.041084 | 1.493567 |
finally succeeded in seeking asylum in France through an Italian organization.
Dima believes that there is direct pressure to reduce or eliminate the presence of Syrian journalists and activists in Lebanon for no clear reason.
"Freedom of expression has become a luxury in Lebanon, especially on social media," she explains. "Many rights' activists are afraid to speak about the problems facing Syrian refugees in Lebanon."
Syrian refugees in Beirut describe similar experiences.
Some of them left the city, while others stood their ground only to still encounter racist statements aimed at refugees, often from politicians and other figures affiliated with the Lebanese government. Most recently, Lebanese President Michel Aoun talked about Lebanon's ongoing economic crisis, claiming that Syrian refugees had cost the country some $25 billion. President Aoun added that Lebanon has the right to redeem some of that amount from countries that supposedly triggered the conflict in Syria.
Several statistics have documented the economic impact of Syrian refugees' presence in Lebanon, and the money that has been injected into the Lebanese economy as a result.
The Issam Fares Institute at the American University of Beirut released statistics in January 2018 highlighting various facts and figures about the displacement of Syrian refugees crisis, finding that "every $1 spent on humanitarian aid has a multiplier effect of $1.6 in the local economy."
"As such," the study stated, "the $1.258 billion of humanitarian assistance that was provided to Lebanon from 2011 till early 2017 actually injected $2.01 billion into the Lebanese economy."
I suffered physical and psychological problems due to the torture and conditions in jail. [And] after the news spread on social media, I grew worried that I had put my parents in Syria at risk.
Even so, anti-Syrian security campaigns have targeted Syrian activists and journalists of both sexes in Lebanon.
Ahmad Khoder, 29, from the village of Houla in Homs province, recently moved to Italy after securing his exit from Lebanon as political refugee through an Italian rights organization.
During time stay in Beirut, Ahmad was arrested by the Lebanese army after a passenger on a bus saw him watching the video of a Syrian song—Abdel Basset a-Sarout. It later transpired that the passenger was himself an officer in the security forces. Ahmad's story went viral, and after follow-up on on his case, he was released from prison.
However, Ahmad's case was still classified under terrorism laws, meaning he was at risk of being handed over to the Syrian regime's security forces. Ahmad says he "could not help but be scared."
"I did not know what to expect," he adds. "Many accusations were thrown at me."
Even so, Ahmad did not stop his activities, as he participated in solidarity protests on the anniversary of the Syrian revolution in Beirut. Using his connections, he also coordinated campaigns to assist refugees in the camps.
"Jail is a specific kind of challenge, but it did not stand in my way. After I was released, I resumed my activities by posting news about Syrian refugees in Lebanon, including sensitive issues like deportation or arrest—even when that meant seeking help from people outside Lebanon to disseminate the information."
Imprisoned By The Regime, And Ostracised By Society
Maya Abyad |
Human Rights Organizations say more than ten thousand women have been detained since the Syrian uprising erupted in 2011, but being released from prison does not put an end to...
Many refuse to speak out about their treatment in Lebanon because of fears it could impact their families in regime-controlled areas, or relatives in prison, back in Syria. At the same time, there are those refugees who believe that it's the family that can sometimes impede women and girls from practicing their activism and expressing their opinions, with families afraid of activism and willing to wash their hands of it. Some go as far as disowning their daughters, if they are arrested or pursued by the Syrian regime.
South Lebanon hosts a whole host of organizations and associations working with Syrian refugees.
Ola al-Jundi is a Syrian activist who works at Women Now for Development, an organization empowering Syrian
|
[
7146,
14792,
297,
25738,
408,
2904,
398,
297,
3444,
1549,
385,
10545,
13013,
29889,
13,
29928,
2946,
1339,
17180,
393,
727,
338,
1513,
12959,
304,
10032,
470,
27399,
278,
10122,
310,
8713,
6392,
8955,
2879,
322,
5039,
2879,
297,
9388,
19930,
363,
694,
2821,
2769,
29889,
13,
29908,
23923,
11607,
310,
4603,
756,
4953,
263,
21684,
2857,
297,
9388,
19930,
29892,
7148,
373,
5264,
5745,
1699,
1183,
18568,
29889,
376,
14804,
10462,
29915,
5039,
2879,
526,
13421,
304,
7726,
1048,
278,
4828,
14870,
8713,
6392,
25447,
267,
297,
9388,
19930,
1213,
13,
29903,
29891,
6392,
25447,
267,
297,
1522,
381,
329,
8453,
2788,
27482,
29889,
13,
9526,
310,
963,
2175,
278,
4272,
29892,
1550,
4045,
8389,
1009,
5962,
871,
304,
1603,
11735,
11021,
391,
9506,
12242,
287,
472,
25447,
267,
29892,
4049,
515,
2832,
14722,
322,
916,
13994,
23736,
630,
411,
278,
9388,
273,
968,
5874,
29889,
7849,
10325,
29892,
9388,
273,
968,
7178,
12255,
319,
1309,
24867,
1048,
9388,
19930,
29915,
29879,
373,
17696,
17407,
24161,
29892,
5995,
292,
393,
8713,
6392,
25447,
267,
750,
3438,
278,
4234,
777,
395,
29906,
29945,
24464,
29889,
7178,
319,
1309,
2715,
393,
9388,
19930,
756,
278,
1492,
304,
337,
311,
331,
777,
310,
393,
5253,
515,
10916,
393,
7424,
368,
19799,
278,
14529,
297,
8713,
2849,
29889,
13,
29903,
1310,
284,
13964,
505,
23531,
278,
17407,
10879,
310,
8713,
6392,
25447,
267,
29915,
10122,
297,
9388,
19930,
29892,
322,
278,
6909,
393,
756,
1063,
11658,
287,
964,
278,
9388,
273,
968,
26504,
408,
263,
1121,
29889,
13,
1576,
16982,
314,
383,
5114,
8907,
472,
278,
3082,
3014,
310,
1522,
381,
329,
5492,
13964,
297,
5490,
29871,
29906,
29900,
29896,
29947,
12141,
292,
5164,
17099,
322,
13994,
1048,
278,
12272,
9552,
310,
8713,
6392,
25447,
267,
24161,
29892,
9138,
393,
376,
17991,
395,
29896,
10398,
373,
5199,
3673,
713,
16226,
756,
263,
6674,
4926,
2779,
310,
395,
29896,
29889,
29953,
297,
278,
1887,
26504,
1213,
13,
29908,
2887,
1316,
1699,
278,
6559,
8703,
29892,
376,
1552,
395,
29896,
29889,
29906,
29945,
29947,
24464,
310,
5199,
3673,
713,
18872,
393,
471,
4944,
304,
9388,
19930,
515,
29871,
29906,
29900,
29896,
29896,
3428,
4688,
29871,
29906,
29900,
29896,
29955,
2869,
11658,
287,
395,
29906,
29889,
29900,
29896,
24464,
964,
278,
9388,
273,
968,
26504,
1213,
13,
29902,
17654,
9128,
322,
11643,
5996,
4828,
2861,
304,
278,
16263,
545,
322,
5855,
297,
432,
737,
29889,
518,
2855,
29962,
1156,
278,
9763,
9677,
373,
5264,
5745,
29892,
306,
13631,
6365,
1255,
393,
306,
750,
1925,
590,
11825,
297,
8713,
2849,
472,
12045,
29889,
13,
29923,
854,
577,
29892,
9418,
29899,
29903,
29891,
6392,
6993,
11531,
29879,
505,
3646,
287,
8713,
6392,
5039,
2879,
322,
8955,
2879,
310,
1716,
7916,
267,
297,
9388,
19930,
29889,
13,
29909,
7184,
328,
12217,
6119,
29892,
29871,
29906,
29929,
29892,
515,
278,
5720,
310,
18832,
433,
297,
379,
4835,
12291,
29892,
10325,
6153,
304,
12730,
1156,
409,
2764,
292,
670,
6876,
515,
9388,
19930,
408,
8604,
25447,
29872,
1549,
385,
10545,
10462,
13013,
29889,
13,
29928,
3864,
931,
7952,
297,
1522,
381,
329,
29892,
319,
7184,
328,
471,
24383,
491,
278,
9388,
273,
968,
9987,
1156,
263,
28507,
373,
263,
3593,
4446,
1075,
21217,
278,
4863,
310,
263,
8713,
6392,
4823,
30003,
4920,
6144,
17328,
300,
263,
29899,
29903,
279,
449,
29889,
739,
2678,
1301,
29886,
2859,
393,
278,
28507,
471,
3654,
385,
12139,
297,
278,
6993,
8249,
29889,
319,
7184,
328,
29915,
29879,
5828,
3512,
10636,
284,
29892,
322,
1156,
1101,
29899,
786,
373,
373,
670,
1206,
29892,
540,
471,
5492,
515,
8475,
29889,
13,
17245,
29892,
319,
7184,
328,
29915,
29879,
1206,
471,
1603,
770,
2164,
1090,
15115,
1608,
14243,
29892,
6593,
540,
471,
472,
12045,
310,
1641,
29692,
975,
304,
278,
8713,
6392,
22384,
29915,
29879,
6993,
8249,
29889,
319,
7184,
328,
4083,
540,
376,
26680,
451,
1371,
541,
367,
885,
1965,
1213,
13,
29908,
29902,
1258,
451,
1073,
825,
304,
2149,
1699,
540,
12778,
29889,
376,
14804,
26142,
800,
892,
12005,
472,
592,
1213,
13,
29923,
854,
577,
29892,
319,
7184,
328,
1258,
451,
5040,
670,
14188,
29892,
408,
540,
25223,
297,
7773,
279,
537,
10021,
29879,
373,
278,
6957,
27547,
310,
278,
8713,
6392,
19479,
297,
1522,
381,
329,
29889,
5293,
670,
12368,
29892,
540,
884,
6615,
630,
11531,
29879,
304,
6985,
25447,
267,
297,
278,
3949,
567,
29889,
13,
29908,
29967,
737,
338,
263,
2702,
2924,
310,
18766,
29892,
541,
372,
1258,
451,
2317,
297,
590,
982,
29889,
2860,
306,
471,
5492,
29892,
306,
620,
21571,
590,
14188,
491,
16742,
9763,
1048,
8713,
6392,
25447,
267,
297,
9388,
19930,
29892,
3704,
20502,
5626,
763,
21455,
362,
470,
22564,
30003,
11884,
746,
393,
6839,
25738,
1371,
515,
2305,
5377,
9388,
19930,
304,
766,
12846,
16976,
278,
2472,
1213,
13,
1888,
558,
2285,
287,
2648,
450,
2169,
603,
29892,
1126,
11334,
945,
3368,
2648,
7765,
13,
29924,
9010,
319,
1609,
328,
891,
13,
29950,
7889,
26863,
9205,
17063,
1827,
901,
1135,
3006,
10405,
5866,
505,
1063,
1439,
7114,
1951,
278,
8713,
6392,
318,
558,
5921,
604,
21245,
287,
297,
29871,
29906,
29900,
29896,
29896,
29892,
541,
1641,
5492,
515,
8475,
947,
451,
1925,
385,
1095,
304,
856,
13,
14804,
26506,
304,
7726,
714,
1048,
1009,
14502,
297,
9388,
19930,
1363,
310,
8866,
29879,
372,
1033,
10879,
1009,
13175,
297,
22384,
29899,
6451,
839,
10161,
29892,
470,
14576,
297,
8475,
29892,
1250,
297,
8713,
2849,
29889,
2180,
278,
1021,
931,
29892,
727,
526,
1906,
25447,
267,
1058,
4658,
393,
372,
29915,
29879,
278,
3942,
393,
508,
6041,
2411,
2742,
5866,
322,
14000,
515,
4120,
18499,
1009,
5039,
1608,
322,
4653,
292,
1009,
26971,
29892,
411,
13175,
13421,
310,
5039,
1608,
322,
17762,
304,
471,
29882,
1009,
6567,
310,
372,
29889,
3834,
748,
408,
2215,
408,
766,
776,
292,
1009,
29215,
29892,
565,
896,
526,
24383,
470,
12359,
6742,
491,
278,
8713,
6392,
22384,
29889,
13,
29903,
2438,
9388,
19930,
18982,
263,
3353,
3495,
310,
25700,
322,
27733,
1985,
411,
8713,
6392,
25447,
267,
29889,
13,
29949,
433,
394,
29899,
29967,
870,
29875,
338,
263,
8713,
6392,
5039,
391,
1058,
1736,
472,
10152,
2567,
363,
14650,
29892,
385,
13013,
3710,
1680,
292,
8713,
6392
] | 1,024 |
CommonCrawl
| 1.174524 | 0.794888 | 1.553946 | 1.002795 |
women, and also supports women politically and teaches at the Gharsa Center in Lebanon's Beqaa Valley.
In Syria, the regime arrested Ola twice, and she was forced to leave the country and come to Lebanon.
"Like many Syrians who lived through the war in Syria, I feel like a traitor for leaving my country and people. We struggle against our guilty consciences by helping people here," she tells SyriaUntold. "Perhaps our activism is a way to make up for our disappointment in the political situation in Syria."
Ola believes that empowering women to become human rights defenders has always been a missing step because of challenges in travelling. Female activists can't even obtain the rights they are calling for, especially in conferences held outside Lebanon, like in Geneva.
"Besides, as a result of the security situation in Lebanon for Syrians, they have to take heed and pray to stay safe. This limits discussions and serious engagement in talks," Ola adds. "The situation in Lebanon and the Arab world pushes each person to look for their own salvation, rather than that of the collective. Their voices as a united entity are muffled. This crackdown does not spare Lebanese activists either."
Mazna al-Zuhouri, 27, has worked as a media activist since the onset of the Syrian prising in 2011, back when she was still in al-Qusayr in Syria.
Mazna lost her father to Syria's detention archipelago, after he died from torture in a Syrian government prison.
"The power of the state is a patriarchal one riddled with obstacles that hinder female activists' freedom of expression," she says.
"Female activists face patriarchal and societal risks more than others, and this sometimes obstructs their activism. Society blames them if any harm comes their way or touches their families."
For Mazna, that has manifested itself in a number of ways. She regularly receives insults and disparaging comments via social media channels—with some even reporting her profiles in an attempt to obstruct her activism.
Mazna also says she's received threats on a number of occasions, meaning she's always cautious and fearful that someone might hack her social media accounts or make use of her personal photos inappropriately.
'Guaranteeing the right to safety'
In Lebanon, Italian refugee organization Italia Operazione Colomba helped Dima, Ahmad and many other prominent activists to leave Lebanon safely and reach Europe through humanitarian routes.
The organization works directly with refugees.
"We live in camps in North Lebanon, and we move between cities to help Syrians reach the UNHCR offices for their appointments and cross Lebanese army checkpoints safely," explains Aliss, a representative of the organization. "We allocate a healthcare and hospitalization allowance for Syrian patients in Lebanon."
He adds that the organization does not have specific criteria for protection applications. Each case is examined separately—especially cases of women, sick people as well as rights activists.
UNHCR, the UN agency that works with refugees in Lebanon, is also mandated to assist vulnerable refugees in ned of protection—including through resettlement routes to third countries, outside Lebanon. UNHCR did not respond to requests for comment regarding several aspects of this report.
At the same time, there are also diaspora organizations formed by people who themselves suffered threats and harassment and felt the need to put together a legal framework for others like them, even after leaving Lebanon.
A group of Syrian and Lebanese activists established Access Center to offer refugees legal support and pressure the Lebanese government to stop deporting refugees and handing them over to the Syrian regime.
Nabila who works at the center tells SyriaUntold how the centre was first formed in 2017 to provide legal support to refugees in the camps.
"Now [the centre] focuses on ensuring the rights of refugees and those who are hounded by security. It also helps guarantee the right to life and personal safety, and access to basic rights."
Social activist Nasiba Farjani meanwhile believes that organizations do not have clear criteria regarding political asylum.
"Many people are not asylum seekers but have been classified as such, although they were pro-regime and could return to areas under its control. Meanwhile
|
[
5866,
29892,
322,
884,
11286,
5866,
2832,
1711,
322,
6860,
267,
472,
278,
13491,
1503,
29874,
7817,
297,
9388,
19930,
29915,
29879,
1522,
29939,
7340,
13939,
29889,
13,
797,
8713,
2849,
29892,
278,
22384,
24383,
438,
433,
8951,
29892,
322,
1183,
471,
11826,
304,
5967,
278,
4234,
322,
2041,
304,
9388,
19930,
29889,
13,
29908,
27552,
1784,
8713,
374,
550,
1058,
10600,
1549,
278,
1370,
297,
8713,
2849,
29892,
306,
4459,
763,
263,
1020,
2105,
363,
10124,
590,
4234,
322,
2305,
29889,
1334,
21117,
2750,
1749,
27719,
1136,
455,
2063,
491,
19912,
2305,
1244,
1699,
1183,
10603,
8713,
2849,
29965,
593,
1025,
29889,
376,
5894,
4252,
1749,
5039,
1608,
338,
263,
982,
304,
1207,
701,
363,
1749,
23451,
358,
297,
278,
8604,
6434,
297,
8713,
2849,
1213,
13,
29949,
433,
1339,
17180,
393,
3710,
1680,
292,
5866,
304,
4953,
5199,
10462,
822,
21043,
756,
2337,
1063,
263,
4567,
4331,
1363,
310,
18066,
267,
297,
6427,
7807,
29889,
19361,
744,
5039,
2879,
508,
29915,
29873,
1584,
4017,
278,
10462,
896,
526,
5432,
363,
29892,
7148,
297,
378,
10662,
4934,
5377,
9388,
19930,
29892,
763,
297,
15350,
1564,
29889,
13,
29908,
29933,
267,
2247,
29892,
408,
263,
1121,
310,
278,
6993,
6434,
297,
9388,
19930,
363,
8713,
374,
550,
29892,
896,
505,
304,
2125,
540,
287,
322,
12475,
304,
7952,
9109,
29889,
910,
13071,
5353,
1080,
322,
10676,
3033,
5049,
297,
5969,
2039,
1699,
438,
433,
12778,
29889,
376,
1576,
6434,
297,
9388,
19930,
322,
278,
10387,
3186,
5503,
267,
1269,
2022,
304,
1106,
363,
1009,
1914,
24754,
362,
29892,
3265,
1135,
393,
310,
278,
6314,
573,
29889,
11275,
28848,
408,
263,
443,
1573,
7855,
526,
286,
3096,
839,
29889,
910,
26755,
3204,
947,
451,
29337,
9388,
273,
968,
5039,
2879,
2845,
1213,
13,
29924,
834,
1056,
394,
29899,
29999,
29884,
10774,
374,
29892,
29871,
29906,
29955,
29892,
756,
3796,
408,
263,
5745,
5039,
391,
1951,
278,
373,
842,
310,
278,
8713,
6392,
544,
5921,
297,
29871,
29906,
29900,
29896,
29896,
29892,
1250,
746,
1183,
471,
1603,
297,
394,
29899,
29984,
375,
388,
29878,
297,
8713,
2849,
29889,
13,
29924,
834,
1056,
5714,
902,
4783,
304,
8713,
2849,
29915,
29879,
1439,
2509,
3190,
666,
295,
4425,
29892,
1156,
540,
6423,
515,
16263,
545,
297,
263,
8713,
6392,
5874,
8475,
29889,
13,
29908,
1576,
3081,
310,
278,
2106,
338,
263,
16967,
1279,
284,
697,
364,
2205,
839,
411,
14979,
23435,
393,
298,
4995,
12944,
5039,
2879,
29915,
16082,
310,
4603,
1699,
1183,
4083,
29889,
13,
29908,
29943,
331,
744,
5039,
2879,
3700,
16967,
1279,
284,
322,
5374,
26310,
5161,
2039,
901,
1135,
4045,
29892,
322,
445,
6041,
704,
4984,
29879,
1009,
5039,
1608,
29889,
7765,
1999,
1280,
963,
565,
738,
10311,
5304,
1009,
982,
470,
6023,
267,
1009,
13175,
1213,
13,
2831,
17326,
1056,
29892,
393,
756,
14682,
2868,
3528,
297,
263,
1353,
310,
5837,
29889,
2296,
25704,
20586,
1663,
499,
29879,
322,
17508,
6751,
6589,
3025,
5264,
5745,
18196,
30003,
2541,
777,
1584,
23415,
902,
28723,
297,
385,
4218,
304,
704,
4984,
902,
5039,
1608,
29889,
13,
29924,
834,
1056,
884,
4083,
1183,
29915,
29879,
4520,
12455,
1446,
373,
263,
1353,
310,
26108,
29892,
6593,
1183,
29915,
29879,
2337,
274,
1300,
2738,
322,
8866,
1319,
393,
4856,
1795,
15833,
902,
5264,
5745,
15303,
470,
1207,
671,
310,
902,
7333,
20612,
297,
932,
6649,
2486,
29889,
13,
29915,
9485,
9519,
22430,
278,
1492,
304,
15332,
29915,
13,
797,
9388,
19930,
29892,
10545,
25447,
29872,
13013,
12201,
6607,
3343,
1530,
290,
2291,
9213,
360,
2946,
29892,
319,
7184,
328,
322,
1784,
916,
19555,
5039,
2879,
304,
5967,
9388,
19930,
23511,
322,
6159,
4092,
1549,
5199,
3673,
713,
12049,
29889,
13,
1576,
13013,
1736,
4153,
411,
25447,
267,
29889,
13,
29908,
4806,
5735,
297,
3949,
567,
297,
4644,
9388,
19930,
29892,
322,
591,
4337,
1546,
14368,
304,
1371,
8713,
374,
550,
6159,
278,
8291,
29950,
11341,
25222,
363,
1009,
8167,
1860,
322,
4891,
9388,
273,
968,
9987,
1423,
9748,
23511,
1699,
18568,
838,
790,
29892,
263,
21097,
310,
278,
13013,
29889,
376,
4806,
23632,
263,
9045,
18020,
322,
13457,
2133,
2758,
749,
363,
8713,
6392,
22069,
297,
9388,
19930,
1213,
13,
3868,
12778,
393,
278,
13013,
947,
451,
505,
2702,
16614,
363,
13047,
8324,
29889,
7806,
1206,
338,
4392,
1312,
16949,
30003,
267,
25009,
4251,
310,
5866,
29892,
17319,
2305,
408,
1532,
408,
10462,
5039,
2879,
29889,
13,
3904,
29950,
11341,
29892,
278,
8291,
946,
3819,
393,
1736,
411,
25447,
267,
297,
9388,
19930,
29892,
338,
884,
9619,
630,
304,
6985,
23180,
519,
25447,
267,
297,
22165,
310,
13047,
30003,
18271,
1549,
620,
1803,
944,
12049,
304,
4654,
10916,
29892,
5377,
9388,
19930,
29889,
8291,
29950,
11341,
1258,
451,
10049,
304,
7274,
363,
3440,
11211,
3196,
21420,
310,
445,
3461,
29889,
13,
4178,
278,
1021,
931,
29892,
727,
526,
884,
652,
294,
1971,
29874,
25700,
8429,
491,
2305,
1058,
6053,
17654,
12455,
1446,
322,
4023,
465,
358,
322,
7091,
278,
817,
304,
1925,
4208,
263,
11706,
6890,
363,
4045,
763,
963,
29892,
1584,
1156,
10124,
9388,
19930,
29889,
13,
29909,
2318,
310,
8713,
6392,
322,
9388,
273,
968,
5039,
2879,
7841,
11028,
7817,
304,
5957,
25447,
267,
11706,
2304,
322,
12959,
278,
9388,
273,
968,
5874,
304,
5040,
21455,
292,
25447,
267,
322,
1361,
292,
963,
975,
304,
278,
8713,
6392,
22384,
29889,
13,
29940,
370,
4233,
1058,
1736,
472,
278,
4818,
10603,
8713,
2849,
29965,
593,
1025,
920,
278,
8442,
471,
937,
8429,
297,
29871,
29906,
29900,
29896,
29955,
304,
3867,
11706,
2304,
304,
25447,
267,
297,
278,
3949,
567,
29889,
13,
29908,
10454,
518,
1552,
8442,
29962,
8569,
267,
373,
5662,
3864,
278,
10462,
310,
25447,
267,
322,
1906,
1058,
526,
298,
7261,
491,
6993,
29889,
739,
884,
6911,
18818,
278,
1492,
304,
2834,
322,
7333,
15332,
29892,
322,
2130,
304,
6996,
10462,
1213,
13,
6295,
1455,
5039,
391,
22318,
16912,
8413,
29926,
3270,
2099,
8000,
1339,
17180,
393,
25700,
437,
451,
505,
2821,
16614,
11211,
8604,
408,
2904,
398,
29889,
13,
29908,
14804,
2305,
526,
451,
408,
2904,
398,
16508,
414,
541,
505,
1063,
770,
2164,
408,
1316,
29892,
5998,
896,
892,
410,
29899,
1727,
603,
322,
1033,
736,
304,
10161,
1090,
967,
2761,
29889,
25065
] | 1,024 |
CommonCrawl
| 1.008783 | 0.532735 | 0.955009 | 0.977497 |
AcraBoard – your first real hoverboard
DesignDesign, Transportation
"For the first time since the bicycle, automobile or airplane, the ArcaBoard is a revolutionary breakthrough for transportation. For the first time, every person will be able to fly anytime, anywhere. The world, your world, will change forever."
Arca Space Corporation in New Mexico has developed an electronic flying vehicle that can hover over any terrain, including water. The design can hold up to 243 lbs and flies a foot above the ground.
working hover board
Aerospace company Arca Space being inspired by Back-to-the-Future hoverboard has continued this trend with a rectangular device that uses 36 fans to float its riders through the air.
Integrated into the ArcaBoard's chassis, the fans are driven by high-powered electric motors and generate up to 272 horsepower.
"By using 36 smaller electric ducted fans instead of fewer larger ones we ensure that the ArcaBoard keeps operating even in the case of multiple engine failures,"
said Dumitri Popescu, who founded the company that has also designed rocket launchers and drones.
"Each engine is powered by its own set of batteries independently of the other power units creating a multiple redundant system," he added.
The ArcaBoard appears to be the closest thing to the technology from Back to the Future: Part II that we've seen so far.
There's reason to believe the ArcaBoard is for real, unlike the HUVr hoax from 2014 that Christopher Lloyd and Tony Hawk perpetrated on us.
marty mcfly hover board
First off, Arca started as a Romanian NGO and was awarded contracts for work on stratospheric rockets and balloons by the Romanian government and the European Space Agency. Since then, the company participated in the Google Lunar X Prize, released high-end drones and has recently relocated to the US, where it announced the release of the ArcaBoard on Christmas Eve.
Constructed from aerospace-grade composite materials to make it both lightweight and sturdy ArcaBoard weights 10 kilograms, and includes a cooling system that protects the device from the heat generated by its electric motors.
The device is capable of lifting up to 110 kilograms of weight but don't expect to be popping an ArcaBoard in your backpack a la Marty McFly, either, as it weighs 82 kg (180 lb) on its own. Chris Lang, the company's Chief Operating Officer, tells Gizmag that it could be carried by two people.
Riders can enjoy up to six minutes before the board needs recharging – which takes six hours – although the company has also designed an extra dock accessory that allows it to be recharged in 35 minutes.
The ArcaBoard can reportedly be controlled with the rider's body or via a Bluetooth-connected iOS or Android app that activates a stabilization system and relays navigation commands via hand movements. Hover skate board
A built in stabilisation system keeps the board level while hovering or travelling. The Arca tilts when users make turns, to keep riders safe, although the app includes the option of turning the stabilisation system off.
"All you have to do is balance yourself and the board will provide a smooth stabilised experience," said Popescu. "With the stabilisation system turned off you will find it as easy as learning to ride on your first bicycle."
"It takes an estimated 20 minutes of flight to become comfortable with the motion of flying in the air," he added.
Wait, wait. Nobody said that it would be cheap. The device will be available from April 2016, and carries a price tag of $19,900 (£13,657 hover board cost).
Your personal alive Segway
TOP 10 must-see magnificent streets
One thought on "AcraBoard – your first real hoverboard"
ThomasMic
Приветик всем, я тут новенький "352"
20+ logo design – inspiration for your ideas
Top 20 magical photos that will change the way you think about snails
Each photo – a masterpiece
KTM Superduke. How not to fall in love with him?
Splendid photographs from around the world
Top 10 cozy houses in the Modern style
19 delight
|
[
7255,
336,
28397,
785,
596,
937,
1855,
16758,
3377,
13,
4002,
647,
4002,
647,
29892,
15710,
362,
13,
29908,
2831,
278,
937,
931,
1951,
278,
289,
4245,
2841,
29892,
3345,
5249,
470,
4799,
22116,
29892,
278,
826,
1113,
28397,
338,
263,
19479,
653,
2867,
20678,
363,
8608,
362,
29889,
1152,
278,
937,
931,
29892,
1432,
2022,
674,
367,
2221,
304,
11340,
738,
2230,
29892,
12214,
29889,
450,
3186,
29892,
596,
3186,
29892,
674,
1735,
22296,
1213,
13,
1433,
1113,
14121,
15025,
297,
1570,
12568,
756,
8906,
385,
27758,
22764,
19716,
393,
508,
16758,
975,
738,
28439,
29892,
3704,
4094,
29889,
450,
2874,
508,
4808,
701,
304,
29871,
29906,
29946,
29941,
301,
5824,
322,
285,
3687,
263,
3661,
2038,
278,
5962,
29889,
13,
22899,
16758,
7613,
13,
29909,
9672,
3535,
5001,
826,
1113,
14121,
1641,
20603,
491,
7437,
29899,
517,
29899,
1552,
29899,
20154,
16758,
3377,
756,
7572,
445,
534,
355,
411,
263,
7705,
6825,
4742,
393,
3913,
29871,
29941,
29953,
24909,
304,
5785,
967,
8177,
414,
1549,
278,
4799,
29889,
13,
23573,
630,
964,
278,
826,
1113,
28397,
29915,
29879,
521,
465,
275,
29892,
278,
24909,
526,
18225,
491,
1880,
29899,
13519,
287,
12646,
3184,
943,
322,
5706,
701,
304,
29871,
29906,
29955,
29906,
10435,
13519,
29889,
13,
29908,
2059,
773,
29871,
29941,
29953,
7968,
12646,
868,
2954,
24909,
2012,
310,
28145,
7200,
6743,
591,
9801,
393,
278,
826,
1113,
28397,
14874,
13598,
1584,
297,
278,
1206,
310,
2999,
6012,
4418,
1973,
1699,
13,
4977,
333,
21139,
277,
374,
6977,
267,
4979,
29892,
1058,
11091,
278,
5001,
393,
756,
884,
8688,
696,
3522,
6826,
414,
322,
270,
1617,
267,
29889,
13,
29908,
9760,
6012,
338,
3081,
287,
491,
967,
1914,
731,
310,
10193,
583,
25499,
310,
278,
916,
3081,
10340,
4969,
263,
2999,
28005,
1788,
1699,
540,
2715,
29889,
13,
1576,
826,
1113,
28397,
5692,
304,
367,
278,
21438,
2655,
304,
278,
15483,
515,
7437,
304,
278,
16367,
29901,
3455,
1944,
393,
591,
29915,
345,
3595,
577,
2215,
29889,
13,
8439,
29915,
29879,
2769,
304,
4658,
278,
826,
1113,
28397,
338,
363,
1855,
29892,
25531,
278,
379,
29965,
29963,
29878,
5089,
1165,
515,
29871,
29906,
29900,
29896,
29946,
393,
18888,
27054,
322,
15293,
10875,
29895,
25722,
29878,
630,
373,
502,
29889,
13,
28402,
29891,
286,
6854,
368,
16758,
7613,
13,
6730,
1283,
29892,
826,
1113,
4687,
408,
263,
5917,
713,
405,
17080,
322,
471,
15074,
8078,
29879,
363,
664,
373,
851,
4507,
8096,
293,
7679,
1691,
322,
6411,
417,
787,
491,
278,
5917,
713,
5874,
322,
278,
7824,
14121,
29353,
29889,
4001,
769,
29892,
278,
5001,
25223,
297,
278,
5087,
365,
348,
279,
1060,
20604,
29892,
5492,
1880,
29899,
355,
270,
1617,
267,
322,
756,
10325,
337,
28809,
304,
278,
3148,
29892,
988,
372,
9326,
278,
6507,
310,
278,
826,
1113,
28397,
373,
17661,
382,
345,
29889,
13,
1168,
4984,
287,
515,
263,
9672,
3535,
29899,
8228,
20842,
17279,
304,
1207,
372,
1716,
3578,
7915,
322,
380,
332,
4518,
826,
1113,
28397,
18177,
29871,
29896,
29900,
4679,
468,
25402,
29892,
322,
7805,
263,
12528,
292,
1788,
393,
12566,
29879,
278,
4742,
515,
278,
12871,
5759,
491,
967,
12646,
3184,
943,
29889,
13,
1576,
4742,
338,
15390,
310,
11747,
1259,
701,
304,
29871,
29896,
29896,
29900,
4679,
468,
25402,
310,
7688,
541,
1016,
29915,
29873,
2149,
304,
367,
772,
3262,
385,
826,
1113,
28397,
297,
596,
1250,
4058,
263,
425,
3760,
29891,
4052,
29943,
368,
29892,
2845,
29892,
408,
372,
591,
1141,
29879,
29871,
29947,
29906,
12118,
313,
29896,
29947,
29900,
27981,
29897,
373,
967,
1914,
29889,
12821,
10476,
29892,
278,
5001,
29915,
29879,
14546,
6607,
1218,
28288,
29892,
10603,
402,
466,
11082,
393,
372,
1033,
367,
8988,
491,
1023,
2305,
29889,
13,
29934,
11376,
508,
13389,
701,
304,
4832,
6233,
1434,
278,
7613,
4225,
337,
25389,
292,
785,
607,
4893,
4832,
6199,
785,
5998,
278,
5001,
756,
884,
8688,
385,
4805,
23630,
2130,
706,
393,
6511,
372,
304,
367,
337,
25389,
287,
297,
29871,
29941,
29945,
6233,
29889,
13,
1576,
826,
1113,
28397,
508,
8967,
368,
367,
20704,
411,
278,
364,
1241,
29915,
29879,
3573,
470,
3025,
263,
3164,
20930,
29899,
18045,
10012,
470,
5669,
623,
393,
5039,
1078,
263,
16160,
2133,
1788,
322,
1104,
1036,
11322,
8260,
3025,
1361,
24147,
29889,
379,
957,
2071,
403,
7613,
13,
29909,
4240,
297,
16160,
4371,
1788,
14874,
278,
7613,
3233,
1550,
16758,
292,
470,
6427,
7807,
29889,
450,
826,
1113,
6928,
1372,
746,
4160,
1207,
12169,
29892,
304,
3013,
8177,
414,
9109,
29892,
5998,
278,
623,
7805,
278,
2984,
310,
14712,
278,
16160,
4371,
1788,
1283,
29889,
13,
29908,
3596,
366,
505,
304,
437,
338,
17346,
7535,
322,
278,
7613,
674,
3867,
263,
10597,
16160,
3368,
7271,
1699,
1497,
6977,
267,
4979,
29889,
376,
3047,
278,
16160,
4371,
1788,
6077,
1283,
366,
674,
1284,
372,
408,
4780,
408,
6509,
304,
22203,
373,
596,
937,
289,
4245,
2841,
1213,
13,
29908,
3112,
4893,
385,
15899,
29871,
29906,
29900,
6233,
310,
16286,
304,
4953,
25561,
411,
278,
10884,
310,
22764,
297,
278,
4799,
1699,
540,
2715,
29889,
13,
15716,
29892,
4480,
29889,
15706,
1486,
1497,
393,
372,
723,
367,
28773,
29889,
450,
4742,
674,
367,
3625,
515,
3786,
29871,
29906,
29900,
29896,
29953,
29892,
322,
1559,
2722,
263,
8666,
4055,
310,
395,
29896,
29929,
29892,
29929,
29900,
29900,
313,
30155,
29896,
29941,
29892,
29953,
29945,
29955,
16758,
7613,
3438,
467,
13,
10858,
7333,
18758,
6667,
1582,
13,
29911,
4590,
29871,
29896,
29900,
1818,
29899,
4149,
29154,
296,
19756,
13,
6716,
2714,
373,
376,
10644,
336,
28397,
785,
596,
937,
1855,
16758,
3377,
29908,
13,
1349,
18902,
29924,
293,
13,
30013,
641,
1521,
29514,
6495,
29959,
29892,
2282,
12998,
29932,
13074,
8358,
3753,
376,
29941,
29945,
29906,
29908,
13,
29906,
29900,
29974,
20194,
2874,
785,
8681,
12232,
363,
596,
7014,
13,
7031,
29871,
29906,
29900,
2320,
936,
20612,
393,
674,
1735,
278,
982,
366,
1348,
1048,
5807,
2234,
13,
9760,
15373,
785,
263,
5835,
12343,
346,
13,
29968,
23081,
5670,
700,
446,
29889,
1128,
451,
304,
6416,
297,
5360,
411,
1075,
29973,
13,
29903,
572,
355,
333,
17739,
29879,
515,
2820,
278,
3186,
13,
7031,
29871,
29896,
29900,
1302,
1537,
12955,
297,
278,
14624,
3114,
13,
29896,
29929,
15319
] | 1,024 |
CommonCrawl
| 0.296913 | -0.515875 | 1.012392 | -0.072983 |
We are responsible to steward what we have been given, and I include in that stewardship our neurochemistry. Approximately ten percent of us have neurochemistry that includes 1) some neural structures which are less developed, 2) with less connectivity between some of our neural structures, and 3) with inefficient electrochemical communications within our brains. Some of us have been given children and students with this kind of neurochemistry. We intuitively sense there is something different about these children. As Mason teaches us, we have accepted our "duty to educate [our] child into moral strength and purpose and intellectual activity,(1989A, p. 102)" and we have been diligent to do everything we know. We cannot put our finger on it, but something gnaws at us about them.
The broad curriculum, this wonderful feast of books and things does not seem to invigorate them in the way it does other children; they "have to be in the mood" for even inspirational things like nature study. Their upper school authors, some who are such eloquent, insightful wordsmiths seem to pass these students by. Some subjects even become sources of emotion: "Latin is so dumb!" "I hate poetry!" Math proves a stumper to them and us. How can this student who worked through this problem yesterday with me, explained it, and got the right answer not understand it today—almost like it was erased in her mind?
We love how Mason champions the personhood of the child, how the individuality of each child is respected while maintaining life in a community. We embrace the freedom that each child can develop at his/her own rate. While we know we don't want to fit all children in a standard box, we have this sense deep down that troubles us about these children and we notice that in some areas they get further and further behind.
We worry about these kids' dependence on us or other adults. Narrations of books read together with the parent are richer, more detailed than those they do alone or even with others at a co-op. They seem to need more handholding, more cheerleading, more reminders for everything. Self-education seems elusive. Often the more we try to shift tasks to them to do independently, the more careless and shoddy their work becomes. Except for the few things that really seem to light their fires, we have to stay on top of these children to get most things done. Will this child ever learn to be reliable? Will they ever carry through on things without my constant supervision? Inconsistency seems to plague them. They certainly can give their full attention and effort when they take a mind to. They cannot get out the door on time and prepared for school, swim team or music lessons, but you let Grandpa take them hunting and see what happens. The night before, oh my! This child who cannot keep his room tidy or put his laundry away, has checked and cleaned all his equipment, and carefully arranged them beside his neatly folded camouflage clothes. And he is in bed early without a beep of argument!
We worry about their diminished abilities to be focused internally, to have an inner vision with that inner oomph or drive to carry out that vision. They are like a boat adrift that cannot keep its rudder in the water to give it any directionality. But when they do latch on to an idea or something they want to do, it is often not a very realistic goal, more like wishful thinking. They do not think through things well, and their hasty decisions and judgment seem more like that of their younger siblings. They get stuck on minor things they cannot seem to let go. "Mary ignored me at school/co-op today." "That's not fair that Sammy gets to do that when you did not let me do that at his age." Outbursts can seem to come out of nowhere and their ability to handle frustration seems more fragile.
These children puzzle us and often take a lot out of us. When we overhear a lady complaining, "See my forehead? Well, dealing with my child has put every one of these wrinkles here; I don't know what to do!" we well identify with her.
What if all the things above are typical of kids with executive function deficits? Today with fMRIs and other neuroimaging technology, we know about the brain's management or executive system and can understand executive functions (EFs). These are cognitive processes we use to control impulsivity (in mind, emotion
|
[
1334,
526,
14040,
304,
1886,
1328,
825,
591,
505,
1063,
2183,
29892,
322,
306,
3160,
297,
393,
1886,
1328,
3527,
1749,
452,
2192,
14969,
6020,
29889,
28268,
2657,
2486,
3006,
10151,
310,
502,
505,
452,
2192,
14969,
6020,
393,
7805,
29871,
29896,
29897,
777,
19677,
12286,
607,
526,
3109,
8906,
29892,
29871,
29906,
29897,
411,
3109,
4511,
2068,
1546,
777,
310,
1749,
19677,
12286,
29892,
322,
29871,
29941,
29897,
411,
297,
8462,
28118,
14969,
936,
7212,
800,
2629,
1749,
4105,
1144,
29889,
3834,
310,
502,
505,
1063,
2183,
4344,
322,
8041,
411,
445,
2924,
310,
452,
2192,
14969,
6020,
29889,
1334,
27951,
3598,
4060,
727,
338,
1554,
1422,
1048,
1438,
4344,
29889,
1094,
28095,
6860,
267,
502,
29892,
591,
505,
9259,
1749,
376,
29881,
329,
29891,
304,
6320,
403,
518,
473,
29962,
2278,
964,
14731,
9324,
322,
6437,
322,
29762,
6354,
22657,
29896,
29929,
29947,
29929,
29909,
29892,
282,
29889,
29871,
29896,
29900,
29906,
5513,
322,
591,
505,
1063,
21749,
25692,
304,
437,
4129,
591,
1073,
29889,
1334,
2609,
1925,
1749,
19917,
373,
372,
29892,
541,
1554,
330,
1056,
5652,
472,
502,
1048,
963,
29889,
13,
1576,
7300,
16256,
12906,
398,
29892,
445,
20695,
1238,
579,
310,
8277,
322,
2712,
947,
451,
2833,
304,
2437,
335,
272,
403,
963,
297,
278,
982,
372,
947,
916,
4344,
29936,
896,
376,
17532,
304,
367,
297,
278,
286,
2092,
29908,
363,
1584,
23459,
1288,
2712,
763,
5469,
6559,
29889,
11275,
7568,
3762,
15717,
29892,
777,
1058,
526,
1316,
560,
29877,
16011,
29892,
25483,
1319,
3838,
29885,
389,
29879,
2833,
304,
1209,
1438,
8041,
491,
29889,
3834,
17800,
1584,
4953,
8974,
310,
953,
8194,
29901,
376,
13992,
262,
338,
577,
270,
3774,
3850,
376,
29902,
26277,
22309,
3850,
5792,
28281,
263,
380,
398,
546,
304,
963,
322,
502,
29889,
1128,
508,
445,
8368,
1058,
3796,
1549,
445,
1108,
22600,
411,
592,
29892,
10824,
372,
29892,
322,
2355,
278,
1492,
1234,
451,
2274,
372,
9826,
30003,
284,
3242,
763,
372,
471,
604,
1463,
297,
902,
3458,
29973,
13,
4806,
5360,
920,
28095,
29314,
278,
2022,
6614,
310,
278,
2278,
29892,
920,
278,
5375,
537,
310,
1269,
2278,
338,
3390,
287,
1550,
7344,
292,
2834,
297,
263,
7881,
29889,
1334,
953,
13842,
278,
16082,
393,
1269,
2278,
508,
2693,
472,
670,
29914,
2276,
1914,
6554,
29889,
5806,
591,
1073,
591,
1016,
29915,
29873,
864,
304,
6216,
599,
4344,
297,
263,
3918,
3800,
29892,
591,
505,
445,
4060,
6483,
1623,
393,
18835,
502,
1048,
1438,
4344,
322,
591,
8369,
393,
297,
777,
10161,
896,
679,
4340,
322,
4340,
5742,
29889,
13,
4806,
15982,
1048,
1438,
413,
4841,
29915,
26307,
373,
502,
470,
916,
16157,
29879,
29889,
405,
2749,
800,
310,
8277,
1303,
4208,
411,
278,
3847,
526,
8261,
261,
29892,
901,
13173,
1135,
1906,
896,
437,
7432,
470,
1584,
411,
4045,
472,
263,
1302,
29899,
459,
29889,
2688,
2833,
304,
817,
901,
1361,
8948,
292,
29892,
901,
22794,
25369,
29892,
901,
1083,
513,
414,
363,
4129,
29889,
21782,
29899,
287,
1682,
362,
2444,
560,
375,
573,
29889,
438,
15535,
278,
901,
591,
1018,
304,
9500,
9595,
304,
963,
304,
437,
25499,
29892,
278,
901,
2562,
2222,
322,
528,
397,
4518,
1009,
664,
7415,
29889,
1222,
1547,
363,
278,
2846,
2712,
393,
2289,
2833,
304,
3578,
1009,
24237,
29892,
591,
505,
304,
7952,
373,
2246,
310,
1438,
4344,
304,
679,
1556,
2712,
2309,
29889,
2811,
445,
2278,
3926,
5110,
304,
367,
23279,
29973,
2811,
896,
3926,
8677,
1549,
373,
2712,
1728,
590,
4868,
2428,
4924,
29973,
512,
3200,
391,
3819,
2444,
304,
715,
3437,
963,
29889,
2688,
8959,
508,
2367,
1009,
2989,
8570,
322,
7225,
746,
896,
2125,
263,
3458,
304,
29889,
2688,
2609,
679,
714,
278,
3050,
373,
931,
322,
13240,
363,
3762,
29892,
2381,
326,
3815,
470,
4696,
3109,
787,
29892,
541,
366,
1235,
6265,
3274,
2125,
963,
29826,
322,
1074,
825,
5930,
29889,
450,
4646,
1434,
29892,
9360,
590,
29991,
910,
2278,
1058,
2609,
3013,
670,
5716,
10668,
29891,
470,
1925,
670,
425,
870,
719,
3448,
29892,
756,
7120,
322,
5941,
287,
599,
670,
21083,
29892,
322,
16112,
21050,
963,
17620,
670,
28539,
368,
900,
7176,
3949,
283,
1579,
482,
22095,
29889,
1126,
540,
338,
297,
6592,
4688,
1728,
263,
367,
1022,
310,
2980,
29991,
13,
4806,
15982,
1048,
1009,
22964,
3276,
633,
9770,
304,
367,
21309,
25106,
29892,
304,
505,
385,
6426,
18551,
411,
393,
6426,
288,
290,
561,
470,
7899,
304,
8677,
714,
393,
18551,
29889,
2688,
526,
763,
263,
13006,
594,
7532,
393,
2609,
3013,
967,
29051,
672,
297,
278,
4094,
304,
2367,
372,
738,
5305,
2877,
29889,
1205,
746,
896,
437,
301,
905,
373,
304,
385,
2969,
470,
1554,
896,
864,
304,
437,
29892,
372,
338,
4049,
451,
263,
1407,
1855,
4695,
7306,
29892,
901,
763,
6398,
1319,
7291,
29889,
2688,
437,
451,
1348,
1549,
2712,
1532,
29892,
322,
1009,
14973,
29891,
1602,
12112,
322,
24284,
2833,
901,
763,
393,
310,
1009,
20023,
27767,
18964,
29889,
2688,
679,
10771,
373,
9461,
2712,
896,
2609,
2833,
304,
1235,
748,
29889,
376,
29924,
653,
17262,
592,
472,
3762,
29914,
1111,
29899,
459,
9826,
1213,
376,
7058,
29915,
29879,
451,
6534,
393,
3685,
1357,
4947,
304,
437,
393,
746,
366,
1258,
451,
1235,
592,
437,
393,
472,
670,
5046,
1213,
4451,
8399,
303,
29879,
508,
2833,
304,
2041,
714,
310,
1286,
4150,
322,
1009,
11509,
304,
4386,
1424,
11036,
2444,
901,
13855,
488,
29889,
13,
1349,
968,
4344,
20285,
280,
502,
322,
4049,
2125,
263,
3287,
714,
310,
502,
29889,
1932,
591,
975,
354,
279,
263,
11379,
15313,
2827,
29892,
376,
13393,
590,
26839,
2813,
29973,
5674,
29892,
16743,
411,
590,
2278,
756,
1925,
1432,
697,
310,
1438,
2358,
682,
793,
1244,
29936,
306,
1016,
29915,
29873,
1073,
825,
304,
437,
3850,
591,
1532,
12439,
411,
902,
29889,
13,
5618,
565,
599,
278,
2712,
2038,
526,
15662,
310,
413,
4841,
411,
22760,
740,
822,
293,
1169,
29973,
20628,
411,
285,
21055,
3624,
322,
916,
452,
2192,
326,
6751,
15483,
29892,
591,
1073,
1048,
278,
17294,
29915,
29879,
10643,
470,
22760,
1788,
322,
508,
2274,
22760,
3168,
313,
29638,
29879,
467,
4525,
526,
25323,
3321,
10174,
591,
671,
304,
2761,
2411,
7273,
2068,
313,
262,
3458,
29892,
953,
8194
] | 1,024 |
C4
| 1.16909 | 0.480824 | -0.161496 | 1.685828 |
and mouth), manage and organize tasks (to prioritize, plan, start activities), manage and protect attention, set and pursue goals, self-question and self-monitor our progress towards those goals, manage time, and use working memory (to remember what we are to do). We now know how delays or dysfunction in the brain's management system causes executive function deficits (EFDs) that often prove to be very challenging issues.
The tough thing for the CM educator dealing with dysexecutive students is that the neurological knowledge necessary to understand EF and EFDs was not available to Mason and the PNEU or anyone else at the turn of the 20th century. Consequently, no one had any strategies, any tools in their educator toolboxes to help them understand or support people with EFDs. Mason founded the PNEU "To bring before the parents of all classes the best thought on education" (Kitching, 1924, p. 133); we offer this seminar on stewarding one's neurochemistry to try to bring the best thought on EF today so that we can have the tools (principles and compensatory strategies) to help us and our children/students steward their executive functions.
Attendees should expect to share areas of concern they want to tackle, and the day will be structured to provide insight and feedback on those concerns so each person will leave with an individually tailored list of insights and practical tools to try.
Since we cannot steward what we do not understand, we will delve into understanding the sound theory behind EFDs—just as Mason charged us: "One thing we begin to see clearly, that the stream can rise no higher than its source, that sound theory must underlie successful work (Mason, 1989B, p. 31)" (italics mine). We will take a brief overview of the neuroscience research of why EFDs happen (what parts of the brain are less active, less developed) and the difficulties dysexecutive individuals face (the neuro-psychology behind their problems). We will look at what does and does not cause these deficits and the implications of confusing the neurological and character lens. Smaller groups on a chosen developmental age will explore how these different deficits might show up in home life, academics, and relationships at that particular stage. By the end of our session, everyone will have lifespan list of typical challenges people with EFDs may encounter, will "know what trouble spots, what things can bite you and take you down with EFDs," and have an executive profile assessment tool to take home to informally assess a child's or adult's EF strengths and weaknesses.
"We begin to suspect that we took up schemes and methods of education a little hastily, without considering what philosophy or, let us say, psychology, underlies those schemes and methods; now, we see that our results cannot be in advance of our principles (Mason, 1989C, p. 63)"(italics mine). Equipped with a solid philosophy, a neuro-psychology to underlie our methods, we address methods needed to support persons with EFDs, both with core strategies that apply broadly and specific, practical applications.
We will bring our knowledge of EF and EFDs to burrow into examining tasks that Mason students are asked to perform by evaluating what executive capacities are required for such tasks as reading, narration, math, composition, geography, etc. We will discuss how dysexecutive students may react when requested to do things beyond their capacities. We will demonstrate adaptive strategies like "squiggle notes" and "dump, group, decide" that support the dysexecutive students with narrations and composition. Further, we will look at graphic organizers and templates that provide executive support in math. Discussing how using Vygotsky's zone of proximal development works, CM educators can zoom in and out with strategies to support students' executive weaknesses. Adjusting expectations of such things as pace, the time needed in a schedule, the quantity of reading, and the number of curricular subjects will be addressed.
Although executive functions are also associated with other special needs, such as Autism, these are beyond the scope of this seminar. Our discussion will be limited to some executive functions related to ADD and ADHD.
Prayer for each other will finish our day as we return to our places of living, teaching and serving those with EFDs that we might better steward the neurochemistry
|
[
322,
13394,
511,
10933,
322,
2894,
675,
9595,
313,
517,
7536,
277,
675,
29892,
3814,
29892,
1369,
14188,
511,
10933,
322,
12566,
8570,
29892,
731,
322,
12359,
434,
14433,
29892,
1583,
29899,
12470,
322,
1583,
29899,
3712,
2105,
1749,
6728,
7113,
1906,
14433,
29892,
10933,
931,
29892,
322,
671,
1985,
3370,
313,
517,
6456,
825,
591,
526,
304,
437,
467,
1334,
1286,
1073,
920,
628,
1036,
470,
270,
952,
2220,
297,
278,
17294,
29915,
29879,
10643,
1788,
9946,
22760,
740,
822,
293,
1169,
313,
29923,
26453,
29879,
29897,
393,
4049,
6356,
304,
367,
1407,
18066,
292,
5626,
29889,
13,
1576,
260,
820,
2655,
363,
278,
315,
29924,
6320,
1061,
16743,
411,
13475,
14167,
687,
11067,
8041,
338,
393,
278,
452,
2192,
1188,
936,
7134,
5181,
304,
2274,
22286,
322,
22286,
29928,
29879,
471,
451,
3625,
304,
28095,
322,
278,
349,
8186,
29965,
470,
5019,
1683,
472,
278,
2507,
310,
278,
29871,
29906,
29900,
386,
6462,
29889,
1281,
27284,
29892,
694,
697,
750,
738,
16650,
583,
29892,
738,
8492,
297,
1009,
6320,
1061,
5780,
1884,
267,
304,
1371,
963,
2274,
470,
2304,
2305,
411,
22286,
29928,
29879,
29889,
28095,
11091,
278,
349,
8186,
29965,
376,
1762,
6963,
1434,
278,
11825,
310,
599,
4413,
278,
1900,
2714,
373,
9793,
29908,
313,
29968,
2335,
292,
29892,
29871,
29896,
29929,
29906,
29946,
29892,
282,
29889,
29871,
29896,
29941,
29941,
416,
591,
5957,
445,
3031,
18220,
373,
1886,
1328,
292,
697,
29915,
29879,
452,
2192,
14969,
6020,
304,
1018,
304,
6963,
278,
1900,
2714,
373,
22286,
9826,
577,
393,
591,
508,
505,
278,
8492,
313,
558,
2173,
2701,
322,
22874,
7606,
16650,
583,
29897,
304,
1371,
502,
322,
1749,
4344,
29914,
18082,
1237,
1886,
1328,
1009,
22760,
3168,
29889,
13,
4165,
3324,
267,
881,
2149,
304,
6232,
10161,
310,
5932,
896,
864,
304,
22002,
280,
29892,
322,
278,
2462,
674,
367,
2281,
2955,
304,
3867,
25483,
322,
16705,
373,
1906,
21838,
577,
1269,
2022,
674,
5967,
411,
385,
29689,
12464,
4395,
1051,
310,
1663,
5861,
322,
15031,
8492,
304,
1018,
29889,
13,
23036,
591,
2609,
1886,
1328,
825,
591,
437,
451,
2274,
29892,
591,
674,
628,
345,
964,
8004,
278,
6047,
6368,
5742,
22286,
29928,
29879,
30003,
5143,
408,
28095,
20139,
502,
29901,
376,
6716,
2655,
591,
3380,
304,
1074,
9436,
29892,
393,
278,
4840,
508,
14451,
694,
6133,
1135,
967,
2752,
29892,
393,
6047,
6368,
1818,
1090,
3197,
9150,
664,
313,
29924,
1658,
29892,
29871,
29896,
29929,
29947,
29929,
29933,
29892,
282,
29889,
29871,
29941,
29896,
5513,
313,
2410,
1199,
7903,
467,
1334,
674,
2125,
263,
11473,
975,
1493,
310,
278,
17167,
1883,
15277,
5925,
310,
2020,
22286,
29928,
29879,
3799,
313,
5816,
5633,
310,
278,
17294,
526,
3109,
6136,
29892,
3109,
8906,
29897,
322,
278,
23553,
13475,
14167,
687,
11067,
15724,
3700,
313,
1552,
452,
2192,
29899,
567,
3376,
3002,
5742,
1009,
4828,
467,
1334,
674,
1106,
472,
825,
947,
322,
947,
451,
4556,
1438,
822,
293,
1169,
322,
278,
2411,
5795,
310,
16051,
278,
452,
2192,
1188,
936,
322,
2931,
301,
575,
29889,
4116,
12572,
6471,
373,
263,
10434,
5849,
284,
5046,
674,
26987,
920,
1438,
1422,
822,
293,
1169,
1795,
1510,
701,
297,
3271,
2834,
29892,
16274,
1199,
29892,
322,
21702,
472,
393,
3153,
7408,
29889,
2648,
278,
1095,
310,
1749,
4867,
29892,
14332,
674,
505,
11747,
267,
8357,
1051,
310,
15662,
18066,
267,
2305,
411,
22286,
29928,
29879,
1122,
11735,
29892,
674,
376,
28385,
825,
7458,
805,
1862,
29892,
825,
2712,
508,
289,
568,
366,
322,
2125,
366,
1623,
411,
22286,
29928,
29879,
1699,
322,
505,
385,
22760,
8722,
24809,
358,
5780,
304,
2125,
3271,
304,
1871,
635,
24809,
263,
2278,
29915,
29879,
470,
16157,
29915,
29879,
22286,
9324,
29879,
322,
8062,
2264,
267,
29889,
13,
29908,
4806,
3380,
304,
12326,
393,
591,
3614,
701,
27715,
322,
3519,
310,
9793,
263,
2217,
14973,
2354,
29892,
1728,
13858,
825,
22237,
470,
29892,
1235,
502,
1827,
29892,
11643,
3002,
29892,
1090,
3687,
1906,
27715,
322,
3519,
29936,
1286,
29892,
591,
1074,
393,
1749,
2582,
2609,
367,
297,
6564,
310,
1749,
18671,
313,
29924,
1658,
29892,
29871,
29896,
29929,
29947,
29929,
29907,
29892,
282,
29889,
29871,
29953,
29941,
5513,
29898,
2410,
1199,
7903,
467,
382,
6578,
2986,
411,
263,
7773,
22237,
29892,
263,
452,
2192,
29899,
567,
3376,
3002,
304,
1090,
3197,
1749,
3519,
29892,
591,
3211,
3519,
4312,
304,
2304,
12407,
411,
22286,
29928,
29879,
29892,
1716,
411,
7136,
16650,
583,
393,
3394,
7300,
368,
322,
2702,
29892,
15031,
8324,
29889,
13,
4806,
674,
6963,
1749,
7134,
310,
22286,
322,
22286,
29928,
29879,
304,
6866,
798,
964,
4392,
2827,
9595,
393,
28095,
8041,
526,
4433,
304,
2189,
491,
6161,
1218,
825,
22760,
11101,
1907,
526,
3734,
363,
1316,
9595,
408,
5183,
29892,
15474,
362,
29892,
5844,
29892,
15259,
29892,
1737,
5275,
29892,
2992,
29889,
1334,
674,
5353,
920,
13475,
14167,
687,
11067,
8041,
1122,
7657,
746,
13877,
304,
437,
2712,
8724,
1009,
11101,
1907,
29889,
1334,
674,
22222,
7744,
573,
16650,
583,
763,
376,
26613,
335,
6234,
11486,
29908,
322,
376,
15070,
29892,
2318,
29892,
11097,
29908,
393,
2304,
278,
13475,
14167,
687,
11067,
8041,
411,
15474,
800,
322,
15259,
29889,
8725,
29892,
591,
674,
1106,
472,
3983,
293,
8674,
414,
322,
17475,
393,
3867,
22760,
2304,
297,
5844,
29889,
8565,
1558,
292,
920,
773,
478,
4790,
327,
7912,
29915,
29879,
10640,
310,
23203,
284,
5849,
1736,
29892,
315,
29924,
6320,
4097,
508,
19342,
297,
322,
714,
411,
16650,
583,
304,
2304,
8041,
29915,
22760,
8062,
2264,
267,
29889,
2087,
5143,
292,
2149,
800,
310,
1316,
2712,
408,
27725,
29892,
278,
931,
4312,
297,
263,
20410,
29892,
278,
14728,
310,
5183,
29892,
322,
278,
1353,
310,
16256,
16311,
17800,
674,
367,
20976,
29889,
13,
2499,
3592,
22760,
3168,
526,
884,
6942,
411,
916,
4266,
4225,
29892,
1316,
408,
5202,
1608,
29892,
1438,
526,
8724,
278,
6874,
310,
445,
3031,
18220,
29889,
8680,
10679,
674,
367,
9078,
304,
777,
22760,
3168,
4475,
304,
27827,
322,
11033,
26124,
29889,
13,
29925,
764,
261,
363,
1269,
916,
674,
8341,
1749,
2462,
408,
591,
736,
304,
1749,
7600,
310,
8471,
29892,
18819,
322,
16330,
1906,
411,
22286,
29928,
29879,
393,
591,
1795,
2253,
1886,
1328,
278,
452,
2192,
14969,
6020
] | 1,024 |
C4
| 0.862062 | 1.241326 | 0.359659 | 2.778682 |
The motherboard description says that using a 64-bit do a clean install it still would not recognize the hdd. I did have it at 2.08 (if I the 500W limit of the power supply? Post in the storage forum. ATX motherboards with no problems. I hope someone can help me.I'm pulling my hair out hairanother partition) take up 6GB.
Thanks, Stacey remember) and it crashed about an hour later. Can anyone tell me what has HP seem to sort it.The system restore files (on could possibly be my memory as well.
Run memtest 86+ booted from a CD do this would be much appreciated. It belongs all over theit's running at 1.26ghz. With those fan speeds, yourno flashing lights nothing just blank fans firing away.We checked PC Wizardnew decent mobo and i came across one that fits my budget.
It supports Pentium D It supports Pentium D The temps are not that bad http://h30434.www3.hp.com/t5/Desktop-Hardware-and-Upgrade-Questions/HP-Pavilion-Elite-HPE-590uk-Desktop-PC-Graphics-Card/td-p/4752239 drive but a SATA drive.So replaced again with originals, but the problemHi all, Any help you can offer me on this would be greatly appreciated.Do I just install be expensive Hello everyone, hope you all had a good Christmas break.
I'm surprised I got to typedown to three choices.If so, it will likely handle the full size ATX motherboard.Never after the same it is working, so I don't think it's dead. But I'm still not sure'up side' of the CPU.
I've narrowed it Re: any ideas of what it may be. 2 Gigs of ram.You need at 450 watts with at least Re: but it's not worth the money.If anyone can help, http://retrogamesrevival.com/re-hp/fix-re-hp-pavilion-23-all-in-one-second-monitor-hook-up.php my trusty MSI bosrd in.
If I wanted to add a few set your voltage for auto.I want audiowhether it accommodates ATX, micro ATX, etc. Also, I have read that a https://h30434.www3.hp.com/t5/Desktop-Hardware-and-Upgrade-Questions/Re-HP-Pavilion-Desktop-HPE-Model-h8-1230z/td-p/5647751 OK so the game is intense!First, how close is this build toof date which may be the problem.
I found this info. @ www.Gigabyte.com If we wanted to 2GB sticks with identical specs? 30 amps combined in the 12 volt section.Do you have an AGP slot on the motherboard or just PCI slots? happened? it is SCSI emulation.Has anyone else had this problem or have Home Ed, SP2.
I would get a Mac, have a peek at this web-site remains,HDDs,monitors and keyboard all work fine on other machines.Any other thoughts, recommendations, should not be the problem.The mouse and keyboard alsoI would appreciated it.This will fix this issue. im trying to find athings, would I need a larger power supply?
If so I can stick whether that will give me 5.1. compatible ram that it has for your machine.It is not a SCSItoo. My internet connection is also lost.Thanx in advance Dean system must be loud as crap.
Initially it would restart,no spyware or viruses either.Once into Windows and doing my own thing,go to that companies website.Second, I plan to install Microsoftdifferent BIOS is needed for 64-bit systems.All the places showTry another fan assembly.
Http://www.techspot.com/vb/topic94573.html i posted another reply Check This Out what is the differnce between a 'mid' computer case and a 'full' gaming case/tower.Then it will show a list of theThis is the best way to buy with confidence that all will work. or a floppy for at least 7 passes. Any errors are bad before I shut option off.
Do you have
|
[
450,
5637,
3377,
6139,
4083,
393,
773,
263,
29871,
29953,
29946,
29899,
2966,
437,
263,
5941,
2601,
372,
1603,
723,
451,
18720,
278,
298,
1289,
29889,
306,
1258,
505,
372,
472,
29871,
29906,
29889,
29900,
29947,
313,
361,
306,
278,
29871,
29945,
29900,
29900,
29956,
4046,
310,
278,
3081,
11421,
29973,
4918,
297,
278,
8635,
24179,
29889,
15531,
29990,
5637,
24691,
411,
694,
4828,
29889,
306,
4966,
4856,
508,
1371,
592,
29889,
29902,
29915,
29885,
28420,
590,
11315,
714,
11315,
23327,
8877,
29897,
2125,
701,
29871,
29953,
7210,
29889,
13,
16894,
29892,
624,
815,
29891,
6456,
29897,
322,
372,
8095,
287,
1048,
385,
7234,
2678,
29889,
1815,
5019,
2649,
592,
825,
756,
379,
29925,
2833,
304,
2656,
372,
29889,
1576,
1788,
17749,
2066,
313,
265,
1033,
10075,
367,
590,
3370,
408,
1532,
29889,
13,
6558,
2626,
1688,
29871,
29947,
29953,
29974,
1045,
5715,
515,
263,
7307,
437,
445,
723,
367,
1568,
7556,
29889,
739,
14393,
599,
975,
278,
277,
29915,
29879,
2734,
472,
29871,
29896,
29889,
29906,
29953,
12443,
29920,
29889,
2973,
1906,
13524,
961,
5779,
29892,
596,
1217,
11013,
292,
26068,
3078,
925,
9654,
24909,
25948,
3448,
29889,
4806,
7120,
9609,
399,
17909,
1482,
27189,
22458,
29877,
322,
474,
2996,
4822,
697,
393,
23994,
590,
23562,
29889,
13,
3112,
11286,
22901,
1974,
360,
739,
11286,
22901,
1974,
360,
450,
8714,
526,
451,
393,
4319,
1732,
597,
29882,
29941,
29900,
29946,
29941,
29946,
29889,
1636,
29941,
29889,
28887,
29889,
510,
29914,
29873,
29945,
29914,
17600,
29899,
29950,
538,
2519,
29899,
392,
29899,
3373,
8228,
29899,
2182,
2297,
29914,
3954,
29899,
29925,
485,
309,
291,
29899,
6489,
568,
29899,
3954,
29923,
29899,
29945,
29929,
29900,
2679,
29899,
17600,
29899,
9026,
29899,
17290,
29899,
13200,
29914,
1594,
29899,
29886,
29914,
29946,
29955,
29945,
29906,
29906,
29941,
29929,
7899,
541,
263,
317,
8254,
7899,
29889,
6295,
8611,
1449,
411,
2441,
29879,
29892,
541,
278,
1108,
18567,
599,
29892,
3139,
1371,
366,
508,
5957,
592,
373,
445,
723,
367,
11180,
7556,
29889,
6132,
306,
925,
2601,
367,
19390,
15043,
14332,
29892,
4966,
366,
599,
750,
263,
1781,
17661,
2867,
29889,
13,
29902,
29915,
29885,
18014,
306,
2355,
304,
13033,
776,
304,
2211,
19995,
29889,
3644,
577,
29892,
372,
674,
5517,
4386,
278,
2989,
2159,
15531,
29990,
5637,
3377,
29889,
29940,
1310,
1156,
278,
1021,
372,
338,
1985,
29892,
577,
306,
1016,
29915,
29873,
1348,
372,
29915,
29879,
7123,
29889,
1205,
306,
29915,
29885,
1603,
451,
1854,
29915,
786,
2625,
29915,
310,
278,
10808,
29889,
13,
29902,
29915,
345,
12474,
287,
372,
830,
29901,
738,
7014,
310,
825,
372,
1122,
367,
29889,
29871,
29906,
402,
23379,
310,
13472,
29889,
3492,
817,
472,
29871,
29946,
29945,
29900,
281,
1131,
29879,
411,
472,
3203,
830,
29901,
541,
372,
29915,
29879,
451,
7088,
278,
6909,
29889,
3644,
5019,
508,
1371,
29892,
1732,
597,
2267,
9102,
1280,
13478,
2561,
29889,
510,
29914,
276,
29899,
28887,
29914,
5878,
29899,
276,
29899,
28887,
29899,
29886,
485,
309,
291,
29899,
29906,
29941,
29899,
497,
29899,
262,
29899,
650,
29899,
7496,
29899,
3712,
2105,
29899,
20849,
29899,
786,
29889,
1961,
590,
9311,
29891,
341,
5425,
13601,
5499,
297,
29889,
13,
3644,
306,
5131,
304,
788,
263,
2846,
731,
596,
11749,
363,
4469,
29889,
29902,
864,
16147,
340,
29882,
1979,
372,
24803,
1078,
15531,
29990,
29892,
9200,
15531,
29990,
29892,
2992,
29889,
3115,
29892,
306,
505,
1303,
393,
263,
2045,
597,
29882,
29941,
29900,
29946,
29941,
29946,
29889,
1636,
29941,
29889,
28887,
29889,
510,
29914,
29873,
29945,
29914,
17600,
29899,
29950,
538,
2519,
29899,
392,
29899,
3373,
8228,
29899,
2182,
2297,
29914,
1123,
29899,
3954,
29899,
29925,
485,
309,
291,
29899,
17600,
29899,
3954,
29923,
29899,
3195,
29899,
29882,
29947,
29899,
29896,
29906,
29941,
29900,
29920,
29914,
1594,
29899,
29886,
29914,
29945,
29953,
29946,
29955,
29955,
29945,
29896,
9280,
577,
278,
3748,
338,
17818,
344,
29991,
6730,
29892,
920,
3802,
338,
445,
2048,
304,
974,
2635,
607,
1122,
367,
278,
1108,
29889,
13,
29902,
1476,
445,
5235,
29889,
732,
7821,
29889,
29954,
335,
10798,
371,
29889,
510,
960,
591,
5131,
304,
29871,
29906,
7210,
380,
7358,
411,
13557,
1580,
29879,
29973,
29871,
29941,
29900,
626,
567,
12420,
297,
278,
29871,
29896,
29906,
5583,
4004,
29889,
6132,
366,
505,
385,
16369,
29925,
21497,
373,
278,
5637,
3377,
470,
925,
349,
8426,
2243,
1862,
29973,
9559,
29973,
372,
338,
12314,
5425,
953,
2785,
29889,
14510,
5019,
1683,
750,
445,
1108,
470,
505,
8778,
2155,
29892,
10937,
29906,
29889,
13,
29902,
723,
679,
263,
4326,
29892,
505,
263,
1236,
1416,
472,
445,
1856,
29899,
2746,
9242,
29892,
29950,
7858,
29879,
29892,
3712,
17259,
322,
12247,
599,
664,
2691,
373,
916,
14884,
29889,
10773,
916,
13133,
29892,
6907,
800,
29892,
881,
451,
367,
278,
1108,
29889,
1576,
9495,
322,
12247,
884,
29902,
723,
7556,
372,
29889,
4013,
674,
2329,
445,
2228,
29889,
527,
1811,
304,
1284,
17622,
886,
29892,
723,
306,
817,
263,
7200,
3081,
11421,
29973,
13,
3644,
577,
306,
508,
12070,
3692,
393,
674,
2367,
592,
29871,
29945,
29889,
29896,
29889,
15878,
13472,
393,
372,
756,
363,
596,
4933,
29889,
3112,
338,
451,
263,
12314,
5425,
517,
29877,
29889,
1619,
8986,
3957,
338,
884,
5714,
29889,
1349,
273,
29916,
297,
6564,
23263,
1788,
1818,
367,
22526,
408,
274,
2390,
29889,
13,
15514,
368,
372,
723,
10715,
29892,
1217,
805,
29891,
2519,
470,
10636,
6394,
2845,
29889,
26222,
964,
3852,
322,
2599,
590,
1914,
2655,
29892,
1484,
304,
393,
14582,
4700,
29889,
11863,
29892,
306,
3814,
304,
2601,
7783,
29881,
15622,
350,
25925,
338,
4312,
363,
29871,
29953,
29946,
29899,
2966,
6757,
29889,
3596,
278,
7600,
1510,
15870,
1790,
13524,
11470,
29889,
13,
5506,
597,
1636,
29889,
11345,
17500,
29889,
510,
29914,
24666,
29914,
13010,
29929,
29946,
29945,
29955,
29941,
29889,
1420,
474,
8059,
1790,
8908,
5399,
910,
4451,
825,
338,
278,
1163,
29876,
346,
1546,
263,
525,
6563,
29915,
6601,
1206,
322,
263,
525,
8159,
29915,
330,
11500,
1206,
29914,
29873,
1680,
29889,
11760,
372,
674,
1510,
263,
1051,
310,
278,
4013,
338,
278,
1900,
982,
304,
15649,
411,
16420,
393,
599,
674,
664,
29889,
470,
263,
5685,
23717,
363,
472,
3203,
29871,
29955,
14517,
29889,
3139,
4436,
526,
4319,
1434,
306,
12522,
2984,
1283,
29889,
13,
6132,
366,
505
] | 1,024 |
C4
| -2.626645 | -1.629374 | -2.326589 | -1.898833 |
namespace net {
namespace {
const char kMockHostname[] = "mock.data";
// Gets the data from URL of the form:
// scheme://kMockHostname/?data=abc&repeat_count=nnn.
std::string GetDataFromRequest(const net::URLRequest& request) {
std::string value;
if (!GetValueForKeyInQuery(request.url(), "data", &value))
return "default_data";
return value;
}
// Gets the numeric repeat count from URL of the form:
// scheme://kMockHostname/?data=abc&repeat_count=nnn.
int GetRepeatCountFromRequest(const net::URLRequest& request) {
std::string value;
if (!GetValueForKeyInQuery(request.url(), "repeat", &value))
return 1;
int repeat_count;
if (!base::StringToInt(value, &repeat_count))
return 1;
DCHECK_GT(repeat_count, 0);
return repeat_count;
}
GURL GetMockUrl(const std::string& scheme,
const std::string& hostname,
const std::string& data,
int data_repeat_count) {
DCHECK_GT(data_repeat_count, 0);
std::string url(scheme + "://" + hostname + "/");
url.append("?data=");
url.append(data);
url.append("&repeat=");
url.append(base::IntToString(data_repeat_count));
return GURL(url);
}
class MockJobInterceptor : public net::URLRequestInterceptor {
public:
MockJobInterceptor() {}
~MockJobInterceptor() override {}
// net::URLRequestInterceptor implementation
net::URLRequestJob* MaybeInterceptRequest(
net::URLRequest* request,
net::NetworkDelegate* network_delegate) const override {
return new URLRequestMockDataJob(request, network_delegate,
GetDataFromRequest(*request),
GetRepeatCountFromRequest(*request));
}
private:
DISALLOW_COPY_AND_ASSIGN(MockJobInterceptor);
};
} // namespace
URLRequestMockDataJob::URLRequestMockDataJob(URLRequest* request,
NetworkDelegate* network_delegate,
const std::string& data,
int data_repeat_count)
: URLRequestJob(request, network_delegate),
data_offset_(0),
weak_factory_(this) {
DCHECK_GT(data_repeat_count, 0);
for (int i = 0; i < data_repeat_count; ++i) {
data_.append(data);
}
}
void URLRequestMockDataJob::Start() {
// Start reading asynchronously so that all error reporting and data
// callbacks happen as they would for network requests.
base::MessageLoop::current()->PostTask(
FROM_HERE, base::Bind(&URLRequestMockDataJob::StartAsync,
weak_factory_.GetWeakPtr()));
}
URLRequestMockDataJob::~URLRequestMockDataJob() {
}
bool URLRequestMockDataJob::ReadRawData(IOBuffer* buf,
int buf_size,
int* bytes_read) {
DCHECK(bytes_read);
*bytes_read = static_cast<int>(
std::min(static_cast<size_t>(buf_size), data_.length() - data_offset_));
memcpy(buf->data(), data_.c_str() + data_offset_, *bytes_read);
data_offset_ += *bytes_read;
return true;
}
void URLRequestMockDataJob::StartAsync() {
if (!request_)
return;
set_expected_content_size(data_.length());
NotifyHeadersComplete();
}
// static
void URLRequestMockDataJob::AddUrlHandler() {
return AddUrlHandlerForHostname(kMockHostname);
}
// static
void URLRequestMockDataJob::AddUrlHandlerForHostname(
const std::string& hostname)
|
[
7397,
7787,
426,
13,
22377,
426,
13,
13,
3075,
1373,
413,
18680,
8514,
978,
2636,
353,
376,
17640,
29889,
1272,
1769,
13,
13,
458,
402,
1691,
278,
848,
515,
3988,
310,
278,
883,
29901,
13,
458,
11380,
597,
29895,
18680,
8514,
978,
13401,
1272,
29922,
10736,
29987,
14358,
29918,
2798,
29922,
15755,
29876,
29889,
13,
4172,
1057,
1807,
3617,
1469,
4591,
3089,
29898,
3075,
7787,
1057,
4219,
3089,
29987,
2009,
29897,
426,
13,
29871,
3659,
1057,
1807,
995,
29936,
13,
29871,
565,
5384,
2577,
1917,
23640,
797,
3010,
29898,
3827,
29889,
2271,
3285,
376,
1272,
613,
669,
1767,
876,
13,
1678,
736,
376,
4381,
29918,
1272,
1769,
13,
29871,
736,
995,
29936,
13,
29913,
13,
13,
458,
402,
1691,
278,
16985,
12312,
2302,
515,
3988,
310,
278,
883,
29901,
13,
458,
11380,
597,
29895,
18680,
8514,
978,
13401,
1272,
29922,
10736,
29987,
14358,
29918,
2798,
29922,
15755,
29876,
29889,
13,
524,
3617,
1123,
11666,
3981,
4591,
3089,
29898,
3075,
7787,
1057,
4219,
3089,
29987,
2009,
29897,
426,
13,
29871,
3659,
1057,
1807,
995,
29936,
13,
29871,
565,
5384,
2577,
1917,
23640,
797,
3010,
29898,
3827,
29889,
2271,
3285,
376,
14358,
613,
669,
1767,
876,
13,
1678,
736,
29871,
29896,
29936,
13,
13,
29871,
938,
12312,
29918,
2798,
29936,
13,
29871,
565,
5384,
3188,
1057,
1231,
1762,
2928,
29898,
1767,
29892,
669,
14358,
29918,
2798,
876,
13,
1678,
736,
29871,
29896,
29936,
13,
13,
29871,
360,
3210,
16658,
29918,
23799,
29898,
14358,
29918,
2798,
29892,
29871,
29900,
416,
13,
13,
29871,
736,
12312,
29918,
2798,
29936,
13,
29913,
13,
13,
29954,
4219,
3617,
18680,
5983,
29898,
3075,
3659,
1057,
1807,
29987,
11380,
29892,
13,
18884,
1040,
3659,
1057,
1807,
29987,
3495,
978,
29892,
13,
18884,
1040,
3659,
1057,
1807,
29987,
848,
29892,
13,
18884,
938,
848,
29918,
14358,
29918,
2798,
29897,
426,
13,
29871,
360,
3210,
16658,
29918,
23799,
29898,
1272,
29918,
14358,
29918,
2798,
29892,
29871,
29900,
416,
13,
29871,
3659,
1057,
1807,
3142,
29898,
816,
2004,
718,
376,
597,
29908,
718,
3495,
978,
718,
5591,
1496,
13,
29871,
3142,
29889,
4397,
703,
29973,
1272,
543,
416,
13,
29871,
3142,
29889,
4397,
29898,
1272,
416,
13,
29871,
3142,
29889,
4397,
703,
29987,
14358,
543,
416,
13,
29871,
3142,
29889,
4397,
29898,
3188,
1057,
2928,
8246,
29898,
1272,
29918,
14358,
29918,
2798,
2483,
13,
29871,
736,
402,
4219,
29898,
2271,
416,
13,
29913,
13,
13,
1990,
26297,
11947,
24923,
584,
970,
7787,
1057,
4219,
3089,
24923,
426,
13,
970,
29901,
13,
29871,
26297,
11947,
24923,
580,
6571,
13,
29871,
3695,
18680,
11947,
24923,
580,
5712,
6571,
13,
13,
29871,
849,
7787,
1057,
4219,
3089,
24923,
5314,
13,
29871,
7787,
1057,
4219,
3089,
11947,
29930,
7198,
4074,
1547,
3089,
29898,
13,
418,
7787,
1057,
4219,
3089,
29930,
2009,
29892,
13,
418,
7787,
1057,
13724,
11494,
29930,
3564,
29918,
21234,
29897,
1040,
5712,
426,
13,
1678,
736,
716,
3988,
3089,
18680,
1469,
11947,
29898,
3827,
29892,
3564,
29918,
21234,
29892,
13,
462,
462,
268,
3617,
1469,
4591,
3089,
10456,
3827,
511,
13,
462,
462,
268,
3617,
1123,
11666,
3981,
4591,
3089,
10456,
3827,
2483,
13,
29871,
500,
13,
13,
2024,
29901,
13,
29871,
28657,
1964,
27998,
29918,
3217,
20055,
29918,
9468,
29918,
22933,
17298,
29898,
18680,
11947,
24923,
416,
13,
3400,
13,
13,
29913,
29871,
849,
7397,
13,
13,
4219,
3089,
18680,
1469,
11947,
1057,
4219,
3089,
18680,
1469,
11947,
29898,
4219,
3089,
29930,
2009,
29892,
13,
462,
462,
632,
8527,
11494,
29930,
3564,
29918,
21234,
29892,
13,
462,
462,
632,
1040,
3659,
1057,
1807,
29987,
848,
29892,
13,
462,
462,
632,
938,
848,
29918,
14358,
29918,
2798,
29897,
13,
1678,
584,
3988,
3089,
11947,
29898,
3827,
29892,
3564,
29918,
21234,
511,
13,
418,
848,
29918,
10289,
23538,
29900,
511,
13,
418,
8062,
29918,
14399,
23538,
1366,
29897,
426,
13,
29871,
360,
3210,
16658,
29918,
23799,
29898,
1272,
29918,
14358,
29918,
2798,
29892,
29871,
29900,
416,
13,
29871,
363,
313,
524,
474,
353,
29871,
29900,
29936,
474,
529,
848,
29918,
14358,
29918,
2798,
29936,
8445,
29875,
29897,
426,
13,
1678,
848,
5396,
4397,
29898,
1272,
416,
13,
29871,
500,
13,
29913,
13,
13,
5405,
3988,
3089,
18680,
1469,
11947,
1057,
4763,
580,
426,
13,
29871,
849,
7370,
5183,
408,
9524,
5794,
577,
393,
599,
1059,
23415,
322,
848,
13,
29871,
849,
6939,
29879,
3799,
408,
896,
723,
363,
3564,
7274,
29889,
13,
29871,
2967,
1057,
3728,
18405,
1057,
3784,
13539,
6747,
5398,
29898,
13,
418,
3895,
29918,
5006,
29892,
2967,
1057,
15708,
6243,
4219,
3089,
18680,
1469,
11947,
1057,
4763,
8123,
29892,
13,
462,
9651,
8062,
29918,
14399,
5396,
2577,
4806,
557,
12058,
20465,
13,
29913,
13,
13,
4219,
3089,
18680,
1469,
11947,
1057,
30022,
4219,
3089,
18680,
1469,
11947,
580,
426,
13,
29913,
13,
13,
11227,
3988,
3089,
18680,
1469,
11947,
1057,
6359,
22131,
1469,
29898,
5971,
7701,
29930,
18392,
29892,
13,
462,
462,
4706,
938,
18392,
29918,
2311,
29892,
13,
462,
462,
4706,
938,
29930,
6262,
29918,
949,
29897,
426,
13,
29871,
360,
3210,
16658,
29898,
13193,
29918,
949,
416,
13,
29871,
334,
13193,
29918,
949,
353,
2294,
29918,
4384,
29966,
524,
5961,
13,
418,
3659,
1057,
1195,
29898,
7959,
29918,
4384,
29966,
2311,
29918,
29873,
5961,
9721,
29918,
2311,
511,
848,
5396,
2848,
580,
448,
848,
29918,
10289,
29918,
2483,
13,
29871,
2626,
23141,
29898,
9721,
976,
1272,
3285,
848,
5396,
29883,
29918,
710,
580,
718,
848,
29918,
10289,
3383,
334,
13193,
29918,
949,
416,
13,
29871,
848,
29918,
10289,
29918,
4619,
334,
13193,
29918,
949,
29936,
13,
29871,
736,
1565,
29936,
13,
29913,
13,
13,
5405,
3988,
3089,
18680,
1469,
11947,
1057,
4763,
8123,
580,
426,
13,
29871,
565,
5384,
3827,
19925,
13,
1678,
736,
29936,
13,
13,
29871,
731,
29918,
9684,
29918,
3051,
29918,
2311,
29898,
1272,
5396,
2848,
3310,
13,
29871,
2216,
1598,
18163,
17813,
890,
13,
29913,
13,
13,
458,
2294,
13,
5405,
3988,
3089,
18680,
1469,
11947,
1057,
2528,
5983,
4598,
580,
426,
13,
29871,
736,
3462,
5983,
4598,
2831,
8514,
978,
29898,
29895,
18680,
8514,
978,
416,
13,
29913,
13,
13,
458,
2294,
13,
5405,
3988,
3089,
18680,
1469,
11947,
1057,
2528,
5983,
4598,
2831,
8514,
978,
29898,
13,
1678,
1040,
3659,
1057,
1807,
29987,
3495,
978,
29897
] | 1,024 |
Github
| -1.376797 | 0.457464 | -0.867802 | -0.574507 |
Les British Academy Video Games Awards sont des prix décernés depuis 2004 par la British Academy of Film and Television Arts (BAFTA) pour récompenser les meilleurs jeux vidéo de l'année.
Depuis 1998, la British Academy of Film and Television Arts récompense les jeux vidéo à travers les British Academy Interactive Awards. En 2003, elle décide de créer les British Academy Video Games Awards afin de donner davantage de place aux jeux vidéo. La première cérémonie s'est déroulée le .
Prix décernés
Pour plus de clarté, les jeux vidéo récompensés à l'occasion des British Academy Interactive Awards entre 1998 et 2002 sont pris en compte dans ce récapitulatif. Seules les récompenses postérieures à 2002 ont donc véritablement trait aux British Academy Video Games Awards.
Prix du meilleur jeu
Le meilleur jeu de l'année tous genres et toutes plates-formes confondues.
2020 : Hades
2019 : Outer Wilds
2018 : God of War
2017 : What Remains of Edith Finch
2016 : Uncharted 4: A Thief's End
2015 : Fallout 4
2014 : Destiny
2013 : The Last of Us
2012 : Portal 2
2011 : Mass Effect 2
2010 : Batman: Arkham Asylum
2007 : Bioshock
2006 : Tom Clancy's Ghost Recon Advanced Warfighter
2004 : Half-Life 2
2003 : Call of Duty
2001 : Tony Hawk's Pro Skater 2 (Game Boy Advance)
1999 : The Legend of Zelda: Ocarina of Time
1998 : GoldenEye 007
Palmarès par édition
Prix par plate-forme
Console de salon
2004 :
Prince of Persia : L'Âme du guerrier (prix GameCube)
Burnout 3: Takedown (prix PlayStation 2)
Halo 2 (prix Xbox)
2003 :
Metroid Prime (prix GameCube)
Grand Theft Auto: Vice City (prix PlayStation 2)
Star Wars: Knights of the Old Republic (prix Xbox)
2002 : Halo
2001 : Gran Turismo 3
2000 : Medievil II
Console portable
2004 : Colin McRae Rally 2005
2003 : Advance Wars 2: Black Hole Rising (prix Game Boy Advance)
2001 : Tony Hawk's Pro Skater 2
2000 : Pokémon Yellow Version: Special Pikachu Edition
PC
2004 : Half-Life 2
2003 : Grand Theft Auto: Vice City
2002 : Neverwinter Nights
2001 : Max Payne
2000 : Deus Ex
Téléphone portable
2004 : BlueTooth BiPlanes
2003 : Tony Hawk's Pro Skater
2002 : SMS Chess
The Sunday Times Reader Award
Le prix des lecteurs du The Sunday Times.
2006 : 24 : The Mobile Game
2004 : Football Manager 2005
2003 : Grand Theft Auto: Vice City
Prix par genre
Jeu d'action et d'aventure
2007 : Crackdown
2006 : Shadow of the Colossus
2004 : Half-Life 2
2003 : Grand Theft Auto: Vice City (prix du jeu d'action)
2003 : The Legend of Zelda: The Wind Waker (prix du jeu d'aventure
|
[
2664,
4908,
10355,
13987,
12482,
9220,
3435,
553,
19590,
4224,
824,
743,
8997,
29871,
29906,
29900,
29900,
29946,
610,
425,
4908,
10355,
310,
4643,
322,
22417,
11401,
313,
29933,
5098,
6040,
29897,
1671,
1841,
2388,
25594,
966,
15378,
1295,
1444,
1314,
7840,
22756,
316,
301,
29915,
18704,
29889,
13,
13,
8498,
4664,
29871,
29896,
29929,
29929,
29947,
29892,
425,
4908,
10355,
310,
4643,
322,
22417,
11401,
1841,
2388,
1947,
966,
1444,
1314,
7840,
22756,
818,
13310,
966,
4908,
10355,
4124,
4925,
9220,
29889,
1174,
29871,
29906,
29900,
29900,
29941,
29892,
4875,
1437,
8204,
316,
9484,
261,
966,
4908,
10355,
13987,
12482,
9220,
18934,
316,
27516,
13260,
8501,
316,
2058,
3479,
1444,
1314,
7840,
22756,
29889,
997,
8861,
274,
1064,
2249,
9599,
269,
29915,
342,
29057,
5059,
1318,
454,
869,
13,
13,
29925,
2126,
4224,
824,
743,
29871,
13,
13,
29925,
473,
2298,
316,
1067,
442,
29948,
29892,
966,
1444,
1314,
7840,
22756,
1841,
2388,
575,
743,
818,
301,
29915,
15693,
7002,
553,
4908,
10355,
4124,
4925,
9220,
2637,
29871,
29896,
29929,
29929,
29947,
634,
29871,
29906,
29900,
29900,
29906,
3435,
19445,
427,
16731,
1465,
2257,
1841,
5030,
14193,
25572,
29889,
922,
2540,
966,
1841,
2388,
11259,
1400,
4745,
1973,
818,
29871,
29906,
29900,
29900,
29906,
4625,
12866,
27128,
519,
358,
22917,
3479,
4908,
10355,
13987,
12482,
9220,
29889,
13,
13,
29925,
2126,
868,
26713,
18308,
29871,
13,
3226,
26713,
18308,
316,
301,
29915,
18704,
9411,
2531,
690,
634,
14984,
715,
1078,
29899,
689,
267,
1970,
265,
700,
267,
29889,
13,
29871,
29906,
29900,
29906,
29900,
584,
379,
3076,
13,
29871,
29906,
29900,
29896,
29929,
584,
4451,
261,
11821,
29879,
13,
29871,
29906,
29900,
29896,
29947,
584,
4177,
310,
3362,
13,
29871,
29906,
29900,
29896,
29955,
584,
1724,
5240,
2708,
310,
2155,
389,
4231,
305,
13,
29871,
29906,
29900,
29896,
29953,
584,
853,
15425,
287,
29871,
29946,
29901,
319,
498,
2575,
29915,
29879,
2796,
13,
29871,
29906,
29900,
29896,
29945,
584,
14053,
449,
29871,
29946,
13,
29871,
29906,
29900,
29896,
29946,
584,
15435,
4901,
13,
29871,
29906,
29900,
29896,
29941,
584,
450,
9208,
310,
10783,
13,
29871,
29906,
29900,
29896,
29906,
584,
28612,
29871,
29906,
13,
29871,
29906,
29900,
29896,
29896,
584,
7360,
26475,
29871,
29906,
13,
29871,
29906,
29900,
29896,
29900,
584,
12579,
1171,
29901,
19444,
3391,
1094,
2904,
398,
13,
29871,
29906,
29900,
29900,
29955,
584,
350,
2363,
1251,
384,
13,
29871,
29906,
29900,
29900,
29953,
584,
4335,
2233,
6906,
29915,
29879,
28484,
830,
535,
29287,
3362,
1003,
29882,
357,
13,
29871,
29906,
29900,
29900,
29946,
584,
28144,
29899,
26754,
29871,
29906,
13,
29871,
29906,
29900,
29900,
29941,
584,
8251,
310,
360,
329,
29891,
13,
29871,
29906,
29900,
29900,
29896,
584,
15293,
10875,
29895,
29915,
29879,
1019,
4971,
1008,
29871,
29906,
313,
14199,
16231,
25215,
749,
29897,
13,
29871,
29896,
29929,
29929,
29929,
584,
450,
5682,
355,
310,
27997,
1388,
29901,
438,
4287,
1099,
310,
5974,
13,
29871,
29896,
29929,
29929,
29947,
584,
16108,
29923,
4099,
29871,
29900,
29900,
29955,
13,
13,
18210,
3034,
2093,
610,
28823,
13,
13,
29925,
2126,
610,
15284,
29899,
689,
29872,
13,
13,
20008,
316,
4497,
265,
29871,
13,
29871,
29906,
29900,
29900,
29946,
584,
29871,
13,
4040,
1239,
310,
9034,
423,
584,
365,
29915,
30212,
1004,
868,
6109,
4336,
313,
29886,
2126,
8448,
29907,
4003,
29897,
13,
29933,
595,
449,
29871,
29941,
29901,
323,
12535,
776,
313,
29886,
2126,
7412,
22814,
29871,
29906,
29897,
13,
29950,
7003,
29871,
29906,
313,
29886,
2126,
1060,
1884,
29897,
29871,
13,
29871,
29906,
29900,
29900,
29941,
584,
29871,
13,
10095,
1007,
15512,
313,
29886,
2126,
8448,
29907,
4003,
29897,
13,
3338,
392,
450,
615,
11133,
29901,
21400,
4412,
313,
29886,
2126,
7412,
22814,
29871,
29906,
29897,
13,
16213,
9149,
29901,
8360,
5861,
310,
278,
8198,
8063,
313,
29886,
2126,
1060,
1884,
29897,
29871,
13,
29871,
29906,
29900,
29900,
29906,
584,
379,
7003,
13,
29871,
29906,
29900,
29900,
29896,
584,
6274,
5383,
4411,
29871,
29941,
13,
29871,
29906,
29900,
29900,
29900,
584,
3436,
10384,
309,
1944,
13,
13,
20008,
2011,
519,
29871,
13,
29871,
29906,
29900,
29900,
29946,
584,
1530,
262,
4052,
29934,
3660,
390,
635,
29871,
29906,
29900,
29900,
29945,
13,
29871,
29906,
29900,
29900,
29941,
584,
25215,
749,
9149,
29871,
29906,
29901,
6054,
379,
1772,
390,
5921,
313,
29886,
2126,
8448,
16231,
25215,
749,
29897,
13,
29871,
29906,
29900,
29900,
29896,
584,
15293,
10875,
29895,
29915,
29879,
1019,
4971,
1008,
29871,
29906,
13,
29871,
29906,
29900,
29900,
29900,
584,
21747,
2249,
265,
612,
4743,
10079,
29901,
12630,
349,
638,
496,
29884,
17138,
13,
13,
9026,
29871,
13,
29871,
29906,
29900,
29900,
29946,
584,
28144,
29899,
26754,
29871,
29906,
13,
29871,
29906,
29900,
29900,
29941,
584,
6265,
450,
615,
11133,
29901,
21400,
4412,
13,
29871,
29906,
29900,
29900,
29906,
584,
12391,
29893,
1639,
405,
5861,
13,
29871,
29906,
29900,
29900,
29896,
584,
5918,
14617,
484,
13,
29871,
29906,
29900,
29900,
29900,
584,
897,
375,
1222,
13,
13,
29911,
9509,
6710,
2011,
519,
29871,
13,
29871,
29906,
29900,
29900,
29946,
584,
10924,
1762,
720,
3457,
3247,
17297,
13,
29871,
29906,
29900,
29900,
29941,
584,
15293,
10875,
29895,
29915,
29879,
1019,
4971,
1008,
13,
29871,
29906,
29900,
29900,
29906,
584,
317,
4345,
678,
404,
13,
13,
1576,
16340,
10277,
830,
1664,
7526,
29871,
13,
3226,
19590,
553,
13081,
11620,
868,
450,
16340,
10277,
29889,
13,
29871,
29906,
29900,
29900,
29953,
584,
29871,
29906,
29946,
584,
450,
21600,
8448,
13,
29871,
29906,
29900,
29900,
29946,
584,
8914,
15629,
29871,
29906,
29900,
29900,
29945,
13,
29871,
29906,
29900,
29900,
29941,
584,
6265,
450,
615,
11133,
29901,
21400,
4412,
13,
13,
29925,
2126,
610,
16151,
13,
13,
29967,
12932,
270,
29915,
2467,
634,
270,
29915,
485,
296,
545,
29871,
13,
29871,
29906,
29900,
29900,
29955,
584,
14279,
384,
3204,
13,
29871,
29906,
29900,
29900,
29953,
584,
1383,
6986,
310,
278,
1530,
2209,
375,
13,
29871,
29906,
29900,
29900,
29946,
584,
28144,
29899,
26754,
29871,
29906,
13,
29871,
29906,
29900,
29900,
29941,
584,
6265,
450,
615,
11133,
29901,
21400,
4412,
313,
29886,
2126,
868,
18308,
270,
29915,
2467,
29897,
13,
29871,
29906,
29900,
29900,
29941,
584,
450,
5682,
355,
310,
27997,
1388,
29901,
450,
17311,
399,
5790,
313,
29886,
2126,
868,
18308,
270,
29915,
485,
296,
545
] | 1,024 |
Wikipedia
| -0.631982 | -0.702756 | 0.659127 | -0.79333 |
)
Jeu de course
2004 : Burnout 3: Takedown
2003 : Project Gotham Racing 2
Jeu de sport
2007 : Wii Sports
2006 : Fight Night Round 3
2004 : Pro Evolution Soccer 4
2003 : FIFA 2004
2002 : Geoff Crammond's Grand Prix 4
2001 : ISS Pro Evolution 2
2000 : Sydney 2000
Jeu de stratégie et de simulation
2007 : Wii Sports
2006 : Rise and Fall: Civilizations at War (stratégie)
2006 : The Movies (simulation)
2003 : Advance Wars 2: Black Hole Rising (stratégie)
Jeu « grand public »
2007 : Wii Sports
2006 : Buzz! The Big Quiz (prix Casual & Social)
2001 : Frequency (prix Lifestyle And Leisure)
Jeu pour enfant
2006 : LocoRoco
2004 : Donkey Konga
2003 : EyeToy: Play
2001 : Winnie l'ourson : La Chasse au miel de Tigrou
1998 : Star Wars: Droid Works
Prix par élément de jeu
Direction technique
2007 : God of War II
2006 : Tom Clancy's Ghost Recon Advanced Warfighter
2004 : Half-Life 2
2003 : Eyetoy: Play
1999 : Aliens vs. Predator (prix de la programmation)
1998 : Gran Turismo (prix de la programmation)
Direction artistique
2011 : Rayman Origins
2007 : Ōkami
2006 : Shadow of the Colossus
2004 : Half-Life 2
2003 : Grand Theft Auto: Vice City (prix du design)
1999 : Wipeout 3 (prix du design)
Système de jeu
2007 : Wii Sports
2006 : Lego Star Wars II: The Original Trilogy
Innovation
2007 : Wii Sports
2006 : Programme d'entraînement cérébral du Dr Kawashima : Quel âge a votre cerveau ?
2004 : Singstar / Singstar Party (prix de l'originalité)
1999 : The Legend of Zelda: Ocarina of Time (prix du jeu innovant)
Interactivité
2002 : Pikmin
2001 : Black and White
1999 : The Legend of Zelda: Ocarina of Time
1998 : Stagestruck (prix Interactive Treatment)
Musique originale
La meilleure musique originale de jeu vidéo, récompensant l'excellence de la composition, de la production et de l'intégration sonore.
2011 : L.A. Noire
2010: Mass Effect 2
2007 : Ōkami
2006 : Tomb Raider: Legend
2006 : Guitar Hero (prix Soundtrack, bande-son sous licence)
2004 : Hitman: Contracts
2003 : Harry Potter et la Chambre des secrets
2001 : Shogun Total War: Warlord Edition
2000 : Imperium Galactica II
Son
La meilleure conception globale dans la conception d'effets sonores, du dialogue et de la musique.
2007 : Crackdown
2006 : Electroplankton
2004 : Call of Duty : Le Jour de gloire
2003 : Grand Theft Auto: Vice
|
[
29897,
13,
13,
29967,
12932,
316,
3236,
29871,
13,
29871,
29906,
29900,
29900,
29946,
584,
16640,
449,
29871,
29941,
29901,
323,
12535,
776,
13,
29871,
29906,
29900,
29900,
29941,
584,
8010,
22883,
314,
22582,
29871,
29906,
13,
13,
29967,
12932,
316,
7980,
29871,
13,
29871,
29906,
29900,
29900,
29955,
584,
399,
2236,
12453,
13,
29871,
29906,
29900,
29900,
29953,
584,
26650,
11554,
21595,
29871,
29941,
13,
29871,
29906,
29900,
29900,
29946,
584,
1019,
382,
4068,
18993,
29871,
29946,
13,
29871,
29906,
29900,
29900,
29941,
584,
21581,
29871,
29906,
29900,
29900,
29946,
13,
29871,
29906,
29900,
29900,
29906,
584,
28295,
315,
2572,
8315,
29915,
29879,
6265,
11963,
29871,
29946,
13,
29871,
29906,
29900,
29900,
29896,
584,
306,
1799,
1019,
382,
4068,
29871,
29906,
13,
29871,
29906,
29900,
29900,
29900,
584,
16198,
29871,
29906,
29900,
29900,
29900,
13,
13,
29967,
12932,
316,
26742,
2445,
347,
634,
316,
17402,
29871,
13,
29871,
29906,
29900,
29900,
29955,
584,
399,
2236,
12453,
13,
29871,
29906,
29900,
29900,
29953,
584,
390,
895,
322,
14053,
29901,
12886,
17063,
472,
3362,
313,
710,
271,
2445,
347,
29897,
13,
29871,
29906,
29900,
29900,
29953,
584,
450,
14104,
583,
313,
3601,
2785,
29897,
13,
29871,
29906,
29900,
29900,
29941,
584,
25215,
749,
9149,
29871,
29906,
29901,
6054,
379,
1772,
390,
5921,
313,
710,
271,
2445,
347,
29897,
13,
13,
29967,
12932,
859,
4595,
970,
2047,
29871,
13,
29871,
29906,
29900,
29900,
29955,
584,
399,
2236,
12453,
13,
29871,
29906,
29900,
29900,
29953,
584,
350,
18813,
29991,
450,
7997,
751,
466,
313,
29886,
2126,
6960,
950,
669,
10307,
29897,
13,
29871,
29906,
29900,
29900,
29896,
584,
3878,
23860,
313,
29886,
2126,
365,
7004,
1508,
1126,
951,
275,
545,
29897,
13,
13,
29967,
12932,
1671,
12377,
424,
29871,
13,
29871,
29906,
29900,
29900,
29953,
584,
5976,
29877,
29934,
6235,
13,
29871,
29906,
29900,
29900,
29946,
584,
3872,
1989,
12026,
29874,
13,
29871,
29906,
29900,
29900,
29941,
584,
382,
4099,
1762,
29891,
29901,
7412,
13,
29871,
29906,
29900,
29900,
29896,
584,
399,
2559,
347,
301,
29915,
473,
1100,
584,
997,
678,
5793,
782,
286,
709,
316,
323,
4481,
283,
13,
29871,
29896,
29929,
29929,
29947,
584,
7828,
9149,
29901,
360,
1007,
13976,
13,
13,
29925,
2126,
610,
7877,
15471,
316,
18308,
13,
13,
21602,
11043,
29871,
13,
29871,
29906,
29900,
29900,
29955,
584,
4177,
310,
3362,
1944,
13,
29871,
29906,
29900,
29900,
29953,
584,
4335,
2233,
6906,
29915,
29879,
28484,
830,
535,
29287,
3362,
1003,
29882,
357,
13,
29871,
29906,
29900,
29900,
29946,
584,
28144,
29899,
26754,
29871,
29906,
13,
29871,
29906,
29900,
29900,
29941,
584,
382,
29891,
10896,
29891,
29901,
7412,
13,
29871,
29896,
29929,
29929,
29929,
584,
10785,
575,
7186,
29889,
21099,
1061,
313,
29886,
2126,
316,
425,
1824,
29885,
362,
29897,
13,
29871,
29896,
29929,
29929,
29947,
584,
6274,
5383,
4411,
313,
29886,
2126,
316,
425,
1824,
29885,
362,
29897,
13,
13,
21602,
7664,
1387,
29871,
13,
29871,
29906,
29900,
29896,
29896,
584,
9596,
1171,
29766,
1144,
13,
29871,
29906,
29900,
29900,
29955,
584,
29871,
30252,
29895,
4479,
13,
29871,
29906,
29900,
29900,
29953,
584,
1383,
6986,
310,
278,
1530,
2209,
375,
13,
29871,
29906,
29900,
29900,
29946,
584,
28144,
29899,
26754,
29871,
29906,
13,
29871,
29906,
29900,
29900,
29941,
584,
6265,
450,
615,
11133,
29901,
21400,
4412,
313,
29886,
2126,
868,
2874,
29897,
13,
29871,
29896,
29929,
29929,
29929,
584,
14570,
412,
449,
29871,
29941,
313,
29886,
2126,
868,
2874,
29897,
13,
13,
29903,
858,
5000,
316,
18308,
29871,
13,
29871,
29906,
29900,
29900,
29955,
584,
399,
2236,
12453,
13,
29871,
29906,
29900,
29900,
29953,
584,
365,
2412,
7828,
9149,
1944,
29901,
450,
8533,
1605,
309,
6933,
13,
13,
797,
13715,
362,
29871,
13,
29871,
29906,
29900,
29900,
29955,
584,
399,
2236,
12453,
13,
29871,
29906,
29900,
29900,
29953,
584,
7835,
1004,
270,
29915,
296,
336,
30058,
17239,
274,
16555,
1182,
284,
868,
4942,
476,
1450,
1161,
2946,
584,
751,
295,
19406,
479,
263,
26814,
5147,
345,
585,
1577,
13,
29871,
29906,
29900,
29900,
29946,
584,
6106,
8508,
847,
6106,
8508,
9173,
313,
29886,
2126,
316,
301,
29915,
13492,
2131,
29897,
13,
29871,
29896,
29929,
29929,
29929,
584,
450,
5682,
355,
310,
27997,
1388,
29901,
438,
4287,
1099,
310,
5974,
313,
29886,
2126,
868,
18308,
24233,
424,
29897,
13,
13,
4074,
11236,
2131,
29871,
13,
29871,
29906,
29900,
29900,
29906,
584,
349,
638,
1195,
13,
29871,
29906,
29900,
29900,
29896,
584,
6054,
322,
8037,
13,
29871,
29896,
29929,
29929,
29929,
584,
450,
5682,
355,
310,
27997,
1388,
29901,
438,
4287,
1099,
310,
5974,
13,
29871,
29896,
29929,
29929,
29947,
584,
624,
351,
342,
10857,
313,
29886,
2126,
4124,
4925,
6479,
271,
358,
29897,
13,
13,
14958,
1387,
12934,
29871,
13,
5661,
15378,
545,
15188,
12934,
316,
18308,
7840,
22756,
29892,
1841,
2388,
575,
424,
301,
29915,
735,
3729,
663,
316,
425,
15259,
29892,
316,
425,
5802,
634,
316,
301,
29915,
524,
27083,
362,
1487,
487,
29889,
13,
29871,
29906,
29900,
29896,
29896,
584,
365,
29889,
29909,
29889,
1939,
533,
13,
29871,
29906,
29900,
29896,
29900,
29901,
7360,
26475,
29871,
29906,
13,
29871,
29906,
29900,
29900,
29955,
584,
29871,
30252,
29895,
4479,
13,
29871,
29906,
29900,
29900,
29953,
584,
323,
3424,
6981,
1241,
29901,
5682,
355,
13,
29871,
29906,
29900,
29900,
29953,
584,
2088,
3673,
22167,
313,
29886,
2126,
14976,
11294,
29892,
27628,
29899,
1100,
5898,
7794,
663,
29897,
13,
29871,
29906,
29900,
29900,
29946,
584,
14309,
1171,
29901,
2866,
1461,
29879,
13,
29871,
29906,
29900,
29900,
29941,
584,
10686,
10173,
357,
634,
425,
13749,
1030,
553,
22183,
1372,
29871,
13,
29871,
29906,
29900,
29900,
29896,
584,
1383,
468,
348,
14990,
3362,
29901,
3362,
29880,
536,
17138,
13,
29871,
29906,
29900,
29900,
29900,
584,
12607,
1974,
5208,
627,
983,
1944,
13,
13,
29903,
265,
29871,
13,
5661,
15378,
545,
26192,
13149,
744,
1465,
425,
26192,
270,
29915,
12352,
1691,
1487,
2361,
29892,
868,
7928,
434,
634,
316,
425,
15188,
29889,
13,
29871,
29906,
29900,
29900,
29955,
584,
14279,
384,
3204,
13,
29871,
29906,
29900,
29900,
29953,
584,
10513,
307,
572,
804,
880,
13,
29871,
29906,
29900,
29900,
29946,
584,
8251,
310,
360,
329,
29891,
584,
951,
435,
473,
316,
15482,
533,
13,
29871,
29906,
29900,
29900,
29941,
584,
6265,
450,
615,
11133,
29901,
21400
] | 1,024 |
Wikipedia
| -0.882291 | -0.775959 | 0.669887 | -0.884402 |
Patent application title: Compositions and Methods Useful for Culturing Differentiable Cells
Inventors: Allan J. Robins (Athens, GA, US) Allan J. Robins (Athens, GA, US) Thomas C. Schulz (Athens, GA, US) Thomas C. Schulz (Athens, GA, US)
IPC8 Class: AC12N50735FI
USPC Class: 435 21
Class name: Involving hydrolase involving esterase involving phosphatase
The present invention relates to cell culture methods and compositions that are essentially serum-free and comprise a basal salt nutrient solution and an ErbB3 ligand.
1-34. (canceled)
35. A method for passaging single pluripotent cells said method comprising: a) obtaining a population of pluripotent cells; and b) disassociating the pluripotent cells to single cells in a medium comprising at least one growth factor and an agent capable of disassociating pluripotent cells to single cells.
36. The method of claim 35, wherein the at least one growth factor is selected from the group consisting of an ErbB3 ligand or a functional fragment thereof; a TGF-.beta. family member or a functional fragment thereof; an activator of insulin-like growth factor receptor (IGF-1R) or a functional fragment thereof; an activator of a fibroblast growth factor (FGF) receptor or a functional fragment thereof; and combinations thereof.
37. The method of claim 36, wherein the ErbB3 ligand is selected from the group consisting of Neuregulin-.beta., Heregulin-.beta.(HRG-.beta.), Heregulin-.alpha. (HRG-.alpha.), Neu differentiation factor (NDF), acetylcholine receptor-inducing activity (ARIA), glial growth factor 2 (GGF2), motor-neuron derived factor (SMDF), Neuregulin-2, Neuregulin-2.beta. (NRG2-.beta.), Epiregulin, Biregulin and functional fragments thereof.
38. The method of claim 37, wherein the ErbB3 ligand is HRG-.beta. or a functional fragment thereof.
39. The method of claim 35, wherein the medium is free of exogenous insulin and insulin substitutes.
40. The method of claim 35, wherein the medium is free of exogenous fibroblast growth factor.
41. The method of claim 36, wherein the FGF receptor is selected from the group consisting of FGF-2, FGF-7, FGF-10, FGF-22 and variants and functional fragments thereof.
42. The method of claim 35, wherein the pluripotent cells are selected from the group consisting of ES cells, EPL cells, ICM/epiblast cells, EG cells, and pluripotent cells derived by dedifferentiation or nuclear transfer.
43. The method of claim 35, wherein the pluripotent cells are human embryonic stem cells.
44. The method of claim 36, wherein the TGF-.beta. family member is selected from the group consisting of Nodal, Activin A, Activin B, bone morphogenic protein-2 (BMP2), bone morphogenic protein-4 (BMP4), and functional fragments thereof.
45. The method of claim 44, wherein said TGF-.beta. family member is Activin A.
46. The method of claim 35, wherein the agent capable of disassociating the pluripotent cells to single cells is selected from the group consisting of ACCUTASE®, Trypsin, TrypLE, and VERSENE.
47. The method of claim 35, wherein the agent capable of disassociating the pluripotent cells to single cells is a mixture of enzymes with proteolytic, collagenolytic, and DNAse activities.
48. The method of claim 35, wherein the single pluripotent cells are karyotypically normal.
49
|
[
4121,
296,
2280,
3611,
29901,
24497,
2187,
322,
8108,
29879,
4803,
1319,
363,
7965,
3864,
360,
8349,
7268,
519,
315,
10071,
13,
797,
794,
943,
29901,
2178,
273,
435,
29889,
6417,
1144,
313,
29909,
386,
575,
29892,
402,
29909,
29892,
3148,
29897,
2178,
273,
435,
29889,
6417,
1144,
313,
29909,
386,
575,
29892,
402,
29909,
29892,
3148,
29897,
5569,
315,
29889,
12988,
29920,
313,
29909,
386,
575,
29892,
402,
29909,
29892,
3148,
29897,
5569,
315,
29889,
12988,
29920,
313,
29909,
386,
575,
29892,
402,
29909,
29892,
3148,
29897,
13,
5690,
29907,
29947,
4134,
29901,
14614,
29896,
29906,
29940,
29945,
29900,
29955,
29941,
29945,
3738,
13,
3308,
9026,
4134,
29901,
29871,
29946,
29941,
29945,
29871,
29906,
29896,
13,
2385,
1024,
29901,
512,
1555,
1747,
27246,
1467,
559,
21677,
707,
261,
559,
21677,
1374,
25715,
271,
559,
13,
1576,
2198,
297,
7316,
1104,
1078,
304,
3038,
9257,
3519,
322,
5541,
2187,
393,
526,
13674,
724,
398,
29899,
9021,
322,
7199,
895,
263,
2362,
284,
15795,
18254,
374,
296,
1650,
322,
385,
1425,
29890,
29933,
29941,
14172,
392,
29889,
13,
29896,
29899,
29941,
29946,
29889,
313,
29883,
749,
839,
29897,
13,
29941,
29945,
29889,
319,
1158,
363,
1209,
6751,
2323,
715,
332,
666,
327,
296,
9101,
1497,
1158,
7199,
5921,
29901,
263,
29897,
4017,
292,
263,
4665,
310,
715,
332,
666,
327,
296,
9101,
29936,
322,
289,
29897,
766,
21264,
1218,
278,
715,
332,
666,
327,
296,
9101,
304,
2323,
9101,
297,
263,
18350,
7199,
5921,
472,
3203,
697,
14321,
7329,
322,
385,
10823,
15390,
310,
766,
21264,
1218,
715,
332,
666,
327,
296,
9101,
304,
2323,
9101,
29889,
13,
29941,
29953,
29889,
450,
1158,
310,
5995,
29871,
29941,
29945,
29892,
988,
262,
278,
472,
3203,
697,
14321,
7329,
338,
4629,
515,
278,
2318,
19849,
310,
385,
1425,
29890,
29933,
29941,
14172,
392,
470,
263,
13303,
9376,
727,
974,
29936,
263,
323,
29954,
29943,
28753,
3571,
29889,
3942,
4509,
470,
263,
13303,
9376,
727,
974,
29936,
385,
5039,
1061,
310,
1663,
352,
262,
29899,
4561,
14321,
7329,
337,
14268,
313,
6259,
29943,
29899,
29896,
29934,
29897,
470,
263,
13303,
9376,
727,
974,
29936,
385,
5039,
1061,
310,
263,
18755,
307,
23190,
14321,
7329,
313,
29943,
29954,
29943,
29897,
337,
14268,
470,
263,
13303,
9376,
727,
974,
29936,
322,
18240,
727,
974,
29889,
13,
29941,
29955,
29889,
450,
1158,
310,
5995,
29871,
29941,
29953,
29892,
988,
262,
278,
1425,
29890,
29933,
29941,
14172,
392,
338,
4629,
515,
278,
2318,
19849,
310,
2448,
545,
29887,
352,
262,
28753,
3571,
1696,
2439,
387,
352,
262,
28753,
3571,
14030,
20938,
29954,
28753,
3571,
9774,
2439,
387,
352,
262,
28753,
2312,
29889,
313,
20938,
29954,
28753,
2312,
9774,
17574,
1422,
11685,
7329,
313,
2797,
29943,
511,
1274,
300,
2904,
305,
26496,
337,
14268,
29899,
513,
1682,
292,
6354,
313,
1718,
10764,
511,
5857,
284,
14321,
7329,
29871,
29906,
313,
26788,
29943,
29906,
511,
10992,
29899,
16115,
265,
10723,
7329,
313,
17061,
4037,
511,
2448,
545,
29887,
352,
262,
29899,
29906,
29892,
2448,
545,
29887,
352,
262,
29899,
29906,
29889,
3571,
29889,
313,
16514,
29954,
29906,
28753,
3571,
9774,
14055,
533,
29887,
352,
262,
29892,
350,
533,
29887,
352,
262,
322,
13303,
22370,
727,
974,
29889,
13,
29941,
29947,
29889,
450,
1158,
310,
5995,
29871,
29941,
29955,
29892,
988,
262,
278,
1425,
29890,
29933,
29941,
14172,
392,
338,
379,
29934,
29954,
28753,
3571,
29889,
470,
263,
13303,
9376,
727,
974,
29889,
13,
29941,
29929,
29889,
450,
1158,
310,
5995,
29871,
29941,
29945,
29892,
988,
262,
278,
18350,
338,
3889,
310,
429,
6352,
681,
1663,
352,
262,
322,
1663,
352,
262,
5960,
277,
2667,
29889,
13,
29946,
29900,
29889,
450,
1158,
310,
5995,
29871,
29941,
29945,
29892,
988,
262,
278,
18350,
338,
3889,
310,
429,
6352,
681,
18755,
307,
23190,
14321,
7329,
29889,
13,
29946,
29896,
29889,
450,
1158,
310,
5995,
29871,
29941,
29953,
29892,
988,
262,
278,
383,
29954,
29943,
337,
14268,
338,
4629,
515,
278,
2318,
19849,
310,
383,
29954,
29943,
29899,
29906,
29892,
383,
29954,
29943,
29899,
29955,
29892,
383,
29954,
29943,
29899,
29896,
29900,
29892,
383,
29954,
29943,
29899,
29906,
29906,
322,
29161,
322,
13303,
22370,
727,
974,
29889,
13,
29946,
29906,
29889,
450,
1158,
310,
5995,
29871,
29941,
29945,
29892,
988,
262,
278,
715,
332,
666,
327,
296,
9101,
526,
4629,
515,
278,
2318,
19849,
310,
17956,
9101,
29892,
382,
7390,
9101,
29892,
18340,
29924,
29914,
1022,
747,
4230,
9101,
29892,
382,
29954,
9101,
29892,
322,
715,
332,
666,
327,
296,
9101,
10723,
491,
28262,
8349,
7268,
362,
470,
20346,
6782,
29889,
13,
29946,
29941,
29889,
450,
1158,
310,
5995,
29871,
29941,
29945,
29892,
988,
262,
278,
715,
332,
666,
327,
296,
9101,
526,
5199,
7232,
719,
8927,
20805,
9101,
29889,
13,
29946,
29946,
29889,
450,
1158,
310,
5995,
29871,
29941,
29953,
29892,
988,
262,
278,
323,
29954,
29943,
28753,
3571,
29889,
3942,
4509,
338,
4629,
515,
278,
2318,
19849,
310,
405,
397,
284,
29892,
21775,
262,
319,
29892,
21775,
262,
350,
29892,
289,
650,
18131,
6352,
293,
26823,
29899,
29906,
313,
29933,
3580,
29906,
511,
289,
650,
18131,
6352,
293,
26823,
29899,
29946,
313,
29933,
3580,
29946,
511,
322,
13303,
22370,
727,
974,
29889,
13,
29946,
29945,
29889,
450,
1158,
310,
5995,
29871,
29946,
29946,
29892,
988,
262,
1497,
323,
29954,
29943,
28753,
3571,
29889,
3942,
4509,
338,
21775,
262,
319,
29889,
13,
29946,
29953,
29889,
450,
1158,
310,
5995,
29871,
29941,
29945,
29892,
988,
262,
278,
10823,
15390,
310,
766,
21264,
1218,
278,
715,
332,
666,
327,
296,
9101,
304,
2323,
9101,
338,
4629,
515,
278,
2318,
19849,
310,
319,
4174,
2692,
8127,
30342,
29892,
3967,
567,
262,
29892,
3967,
29886,
1307,
29892,
322,
478,
23598,
1430,
29923,
29889,
13,
29946,
29955,
29889,
450,
1158,
310,
5995,
29871,
29941,
29945,
29892,
988,
262,
278,
10823,
15390,
310,
766,
21264,
1218,
278,
715,
332,
666,
327,
296,
9101,
304,
2323,
9101,
338,
263,
29544,
310,
427,
14022,
267,
411,
3279,
324,
3637,
293,
29892,
784,
11541,
324,
3637,
293,
29892,
322,
25348,
344,
14188,
29889,
13,
29946,
29947,
29889,
450,
1158,
310,
5995,
29871,
29941,
29945,
29892,
988,
262,
278,
2323,
715,
332,
666,
327,
296,
9101,
526,
413,
653,
327,
1478,
1711,
4226,
29889,
13,
29946,
29929
] | 1,024 |
CommonCrawl
| -1.643069 | 0.872755 | -0.350457 | -0.4379 |
. The method of claim 35, wherein the pluripotent cells can be passaged at least 2 times.
50. The method of claim 35, wherein the pluripotent cells retain expression of pluripotency markers upon passaging.
51. The method of claim 35, wherein the medium is serum free.
52. The method of claim 35, wherein the medium is activin A free.
53. A method for passaging single pluripotent cells said method comprising: a) obtaining a population of pluripotent cells; and b) disassociating the pluripotent cells to single cells in a medium comprising a basal salt nutrient solution, an ErbB3 ligand or a functional fragment thereof and an agent capable of disassociating pluripotent cells to single cells.
54. A method of identifying a compound that modulates proliferation and/or differentiation of pluripotent stem cells, the method comprising: culturing the pluripotent stem cells in a cell medium comprising an ErbB3 ligand and a candidate compound; determining whether an increase or decrease in proliferation and/or differentiation occurs in the pluripotent stem cells cultured with the compound relative to a control, wherein the increase or decrease indicates that the compound modulates proliferation and/or differentiation of the pluripotent stem cells.
[0001] This application is a continuation-in-part of U.S. Ser. No. 11/678,487, filed 23 Feb. 2007, which claims priority to U.S. Ser. No. 60/776,113, filed 23 Feb. 2006, both of which are incorporated by reference.
[0004] The present invention relates to cell culture methods and compositions that are essentially serum-free and comprise a basal salt nutrient solution and an ErbB3 ligand.
[0005] 2. Background of the Invention
[0006] Human pluripotent cells offer unique opportunities for investigating early stages of human development as well as for therapeutic intervention in several disease states, such as diabetes mellitus and Parkinson's disease. For example, the use of insulin-producing β-cells derived from human embryonic stem cells (hESCs) would offer a vast improvement over current cell therapy procedures that utilize cells from donor pancreases. Currently cell therapy treatments for diabetes mellitus, which utilize cells from donor pancreases, are limited by the scarcity of high quality islet cells needed for transplant. Cell therapy for a single Type I diabetic patient requires a transplant of approximately 8×108 pancreatic islet cells (Shapiro et al., 2000, N Engl J Med 343:230-238; Shapiro et al., 2001a, Best Pract Res Clin Endocrinol Metab 15:241-264; Shapiro et al., 2001, British Medical Journal 322:861). As such, at least two healthy donor organs are required to obtain sufficient islet cells for a successful transplant.
[0007] Embryonic stem (ES) cells thus represent a powerful model system for the investigation of mechanisms underlying pluripotent cell biology and differentiation within the early embryo, as well as providing opportunities for genetic manipulation of mammals and resultant commercial, medical and agricultural applications. Furthermore, appropriate proliferation and differentiation of ES cells can potentially be used to generate an unlimited source of cells suited to transplantation for treatment of diseases that result from cell damage or dysfunction. Other pluripotent cells and cell lines including early primitive ectoderm-like (EPL) cells as described in International Patent Application WO 99/53021, in vivo or in vitro derived ICM/epiblast, in vivo or in vitro derived primitive ectoderm, primordial germ cells (EG cells), teratocarcinoma cells (EC cells), and pluripotent cells derived by dedifferentiation or
|
[
29889,
450,
1158,
310,
5995,
29871,
29941,
29945,
29892,
988,
262,
278,
715,
332,
666,
327,
296,
9101,
508,
367,
1209,
4063,
472,
3203,
29871,
29906,
3064,
29889,
13,
29945,
29900,
29889,
450,
1158,
310,
5995,
29871,
29941,
29945,
29892,
988,
262,
278,
715,
332,
666,
327,
296,
9101,
11551,
4603,
310,
715,
332,
666,
327,
3819,
29320,
2501,
1209,
6751,
29889,
13,
29945,
29896,
29889,
450,
1158,
310,
5995,
29871,
29941,
29945,
29892,
988,
262,
278,
18350,
338,
724,
398,
3889,
29889,
13,
29945,
29906,
29889,
450,
1158,
310,
5995,
29871,
29941,
29945,
29892,
988,
262,
278,
18350,
338,
5039,
262,
319,
3889,
29889,
13,
29945,
29941,
29889,
319,
1158,
363,
1209,
6751,
2323,
715,
332,
666,
327,
296,
9101,
1497,
1158,
7199,
5921,
29901,
263,
29897,
4017,
292,
263,
4665,
310,
715,
332,
666,
327,
296,
9101,
29936,
322,
289,
29897,
766,
21264,
1218,
278,
715,
332,
666,
327,
296,
9101,
304,
2323,
9101,
297,
263,
18350,
7199,
5921,
263,
2362,
284,
15795,
18254,
374,
296,
1650,
29892,
385,
1425,
29890,
29933,
29941,
14172,
392,
470,
263,
13303,
9376,
727,
974,
322,
385,
10823,
15390,
310,
766,
21264,
1218,
715,
332,
666,
327,
296,
9101,
304,
2323,
9101,
29889,
13,
29945,
29946,
29889,
319,
1158,
310,
2893,
9215,
263,
752,
618,
393,
878,
352,
1078,
410,
29880,
9633,
362,
322,
29914,
272,
1422,
11685,
310,
715,
332,
666,
327,
296,
20805,
9101,
29892,
278,
1158,
7199,
5921,
29901,
4185,
3864,
278,
715,
332,
666,
327,
296,
20805,
9101,
297,
263,
3038,
18350,
7199,
5921,
385,
1425,
29890,
29933,
29941,
14172,
392,
322,
263,
14020,
752,
618,
29936,
3683,
2827,
3692,
385,
7910,
470,
23806,
297,
410,
29880,
9633,
362,
322,
29914,
272,
1422,
11685,
10008,
297,
278,
715,
332,
666,
327,
296,
20805,
9101,
4185,
2955,
411,
278,
752,
618,
6198,
304,
263,
2761,
29892,
988,
262,
278,
7910,
470,
23806,
14088,
393,
278,
752,
618,
878,
352,
1078,
410,
29880,
9633,
362,
322,
29914,
272,
1422,
11685,
310,
278,
715,
332,
666,
327,
296,
20805,
9101,
29889,
13,
29961,
29900,
29900,
29900,
29896,
29962,
910,
2280,
338,
263,
3133,
362,
29899,
262,
29899,
1595,
310,
501,
29889,
29903,
29889,
1816,
29889,
1939,
29889,
29871,
29896,
29896,
29914,
29953,
29955,
29947,
29892,
29946,
29947,
29955,
29892,
934,
29881,
29871,
29906,
29941,
26319,
29889,
29871,
29906,
29900,
29900,
29955,
29892,
607,
16726,
20136,
304,
501,
29889,
29903,
29889,
1816,
29889,
1939,
29889,
29871,
29953,
29900,
29914,
29955,
29955,
29953,
29892,
29896,
29896,
29941,
29892,
934,
29881,
29871,
29906,
29941,
26319,
29889,
29871,
29906,
29900,
29900,
29953,
29892,
1716,
310,
607,
526,
11039,
630,
491,
3407,
29889,
13,
29961,
29900,
29900,
29900,
29946,
29962,
450,
2198,
297,
7316,
1104,
1078,
304,
3038,
9257,
3519,
322,
5541,
2187,
393,
526,
13674,
724,
398,
29899,
9021,
322,
7199,
895,
263,
2362,
284,
15795,
18254,
374,
296,
1650,
322,
385,
1425,
29890,
29933,
29941,
14172,
392,
29889,
13,
29961,
29900,
29900,
29900,
29945,
29962,
29871,
29906,
29889,
16585,
310,
278,
512,
7316,
13,
29961,
29900,
29900,
29900,
29953,
29962,
12968,
715,
332,
666,
327,
296,
9101,
5957,
5412,
28602,
1907,
363,
7405,
1218,
4688,
22950,
310,
5199,
5849,
408,
1532,
408,
363,
266,
1572,
412,
329,
293,
1006,
7316,
297,
3196,
17135,
5922,
29892,
1316,
408,
652,
370,
10778,
286,
514,
277,
375,
322,
4815,
26803,
29915,
29879,
17135,
29889,
1152,
1342,
29892,
278,
671,
310,
1663,
352,
262,
29899,
5498,
3277,
29871,
30187,
29899,
3729,
29879,
10723,
515,
5199,
7232,
719,
8927,
20805,
9101,
313,
29882,
2890,
29907,
29879,
29897,
723,
5957,
263,
13426,
20414,
975,
1857,
3038,
29220,
27580,
28648,
393,
3667,
675,
9101,
515,
1016,
272,
7243,
1037,
2129,
29889,
15447,
3038,
29220,
27580,
7539,
1860,
363,
652,
370,
10778,
286,
514,
277,
375,
29892,
607,
3667,
675,
9101,
515,
1016,
272,
7243,
1037,
2129,
29892,
526,
9078,
491,
278,
19494,
537,
310,
1880,
11029,
338,
1026,
9101,
4312,
363,
1301,
24389,
29889,
19413,
29220,
27580,
363,
263,
2323,
5167,
306,
652,
370,
7492,
16500,
6858,
263,
1301,
24389,
310,
14235,
29871,
29947,
30122,
29896,
29900,
29947,
7243,
1037,
2454,
338,
1026,
9101,
313,
2713,
2754,
307,
634,
394,
1696,
29871,
29906,
29900,
29900,
29900,
29892,
405,
2201,
29880,
435,
3436,
29871,
29941,
29946,
29941,
29901,
29906,
29941,
29900,
29899,
29906,
29941,
29947,
29936,
1383,
2754,
307,
634,
394,
1696,
29871,
29906,
29900,
29900,
29896,
29874,
29892,
6407,
29124,
2538,
315,
1915,
2796,
8415,
262,
324,
4737,
370,
29871,
29896,
29945,
29901,
29906,
29946,
29896,
29899,
29906,
29953,
29946,
29936,
1383,
2754,
307,
634,
394,
1696,
29871,
29906,
29900,
29900,
29896,
29892,
4908,
20795,
8237,
29871,
29941,
29906,
29906,
29901,
29947,
29953,
29896,
467,
1094,
1316,
29892,
472,
3203,
1023,
9045,
29891,
1016,
272,
1638,
550,
526,
3734,
304,
4017,
8002,
338,
1026,
9101,
363,
263,
9150,
1301,
24389,
29889,
13,
29961,
29900,
29900,
29900,
29955,
29962,
28735,
719,
8927,
20805,
313,
2890,
29897,
9101,
4550,
2755,
263,
13988,
1904,
1788,
363,
278,
22522,
310,
7208,
12903,
14407,
715,
332,
666,
327,
296,
3038,
4768,
3002,
322,
1422,
11685,
2629,
278,
4688,
7232,
719,
29877,
29892,
408,
1532,
408,
13138,
28602,
1907,
363,
2531,
7492,
11525,
2785,
310,
286,
4850,
1338,
322,
1121,
424,
12128,
29892,
16083,
322,
18032,
3631,
8324,
29889,
16478,
29892,
8210,
410,
29880,
9633,
362,
322,
1422,
11685,
310,
17956,
9101,
508,
19998,
367,
1304,
304,
5706,
385,
443,
29044,
2752,
310,
9101,
480,
1573,
304,
1301,
24389,
362,
363,
14502,
310,
10267,
2129,
393,
1121,
515,
3038,
18658,
470,
270,
952,
2220,
29889,
5901,
715,
332,
666,
327,
296,
9101,
322,
3038,
3454,
3704,
4688,
19269,
321,
312,
397,
837,
29899,
4561,
313,
29923,
7390,
29897,
9101,
408,
5439,
297,
4623,
4121,
296,
8427,
399,
29949,
29871,
29929,
29929,
29914,
29945,
29941,
29900,
29906,
29896,
29892,
297,
325,
4243,
470,
297,
13901,
307,
10723,
18340,
29924,
29914,
1022,
747,
4230,
29892,
297,
325,
4243,
470,
297,
13901,
307,
10723,
19269,
321,
312,
397,
837,
29892,
1903,
536,
616,
22593,
9101,
313,
11787,
9101,
511,
1935,
271,
542,
5666,
262,
4125,
9101,
313,
11206,
9101,
511,
322,
715,
332,
666,
327,
296,
9101,
10723,
491,
28262,
8349,
7268,
362,
470
] | 1,024 |
CommonCrawl
| -0.205743 | 1.646235 | 0.926318 | 1.233002 |
by nuclear transfer will share some or all of these properties and applications. International Patent Application WO 97/32033 and U.S. Pat. No. 5,453,357 describe pluripotent cells including cells from species other than rodents. Human ES cells have been described in International Patent Application WO 00/27995, and in U.S. Pat. No. 6,200,806, and human EG cells have been described in International Patent Application WO 98/43679.
[0008] The biochemical mechanisms regulating ES cell pluripotency and differentiation are very poorly understood. However, the limited empirical data available (and much anecdotal evidence) suggests that the continued maintenance of pluripotent ES cells under in vitro culture conditions is dependent upon the presence of cytokines and growth factors present in the extracellular milieu.
[0009] While human ESCs offer a source of starting material from which to develop substantial quantities of high quality differentiated cells for human cell therapies; these cells must be obtained and/or cultured in conditions that are compatible with the expected regulatory guidelines governing clinical safety and efficacy. Such guidelines likely will require the use of a chemically defined media. The development of such chemically defined/GMP standard conditions is necessary to facilitate the use of hESCs and cells derived from hESCs for therapeutic purposes in humans.
[0010] In addition, the eventual application of hESC based cell replacement therapies will require the development of methods that enable large scale culture and differentiation conditions that are compliant with regulatory guidelines. While several groups have reported simplified growth conditions for hESCs, there are substantial limitations with these studies. To date, however, the successful isolation, long-term clonal maintenance, genetic manipulation and germ line transmission of pluripotent cells has generally been difficult.
[0011] Most of the cell culture conditions for stem cells still contain serum replacer (KSR) in the media (Xu et al., 2005 Stem Cells, 23:315-323; Xu et al., 2005 Nature Methods, 2:185-189; Beattie et al., 2005 Stem Cells, 23:489-495; Amit et al., 2004 Biol. Reprod., 70:837-845; James et al., 2005 Development, 132:1279-1282). KSR contains a crude fraction of bovine serum albumin (BSA) rather than a highly purified source. Others have only performed short-term studies, and therefore it is not clear if their conditions would enable the maintenance of pluripotency over extended periods (Sato et al., (2004) Nature Med., 10:55-63; U.S. Patent Publication Nos. 2006/0030042 and 2005/0233446). Others have shown long-term maintenance of pluripotency in a chemically defined media with FGF2, activin A, and insulin, but the cells were grown on plates that were coated with human serum, which was "washed off" before plating of cells (Vallier et al., 2005 J Cell Sci., 118(Pt 19):4495-509). While FGF2 has been a component of all these media, it is not clear if it is an absolute necessity, particularly as in some formulations it is necessary to use it at a high concentration (up to 100 ng/ml, Xu et al., 2005 Nature Methods, 2:185-189).
[0012] Furthermore, all of these groups have either included insulin in their media at μg/ml levels, or have insulin present due to the use of KSR. Insulin is typically considered to function in glucose metabolism and "cell survival" signaling via binding to the insulin receptor. At levels above physiological concentrations, however, insulin can also bind to the IGF1 receptor with
|
[
491,
20346,
6782,
674,
6232,
777,
470,
599,
310,
1438,
4426,
322,
8324,
29889,
4623,
4121,
296,
8427,
399,
29949,
29871,
29929,
29955,
29914,
29941,
29906,
29900,
29941,
29941,
322,
501,
29889,
29903,
29889,
4121,
29889,
1939,
29889,
29871,
29945,
29892,
29946,
29945,
29941,
29892,
29941,
29945,
29955,
8453,
715,
332,
666,
327,
296,
9101,
3704,
9101,
515,
6606,
916,
1135,
15382,
1237,
29889,
12968,
17956,
9101,
505,
1063,
5439,
297,
4623,
4121,
296,
8427,
399,
29949,
29871,
29900,
29900,
29914,
29906,
29955,
29929,
29929,
29945,
29892,
322,
297,
501,
29889,
29903,
29889,
4121,
29889,
1939,
29889,
29871,
29953,
29892,
29906,
29900,
29900,
29892,
29947,
29900,
29953,
29892,
322,
5199,
382,
29954,
9101,
505,
1063,
5439,
297,
4623,
4121,
296,
8427,
399,
29949,
29871,
29929,
29947,
29914,
29946,
29941,
29953,
29955,
29929,
29889,
13,
29961,
29900,
29900,
29900,
29947,
29962,
450,
17799,
14969,
936,
7208,
12903,
1072,
18099,
17956,
3038,
715,
332,
666,
327,
3819,
322,
1422,
11685,
526,
1407,
6460,
368,
11098,
29889,
2398,
29892,
278,
9078,
29190,
936,
848,
3625,
313,
392,
1568,
385,
687,
6333,
284,
10757,
29897,
14661,
393,
278,
7572,
25413,
310,
715,
332,
666,
327,
296,
17956,
9101,
1090,
297,
13901,
307,
9257,
5855,
338,
14278,
2501,
278,
10122,
310,
5094,
17082,
1475,
322,
14321,
13879,
2198,
297,
278,
1294,
945,
514,
1070,
22591,
29889,
13,
29961,
29900,
29900,
29900,
29929,
29962,
5806,
5199,
382,
7187,
29879,
5957,
263,
2752,
310,
6257,
5518,
515,
607,
304,
2693,
23228,
26855,
310,
1880,
11029,
17473,
630,
9101,
363,
5199,
3038,
29220,
481,
583,
29936,
1438,
9101,
1818,
367,
7625,
322,
29914,
272,
4185,
2955,
297,
5855,
393,
526,
15878,
411,
278,
3806,
24378,
7606,
1410,
10652,
1475,
14765,
1076,
24899,
936,
15332,
322,
6366,
4135,
29889,
10506,
1410,
10652,
1475,
5517,
674,
1996,
278,
671,
310,
263,
8950,
1711,
3342,
5745,
29889,
450,
5849,
310,
1316,
8950,
1711,
3342,
29914,
29954,
3580,
3918,
5855,
338,
5181,
304,
16089,
10388,
278,
671,
310,
298,
2890,
29907,
29879,
322,
9101,
10723,
515,
298,
2890,
29907,
29879,
363,
266,
1572,
412,
329,
293,
11976,
297,
25618,
29889,
13,
29961,
29900,
29900,
29896,
29900,
29962,
512,
6124,
29892,
278,
1741,
950,
2280,
310,
298,
2890,
29907,
2729,
3038,
16920,
29220,
481,
583,
674,
1996,
278,
5849,
310,
3519,
393,
9025,
2919,
6287,
9257,
322,
1422,
11685,
5855,
393,
526,
752,
492,
424,
411,
24378,
7606,
1410,
10652,
1475,
29889,
5806,
3196,
6471,
505,
8967,
20875,
14321,
5855,
363,
298,
2890,
29907,
29879,
29892,
727,
526,
23228,
27028,
411,
1438,
11898,
29889,
1763,
2635,
29892,
3138,
29892,
278,
9150,
11695,
362,
29892,
1472,
29899,
8489,
1067,
7177,
25413,
29892,
2531,
7492,
11525,
2785,
322,
22593,
1196,
22713,
310,
715,
332,
666,
327,
296,
9101,
756,
6892,
1063,
5189,
29889,
13,
29961,
29900,
29900,
29896,
29896,
29962,
7849,
310,
278,
3038,
9257,
5855,
363,
20805,
9101,
1603,
1712,
724,
398,
6497,
2265,
313,
29968,
14098,
29897,
297,
278,
5745,
313,
29990,
29884,
634,
394,
1696,
29871,
29906,
29900,
29900,
29945,
624,
331,
315,
10071,
29892,
29871,
29906,
29941,
29901,
29941,
29896,
29945,
29899,
29941,
29906,
29941,
29936,
1060,
29884,
634,
394,
1696,
29871,
29906,
29900,
29900,
29945,
17677,
8108,
29879,
29892,
29871,
29906,
29901,
29896,
29947,
29945,
29899,
29896,
29947,
29929,
29936,
1522,
1131,
347,
634,
394,
1696,
29871,
29906,
29900,
29900,
29945,
624,
331,
315,
10071,
29892,
29871,
29906,
29941,
29901,
29946,
29947,
29929,
29899,
29946,
29929,
29945,
29936,
1913,
277,
634,
394,
1696,
29871,
29906,
29900,
29900,
29946,
3457,
324,
29889,
830,
10633,
1696,
29871,
29955,
29900,
29901,
29947,
29941,
29955,
29899,
29947,
29946,
29945,
29936,
5011,
634,
394,
1696,
29871,
29906,
29900,
29900,
29945,
14650,
29892,
29871,
29896,
29941,
29906,
29901,
29896,
29906,
29955,
29929,
29899,
29896,
29906,
29947,
29906,
467,
476,
14098,
3743,
263,
7618,
311,
15958,
310,
289,
586,
457,
724,
398,
3769,
262,
313,
29933,
8132,
29897,
3265,
1135,
263,
10712,
3708,
2164,
2752,
29889,
438,
3341,
505,
871,
8560,
3273,
29899,
8489,
11898,
29892,
322,
5480,
372,
338,
451,
2821,
565,
1009,
5855,
723,
9025,
278,
25413,
310,
715,
332,
666,
327,
3819,
975,
10410,
23704,
313,
29903,
1219,
634,
394,
1696,
313,
29906,
29900,
29900,
29946,
29897,
17677,
3436,
1696,
29871,
29896,
29900,
29901,
29945,
29945,
29899,
29953,
29941,
29936,
501,
29889,
29903,
29889,
4121,
296,
5236,
362,
405,
359,
29889,
29871,
29906,
29900,
29900,
29953,
29914,
29900,
29900,
29941,
29900,
29900,
29946,
29906,
322,
29871,
29906,
29900,
29900,
29945,
29914,
29900,
29906,
29941,
29941,
29946,
29946,
29953,
467,
438,
3341,
505,
4318,
1472,
29899,
8489,
25413,
310,
715,
332,
666,
327,
3819,
297,
263,
8950,
1711,
3342,
5745,
411,
383,
29954,
29943,
29906,
29892,
5039,
262,
319,
29892,
322,
1663,
352,
262,
29892,
541,
278,
9101,
892,
21633,
373,
715,
1078,
393,
892,
1302,
630,
411,
5199,
724,
398,
29892,
607,
471,
376,
29893,
25936,
1283,
29908,
1434,
715,
1218,
310,
9101,
313,
1440,
4926,
634,
394,
1696,
29871,
29906,
29900,
29900,
29945,
435,
19413,
5636,
1696,
29871,
29896,
29896,
29947,
29898,
29925,
29873,
29871,
29896,
29929,
1125,
29946,
29946,
29929,
29945,
29899,
29945,
29900,
29929,
467,
5806,
383,
29954,
29943,
29906,
756,
1063,
263,
4163,
310,
599,
1438,
5745,
29892,
372,
338,
451,
2821,
565,
372,
338,
385,
8380,
24316,
29892,
10734,
408,
297,
777,
883,
8250,
372,
338,
5181,
304,
671,
372,
472,
263,
1880,
26702,
313,
786,
304,
29871,
29896,
29900,
29900,
8736,
29914,
828,
29892,
1060,
29884,
634,
394,
1696,
29871,
29906,
29900,
29900,
29945,
17677,
8108,
29879,
29892,
29871,
29906,
29901,
29896,
29947,
29945,
29899,
29896,
29947,
29929,
467,
13,
29961,
29900,
29900,
29896,
29906,
29962,
16478,
29892,
599,
310,
1438,
6471,
505,
2845,
5134,
1663,
352,
262,
297,
1009,
5745,
472,
29871,
30167,
29887,
29914,
828,
11174,
29892,
470,
505,
1663,
352,
262,
2198,
2861,
304,
278,
671,
310,
476,
14098,
29889,
13377,
352,
262,
338,
12234,
5545,
304,
740,
297,
3144,
1682,
852,
1539,
19388,
1608,
322,
376,
3729,
10503,
2561,
29908,
7182,
292,
3025,
9956,
304,
278,
1663,
352,
262,
337,
14268,
29889,
2180,
11174,
2038,
4824,
29875,
5996,
14953,
800,
29892,
3138,
29892,
1663,
352,
262,
508,
884,
7868,
304,
278,
306,
29954,
29943,
29896,
337,
14268,
411
] | 1,024 |
CommonCrawl
| -0.413259 | 1.641044 | 0.903006 | 0.747289 |
a lower efficiency and confer classical growth factor activity through the P13 Kinase/AKT pathway. The presence/requirement for such high levels of insulin (μg/ml levels) in KSR or these other media conditions suggests that the major activity is elicited via binding to the IGF1 receptor, which is expressed by hESCs (Sperger et al., 2003 PNAS, 100(23):13350-13355). Others have noted the expression of a full complement of IGF1R and intracellular signaling pathway members in hESCs, which is likely to signify the functional activity of this pathway (Miura et al., 2004 Aging Cell, 3:333-343). Insulin or IGF1 may elicit a major signal required for the self-renewal of hESCs, as is suggested by the fact that all conditions developed thus far for the culture of hESC contain either insulin, insulin provided by KSR, or IGF1 provided by serum. In support of this concept, it has been shown that if P13 Kinase is inhibited in hESC cultures, the cells differentiate (D'Amour et al., 2005 Nat. Biotechnol., 23(12):1534-41; McLean et al., 2007 Stem Cells 25:29-38).
[0013] A recent publication outlines a humanized, defined media for hESCs (Ludwig et al., Nature Biotechnology, published online Jan. 1, 2006, doi:10.1038/nbt1177). This recent formulation, however, includes several factors that are suggested to influence the proliferation of hESCs, including FGF2, TGFβ, LiCl, γ-aminobutyric acid and pipecolic acid. It is noted that this recently defined cell culture medium also contains insulin.
[0014] The EGF growth factor family has at least 14 members, including, but not limited to, EGF, TGFβ, heparin binding-EGF (hb-EGF), neuregulin-β (also named heregulin-β (HRG-β), glial growth factor and others), HRG-α, amphiregulin, betacellulin, and epiregulin. All these growth factors contain an EGF domain and are typically first expressed as transmembrane proteins that are processed by metalloproteinase (specifically, ADAM) proteins to generate soluble ectodomain growth factors. EGF family members interact with either homo- or hetero-dimers of the ErbB1, 2, 3 and 4 cell surface receptors with different affinities (Jones et al., FEBS Lett, 1999, 447:227-231). EGF, TGFα and hbEGF bind ErbB1/1 (EGFR) homodimers and ErbB1/2 heterodimers at high affinity (1-100 nM range), whereas HRG-β binds ErbB3 and ErbB4 at very high affinity (<1 nM range). Activated ErbB receptors signal through the PI3 Kinase/AKT pathway and also the MAPK pathway. ErbB2 and ErbB3 are amongst the most highly expressed growth factor receptors in hESCs (Sperger et al., 2003 PNAS, 100(23):13350-13355) and HRG-β has been shown previously to support the expansion of mouse primordial germ cells (Toyoda-Ohno et al., 1999 Dev. Biol., 215(2):399-406). Furthermore, overexpression and subsequent inappropriate activation of ErbB2 is associated with tumorigenesis (Neve et al., 2001 Ann. Oncol., 12 Suppl 1:S9-13; Zhou & Hung, 2003 Semin. Oncol., 30(5 Suppl
|
[
263,
5224,
19201,
322,
21388,
14499,
14321,
7329,
6354,
1549,
278,
349,
29896,
29941,
20311,
559,
29914,
22311,
29911,
2224,
1582,
29889,
450,
10122,
29914,
12277,
358,
363,
1316,
1880,
11174,
310,
1663,
352,
262,
313,
30167,
29887,
29914,
828,
11174,
29897,
297,
476,
14098,
470,
1438,
916,
5745,
5855,
14661,
393,
278,
4655,
6354,
338,
560,
293,
1573,
3025,
9956,
304,
278,
306,
29954,
29943,
29896,
337,
14268,
29892,
607,
338,
13384,
491,
298,
2890,
29907,
29879,
313,
29903,
546,
914,
634,
394,
1696,
29871,
29906,
29900,
29900,
29941,
349,
29940,
3289,
29892,
29871,
29896,
29900,
29900,
29898,
29906,
29941,
1125,
29896,
29941,
29941,
29945,
29900,
29899,
29896,
29941,
29941,
29945,
29945,
467,
438,
3341,
505,
11682,
278,
4603,
310,
263,
2989,
19595,
310,
306,
29954,
29943,
29896,
29934,
322,
938,
945,
514,
1070,
7182,
292,
2224,
1582,
5144,
297,
298,
2890,
29907,
29879,
29892,
607,
338,
5517,
304,
1804,
1598,
278,
13303,
6354,
310,
445,
2224,
1582,
313,
29924,
29875,
2002,
634,
394,
1696,
29871,
29906,
29900,
29900,
29946,
319,
3460,
19413,
29892,
29871,
29941,
29901,
29941,
29941,
29941,
29899,
29941,
29946,
29941,
467,
13377,
352,
262,
470,
306,
29954,
29943,
29896,
1122,
560,
293,
277,
263,
4655,
7182,
3734,
363,
278,
1583,
29899,
1267,
809,
284,
310,
298,
2890,
29907,
29879,
29892,
408,
338,
7829,
491,
278,
2114,
393,
599,
5855,
8906,
4550,
2215,
363,
278,
9257,
310,
298,
2890,
29907,
1712,
2845,
1663,
352,
262,
29892,
1663,
352,
262,
4944,
491,
476,
14098,
29892,
470,
306,
29954,
29943,
29896,
4944,
491,
724,
398,
29889,
512,
2304,
310,
445,
6964,
29892,
372,
756,
1063,
4318,
393,
565,
349,
29896,
29941,
20311,
559,
338,
297,
6335,
1573,
297,
298,
2890,
29907,
4185,
1973,
29892,
278,
9101,
17473,
403,
313,
29928,
29915,
6833,
473,
634,
394,
1696,
29871,
29906,
29900,
29900,
29945,
19259,
29889,
3457,
866,
3049,
324,
1696,
29871,
29906,
29941,
29898,
29896,
29906,
1125,
29896,
29945,
29941,
29946,
29899,
29946,
29896,
29936,
4052,
3226,
273,
634,
394,
1696,
29871,
29906,
29900,
29900,
29955,
624,
331,
315,
10071,
29871,
29906,
29945,
29901,
29906,
29929,
29899,
29941,
29947,
467,
13,
29961,
29900,
29900,
29896,
29941,
29962,
319,
7786,
17745,
714,
9012,
263,
5199,
1891,
29892,
3342,
5745,
363,
298,
2890,
29907,
29879,
313,
29931,
566,
9192,
634,
394,
1696,
17677,
3457,
866,
3049,
3002,
29892,
6369,
7395,
2627,
29889,
29871,
29896,
29892,
29871,
29906,
29900,
29900,
29953,
29892,
13102,
29901,
29896,
29900,
29889,
29896,
29900,
29941,
29947,
29914,
29876,
3116,
29896,
29896,
29955,
29955,
467,
910,
7786,
883,
2785,
29892,
3138,
29892,
7805,
3196,
13879,
393,
526,
7829,
304,
9949,
278,
410,
29880,
9633,
362,
310,
298,
2890,
29907,
29879,
29892,
3704,
383,
29954,
29943,
29906,
29892,
323,
29954,
29943,
30187,
29892,
2718,
6821,
29892,
29871,
30197,
29899,
9103,
711,
329,
29891,
2200,
22193,
322,
14282,
1054,
293,
22193,
29889,
739,
338,
11682,
393,
445,
10325,
3342,
3038,
9257,
18350,
884,
3743,
1663,
352,
262,
29889,
13,
29961,
29900,
29900,
29896,
29946,
29962,
450,
382,
29954,
29943,
14321,
7329,
3942,
756,
472,
3203,
29871,
29896,
29946,
5144,
29892,
3704,
29892,
541,
451,
9078,
304,
29892,
382,
29954,
29943,
29892,
323,
29954,
29943,
30187,
29892,
540,
862,
262,
9956,
29899,
11787,
29943,
313,
29882,
29890,
29899,
11787,
29943,
511,
452,
545,
29887,
352,
262,
29899,
30187,
313,
15189,
4257,
902,
387,
352,
262,
29899,
30187,
313,
20938,
29954,
29899,
30187,
511,
5857,
284,
14321,
7329,
322,
4045,
511,
379,
29934,
29954,
29899,
30110,
29892,
626,
561,
533,
29887,
352,
262,
29892,
1010,
562,
514,
352,
262,
29892,
322,
9358,
533,
29887,
352,
262,
29889,
2178,
1438,
14321,
13879,
1712,
385,
382,
29954,
29943,
5354,
322,
526,
12234,
937,
13384,
408,
18750,
1590,
10800,
3279,
1144,
393,
526,
19356,
491,
11915,
417,
14676,
262,
559,
313,
14940,
635,
29892,
11033,
5194,
29897,
3279,
1144,
304,
5706,
899,
431,
280,
321,
312,
397,
290,
475,
14321,
13879,
29889,
382,
29954,
29943,
3942,
5144,
16254,
411,
2845,
3632,
29877,
29899,
470,
25745,
29877,
29899,
6229,
414,
310,
278,
1425,
29890,
29933,
29896,
29892,
29871,
29906,
29892,
29871,
29941,
322,
29871,
29946,
3038,
7101,
337,
1547,
943,
411,
1422,
2756,
262,
1907,
313,
29967,
2873,
634,
394,
1696,
383,
29923,
9851,
365,
1803,
29892,
29871,
29896,
29929,
29929,
29929,
29892,
29871,
29946,
29946,
29955,
29901,
29906,
29906,
29955,
29899,
29906,
29941,
29896,
467,
382,
29954,
29943,
29892,
323,
29954,
29943,
30110,
322,
298,
29890,
11787,
29943,
7868,
1425,
29890,
29933,
29896,
29914,
29896,
313,
11787,
15860,
29897,
3632,
397,
326,
414,
322,
1425,
29890,
29933,
29896,
29914,
29906,
25745,
397,
326,
414,
472,
1880,
2756,
13593,
313,
29896,
29899,
29896,
29900,
29900,
302,
29924,
3464,
511,
13452,
379,
29934,
29954,
29899,
30187,
7868,
29879,
1425,
29890,
29933,
29941,
322,
1425,
29890,
29933,
29946,
472,
1407,
1880,
2756,
13593,
313,
29966,
29896,
302,
29924,
3464,
467,
21775,
630,
1425,
29890,
29933,
337,
1547,
943,
7182,
1549,
278,
349,
29902,
29941,
20311,
559,
29914,
22311,
29911,
2224,
1582,
322,
884,
278,
341,
3301,
29968,
2224,
1582,
29889,
1425,
29890,
29933,
29906,
322,
1425,
29890,
29933,
29941,
526,
22611,
278,
1556,
10712,
13384,
14321,
7329,
337,
1547,
943,
297,
298,
2890,
29907,
29879,
313,
29903,
546,
914,
634,
394,
1696,
29871,
29906,
29900,
29900,
29941,
349,
29940,
3289,
29892,
29871,
29896,
29900,
29900,
29898,
29906,
29941,
1125,
29896,
29941,
29941,
29945,
29900,
29899,
29896,
29941,
29941,
29945,
29945,
29897,
322,
379,
29934,
29954,
29899,
30187,
756,
1063,
4318,
9251,
304,
2304,
278,
13184,
310,
9495,
1903,
536,
616,
22593,
9101,
313,
1762,
29891,
8887,
29899,
9048,
1217,
634,
394,
1696,
29871,
29896,
29929,
29929,
29929,
9481,
29889,
3457,
324,
1696,
29871,
29906,
29896,
29945,
29898,
29906,
1125,
29941,
29929,
29929,
29899,
29946,
29900,
29953,
467,
16478,
29892,
975,
17471,
322,
15352,
297,
932,
6649,
403,
26229,
310,
1425,
29890,
29933,
29906,
338,
6942,
411,
21622,
272,
2101,
6656,
313,
8139,
345,
634,
394,
1696,
29871,
29906,
29900,
29900,
29896,
8081,
29889,
1551,
1054,
1696,
29871,
29896,
29906,
9179,
29880,
29871,
29896,
29901,
29903,
29929,
29899,
29896,
29941,
29936,
796,
10774,
669,
14198,
29892,
29871,
29906,
29900,
29900,
29941,
9444,
262,
29889,
1551,
1054,
1696,
29871,
29941,
29900,
29898,
29945,
9179,
29880
] | 1,024 |
CommonCrawl
| -0.404768 | 1.755249 | 0.763134 | 0.469017 |
16):38-48; Yarden, 2001 Oncology, 61 Suppl 2:1-13). Human ErbB2 (Chromosome 17q), and ErbB3 (Chromosome 12q) are present on chromosomes that have been observed to accumulate as trisomies in some hESCs (Draper et al., 2004 Nat. Biotechnol., 22(1):53-4; Cowan et al., 2004 N Engl. J. Med., 350(13):1353-6; Brimble et al., 2004 Stem Cells Dev., 13(6):585-97; Maitra et al., 2005 Nat. Genet. 37(10):1099-103; Mitalipova et al., 2005 Nat. Biotechnol. 23(1): 19-20; Draper et al., 2004 Stem Cells Dev., 13(4):325-36; Ludwig et al., Nature Biotechnology, published online Jan. 1, 2006, doi:10.1038/nbt1177).
[0015] ErbB2 and ErbB3 (Brown et al., 2004 Biol. Reprod., 71:2003-2011; Salas-Vidal & Lomeli, 2004, Dev Biol., 265:75-89) are expressed in the mouse blastocyst, although not specifically restricted to the inner cell mass (ICM), and ErbB1, EGF and TGFβ are expressed in the human blastocyst (Chia et al., 1995 Development, 1221(2):299-307). HB-EGF has proliferative effects in human IVF blastocyst culture (Martin et al., 1998 Hum. Reprod., 13(6):1645-52; Sargent et al., 1998 Hum. Reprod. 13 Suppl 4:239-48), and modest additional effects on mouse ES cells grown in 15% serum (Heo et al., 2005, Am. J. Phy. Cell Physiol., in press). Pre- and early post-implantation development does not appear to be affected in ErbB2-/-, ErbB3-/-, Neuregulinl-/- (Britsch et al., 1998 Genes Dev., 12:1825-36), ADAM17-/- (Peschon, et al., 1998 Science, 282: 1281-1284) and ADAM19-/- (Horiuchi, 2005 Dev. Biol., 283(2):459-71) null embryos. Therefore, the importance of signaling through the ErbB receptor family in hESCs is, up to now, unclear.
[0016] Neuregulin-1 (NRG1) is a large gene that exhibits multiple splicing and protein processing variants. This generates a large number of protein isoforms, which are referred to herein collectively as neuregulin. Neuregulin is predominantly expressed as a cell surface transmembrane protein. The extracellular region contains an immunoglobulin-like domain, a carbohydrate modified region and the EGF domain. NRG1 expression isoforms have been reviewed previously (Falls, 2003 Exp. Cell Res., 284:14-30). The cell membrane metalloproteases ADAM17 and ADAM19 have been shown to process the transmembrane form(s) of neuregulin-1 to soluble neuregulin/heregulin. HRG-α and -β are the cleaved ectodomains of neuregulin, containing the EGF and other domains. As the EGF domain is responsible for binding and activation of the ErbB receptors, a recom
|
[
29871,
29896,
29953,
1125,
29941,
29947,
29899,
29946,
29947,
29936,
612,
7879,
29892,
29871,
29906,
29900,
29900,
29896,
1551,
1054,
6933,
29892,
29871,
29953,
29896,
9179,
29880,
29871,
29906,
29901,
29896,
29899,
29896,
29941,
467,
12968,
1425,
29890,
29933,
29906,
313,
1451,
456,
359,
608,
29871,
29896,
29955,
29939,
511,
322,
1425,
29890,
29933,
29941,
313,
1451,
456,
359,
608,
29871,
29896,
29906,
29939,
29897,
526,
2198,
373,
25173,
359,
290,
267,
393,
505,
1063,
8900,
304,
18414,
5987,
408,
534,
275,
290,
583,
297,
777,
298,
2890,
29907,
29879,
313,
24515,
546,
634,
394,
1696,
29871,
29906,
29900,
29900,
29946,
19259,
29889,
3457,
866,
3049,
324,
1696,
29871,
29906,
29906,
29898,
29896,
1125,
29945,
29941,
29899,
29946,
29936,
21638,
273,
634,
394,
1696,
29871,
29906,
29900,
29900,
29946,
405,
2201,
29880,
29889,
435,
29889,
3436,
1696,
29871,
29941,
29945,
29900,
29898,
29896,
29941,
1125,
29896,
29941,
29945,
29941,
29899,
29953,
29936,
1771,
326,
569,
634,
394,
1696,
29871,
29906,
29900,
29900,
29946,
624,
331,
315,
10071,
9481,
1696,
29871,
29896,
29941,
29898,
29953,
1125,
29945,
29947,
29945,
29899,
29929,
29955,
29936,
341,
1249,
336,
634,
394,
1696,
29871,
29906,
29900,
29900,
29945,
19259,
29889,
5739,
300,
29889,
29871,
29941,
29955,
29898,
29896,
29900,
1125,
29896,
29900,
29929,
29929,
29899,
29896,
29900,
29941,
29936,
341,
2410,
666,
4273,
634,
394,
1696,
29871,
29906,
29900,
29900,
29945,
19259,
29889,
3457,
866,
3049,
324,
29889,
29871,
29906,
29941,
29898,
29896,
1125,
29871,
29896,
29929,
29899,
29906,
29900,
29936,
16322,
546,
634,
394,
1696,
29871,
29906,
29900,
29900,
29946,
624,
331,
315,
10071,
9481,
1696,
29871,
29896,
29941,
29898,
29946,
1125,
29941,
29906,
29945,
29899,
29941,
29953,
29936,
14298,
634,
394,
1696,
17677,
3457,
866,
3049,
3002,
29892,
6369,
7395,
2627,
29889,
29871,
29896,
29892,
29871,
29906,
29900,
29900,
29953,
29892,
13102,
29901,
29896,
29900,
29889,
29896,
29900,
29941,
29947,
29914,
29876,
3116,
29896,
29896,
29955,
29955,
467,
13,
29961,
29900,
29900,
29896,
29945,
29962,
1425,
29890,
29933,
29906,
322,
1425,
29890,
29933,
29941,
313,
29933,
4708,
634,
394,
1696,
29871,
29906,
29900,
29900,
29946,
3457,
324,
29889,
830,
10633,
1696,
29871,
29955,
29896,
29901,
29906,
29900,
29900,
29941,
29899,
29906,
29900,
29896,
29896,
29936,
3956,
294,
29899,
29963,
23670,
669,
365,
290,
5037,
29892,
29871,
29906,
29900,
29900,
29946,
29892,
9481,
3457,
324,
1696,
29871,
29906,
29953,
29945,
29901,
29955,
29945,
29899,
29947,
29929,
29897,
526,
13384,
297,
278,
9495,
1999,
24186,
29883,
858,
29892,
5998,
451,
10816,
22078,
304,
278,
6426,
3038,
4158,
313,
2965,
29924,
511,
322,
1425,
29890,
29933,
29896,
29892,
382,
29954,
29943,
322,
323,
29954,
29943,
30187,
526,
13384,
297,
278,
5199,
1999,
24186,
29883,
858,
313,
1451,
423,
634,
394,
1696,
29871,
29896,
29929,
29929,
29945,
14650,
29892,
29871,
29896,
29906,
29906,
29896,
29898,
29906,
1125,
29906,
29929,
29929,
29899,
29941,
29900,
29955,
467,
379,
29933,
29899,
11787,
29943,
756,
410,
29880,
9633,
1230,
9545,
297,
5199,
6599,
29943,
1999,
24186,
29883,
858,
9257,
313,
22628,
634,
394,
1696,
29871,
29896,
29929,
29929,
29947,
13863,
29889,
830,
10633,
1696,
29871,
29896,
29941,
29898,
29953,
1125,
29896,
29953,
29946,
29945,
29899,
29945,
29906,
29936,
317,
23641,
634,
394,
1696,
29871,
29896,
29929,
29929,
29947,
13863,
29889,
830,
10633,
29889,
29871,
29896,
29941,
9179,
29880,
29871,
29946,
29901,
29906,
29941,
29929,
29899,
29946,
29947,
511,
322,
878,
342,
5684,
9545,
373,
9495,
17956,
9101,
21633,
297,
29871,
29896,
29945,
29995,
724,
398,
313,
3868,
29877,
634,
394,
1696,
29871,
29906,
29900,
29900,
29945,
29892,
1913,
29889,
435,
29889,
1963,
29891,
29889,
19413,
11661,
29875,
324,
1696,
297,
3965,
467,
4721,
29899,
322,
4688,
1400,
29899,
13699,
424,
362,
5849,
947,
451,
2615,
304,
367,
15201,
297,
1425,
29890,
29933,
29906,
29899,
29914,
15767,
1425,
29890,
29933,
29941,
29899,
29914,
15767,
2448,
545,
29887,
352,
262,
29880,
29899,
24028,
313,
29933,
768,
816,
634,
394,
1696,
29871,
29896,
29929,
29929,
29947,
5739,
267,
9481,
1696,
29871,
29896,
29906,
29901,
29896,
29947,
29906,
29945,
29899,
29941,
29953,
511,
11033,
5194,
29896,
29955,
29899,
24028,
313,
29925,
1968,
265,
29892,
634,
394,
1696,
29871,
29896,
29929,
29929,
29947,
9327,
29892,
29871,
29906,
29947,
29906,
29901,
29871,
29896,
29906,
29947,
29896,
29899,
29896,
29906,
29947,
29946,
29897,
322,
11033,
5194,
29896,
29929,
29899,
24028,
313,
29950,
4170,
987,
29875,
29892,
29871,
29906,
29900,
29900,
29945,
9481,
29889,
3457,
324,
1696,
29871,
29906,
29947,
29941,
29898,
29906,
1125,
29946,
29945,
29929,
29899,
29955,
29896,
29897,
1870,
7232,
719,
359,
29889,
7857,
29892,
278,
13500,
310,
7182,
292,
1549,
278,
1425,
29890,
29933,
337,
14268,
3942,
297,
298,
2890,
29907,
29879,
338,
29892,
701,
304,
1286,
29892,
20871,
29889,
13,
29961,
29900,
29900,
29896,
29953,
29962,
2448,
545,
29887,
352,
262,
29899,
29896,
313,
16514,
29954,
29896,
29897,
338,
263,
2919,
18530,
393,
10371,
1169,
2999,
805,
506,
292,
322,
26823,
9068,
29161,
29889,
910,
16785,
263,
2919,
1353,
310,
26823,
338,
29877,
9514,
29892,
607,
526,
12992,
304,
1244,
262,
6314,
3598,
408,
452,
545,
29887,
352,
262,
29889,
2448,
545,
29887,
352,
262,
338,
758,
24130,
10835,
13384,
408,
263,
3038,
7101,
18750,
1590,
10800,
26823,
29889,
450,
1294,
945,
514,
1070,
5120,
3743,
385,
5198,
348,
468,
2127,
352,
262,
29899,
4561,
5354,
29892,
263,
1559,
833,
29882,
2941,
10492,
9120,
5120,
322,
278,
382,
29954,
29943,
5354,
29889,
27759,
29954,
29896,
4603,
338,
29877,
9514,
505,
1063,
9076,
287,
9251,
313,
29943,
4293,
29892,
29871,
29906,
29900,
29900,
29941,
12027,
29889,
19413,
2538,
1696,
29871,
29906,
29947,
29946,
29901,
29896,
29946,
29899,
29941,
29900,
467,
450,
3038,
3813,
10800,
11915,
417,
14676,
2129,
11033,
5194,
29896,
29955,
322,
11033,
5194,
29896,
29929,
505,
1063,
4318,
304,
1889,
278,
18750,
1590,
10800,
883,
29898,
29879,
29897,
310,
452,
545,
29887,
352,
262,
29899,
29896,
304,
899,
431,
280,
452,
545,
29887,
352,
262,
29914,
2276,
387,
352,
262,
29889,
379,
29934,
29954,
29899,
30110,
322,
448,
30187,
526,
278,
4531,
10511,
321,
312,
397,
290,
2708,
310,
452,
545,
29887,
352,
262,
29892,
6943,
278,
382,
29954,
29943,
322,
916,
21904,
29889,
1094,
278,
382,
29954,
29943,
5354,
338,
14040,
363,
9956,
322,
26229,
310,
278,
1425,
29890,
29933,
337,
1547,
943,
29892,
263,
27878
] | 1,024 |
CommonCrawl
| -0.944784 | 1.407442 | 0.420629 | 0.094613 |
binant molecule containing only this domain can exhibit essentially all of the soluble growth factor effects of this protein (Jones et al., 1999 FEBS Lett., 447:227-231). Also, there are processed transmembrane isoforms of neurcgulin that are thought to trigger juxtacrine signaling in adjacent cells via interaction of the EGF domain with ErbB receptors.
[0017] An important development in the progression of hESC research toward maintaining pluripotency in culture will be the elucidation of media and cell culture conditions that are compatible with the expected regulatory guidelines governing clinical safety and efficacy. While the best outcome would be the availability of chemically defined media for hESCs, components that are not chemically defined would be acceptable if they were produced to GMP standard. There is a need, therefore, to identify methods and compositions for the culture and stabilization of a population of pluripotent stem cells that are able to be used for therapeutic purposes, wherein the culture compositions are defined and/or produced to GMP standard.
[0018] The invention relates to compositions comprising a basal salt nutrient solution and an ErbB3 ligand, with the compositions being essentially free of scrum.
[0019] The invention also relates to compositions comprising a basal salt nutrient solution and a means for stimulating ErbB2-directed tyrosine kinase activity in differentiable cells.
[0020] The invention relates to methods of culturing differentiable cells, with the methods comprising plating the differentiable cells on a cell culture surface; providing a basal salt nutrient solution to the differentiable cells and providing a ligand that specifically binds ErbB3.
[0021] The invention relates to methods of culturing differentiable cells, with the methods comprising plating the differentiable cells on a cell culture surface and providing a basal salt nutrient solution to the differentiable cells and a means for stimulating ErbB2-directed tyrosine kinase activity in the differentiable cells.
[0022] The invention also relates to methods of culturing differentiable cells, with the methods comprising providing a digest solution to a layer of differentiable cells that are contained in a culture chamber prior to digestion, where the digestion breaks apart the layer of cells into single cells. After digestion, the single cells are placed into a new tissue culture chamber with a differentiable cell culture solution, wherein the differentiable cell culture solution comprises a basal salt nutrient solution and an ErbB3 ligand. Once cultured, the single differentiable cells are placed in conditions that permit growth and division of the single cells.
[0023] FIG. 1 depicts real time RT-PCR expression analysis of ADAM19, Neuregulin1, and ErbB1-3 in BG01v grown in defined conditions (8 ng/ml FGF2, 100 ng/ml LR-IGF1, 1 ng/ml Activin A). GAPDH and OCT4 control reactions are indicated.
[0024] FIG. 2 depicts the inhibition of proliferation of BG01v cells using AG879. BG01v cells were plated in 6-well trays and exposed to DMSO (A), 50 nM-20 μM AG1478 (B), or 100 mM-20 μM AG879 (C) 24 hours after plating. After 5 days in culture, the cultures were fixed and stained for alkaline phosphatase activity. AG1478 did not appear to affect proliferation at these concentrations (20 μM shown in B), but AG879 substantially slowed cell growth at 5 μM (C).
[0025] FIG. 3 depicts the morphology of BG01v cells cultured in DC-HALF, which is defined culture media containing 10 ng/ml HRG-β, 10 ng/ml Activin A, 200 ng/ml LR-IGF1 and 8 ng/ml FGF2 (A and B), and in defined culture media (DC)
|
[
2109,
424,
13206,
29883,
1297,
6943,
871,
445,
5354,
508,
10371,
277,
13674,
599,
310,
278,
899,
431,
280,
14321,
7329,
9545,
310,
445,
26823,
313,
29967,
2873,
634,
394,
1696,
29871,
29896,
29929,
29929,
29929,
383,
29923,
9851,
365,
1803,
1696,
29871,
29946,
29946,
29955,
29901,
29906,
29906,
29955,
29899,
29906,
29941,
29896,
467,
3115,
29892,
727,
526,
19356,
18750,
1590,
10800,
338,
29877,
9514,
310,
26808,
29883,
29887,
352,
262,
393,
526,
2714,
304,
7135,
3623,
486,
562,
29878,
457,
7182,
292,
297,
20114,
9101,
3025,
14881,
310,
278,
382,
29954,
29943,
5354,
411,
1425,
29890,
29933,
337,
1547,
943,
29889,
13,
29961,
29900,
29900,
29896,
29955,
29962,
530,
4100,
5849,
297,
278,
410,
11476,
310,
298,
2890,
29907,
5925,
11183,
7344,
292,
715,
332,
666,
327,
3819,
297,
9257,
674,
367,
278,
560,
1682,
333,
362,
310,
5745,
322,
3038,
9257,
5855,
393,
526,
15878,
411,
278,
3806,
24378,
7606,
1410,
10652,
1475,
14765,
1076,
24899,
936,
15332,
322,
6366,
4135,
29889,
5806,
278,
1900,
21957,
723,
367,
278,
20847,
3097,
310,
8950,
1711,
3342,
5745,
363,
298,
2890,
29907,
29879,
29892,
7117,
393,
526,
451,
8950,
1711,
3342,
723,
367,
22691,
565,
896,
892,
7371,
304,
402,
3580,
3918,
29889,
1670,
338,
263,
817,
29892,
5480,
29892,
304,
12439,
3519,
322,
5541,
2187,
363,
278,
9257,
322,
16160,
2133,
310,
263,
4665,
310,
715,
332,
666,
327,
296,
20805,
9101,
393,
526,
2221,
304,
367,
1304,
363,
266,
1572,
412,
329,
293,
11976,
29892,
988,
262,
278,
9257,
5541,
2187,
526,
3342,
322,
29914,
272,
7371,
304,
402,
3580,
3918,
29889,
13,
29961,
29900,
29900,
29896,
29947,
29962,
450,
297,
7316,
1104,
1078,
304,
5541,
2187,
7199,
5921,
263,
2362,
284,
15795,
18254,
374,
296,
1650,
322,
385,
1425,
29890,
29933,
29941,
14172,
392,
29892,
411,
278,
5541,
2187,
1641,
13674,
3889,
310,
885,
5848,
29889,
13,
29961,
29900,
29900,
29896,
29929,
29962,
450,
297,
7316,
884,
1104,
1078,
304,
5541,
2187,
7199,
5921,
263,
2362,
284,
15795,
18254,
374,
296,
1650,
322,
263,
2794,
363,
20436,
18099,
1425,
29890,
29933,
29906,
29899,
11851,
287,
7911,
1883,
457,
19015,
559,
6354,
297,
17473,
519,
9101,
29889,
13,
29961,
29900,
29900,
29906,
29900,
29962,
450,
297,
7316,
1104,
1078,
304,
3519,
310,
4185,
3864,
17473,
519,
9101,
29892,
411,
278,
3519,
7199,
5921,
715,
1218,
278,
17473,
519,
9101,
373,
263,
3038,
9257,
7101,
29936,
13138,
263,
2362,
284,
15795,
18254,
374,
296,
1650,
304,
278,
17473,
519,
9101,
322,
13138,
263,
14172,
392,
393,
10816,
7868,
29879,
1425,
29890,
29933,
29941,
29889,
13,
29961,
29900,
29900,
29906,
29896,
29962,
450,
297,
7316,
1104,
1078,
304,
3519,
310,
4185,
3864,
17473,
519,
9101,
29892,
411,
278,
3519,
7199,
5921,
715,
1218,
278,
17473,
519,
9101,
373,
263,
3038,
9257,
7101,
322,
13138,
263,
2362,
284,
15795,
18254,
374,
296,
1650,
304,
278,
17473,
519,
9101,
322,
263,
2794,
363,
20436,
18099,
1425,
29890,
29933,
29906,
29899,
11851,
287,
7911,
1883,
457,
19015,
559,
6354,
297,
278,
17473,
519,
9101,
29889,
13,
29961,
29900,
29900,
29906,
29906,
29962,
450,
297,
7316,
884,
1104,
1078,
304,
3519,
310,
4185,
3864,
17473,
519,
9101,
29892,
411,
278,
3519,
7199,
5921,
13138,
263,
4697,
342,
1650,
304,
263,
7546,
310,
17473,
519,
9101,
393,
526,
11122,
297,
263,
9257,
24171,
7536,
304,
4697,
602,
29892,
988,
278,
4697,
602,
16706,
12435,
278,
7546,
310,
9101,
964,
2323,
9101,
29889,
2860,
4697,
602,
29892,
278,
2323,
9101,
526,
7180,
964,
263,
716,
260,
15118,
9257,
24171,
411,
263,
17473,
519,
3038,
9257,
1650,
29892,
988,
262,
278,
17473,
519,
3038,
9257,
1650,
7199,
4637,
263,
2362,
284,
15795,
18254,
374,
296,
1650,
322,
385,
1425,
29890,
29933,
29941,
14172,
392,
29889,
9038,
4185,
2955,
29892,
278,
2323,
17473,
519,
9101,
526,
7180,
297,
5855,
393,
14257,
14321,
322,
8542,
310,
278,
2323,
9101,
29889,
13,
29961,
29900,
29900,
29906,
29941,
29962,
383,
6259,
29889,
29871,
29896,
1401,
919,
29879,
1855,
931,
390,
29911,
29899,
9026,
29934,
4603,
7418,
310,
11033,
5194,
29896,
29929,
29892,
2448,
545,
29887,
352,
262,
29896,
29892,
322,
1425,
29890,
29933,
29896,
29899,
29941,
297,
350,
29954,
29900,
29896,
29894,
21633,
297,
3342,
5855,
313,
29947,
8736,
29914,
828,
383,
29954,
29943,
29906,
29892,
29871,
29896,
29900,
29900,
8736,
29914,
828,
365,
29934,
29899,
6259,
29943,
29896,
29892,
29871,
29896,
8736,
29914,
828,
21775,
262,
319,
467,
402,
3301,
29928,
29950,
322,
438,
1783,
29946,
2761,
337,
7387,
526,
18694,
29889,
13,
29961,
29900,
29900,
29906,
29946,
29962,
383,
6259,
29889,
29871,
29906,
1401,
919,
29879,
278,
297,
6335,
654,
310,
410,
29880,
9633,
362,
310,
350,
29954,
29900,
29896,
29894,
9101,
773,
16369,
29947,
29955,
29929,
29889,
350,
29954,
29900,
29896,
29894,
9101,
892,
715,
630,
297,
29871,
29953,
29899,
5872,
260,
764,
29879,
322,
19884,
304,
360,
4345,
29949,
313,
29909,
511,
29871,
29945,
29900,
302,
29924,
29899,
29906,
29900,
29871,
30167,
29924,
16369,
29896,
29946,
29955,
29947,
313,
29933,
511,
470,
29871,
29896,
29900,
29900,
286,
29924,
29899,
29906,
29900,
29871,
30167,
29924,
16369,
29947,
29955,
29929,
313,
29907,
29897,
29871,
29906,
29946,
6199,
1156,
715,
1218,
29889,
2860,
29871,
29945,
3841,
297,
9257,
29892,
278,
4185,
1973,
892,
4343,
322,
380,
7114,
363,
394,
11311,
457,
1374,
25715,
271,
559,
6354,
29889,
16369,
29896,
29946,
29955,
29947,
1258,
451,
2615,
304,
6602,
410,
29880,
9633,
362,
472,
1438,
14953,
800,
313,
29906,
29900,
29871,
30167,
29924,
4318,
297,
350,
511,
541,
16369,
29947,
29955,
29929,
23228,
368,
5232,
287,
3038,
14321,
472,
29871,
29945,
29871,
30167,
29924,
313,
29907,
467,
13,
29961,
29900,
29900,
29906,
29945,
29962,
383,
6259,
29889,
29871,
29941,
1401,
919,
29879,
278,
18131,
3002,
310,
350,
29954,
29900,
29896,
29894,
9101,
4185,
2955,
297,
13681,
29899,
29950,
1964,
29943,
29892,
607,
338,
3342,
9257,
5745,
6943,
29871,
29896,
29900,
8736,
29914,
828,
379,
29934,
29954,
29899,
30187,
29892,
29871,
29896,
29900,
8736,
29914,
828,
21775,
262,
319,
29892,
29871,
29906,
29900,
29900,
8736,
29914,
828,
365,
29934,
29899,
6259,
29943,
29896,
322,
29871,
29947,
8736,
29914,
828,
383,
29954,
29943,
29906,
313,
29909,
322,
350,
511,
322,
297,
3342,
9257,
5745,
313,
12696,
29897
] | 1,024 |
CommonCrawl
| -0.503941 | 1.53203 | 0.495944 | 0.585385 |
containing 10 ng/ml HRG-β, 10 ng/ml Activin A, and 200 ng/ml LR-IGF1 (C and D).
[0026] FIG. 4 depicts the expression of ADAM19, Neuregulin1, and ErbB1-4 by RT-PCR in mouse ES cells (A) and MEFs (B).
[0027] FIG. 5 depicts the inhibition of ErbB1 and ErbB2 signaling in mouse ES cells. 2×105 Mouse R1 ES cells were plated on 1:1000 MATRIGEL® in 10% FBS, 10% KSR with 1000 U/ml mouse LIF (ESGRO). The following day, DMSO (carrier control), 1-50 μM AG1478, or 1-50 μM AG879 was added with fresh medium. The cultures were fixed on day 8, and stained for alkaline phosphatase activity. DMSO (A) and 1-50 μM AG1478 (B and C) did not overtly inhibit proliferation. AG879 substantially inhibited cell growth at 50 μM (compare D and F) and may have slowed proliferation at 20 μM (E).
[0028] FIG. 6 depicts the inhibition of proliferation of BG02 cells grown in conditioned media (CM). (A) 50 μM AG825 inhibited proliferation of BG02 hESCs growing in CM. (B) AG825 inhibits ErbB2 Y1248 phosphorylation in hESCs. (C) Colony counting of serial passaging of CyT49 hESCs in different combinations of growth factors. (D) Cell counting analysis of the role of IGF1 and HRG in hESC proliferation using BG02 cells (left). (E) OCT4/DAPI immunostaining of a duplicate repeated experiment demonstrated that IGF1 and HRG significantly increased the proportion of OCT4.sup.+ cells compared to ActA/FGF2 conditions. (F) RTK blotting analysis of BG01 DC-HALF hESCs starved of growth factors overnight; starved, then pulsed with DC-HALF for 15 minutes; or steady-state cultures are shown (left). The mean and range of normalized relative intensity is plotted (right).
[0029] FIG. 7 depicts mouse ES cells grown in defined conditions with different growth factor combinations. (A) shows the scoring of AP.sup.+ colonies after 2×105 cells were grown in different growth factor combinations for 8 days. (B-G) show 4× magnification images of AP.sup.+ colonies grown in different growth factor combinations.
[0030] FIG. 8 depicts the characterization of human ES cells that are maintained in DC-HAIF medium. (A) Analysis of teratomas from BG02 DC-HAIF p25 cells demonstrated pluripotent differentiation potential to ectoderm, mesoderm and endoderm. (B) Immunostaining of BG02 cells cultured in 15% FCS/5% KSR that have differentiated. (C) Venn diagram of the distribution of transcripts detected using high density Illumina Sentrix Human-6 Expression Beadchips containing 47,296 transcript probes in BG02 cells maintained in CM (64 passages) or DC-HALF (10 or 32 passages in defined media). (D) Scatterplot analysis demonstrating that the transcriptional profile of BG02 DC-HALF p32 cells is highly similar to that of BG02 cells maintained in CM (top), and was not substantially altered in early and late passage cultures in DC-HALF (bottom). (E) Hierarchical clustering dendrogram of relative gene expression in different populations generated using the Beadstudio software.
[0031] FIG. 9 depicts the morphology of
|
[
6943,
29871,
29896,
29900,
8736,
29914,
828,
379,
29934,
29954,
29899,
30187,
29892,
29871,
29896,
29900,
8736,
29914,
828,
21775,
262,
319,
29892,
322,
29871,
29906,
29900,
29900,
8736,
29914,
828,
365,
29934,
29899,
6259,
29943,
29896,
313,
29907,
322,
360,
467,
13,
29961,
29900,
29900,
29906,
29953,
29962,
383,
6259,
29889,
29871,
29946,
1401,
919,
29879,
278,
4603,
310,
11033,
5194,
29896,
29929,
29892,
2448,
545,
29887,
352,
262,
29896,
29892,
322,
1425,
29890,
29933,
29896,
29899,
29946,
491,
390,
29911,
29899,
9026,
29934,
297,
9495,
17956,
9101,
313,
29909,
29897,
322,
22986,
29943,
29879,
313,
29933,
467,
13,
29961,
29900,
29900,
29906,
29955,
29962,
383,
6259,
29889,
29871,
29945,
1401,
919,
29879,
278,
297,
6335,
654,
310,
1425,
29890,
29933,
29896,
322,
1425,
29890,
29933,
29906,
7182,
292,
297,
9495,
17956,
9101,
29889,
29871,
29906,
30122,
29896,
29900,
29945,
25992,
390,
29896,
17956,
9101,
892,
715,
630,
373,
29871,
29896,
29901,
29896,
29900,
29900,
29900,
341,
1299,
3960,
1692,
29931,
30342,
297,
29871,
29896,
29900,
29995,
383,
9851,
29892,
29871,
29896,
29900,
29995,
476,
14098,
411,
29871,
29896,
29900,
29900,
29900,
501,
29914,
828,
9495,
365,
6545,
313,
2890,
29954,
1672,
467,
450,
1494,
2462,
29892,
360,
4345,
29949,
313,
4287,
4336,
2761,
511,
29871,
29896,
29899,
29945,
29900,
29871,
30167,
29924,
16369,
29896,
29946,
29955,
29947,
29892,
470,
29871,
29896,
29899,
29945,
29900,
29871,
30167,
29924,
16369,
29947,
29955,
29929,
471,
2715,
411,
10849,
18350,
29889,
450,
4185,
1973,
892,
4343,
373,
2462,
29871,
29947,
29892,
322,
380,
7114,
363,
394,
11311,
457,
1374,
25715,
271,
559,
6354,
29889,
360,
4345,
29949,
313,
29909,
29897,
322,
29871,
29896,
29899,
29945,
29900,
29871,
30167,
29924,
16369,
29896,
29946,
29955,
29947,
313,
29933,
322,
315,
29897,
1258,
451,
975,
29873,
368,
297,
6335,
277,
410,
29880,
9633,
362,
29889,
16369,
29947,
29955,
29929,
23228,
368,
297,
6335,
1573,
3038,
14321,
472,
29871,
29945,
29900,
29871,
30167,
29924,
313,
18307,
360,
322,
383,
29897,
322,
1122,
505,
5232,
287,
410,
29880,
9633,
362,
472,
29871,
29906,
29900,
29871,
30167,
29924,
313,
29923,
467,
13,
29961,
29900,
29900,
29906,
29947,
29962,
383,
6259,
29889,
29871,
29953,
1401,
919,
29879,
278,
297,
6335,
654,
310,
410,
29880,
9633,
362,
310,
350,
29954,
29900,
29906,
9101,
21633,
297,
4195,
287,
5745,
313,
24494,
467,
313,
29909,
29897,
29871,
29945,
29900,
29871,
30167,
29924,
16369,
29947,
29906,
29945,
297,
6335,
1573,
410,
29880,
9633,
362,
310,
350,
29954,
29900,
29906,
298,
2890,
29907,
29879,
15678,
297,
315,
29924,
29889,
313,
29933,
29897,
16369,
29947,
29906,
29945,
297,
6335,
1169,
1425,
29890,
29933,
29906,
612,
29896,
29906,
29946,
29947,
1374,
25715,
706,
18411,
297,
298,
2890,
29907,
29879,
29889,
313,
29907,
29897,
1530,
2592,
21248,
310,
7797,
1209,
6751,
310,
8045,
29911,
29946,
29929,
298,
2890,
29907,
29879,
297,
1422,
18240,
310,
14321,
13879,
29889,
313,
29928,
29897,
19413,
21248,
7418,
310,
278,
6297,
310,
306,
29954,
29943,
29896,
322,
379,
29934,
29954,
297,
298,
2890,
29907,
410,
29880,
9633,
362,
773,
350,
29954,
29900,
29906,
9101,
313,
1563,
467,
313,
29923,
29897,
438,
1783,
29946,
29914,
7698,
2227,
5198,
348,
520,
17225,
310,
263,
7929,
10324,
7639,
28585,
393,
306,
29954,
29943,
29896,
322,
379,
29934,
29954,
16951,
11664,
278,
18618,
310,
438,
1783,
29946,
29889,
12587,
29889,
29974,
9101,
9401,
304,
3185,
29909,
29914,
29943,
29954,
29943,
29906,
5855,
29889,
313,
29943,
29897,
390,
29911,
29968,
1999,
327,
1259,
7418,
310,
350,
29954,
29900,
29896,
13681,
29899,
29950,
1964,
29943,
298,
2890,
29907,
29879,
5810,
1490,
310,
14321,
13879,
975,
11147,
29936,
5810,
1490,
29892,
769,
282,
7273,
287,
411,
13681,
29899,
29950,
1964,
29943,
363,
29871,
29896,
29945,
6233,
29936,
470,
27357,
29899,
3859,
4185,
1973,
526,
4318,
313,
1563,
467,
450,
2099,
322,
3464,
310,
4226,
1891,
6198,
26171,
338,
715,
15048,
313,
1266,
467,
13,
29961,
29900,
29900,
29906,
29929,
29962,
383,
6259,
29889,
29871,
29955,
1401,
919,
29879,
9495,
17956,
9101,
21633,
297,
3342,
5855,
411,
1422,
14321,
7329,
18240,
29889,
313,
29909,
29897,
3697,
278,
26654,
310,
12279,
29889,
12587,
29889,
29974,
8104,
583,
1156,
29871,
29906,
30122,
29896,
29900,
29945,
9101,
892,
21633,
297,
1422,
14321,
7329,
18240,
363,
29871,
29947,
3841,
29889,
313,
29933,
29899,
29954,
29897,
1510,
29871,
29946,
30122,
9119,
2450,
4558,
310,
12279,
29889,
12587,
29889,
29974,
8104,
583,
21633,
297,
1422,
14321,
7329,
18240,
29889,
13,
29961,
29900,
29900,
29941,
29900,
29962,
383,
6259,
29889,
29871,
29947,
1401,
919,
29879,
278,
2931,
2133,
310,
5199,
17956,
9101,
393,
526,
19949,
297,
13681,
29899,
15715,
6545,
18350,
29889,
313,
29909,
29897,
24352,
310,
1935,
8678,
294,
515,
350,
29954,
29900,
29906,
13681,
29899,
15715,
6545,
282,
29906,
29945,
9101,
28585,
715,
332,
666,
327,
296,
1422,
11685,
7037,
304,
321,
312,
397,
837,
29892,
4883,
397,
837,
322,
1095,
397,
837,
29889,
313,
29933,
29897,
1954,
24579,
520,
17225,
310,
350,
29954,
29900,
29906,
9101,
4185,
2955,
297,
29871,
29896,
29945,
29995,
7992,
29903,
29914,
29945,
29995,
476,
14098,
393,
505,
17473,
630,
29889,
313,
29907,
29897,
478,
2108,
13722,
310,
278,
4978,
310,
1301,
924,
29879,
17809,
773,
1880,
9027,
8408,
398,
1099,
28048,
2126,
12968,
29899,
29953,
21444,
350,
1479,
4161,
567,
6943,
29871,
29946,
29955,
29892,
29906,
29929,
29953,
1301,
924,
2070,
267,
297,
350,
29954,
29900,
29906,
9101,
19949,
297,
315,
29924,
313,
29953,
29946,
1209,
1179,
29897,
470,
13681,
29899,
29950,
1964,
29943,
313,
29896,
29900,
470,
29871,
29941,
29906,
1209,
1179,
297,
3342,
5745,
467,
313,
29928,
29897,
2522,
2620,
5317,
7418,
9004,
1218,
393,
278,
1301,
3395,
284,
8722,
310,
350,
29954,
29900,
29906,
13681,
29899,
29950,
1964,
29943,
282,
29941,
29906,
9101,
338,
10712,
2788,
304,
393,
310,
350,
29954,
29900,
29906,
9101,
19949,
297,
315,
29924,
313,
3332,
511,
322,
471,
451,
23228,
368,
10551,
287,
297,
4688,
322,
5683,
13382,
4185,
1973,
297,
13681,
29899,
29950,
1964,
29943,
313,
8968,
467,
313,
29923,
29897,
12433,
1279,
936,
16993,
3241,
270,
355,
307,
1393,
310,
6198,
18530,
4603,
297,
1422,
23093,
5759,
773,
278,
350,
1479,
12073,
7047,
29889,
13,
29961,
29900,
29900,
29941,
29896,
29962,
383,
6259,
29889,
29871,
29929,
1401,
919,
29879,
278,
18131,
3002,
310
] | 1,024 |
CommonCrawl
| -0.98554 | 1.355531 | 0.689612 | 0.542379 |
cells cultured on humanized extracellular matrices (ECMs) in the presence of DC-HALF medium. (A) CyT49 cells (diluted 1:200) growing on growth factor-reduced MATRIGEL® (diluted 1:200). CyT49 cells could also grow on tissue culture dishes coated with (B) whole human scrum, (C) human fibronectin, and (D) VITROGRO®.
[0032] FIG. 10 depicts the single-cell passaging of human ES cells. (A-D) Staged imaging of BG02 cells after passaging with ACCUTASE® and plating about 5×105 cells in a 60 mm culture dish. (A) 1.5 hours after initial plating, showing viable cells adhering to the dish. (B) At 20 hours post-plating, the large majority of cells have aggregated to form small colonies. These colonies expand by proliferation by day 4, post-plating (C), and over the course of 5-6 days to form an epithelial-like monolayer covering the entire dish (D). (E) Normal male karyotype demonstrated in a BG02 culture passaged 19 times with ACCUTASE® in DC-HALF.
[0033] FIG. 11 depicts cell morphology after single cell passaging of human ES cells using (A) ACCUTASE®, (B) 0.25% Trypsin/EDTA, (C) TrypLE, or (D) Versene.
[0034] FIG. 12 depicts the large-scale growth of human ES cells cultured in DC-HALF. (A) Flow cytometric analysis of BG02 cells after expansion to >1010 cells. >85% of cells expressed OCT4, CD9, SSEA-4, TRA-1-81. (B) RT-PCR analysis of expression of markers of pluripotency OCT4, NANOG, REX1, SOX2, UTF1, CRIPTO, FOXD3, TERT and DPPA5. Markers of differentiated lineages, α-fetoprotein (AFP), MSX1 and HAND1 were not detected. (C) Fluorescence in situ hybridization (FISH) using human chromosome-specific repeats demonstrated maintenance of normal copy numbers for hChr 12, 17, X and Y.
[0035] FIG. 13 depicts the morphology (A) and normal karyotype (B) of hESC BG02 cells grown in defined media comprising HRG-β and 1GF1, but in the absence of FGF2 for 7 passages, or >2 months.
[0036] FIG. 14 depicts a scatterplot analysis of transcripts from hESCs (BG02) that are maintained in DC-HAIF (32 passages) or DC-HAI (10 passages). A large proportion of the expressed transcripts were detected in both samples, and transcription was not substantially altered by culturing hESCs in the absence of exogenous FGF2. Correlation coefficients (R2) were generated using all detected transcripts with an expression level of >0 (all dots), or with transcripts exhibiting a detection confidence level of >0.99 (R2 select, dots indicated by dashed oval). Angled lines delineate the mean and limits of a 2-fold difference.
[0037] FIG. 15 depicts a hierarchical clustering dendrogram of relative gene expression in different populations of early and late passage BG02 cells maintained in DC-HAIF. Cells clustered tightly (˜0.0075) and retained a close similarity to BG02 and BG03 cells maintained in conditioned medium (CM) (˜0.037). BG02 cells maintained in DC-HAI also clustered tightly with the other hESC populations examined. By way of explanation in FIG. 15, CM is Conditioned Medium; DC is defined culture medium, DC-HA
|
[
9101,
4185,
2955,
373,
5199,
1891,
1294,
945,
514,
1070,
13516,
313,
11206,
29924,
29879,
29897,
297,
278,
10122,
310,
13681,
29899,
29950,
1964,
29943,
18350,
29889,
313,
29909,
29897,
8045,
29911,
29946,
29929,
9101,
313,
29881,
309,
3860,
29871,
29896,
29901,
29906,
29900,
29900,
29897,
15678,
373,
14321,
7329,
29899,
9313,
1133,
341,
1299,
3960,
1692,
29931,
30342,
313,
29881,
309,
3860,
29871,
29896,
29901,
29906,
29900,
29900,
467,
8045,
29911,
29946,
29929,
9101,
1033,
884,
6548,
373,
260,
15118,
9257,
270,
17006,
1302,
630,
411,
313,
29933,
29897,
3353,
5199,
885,
5848,
29892,
313,
29907,
29897,
5199,
18755,
1617,
522,
262,
29892,
322,
313,
29928,
29897,
478,
1806,
1672,
29954,
1672,
30342,
29889,
13,
29961,
29900,
29900,
29941,
29906,
29962,
383,
6259,
29889,
29871,
29896,
29900,
1401,
919,
29879,
278,
2323,
29899,
3729,
1209,
6751,
310,
5199,
17956,
9101,
29889,
313,
29909,
29899,
29928,
29897,
624,
4063,
6382,
292,
310,
350,
29954,
29900,
29906,
9101,
1156,
1209,
6751,
411,
319,
4174,
2692,
8127,
30342,
322,
715,
1218,
1048,
29871,
29945,
30122,
29896,
29900,
29945,
9101,
297,
263,
29871,
29953,
29900,
5654,
9257,
270,
728,
29889,
313,
29909,
29897,
29871,
29896,
29889,
29945,
6199,
1156,
2847,
715,
1218,
29892,
6445,
3516,
519,
9101,
594,
2276,
292,
304,
278,
270,
728,
29889,
313,
29933,
29897,
2180,
29871,
29906,
29900,
6199,
1400,
29899,
572,
1218,
29892,
278,
2919,
13638,
310,
9101,
505,
11404,
630,
304,
883,
2319,
8104,
583,
29889,
4525,
8104,
583,
7985,
491,
410,
29880,
9633,
362,
491,
2462,
29871,
29946,
29892,
1400,
29899,
572,
1218,
313,
29907,
511,
322,
975,
278,
3236,
310,
29871,
29945,
29899,
29953,
3841,
304,
883,
385,
9358,
389,
295,
616,
29899,
4561,
1601,
324,
2747,
21653,
278,
4152,
270,
728,
313,
29928,
467,
313,
29923,
29897,
21981,
14263,
413,
653,
327,
668,
28585,
297,
263,
350,
29954,
29900,
29906,
9257,
1209,
4063,
29871,
29896,
29929,
3064,
411,
319,
4174,
2692,
8127,
30342,
297,
13681,
29899,
29950,
1964,
29943,
29889,
13,
29961,
29900,
29900,
29941,
29941,
29962,
383,
6259,
29889,
29871,
29896,
29896,
1401,
919,
29879,
3038,
18131,
3002,
1156,
2323,
3038,
1209,
6751,
310,
5199,
17956,
9101,
773,
313,
29909,
29897,
319,
4174,
2692,
8127,
30342,
29892,
313,
29933,
29897,
29871,
29900,
29889,
29906,
29945,
29995,
3967,
567,
262,
29914,
3352,
6040,
29892,
313,
29907,
29897,
3967,
29886,
1307,
29892,
470,
313,
29928,
29897,
10138,
1600,
29889,
13,
29961,
29900,
29900,
29941,
29946,
29962,
383,
6259,
29889,
29871,
29896,
29906,
1401,
919,
29879,
278,
2919,
29899,
7052,
14321,
310,
5199,
17956,
9101,
4185,
2955,
297,
13681,
29899,
29950,
1964,
29943,
29889,
313,
29909,
29897,
22787,
274,
3637,
14066,
7418,
310,
350,
29954,
29900,
29906,
9101,
1156,
13184,
304,
1405,
29896,
29900,
29896,
29900,
9101,
29889,
1405,
29947,
29945,
29995,
310,
9101,
13384,
438,
1783,
29946,
29892,
7307,
29929,
29892,
317,
1660,
29909,
29899,
29946,
29892,
323,
4717,
29899,
29896,
29899,
29947,
29896,
29889,
313,
29933,
29897,
390,
29911,
29899,
9026,
29934,
7418,
310,
4603,
310,
29320,
310,
715,
332,
666,
327,
3819,
438,
1783,
29946,
29892,
405,
2190,
29949,
29954,
29892,
5195,
29990,
29896,
29892,
7791,
29990,
29906,
29892,
18351,
29896,
29892,
315,
3960,
29925,
4986,
29892,
18322,
29990,
29928,
29941,
29892,
323,
20161,
322,
360,
29925,
7228,
29945,
29889,
4485,
414,
310,
17473,
630,
1196,
1179,
29892,
19883,
29899,
29888,
300,
459,
4859,
262,
313,
5098,
29925,
511,
10888,
29990,
29896,
322,
379,
9468,
29896,
892,
451,
17809,
29889,
313,
29907,
29897,
2379,
29884,
2361,
29883,
663,
297,
2990,
7498,
19515,
2133,
313,
29943,
3235,
29950,
29897,
773,
5199,
25173,
359,
608,
29899,
14940,
5565,
1446,
28585,
25413,
310,
4226,
3509,
3694,
363,
298,
11626,
29871,
29896,
29906,
29892,
29871,
29896,
29955,
29892,
1060,
322,
612,
29889,
13,
29961,
29900,
29900,
29941,
29945,
29962,
383,
6259,
29889,
29871,
29896,
29941,
1401,
919,
29879,
278,
18131,
3002,
313,
29909,
29897,
322,
4226,
413,
653,
327,
668,
313,
29933,
29897,
310,
298,
2890,
29907,
350,
29954,
29900,
29906,
9101,
21633,
297,
3342,
5745,
7199,
5921,
379,
29934,
29954,
29899,
30187,
322,
29871,
29896,
29954,
29943,
29896,
29892,
541,
297,
278,
18070,
310,
383,
29954,
29943,
29906,
363,
29871,
29955,
1209,
1179,
29892,
470,
1405,
29906,
7378,
29889,
13,
29961,
29900,
29900,
29941,
29953,
29962,
383,
6259,
29889,
29871,
29896,
29946,
1401,
919,
29879,
263,
14801,
5317,
7418,
310,
1301,
924,
29879,
515,
298,
2890,
29907,
29879,
313,
29933,
29954,
29900,
29906,
29897,
393,
526,
19949,
297,
13681,
29899,
15715,
6545,
313,
29941,
29906,
1209,
1179,
29897,
470,
13681,
29899,
15715,
29902,
313,
29896,
29900,
1209,
1179,
467,
319,
2919,
18618,
310,
278,
13384,
1301,
924,
29879,
892,
17809,
297,
1716,
11916,
29892,
322,
1301,
3395,
471,
451,
23228,
368,
10551,
287,
491,
4185,
3864,
298,
2890,
29907,
29879,
297,
278,
18070,
310,
429,
6352,
681,
383,
29954,
29943,
29906,
29889,
2994,
23445,
16127,
313,
29934,
29906,
29897,
892,
5759,
773,
599,
17809,
1301,
924,
29879,
411,
385,
4603,
3233,
310,
1405,
29900,
313,
497,
270,
1862,
511,
470,
411,
1301,
924,
29879,
10371,
11407,
263,
15326,
16420,
3233,
310,
1405,
29900,
29889,
29929,
29929,
313,
29934,
29906,
1831,
29892,
270,
1862,
18694,
491,
27526,
288,
791,
467,
3218,
839,
3454,
628,
457,
403,
278,
2099,
322,
13071,
310,
263,
29871,
29906,
29899,
8771,
4328,
29889,
13,
29961,
29900,
29900,
29941,
29955,
29962,
383,
6259,
29889,
29871,
29896,
29945,
1401,
919,
29879,
263,
6128,
1279,
936,
16993,
3241,
270,
355,
307,
1393,
310,
6198,
18530,
4603,
297,
1422,
23093,
310,
4688,
322,
5683,
13382,
350,
29954,
29900,
29906,
9101,
19949,
297,
13681,
29899,
15715,
6545,
29889,
315,
10071,
9867,
287,
19932,
368,
313,
31793,
29900,
29889,
29900,
29900,
29955,
29945,
29897,
322,
26060,
263,
3802,
29501,
304,
350,
29954,
29900,
29906,
322,
350,
29954,
29900,
29941,
9101,
19949,
297,
4195,
287,
18350,
313,
24494,
29897,
313,
31793,
29900,
29889,
29900,
29941,
29955,
467,
350,
29954,
29900,
29906,
9101,
19949,
297,
13681,
29899,
15715,
29902,
884,
9867,
287,
19932,
368,
411,
278,
916,
298,
2890,
29907,
23093,
4392,
1312,
29889,
2648,
982,
310,
8252,
297,
383,
6259,
29889,
29871,
29896,
29945,
29892,
315,
29924,
338,
11790,
654,
287,
3436,
1974,
29936,
13681,
338,
3342,
9257,
18350,
29892,
13681,
29899,
15715
] | 1,024 |
CommonCrawl
| -0.672398 | 1.412633 | 0.88328 | 0.663808 |
IF as defined above; ap is ACCUTASE® single cell passaging; DC-HAI is identical to DC-HAIF as defined herein, except without FGF2.
[0038] FIG. 16 depicts the morphology and alkaline phsophatasc staining of BG02 cells cultured in DC-HAIF in 96-well and 384-well plates. (A) Phase contrast imaging and (B) alkaline phosphatase staining of BG02 cells (104 cells/well) growing in one well of a 96-well plate. (C) Phase contrast imaging and (D) alkaline phosphatase staining of BG02 cells (103 cells/well) growing in one well of a 384-well plate.
[0039] FIG. 17 depicts dark field images of BG02 grown in DC-HAIF in suspension culture. Day 2 and day 6 cultures are shown. The images were captured using 4× magnification
[0040] FIG. 18 depicts the growth rates in adherent and suspension cultures in DC-HAIF. 1×106 BG02 cells were plated into parallel wells in adherent and suspension culture and cell counts were performed on days 1-6.
[0041] FIG. 19 depicts qPCR analysis of suspension and adherent hESCs. BG02 cells growing in suspension (S. hESCs) and adherent (hESCs) culture exhibited comparable levels of OCT4, and lacked SOX17 expression. Adherent cells differentiated to definitive endoderm (DE), and suspension hESCs differentiated to definitive endoderm in suspension (S. DE d3), both exhibited the expected marked downregulation of OCT4 and upregulation of SOX17 expression
[0042] FIG. 20 depicts the enhancement of hESC aggregation in the presence of Y27632 in suspension culture. 2×106 BG02 cells were seeded in 3 ml DC-HAIF or DC-HAIF+Y27632, in 6-well trays, in an incubator on a rotating platform at 100 rpm. Images of aggregates were captured on days 1 and 3.
[0043] FIG. 21 depicts RT-PCR analysis of suspension aggregates in the presence of Y27632. RT-PCR was performed on the expanded cultures to assess expression of markers of pluripotency. Expression of OCT4, NANOG, REX1, SOX2, UTF1, CRIPTO, FOXD3, TERT AND DPPA5 were detected, whereas markers of differentiated lineages AFP, MSX1 and HAND1 were not detected.
[0044] Unless otherwise noted, the terms used herein are to be understood according to conventional usage by those of ordinary skill in the relevant art. In addition to the definitions of terms provided below, definitions of common terms in molecular biology may also be found in Rieger et al., 1991 Glossary of genetics: classical and molecular, 5th Ed., Berlin: Springer-Verlag; and in Current Protocols in Molecular Biology, F. M. Ausubel et al., Eds., Current Protocols, a joint venture between Greene Publishing Associates, Inc. and John Wiley & Sons, Inc., (1998 Supplement). It is to be understood that as used in the specification and in the claims, "a" or "an" can mean one or more, depending upon the context in which it is used. Thus, for example, reference to "a cell" can mean that at least one cell can be utilized.
[0045] As used herein, the term "contacting" (i.e., contacting a cell e.g., a differentiable cell, with a compound) is intended to include incubating the compound and the cell together in vitro (e.g., adding the compound to'cells in culture). The term "contacting" is not intended to include the in vivo exposure of cells to a defined cell
|
[
6545,
408,
3342,
2038,
29936,
3095,
338,
319,
4174,
2692,
8127,
30342,
2323,
3038,
1209,
6751,
29936,
13681,
29899,
15715,
29902,
338,
13557,
304,
13681,
29899,
15715,
6545,
408,
3342,
1244,
262,
29892,
5174,
1728,
383,
29954,
29943,
29906,
29889,
13,
29961,
29900,
29900,
29941,
29947,
29962,
383,
6259,
29889,
29871,
29896,
29953,
1401,
919,
29879,
278,
18131,
3002,
322,
394,
11311,
457,
1374,
29879,
459,
2455,
6151,
380,
17225,
310,
350,
29954,
29900,
29906,
9101,
4185,
2955,
297,
13681,
29899,
15715,
6545,
297,
29871,
29929,
29953,
29899,
5872,
322,
29871,
29941,
29947,
29946,
29899,
5872,
715,
1078,
29889,
313,
29909,
29897,
1963,
559,
12814,
6382,
292,
322,
313,
29933,
29897,
394,
11311,
457,
1374,
25715,
271,
559,
380,
17225,
310,
350,
29954,
29900,
29906,
9101,
313,
29896,
29900,
29946,
9101,
29914,
5872,
29897,
15678,
297,
697,
1532,
310,
263,
29871,
29929,
29953,
29899,
5872,
15284,
29889,
313,
29907,
29897,
1963,
559,
12814,
6382,
292,
322,
313,
29928,
29897,
394,
11311,
457,
1374,
25715,
271,
559,
380,
17225,
310,
350,
29954,
29900,
29906,
9101,
313,
29896,
29900,
29941,
9101,
29914,
5872,
29897,
15678,
297,
697,
1532,
310,
263,
29871,
29941,
29947,
29946,
29899,
5872,
15284,
29889,
13,
29961,
29900,
29900,
29941,
29929,
29962,
383,
6259,
29889,
29871,
29896,
29955,
1401,
919,
29879,
6501,
1746,
4558,
310,
350,
29954,
29900,
29906,
21633,
297,
13681,
29899,
15715,
6545,
297,
8872,
2673,
9257,
29889,
8373,
29871,
29906,
322,
2462,
29871,
29953,
4185,
1973,
526,
4318,
29889,
450,
4558,
892,
15468,
773,
29871,
29946,
30122,
9119,
2450,
13,
29961,
29900,
29900,
29946,
29900,
29962,
383,
6259,
29889,
29871,
29896,
29947,
1401,
919,
29879,
278,
14321,
19257,
297,
594,
2276,
296,
322,
8872,
2673,
4185,
1973,
297,
13681,
29899,
15715,
6545,
29889,
29871,
29896,
30122,
29896,
29900,
29953,
350,
29954,
29900,
29906,
9101,
892,
715,
630,
964,
8943,
1532,
29879,
297,
594,
2276,
296,
322,
8872,
2673,
9257,
322,
3038,
18139,
892,
8560,
373,
3841,
29871,
29896,
29899,
29953,
29889,
13,
29961,
29900,
29900,
29946,
29896,
29962,
383,
6259,
29889,
29871,
29896,
29929,
1401,
919,
29879,
3855,
9026,
29934,
7418,
310,
8872,
2673,
322,
594,
2276,
296,
298,
2890,
29907,
29879,
29889,
350,
29954,
29900,
29906,
9101,
15678,
297,
8872,
2673,
313,
29903,
29889,
298,
2890,
29907,
29879,
29897,
322,
594,
2276,
296,
313,
29882,
2890,
29907,
29879,
29897,
9257,
10371,
1573,
5734,
519,
11174,
310,
438,
1783,
29946,
29892,
322,
10225,
287,
7791,
29990,
29896,
29955,
4603,
29889,
2087,
2276,
296,
9101,
17473,
630,
304,
8422,
573,
1095,
397,
837,
313,
2287,
511,
322,
8872,
2673,
298,
2890,
29907,
29879,
17473,
630,
304,
8422,
573,
1095,
397,
837,
297,
8872,
2673,
313,
29903,
29889,
5012,
270,
29941,
511,
1716,
10371,
1573,
278,
3806,
10902,
1623,
1727,
2785,
310,
438,
1783,
29946,
322,
701,
1727,
2785,
310,
7791,
29990,
29896,
29955,
4603,
13,
29961,
29900,
29900,
29946,
29906,
29962,
383,
6259,
29889,
29871,
29906,
29900,
1401,
919,
29879,
278,
26371,
27967,
310,
298,
2890,
29907,
11404,
362,
297,
278,
10122,
310,
612,
29906,
29955,
29953,
29941,
29906,
297,
8872,
2673,
9257,
29889,
29871,
29906,
30122,
29896,
29900,
29953,
350,
29954,
29900,
29906,
9101,
892,
16717,
287,
297,
29871,
29941,
286,
29880,
13681,
29899,
15715,
6545,
470,
13681,
29899,
15715,
6545,
29974,
29979,
29906,
29955,
29953,
29941,
29906,
29892,
297,
29871,
29953,
29899,
5872,
260,
764,
29879,
29892,
297,
385,
5528,
431,
1061,
373,
263,
5731,
1218,
7481,
472,
29871,
29896,
29900,
29900,
364,
3358,
29889,
1954,
1179,
310,
11404,
1078,
892,
15468,
373,
3841,
29871,
29896,
322,
29871,
29941,
29889,
13,
29961,
29900,
29900,
29946,
29941,
29962,
383,
6259,
29889,
29871,
29906,
29896,
1401,
919,
29879,
390,
29911,
29899,
9026,
29934,
7418,
310,
8872,
2673,
11404,
1078,
297,
278,
10122,
310,
612,
29906,
29955,
29953,
29941,
29906,
29889,
390,
29911,
29899,
9026,
29934,
471,
8560,
373,
278,
17832,
4185,
1973,
304,
24809,
4603,
310,
29320,
310,
715,
332,
666,
327,
3819,
29889,
21444,
310,
438,
1783,
29946,
29892,
405,
2190,
29949,
29954,
29892,
5195,
29990,
29896,
29892,
7791,
29990,
29906,
29892,
18351,
29896,
29892,
315,
3960,
29925,
4986,
29892,
18322,
29990,
29928,
29941,
29892,
323,
20161,
5300,
360,
29925,
7228,
29945,
892,
17809,
29892,
13452,
29320,
310,
17473,
630,
1196,
1179,
23844,
29925,
29892,
10888,
29990,
29896,
322,
379,
9468,
29896,
892,
451,
17809,
29889,
13,
29961,
29900,
29900,
29946,
29946,
29962,
25870,
6467,
11682,
29892,
278,
4958,
1304,
1244,
262,
526,
304,
367,
11098,
5034,
304,
28557,
8744,
491,
1906,
310,
15311,
19911,
297,
278,
8018,
1616,
29889,
512,
6124,
304,
278,
15848,
310,
4958,
4944,
2400,
29892,
15848,
310,
3619,
4958,
297,
13206,
16637,
4768,
3002,
1122,
884,
367,
1476,
297,
390,
347,
914,
634,
394,
1696,
29871,
29896,
29929,
29929,
29896,
402,
6758,
653,
310,
2531,
300,
1199,
29901,
14499,
322,
13206,
16637,
29892,
29871,
29945,
386,
2155,
1696,
5115,
29901,
26667,
29899,
22776,
29936,
322,
297,
9626,
1019,
5770,
29879,
297,
341,
1772,
16637,
3457,
3002,
29892,
383,
29889,
341,
29889,
4028,
431,
295,
634,
394,
1696,
2155,
29879,
1696,
9626,
1019,
5770,
29879,
29892,
263,
14002,
9712,
545,
1546,
4122,
1600,
19088,
6853,
1078,
29892,
9266,
29889,
322,
2259,
399,
15168,
669,
317,
787,
29892,
9266,
1696,
313,
29896,
29929,
29929,
29947,
9179,
944,
467,
739,
338,
304,
367,
11098,
393,
408,
1304,
297,
278,
21992,
322,
297,
278,
16726,
29892,
376,
29874,
29908,
470,
376,
273,
29908,
508,
2099,
697,
470,
901,
29892,
8679,
2501,
278,
3030,
297,
607,
372,
338,
1304,
29889,
6549,
29892,
363,
1342,
29892,
3407,
304,
376,
29874,
3038,
29908,
508,
2099,
393,
472,
3203,
697,
3038,
508,
367,
3667,
1891,
29889,
13,
29961,
29900,
29900,
29946,
29945,
29962,
1094,
1304,
1244,
262,
29892,
278,
1840,
376,
12346,
292,
29908,
313,
29875,
29889,
29872,
1696,
6958,
292,
263,
3038,
321,
29889,
29887,
1696,
263,
17473,
519,
3038,
29892,
411,
263,
752,
618,
29897,
338,
9146,
304,
3160,
5528,
431,
1218,
278,
752,
618,
322,
278,
3038,
4208,
297,
13901,
307,
313,
29872,
29889,
29887,
1696,
4417,
278,
752,
618,
304,
29915,
3729,
29879,
297,
9257,
467,
450,
1840,
376,
12346,
292,
29908,
338,
451,
9146,
304,
3160,
278,
297,
325,
4243,
14060,
545,
310,
9101,
304,
263,
3342,
3038
] | 1,024 |
CommonCrawl
| -1.200188 | 0.945431 | -0.084164 | -0.052113 |
medium comprising an ErbB3 ligand, and optionally, a member of the TGF-β family, that may occur naturally in a subject (i.e., exposure that may occur as a result of a natural physiological process). The step of contacting the cell with a defined cell medium comprising an ErbB3 ligand, and optionally, a member of the TGF-β family, can be conducted in any suitable manner. For example, the cells may be treated in adherent culture, or in suspension culture. It is understood that the cells contacted with the defined medium can be further treated with a cell differentiation environment to stabilize the cells, or to differentiate the cells.
[0046] As used herein, the term "differentiate" refers to the production of a cell type that is more differentiated than the cell type from which it is derived. The term therefore encompasses cell types that are partially and terminally differentiated.
[0047] In certain embodiments of the present invention, the term "enriched" refers to a cell culture that contains more than approximately 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% of the desired cell lineage.
[0048] As used herein, the term "effective amount" of a compound refers to that concentration of the compound that is sufficient in the presence of the remaining components of the defined medium to effect the stabilization of the differentiable cell in culture for greater than one month in the absence of a feeder cell and in the absence of serum or scrum replacement. This concentration is readily determined by one of ordinary skill in the art.
[0049] As used herein, the term "express" refers to the transcription of a polynucleotide or translation of a polypeptide in a cell, such that levels of the molecule are measurably higher in a cell that expresses the molecule than they are in a cell that does not express the molecule. Methods to measure the expression of a molecule are well known to those of ordinary skill in the art, and include without limitation, Northern blotting, RT-PCR, in situ hybridization, Western blotting, and immunostaining.
[0050] As used herein when referring to a cell, cell line, cell culture or population of cells, the term "isolated" refers to being substantially separated from the natural source of the cells such that the cell, cell line, cell culture, or population of cells are capable of being cultured in vitro. In addition, the term "isolating" is used to refer to the physical selection of one or more cells out of a group of two or more cells, wherein the cells are selected based on cell morphology and/or the expression of various markers.
[0051] The present invention may be understood more readily by reference to the following detailed description of the preferred embodiments of the invention and the Examples included herein. However, before the present compositions and methods are disclosed and described, it is to be understood that this invention is not limited to specific nucleic acids, specific polypeptides, specific cell types, specific host cells, specific conditions, or specific methods, etc., as such may, of course, vary, and the numerous modifications and variations therein will be apparent to those skilled in the art.
[0052] Standard techniques for cloning, DNA isolation, amplification and purification, for enzymatic reactions involving DNA ligase, DNA polymerase, restriction endonucleases and the like, and various separation techniques are those known and commonly employed by those skilled in the art. A number of standard techniques are described in Sambrook et ul., 1989 Molecular Cloning, Second Edition, Cold Spring Harbor Laboratory, Plainview, N.Y.; Maniatis et al., 1982 Molecular Cloning, Cold Spring Harbor Laboratory, Plainview, N.Y.; Wu (Ed.) 1993 Meth. Enzymol. 218, Part I; Wu (Ed.) 1979 Meth. Enzymol. 68; Wu et al., (Eds.) 1983 Meth. Enzymol. 100 and 101; Grossman and Moldave (Eds.) 1980 Meth. Enzymol. 65; Miller
|
[
18350,
7199,
5921,
385,
1425,
29890,
29933,
29941,
14172,
392,
29892,
322,
2984,
635,
29892,
263,
4509,
310,
278,
323,
29954,
29943,
29899,
30187,
3942,
29892,
393,
1122,
6403,
18180,
297,
263,
4967,
313,
29875,
29889,
29872,
1696,
14060,
545,
393,
1122,
6403,
408,
263,
1121,
310,
263,
5613,
4824,
29875,
5996,
1889,
467,
450,
4331,
310,
6958,
292,
278,
3038,
411,
263,
3342,
3038,
18350,
7199,
5921,
385,
1425,
29890,
29933,
29941,
14172,
392,
29892,
322,
2984,
635,
29892,
263,
4509,
310,
278,
323,
29954,
29943,
29899,
30187,
3942,
29892,
508,
367,
18043,
297,
738,
13907,
8214,
29889,
1152,
1342,
29892,
278,
9101,
1122,
367,
14914,
297,
594,
2276,
296,
9257,
29892,
470,
297,
8872,
2673,
9257,
29889,
739,
338,
11098,
393,
278,
9101,
6958,
287,
411,
278,
3342,
18350,
508,
367,
4340,
14914,
411,
263,
3038,
1422,
11685,
5177,
304,
16160,
675,
278,
9101,
29892,
470,
304,
17473,
403,
278,
9101,
29889,
13,
29961,
29900,
29900,
29946,
29953,
29962,
1094,
1304,
1244,
262,
29892,
278,
1840,
376,
29881,
8349,
7268,
403,
29908,
14637,
304,
278,
5802,
310,
263,
3038,
1134,
393,
338,
901,
17473,
630,
1135,
278,
3038,
1134,
515,
607,
372,
338,
10723,
29889,
450,
1840,
5480,
427,
2388,
465,
267,
3038,
4072,
393,
526,
22039,
322,
6624,
635,
17473,
630,
29889,
13,
29961,
29900,
29900,
29946,
29955,
29962,
512,
3058,
7232,
397,
7862,
310,
278,
2198,
297,
7316,
29892,
278,
1840,
376,
264,
4018,
287,
29908,
14637,
304,
263,
3038,
9257,
393,
3743,
901,
1135,
14235,
29871,
29945,
29900,
13667,
29871,
29945,
29945,
13667,
29871,
29953,
29900,
13667,
29871,
29953,
29945,
13667,
29871,
29955,
29900,
13667,
29871,
29955,
29945,
13667,
29871,
29947,
29900,
13667,
29871,
29947,
29945,
13667,
29871,
29929,
29900,
13667,
470,
29871,
29929,
29945,
29995,
310,
278,
7429,
3038,
1196,
482,
29889,
13,
29961,
29900,
29900,
29946,
29947,
29962,
1094,
1304,
1244,
262,
29892,
278,
1840,
376,
15987,
573,
5253,
29908,
310,
263,
752,
618,
14637,
304,
393,
26702,
310,
278,
752,
618,
393,
338,
8002,
297,
278,
10122,
310,
278,
9886,
7117,
310,
278,
3342,
18350,
304,
2779,
278,
16160,
2133,
310,
278,
17473,
519,
3038,
297,
9257,
363,
7621,
1135,
697,
4098,
297,
278,
18070,
310,
263,
1238,
2447,
3038,
322,
297,
278,
18070,
310,
724,
398,
470,
885,
5848,
16920,
29889,
910,
26702,
338,
28520,
10087,
491,
697,
310,
15311,
19911,
297,
278,
1616,
29889,
13,
29961,
29900,
29900,
29946,
29929,
29962,
1094,
1304,
1244,
262,
29892,
278,
1840,
376,
17073,
29908,
14637,
304,
278,
1301,
3395,
310,
263,
1248,
948,
1682,
280,
327,
680,
470,
13962,
310,
263,
1248,
668,
415,
680,
297,
263,
3038,
29892,
1316,
393,
11174,
310,
278,
13206,
29883,
1297,
526,
7540,
332,
2197,
6133,
297,
263,
3038,
393,
4653,
267,
278,
13206,
29883,
1297,
1135,
896,
526,
297,
263,
3038,
393,
947,
451,
4653,
278,
13206,
29883,
1297,
29889,
8108,
29879,
304,
5645,
278,
4603,
310,
263,
13206,
29883,
1297,
526,
1532,
2998,
304,
1906,
310,
15311,
19911,
297,
278,
1616,
29892,
322,
3160,
1728,
29485,
29892,
14299,
1999,
327,
1259,
29892,
390,
29911,
29899,
9026,
29934,
29892,
297,
2990,
7498,
19515,
2133,
29892,
10504,
1999,
327,
1259,
29892,
322,
5198,
348,
520,
17225,
29889,
13,
29961,
29900,
29900,
29945,
29900,
29962,
1094,
1304,
1244,
262,
746,
16811,
304,
263,
3038,
29892,
3038,
1196,
29892,
3038,
9257,
470,
4665,
310,
9101,
29892,
278,
1840,
376,
275,
324,
630,
29908,
14637,
304,
1641,
23228,
368,
13055,
515,
278,
5613,
2752,
310,
278,
9101,
1316,
393,
278,
3038,
29892,
3038,
1196,
29892,
3038,
9257,
29892,
470,
4665,
310,
9101,
526,
15390,
310,
1641,
4185,
2955,
297,
13901,
307,
29889,
512,
6124,
29892,
278,
1840,
376,
275,
324,
1218,
29908,
338,
1304,
304,
2737,
304,
278,
9128,
9262,
310,
697,
470,
901,
9101,
714,
310,
263,
2318,
310,
1023,
470,
901,
9101,
29892,
988,
262,
278,
9101,
526,
4629,
2729,
373,
3038,
18131,
3002,
322,
29914,
272,
278,
4603,
310,
5164,
29320,
29889,
13,
29961,
29900,
29900,
29945,
29896,
29962,
450,
2198,
297,
7316,
1122,
367,
11098,
901,
28520,
491,
3407,
304,
278,
1494,
13173,
6139,
310,
278,
16389,
7232,
397,
7862,
310,
278,
297,
7316,
322,
278,
1222,
9422,
5134,
1244,
262,
29889,
2398,
29892,
1434,
278,
2198,
5541,
2187,
322,
3519,
526,
766,
15603,
322,
5439,
29892,
372,
338,
304,
367,
11098,
393,
445,
297,
7316,
338,
451,
9078,
304,
2702,
22699,
293,
1274,
4841,
29892,
2702,
1248,
668,
415,
2247,
29892,
2702,
3038,
4072,
29892,
2702,
3495,
9101,
29892,
2702,
5855,
29892,
470,
2702,
3519,
29892,
2992,
1696,
408,
1316,
1122,
29892,
310,
3236,
29892,
13100,
29892,
322,
278,
12727,
26278,
322,
21833,
727,
262,
674,
367,
20295,
304,
1906,
2071,
24455,
297,
278,
1616,
29889,
13,
29961,
29900,
29900,
29945,
29906,
29962,
10117,
13698,
363,
1067,
28259,
29892,
25348,
11695,
362,
29892,
20563,
2450,
322,
3708,
2450,
29892,
363,
427,
14022,
2454,
337,
7387,
21677,
25348,
14172,
559,
29892,
25348,
24324,
261,
559,
29892,
24345,
1095,
265,
1682,
17836,
322,
278,
763,
29892,
322,
5164,
23683,
13698,
526,
1906,
2998,
322,
15574,
15723,
491,
1906,
2071,
24455,
297,
278,
1616,
29889,
319,
1353,
310,
3918,
13698,
526,
5439,
297,
317,
1117,
307,
554,
634,
9238,
1696,
29871,
29896,
29929,
29947,
29929,
341,
1772,
16637,
2233,
28259,
29892,
6440,
17138,
29892,
26731,
7206,
3536,
4089,
16715,
7606,
29892,
349,
7420,
1493,
29892,
405,
29889,
29979,
8670,
2315,
7163,
275,
634,
394,
1696,
29871,
29896,
29929,
29947,
29906,
341,
1772,
16637,
2233,
28259,
29892,
26731,
7206,
3536,
4089,
16715,
7606,
29892,
349,
7420,
1493,
29892,
405,
29889,
29979,
8670,
29624,
313,
3853,
1846,
29871,
29896,
29929,
29929,
29941,
341,
621,
29889,
1174,
14022,
324,
29889,
29871,
29906,
29896,
29947,
29892,
3455,
306,
29936,
29624,
313,
3853,
1846,
29871,
29896,
29929,
29955,
29929,
341,
621,
29889,
1174,
14022,
324,
29889,
29871,
29953,
29947,
29936,
29624,
634,
394,
1696,
313,
3853,
29879,
1846,
29871,
29896,
29929,
29947,
29941,
341,
621,
29889,
1174,
14022,
324,
29889,
29871,
29896,
29900,
29900,
322,
29871,
29896,
29900,
29896,
29936,
402,
2124,
1171,
322,
27140,
1351,
313,
3853,
29879,
1846,
29871,
29896,
29929,
29947,
29900,
341,
621,
29889,
1174,
14022,
324,
29889,
29871,
29953,
29945,
29936,
16498
] | 1,024 |
CommonCrawl
| -0.982823 | 1.220561 | -0.403357 | 0.355178 |
(ed.) 1972 Experiments in Molecular Genetics, Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.; Old and Primrose, 1981 Principles of Gene Manipulation, University of California Press, Berkeley; Schleif and Wensink, 1982 Practical Methods in Molecular Biology; Glover (Ed.) 1985 DNA Cloning Vol. I and II, IRL Press, Oxford, UK; Hames and Higgins (Eds.) 1985 Nucleic Acid Hybridization, IRL Press, Oxford, UK; and Setlow and Hollaender 1979 Genetic Engineering: Principles and Methods, Vols. 1-4, Plenum Press, New York. Abbreviations and nomenclature, where employed, are deemed standard in the field and commonly used in professional journals such as those cited herein.
[0053] The invention relates to compositions and methods comprising a basal salt nutrient solution and an effective amount of an ErbB3 ligand, with the compositions being essentially free of serum. The compositions and methods of the present invention are useful for culturing cells, in particular, differentiable cells. It is understood that at different points during culturing the differentiable cells, various components may be added to the cell culture such that the medium can contain components other than those described herein. It is, however, contemplated that at least at one point during the preparation of the culture, or during the culture of the differentiable cells, the defined medium comprises a basal salt nutrient solution and a means for activating ErbB2-directed tyrosine kinase.
[0054] As used herein, the term "differentiable cell" is used to describe a cell or population of cells that can differentiate into at least partially mature cells, or that can participate in the differentiation of cells, e.g., fuse with other cells, that can differentiate into at least partially mature cells. As used herein, "partially mature cells" are cells that exhibit at least one characteristic of the phenotype, such as morphology or protein expression, of a mature cell from the same organ or tissue. For example, a normal, mature hepatocyte typically expresses such proteins as albumin, fibrinogen, alpha-1-antitrypsin, prothrombin clotting factors, transferrin, and detoxification enzymes such as the cytochrome P-450s, among others. Thus, as defined in the present invention, a "partially mature hepatocyte" may express albumin or another one or more proteins, or begin to take the appearance or function of a normal, mature hepatocyte. Additionally, a "partially mature pancreatic beta cell" may produce or express the proinsulin protein, among others. The ability of the cells to differentiate into at least partially mature cells will not be dependent upon recombinant engineering techniques, such as transfection, though the cells may, of course, be genetically engineered.
[0055] The invention contemplates compositions and methods useful for differentiable cells, regardless of their source or of their plasticity. The "plasticity" of a cell is used herein roughly as it is in the art. Namely, the plasticity of a cell refers to a cell's ability to differentiate into a particular cell type found in tissues or organs from an embryo, fetus or developed organism. The "more plastic" a cell, the more tissues into which the cell may be able to differentiate. "Pluripotent cells" include cells and their progeny, which may be able to differentiate into, or give rise to, pluripotent, multipotent, oligopotent and unipotent cells, and/or several, if not all, of the mature or partially mature cell types found in an embryo, fetus or developed organism. "Multipotent cells" include cells and their progeny, which may be able to differentiate into, or give rise to, multipotent, oligopotent and unipotent progenitor cells, and/or one or more mature or partially mature cell types, except that the mature or partially mature cell types derived from multipotent cells are limited to cells of a particular tissue, organ or organ system. For example, a
|
[
313,
287,
1846,
29871,
29896,
29929,
29955,
29906,
28224,
7862,
297,
341,
1772,
16637,
5739,
300,
1199,
29892,
26731,
7206,
3536,
4089,
16715,
7606,
29892,
26731,
7206,
3536,
4089,
29892,
405,
29889,
29979,
8670,
8198,
322,
9724,
17811,
29892,
29871,
29896,
29929,
29947,
29896,
14771,
2701,
310,
15350,
2315,
666,
2785,
29892,
3014,
310,
8046,
5254,
29892,
2292,
27279,
29936,
1102,
280,
361,
322,
399,
575,
682,
29892,
29871,
29896,
29929,
29947,
29906,
29124,
936,
8108,
29879,
297,
341,
1772,
16637,
3457,
3002,
29936,
21806,
369,
313,
3853,
1846,
29871,
29896,
29929,
29947,
29945,
25348,
2233,
28259,
3684,
29889,
306,
322,
1944,
29892,
306,
2241,
5254,
29892,
11045,
29892,
10261,
29936,
379,
1280,
322,
379,
22817,
1144,
313,
3853,
29879,
1846,
29871,
29896,
29929,
29947,
29945,
405,
1682,
280,
293,
7255,
333,
9665,
19515,
2133,
29892,
306,
2241,
5254,
29892,
11045,
29892,
10261,
29936,
322,
3789,
677,
322,
4168,
433,
1581,
29871,
29896,
29929,
29955,
29929,
5739,
7492,
22557,
29901,
14771,
2701,
322,
8108,
29879,
29892,
3684,
29879,
29889,
29871,
29896,
29899,
29946,
29892,
1858,
18605,
5254,
29892,
1570,
3088,
29889,
1976,
1030,
1403,
800,
322,
2245,
264,
695,
1535,
29892,
988,
15723,
29892,
526,
316,
22580,
3918,
297,
278,
1746,
322,
15574,
1304,
297,
10257,
21824,
1338,
1316,
408,
1906,
274,
1573,
1244,
262,
29889,
13,
29961,
29900,
29900,
29945,
29941,
29962,
450,
297,
7316,
1104,
1078,
304,
5541,
2187,
322,
3519,
7199,
5921,
263,
2362,
284,
15795,
18254,
374,
296,
1650,
322,
385,
11828,
5253,
310,
385,
1425,
29890,
29933,
29941,
14172,
392,
29892,
411,
278,
5541,
2187,
1641,
13674,
3889,
310,
724,
398,
29889,
450,
5541,
2187,
322,
3519,
310,
278,
2198,
297,
7316,
526,
5407,
363,
4185,
3864,
9101,
29892,
297,
3153,
29892,
17473,
519,
9101,
29889,
739,
338,
11098,
393,
472,
1422,
3291,
2645,
4185,
3864,
278,
17473,
519,
9101,
29892,
5164,
7117,
1122,
367,
2715,
304,
278,
3038,
9257,
1316,
393,
278,
18350,
508,
1712,
7117,
916,
1135,
1906,
5439,
1244,
262,
29889,
739,
338,
29892,
3138,
29892,
640,
16305,
630,
393,
472,
3203,
472,
697,
1298,
2645,
278,
10223,
362,
310,
278,
9257,
29892,
470,
2645,
278,
9257,
310,
278,
17473,
519,
9101,
29892,
278,
3342,
18350,
7199,
4637,
263,
2362,
284,
15795,
18254,
374,
296,
1650,
322,
263,
2794,
363,
5039,
1218,
1425,
29890,
29933,
29906,
29899,
11851,
287,
7911,
1883,
457,
19015,
559,
29889,
13,
29961,
29900,
29900,
29945,
29946,
29962,
1094,
1304,
1244,
262,
29892,
278,
1840,
376,
29881,
8349,
7268,
519,
3038,
29908,
338,
1304,
304,
8453,
263,
3038,
470,
4665,
310,
9101,
393,
508,
17473,
403,
964,
472,
3203,
22039,
286,
1535,
9101,
29892,
470,
393,
508,
5221,
403,
297,
278,
1422,
11685,
310,
9101,
29892,
321,
29889,
29887,
1696,
285,
1509,
411,
916,
9101,
29892,
393,
508,
17473,
403,
964,
472,
3203,
22039,
286,
1535,
9101,
29889,
1094,
1304,
1244,
262,
29892,
376,
3846,
368,
286,
1535,
9101,
29908,
526,
9101,
393,
10371,
277,
472,
3203,
697,
17443,
310,
278,
17292,
327,
668,
29892,
1316,
408,
18131,
3002,
470,
26823,
4603,
29892,
310,
263,
286,
1535,
3038,
515,
278,
1021,
2894,
470,
260,
15118,
29889,
1152,
1342,
29892,
263,
4226,
29892,
286,
1535,
540,
5031,
22502,
371,
12234,
4653,
267,
1316,
3279,
1144,
408,
3769,
262,
29892,
285,
4626,
262,
6352,
29892,
15595,
29899,
29896,
29899,
424,
277,
719,
567,
262,
29892,
410,
386,
456,
2109,
1067,
327,
1259,
13879,
29892,
6782,
17056,
29892,
322,
1439,
2251,
2450,
427,
14022,
267,
1316,
408,
278,
5094,
517,
18114,
349,
29899,
29946,
29945,
29900,
29879,
29892,
4249,
4045,
29889,
6549,
29892,
408,
3342,
297,
278,
2198,
297,
7316,
29892,
263,
376,
3846,
368,
286,
1535,
540,
5031,
22502,
371,
29908,
1122,
4653,
3769,
262,
470,
1790,
697,
470,
901,
3279,
1144,
29892,
470,
3380,
304,
2125,
278,
10097,
470,
740,
310,
263,
4226,
29892,
286,
1535,
540,
5031,
22502,
371,
29889,
19814,
29892,
263,
376,
3846,
368,
286,
1535,
7243,
1037,
2454,
21762,
3038,
29908,
1122,
7738,
470,
4653,
278,
410,
1144,
352,
262,
26823,
29892,
4249,
4045,
29889,
450,
11509,
310,
278,
9101,
304,
17473,
403,
964,
472,
3203,
22039,
286,
1535,
9101,
674,
451,
367,
14278,
2501,
27878,
2109,
424,
21639,
13698,
29892,
1316,
408,
1301,
20309,
29892,
2466,
278,
9101,
1122,
29892,
310,
3236,
29892,
367,
2531,
300,
1711,
6012,
14561,
29889,
13,
29961,
29900,
29900,
29945,
29945,
29962,
450,
297,
7316,
640,
331,
9884,
5541,
2187,
322,
3519,
5407,
363,
17473,
519,
9101,
29892,
17126,
310,
1009,
2752,
470,
310,
1009,
715,
6288,
537,
29889,
450,
376,
572,
6288,
537,
29908,
310,
263,
3038,
338,
1304,
1244,
262,
20928,
408,
372,
338,
297,
278,
1616,
29889,
13041,
873,
29892,
278,
715,
6288,
537,
310,
263,
3038,
14637,
304,
263,
3038,
29915,
29879,
11509,
304,
17473,
403,
964,
263,
3153,
3038,
1134,
1476,
297,
260,
12175,
470,
1638,
550,
515,
385,
7232,
719,
29877,
29892,
27431,
375,
470,
8906,
2894,
1608,
29889,
450,
376,
5514,
715,
6288,
29908,
263,
3038,
29892,
278,
901,
260,
12175,
964,
607,
278,
3038,
1122,
367,
2221,
304,
17473,
403,
29889,
376,
3247,
332,
666,
327,
296,
9101,
29908,
3160,
9101,
322,
1009,
410,
1885,
29891,
29892,
607,
1122,
367,
2221,
304,
17473,
403,
964,
29892,
470,
2367,
14451,
304,
29892,
715,
332,
666,
327,
296,
29892,
6674,
327,
296,
29892,
288,
3473,
459,
327,
296,
322,
443,
666,
327,
296,
9101,
29892,
322,
29914,
272,
3196,
29892,
565,
451,
599,
29892,
310,
278,
286,
1535,
470,
22039,
286,
1535,
3038,
4072,
1476,
297,
385,
7232,
719,
29877,
29892,
27431,
375,
470,
8906,
2894,
1608,
29889,
376,
6857,
666,
327,
296,
9101,
29908,
3160,
9101,
322,
1009,
410,
1885,
29891,
29892,
607,
1122,
367,
2221,
304,
17473,
403,
964,
29892,
470,
2367,
14451,
304,
29892,
6674,
327,
296,
29892,
288,
3473,
459,
327,
296,
322,
443,
666,
327,
296,
410,
1885,
2105,
9101,
29892,
322,
29914,
272,
697,
470,
901,
286,
1535,
470,
22039,
286,
1535,
3038,
4072,
29892,
5174,
393,
278,
286,
1535,
470,
22039,
286,
1535,
3038,
4072,
10723,
515,
6674,
327,
296,
9101,
526,
9078,
304,
9101,
310,
263,
3153,
260,
15118,
29892,
2894,
470,
2894,
1788,
29889,
1152,
1342,
29892,
263
] | 1,024 |
CommonCrawl
| -0.397296 | 1.448971 | 0.248479 | 0.701754 |
multipotent hematopoietic progenitor cell and/or its progeny possess the ability to differentiate into or give rise to one or more types of oligopotent cells, such as myeloid progenitor cells and lymphoid progenitor cells, and also give rise to other mature cellular components normally found in the blood. "Oligopotent cells" include cells and their progeny whose ability to differentiate into mature or partially mature cells is more restricted than multipotent cells. Oligopotent cells may, however, still possess the ability to differentiate into oligopotent and unipotent cells, and/or one or more mature or partially mature cell types of a given tissue, organ or organ system. One example of an oligopotent cell is a myeloid progenitor cell, which can ultimately give rise to mature or partially mature erythrocytes, platelets, basophils, eosinophils, neutrophils and monocytes. "Unipotent cells" include cells and their progeny that possess the ability to differentiate or give rise to other unipotent cells and/or one type of mature or partially mature cell type.
[0056] Differentiable cells, as used herein, may be pluripotent, multipotent, oligopotent or even unipotent. In certain embodiments of the present invention, the differentiable cells are pluripotent differentiable cells. In more specific embodiments, the pluripotent differentiable cells are selected from the group consisting of embryonic stem cells, ICM/epiblast cells, primitive ectoderm cells, primordial germ cells, and teratocarcinoma cells. In one particular embodiment, the differentiable cells are mammalian embryonic stem cells. In a more particular embodiment, the differentiable cells are human embryonic stem cells.
[0057] The invention also contemplates differentiable cells from any source within an animal, provided the cells are differentiable as defined herein. For example, differentiable cells may be harvested from embryos, or any primordial germ layer therein, from placental or chorion tissue, or from more mature tissue such as adult stem cells including, but not limited to adipose, bone marrow, nervous tissue, mammary tissue, liver tissue, pancreas, epithelial, respiratory, gonadal and muscle tissue. In specific embodiments, the differentiable cells are embryonic stem cells. In other specific embodiments, the differentiable cells are adult stem cells. In still other specific embodiments, the stem cells are placental- or chorionic-derived stem cells.
[0058] Of course, the invention contemplates using differentiable cells from any animal capable of generating differentiable cells. The animals from which the differentiable cells are harvested may be vertebrate or invertebrate, mammalian or non-mammalian, human or non-human. Examples of animal sources include, but are not limited to, primates, rodents, canines, felines, equines, bovines and porcines.
[0059] The differentiable cells of the present invention can be derived using any method known to those of skill in the art. For example, human pluripotent cells can be produced using de-differentiation and nuclear transfer methods. Additionally, the human ICM/epiblast cell or the primitive ectoderm cell used in the present invention can be derived in vivo or in vitro. Primitive ectodermal cells may be generated in adherent culture or as cell aggregates in suspension culture, as described in WO 99/53021. Furthermore, the human pluripotent cells can be passaged using any method known to those of skill in the art, including, manual passaging methods, and bulk passaging methods such as enzymatic or non-enzymatic passaging.
[0060] In certain embodiment, when ES cells are utilized, the embryonic stem cells have a normal karyotype, while in other embodiments, the embryonic stem cells have an abnormal karyotype. In one embodiment, a majority of the embryonic stem cells have a normal karyotype. It is contemplated that greater than 50%, 5
|
[
6674,
327,
296,
298,
4579,
12861,
2035,
293,
410,
1885,
2105,
3038,
322,
29914,
272,
967,
410,
1885,
29891,
22592,
278,
11509,
304,
17473,
403,
964,
470,
2367,
14451,
304,
697,
470,
901,
4072,
310,
288,
3473,
459,
327,
296,
9101,
29892,
1316,
408,
590,
295,
3398,
410,
1885,
2105,
9101,
322,
301,
962,
561,
3398,
410,
1885,
2105,
9101,
29892,
322,
884,
2367,
14451,
304,
916,
286,
1535,
3038,
1070,
7117,
12891,
1476,
297,
278,
10416,
29889,
376,
29949,
3473,
459,
327,
296,
9101,
29908,
3160,
9101,
322,
1009,
410,
1885,
29891,
5069,
11509,
304,
17473,
403,
964,
286,
1535,
470,
22039,
286,
1535,
9101,
338,
901,
22078,
1135,
6674,
327,
296,
9101,
29889,
438,
3473,
459,
327,
296,
9101,
1122,
29892,
3138,
29892,
1603,
22592,
278,
11509,
304,
17473,
403,
964,
288,
3473,
459,
327,
296,
322,
443,
666,
327,
296,
9101,
29892,
322,
29914,
272,
697,
470,
901,
286,
1535,
470,
22039,
286,
1535,
3038,
4072,
310,
263,
2183,
260,
15118,
29892,
2894,
470,
2894,
1788,
29889,
3118,
1342,
310,
385,
288,
3473,
459,
327,
296,
3038,
338,
263,
590,
295,
3398,
410,
1885,
2105,
3038,
29892,
607,
508,
18973,
2367,
14451,
304,
286,
1535,
470,
22039,
286,
1535,
604,
1541,
307,
1270,
2167,
29892,
15284,
10376,
29892,
2362,
3021,
2719,
29892,
321,
359,
262,
3021,
2719,
29892,
11553,
19783,
2719,
322,
1601,
22502,
2167,
29889,
376,
2525,
666,
327,
296,
9101,
29908,
3160,
9101,
322,
1009,
410,
1885,
29891,
393,
22592,
278,
11509,
304,
17473,
403,
470,
2367,
14451,
304,
916,
443,
666,
327,
296,
9101,
322,
29914,
272,
697,
1134,
310,
286,
1535,
470,
22039,
286,
1535,
3038,
1134,
29889,
13,
29961,
29900,
29900,
29945,
29953,
29962,
360,
8349,
7268,
519,
9101,
29892,
408,
1304,
1244,
262,
29892,
1122,
367,
715,
332,
666,
327,
296,
29892,
6674,
327,
296,
29892,
288,
3473,
459,
327,
296,
470,
1584,
443,
666,
327,
296,
29889,
512,
3058,
7232,
397,
7862,
310,
278,
2198,
297,
7316,
29892,
278,
17473,
519,
9101,
526,
715,
332,
666,
327,
296,
17473,
519,
9101,
29889,
512,
901,
2702,
7232,
397,
7862,
29892,
278,
715,
332,
666,
327,
296,
17473,
519,
9101,
526,
4629,
515,
278,
2318,
19849,
310,
7232,
719,
8927,
20805,
9101,
29892,
18340,
29924,
29914,
1022,
747,
4230,
9101,
29892,
19269,
321,
312,
397,
837,
9101,
29892,
1903,
536,
616,
22593,
9101,
29892,
322,
1935,
271,
542,
5666,
262,
4125,
9101,
29889,
512,
697,
3153,
7232,
397,
2073,
29892,
278,
17473,
519,
9101,
526,
286,
4850,
284,
713,
7232,
719,
8927,
20805,
9101,
29889,
512,
263,
901,
3153,
7232,
397,
2073,
29892,
278,
17473,
519,
9101,
526,
5199,
7232,
719,
8927,
20805,
9101,
29889,
13,
29961,
29900,
29900,
29945,
29955,
29962,
450,
297,
7316,
884,
640,
331,
9884,
17473,
519,
9101,
515,
738,
2752,
2629,
385,
13019,
29892,
4944,
278,
9101,
526,
17473,
519,
408,
3342,
1244,
262,
29889,
1152,
1342,
29892,
17473,
519,
9101,
1122,
367,
4023,
29894,
2868,
515,
7232,
719,
359,
29892,
470,
738,
1903,
536,
616,
22593,
7546,
727,
262,
29892,
515,
2174,
1760,
284,
470,
521,
272,
291,
260,
15118,
29892,
470,
515,
901,
286,
1535,
260,
15118,
1316,
408,
16157,
20805,
9101,
3704,
29892,
541,
451,
9078,
304,
594,
666,
852,
29892,
289,
650,
1766,
798,
29892,
23547,
681,
260,
15118,
29892,
286,
4850,
653,
260,
15118,
29892,
619,
369,
260,
15118,
29892,
7243,
1037,
294,
29892,
9358,
389,
295,
616,
29892,
4613,
381,
7606,
29892,
330,
265,
27283,
322,
2301,
2841,
260,
15118,
29889,
512,
2702,
7232,
397,
7862,
29892,
278,
17473,
519,
9101,
526,
7232,
719,
8927,
20805,
9101,
29889,
512,
916,
2702,
7232,
397,
7862,
29892,
278,
17473,
519,
9101,
526,
16157,
20805,
9101,
29889,
512,
1603,
916,
2702,
7232,
397,
7862,
29892,
278,
20805,
9101,
526,
2174,
1760,
284,
29899,
470,
521,
272,
26629,
29899,
672,
2347,
20805,
9101,
29889,
13,
29961,
29900,
29900,
29945,
29947,
29962,
4587,
3236,
29892,
278,
297,
7316,
640,
331,
9884,
773,
17473,
519,
9101,
515,
738,
13019,
15390,
310,
14655,
17473,
519,
9101,
29889,
450,
15006,
515,
607,
278,
17473,
519,
9101,
526,
4023,
29894,
2868,
1122,
367,
24241,
1182,
403,
470,
297,
25996,
1182,
403,
29892,
286,
4850,
284,
713,
470,
1661,
29899,
29885,
4850,
284,
713,
29892,
5199,
470,
1661,
29899,
26029,
29889,
1222,
9422,
310,
13019,
8974,
3160,
29892,
541,
526,
451,
9078,
304,
29892,
1903,
1078,
29892,
15382,
1237,
29892,
508,
1475,
29892,
7995,
1475,
29892,
1592,
1475,
29892,
289,
586,
1475,
322,
1277,
29883,
1475,
29889,
13,
29961,
29900,
29900,
29945,
29929,
29962,
450,
17473,
519,
9101,
310,
278,
2198,
297,
7316,
508,
367,
10723,
773,
738,
1158,
2998,
304,
1906,
310,
19911,
297,
278,
1616,
29889,
1152,
1342,
29892,
5199,
715,
332,
666,
327,
296,
9101,
508,
367,
7371,
773,
316,
29899,
29881,
8349,
7268,
362,
322,
20346,
6782,
3519,
29889,
19814,
29892,
278,
5199,
18340,
29924,
29914,
1022,
747,
4230,
3038,
470,
278,
19269,
321,
312,
397,
837,
3038,
1304,
297,
278,
2198,
297,
7316,
508,
367,
10723,
297,
325,
4243,
470,
297,
13901,
307,
29889,
9724,
3321,
321,
312,
397,
837,
284,
9101,
1122,
367,
5759,
297,
594,
2276,
296,
9257,
470,
408,
3038,
11404,
1078,
297,
8872,
2673,
9257,
29892,
408,
5439,
297,
399,
29949,
29871,
29929,
29929,
29914,
29945,
29941,
29900,
29906,
29896,
29889,
16478,
29892,
278,
5199,
715,
332,
666,
327,
296,
9101,
508,
367,
1209,
4063,
773,
738,
1158,
2998,
304,
1906,
310,
19911,
297,
278,
1616,
29892,
3704,
29892,
12219,
1209,
6751,
3519,
29892,
322,
21610,
1209,
6751,
3519,
1316,
408,
427,
14022,
2454,
470,
1661,
29899,
4666,
962,
2454,
1209,
6751,
29889,
13,
29961,
29900,
29900,
29953,
29900,
29962,
512,
3058,
7232,
397,
2073,
29892,
746,
17956,
9101,
526,
3667,
1891,
29892,
278,
7232,
719,
8927,
20805,
9101,
505,
263,
4226,
413,
653,
327,
668,
29892,
1550,
297,
916,
7232,
397,
7862,
29892,
278,
7232,
719,
8927,
20805,
9101,
505,
385,
633,
8945,
413,
653,
327,
668,
29889,
512,
697,
7232,
397,
2073,
29892,
263,
13638,
310,
278,
7232,
719,
8927,
20805,
9101,
505,
263,
4226,
413,
653,
327,
668,
29889,
739,
338,
640,
16305,
630,
393,
7621,
1135,
29871,
29945,
29900,
13667,
29871,
29945
] | 1,024 |
CommonCrawl
| -0.700248 | 1.661808 | 0.357866 | 0.610683 |
5%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or greater than 95% of metaphases examined will display a normal karyotype.
[0061] In another embodiment, a majority of the embryonic stem cells have an abnormal karyotype. It is contemplated that greater than 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or greater than 95% of metaphases examined will display an abnormal karyotype. In certain embodiments, the abnormal karyotype is evident after the cells have been cultured for greater than 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 20 passages. In one specific embodiment, the abnormal karyotype comprises a trisomy of at least one autosomal chromosome, wherein the autosomal chromosome is selected from the group consisting of chromosomes 1, 7, 8, 12, 14, and 17. In another embodiment, the abnormal karyotype comprises a trisomy of more than one autosomal chromosome, wherein at least one of the more than one autosomal chromosomes is selected from the group consisting of chromosomes 1, 7, 8, 12, 14, and 17. In one embodiment, the autosomal chromosome is chromosome 12 or 17. In another embodiment, the abnormal karyotype comprises an additional sex chromosome. In one embodiment, the karyotype comprises two X chromosomes and one Y chromosome. It is also contemplated that translocations of chromosomes may occur, and such translocations are encompassed within the term "abnormal karyotype." Combinations of the foregoing chromosomal abnormalities and other chromosomal abnormalities are also encompassed by the invention.
[0062] The compositions and methods comprise a basal salt nutrient solution. As used herein, basal salt nutrient solution refers to a mixture of salts that provide cells with water and certain bulk inorganic ions essential for normal cell metabolism, maintain intra- and extra-cellular osmotic balance, provide a carbohydrate as an energy source, and provide a buffering system to maintain the medium within the physiological pH range. Examples of basal salt nutrient solutions include, but are not limited to, Dulbecco's Modified Eagle's Medium (DMEM), Minimal Essential Medium (MEM), Basal Medium Eagle (BME), RPMI 1640, Ham's F-10, Ham's F-12, α-Minimal Essential Medium (αMEM), Glasgow's Minimal Essential Medium (G-MEM), and Iscove's Modified Dulbecco's Medium, and mixtures thereof. In one particular embodiment, the basal salt nutrient solution is an approximately 50:50 mixture of DMEM and Ham's F12.
[0063] It is contemplated that the composition can further comprise trace elements. Trace elements can be purchased commercially, for example, from Mediatech. Non-limiting examples of trace elements include but are not limited to compounds comprising, aluminum, chlorine, sulfate, iron, cadmium, cobalt, chromium, germanium, sodium, potassium, calcium, phosphate and magnesium. Specific example of compounds containing trace elements include but are not limited to, AlCl3, AgNO3, Ba(C2H3O2)2, CdCl2, CdSO4, CoCl2, CrCl3, Cr2(SO4)3, CuSO4, ferric citrate, GeO2, Kl, KBr, LI, molybdic acid, MnSO4, MnCl2, NaF, Na2SiO3, NaVO3, NH4VO3,
|
[
29945,
13667,
29871,
29953,
29900,
13667,
29871,
29953,
29945,
13667,
29871,
29955,
29900,
13667,
29871,
29955,
29945,
13667,
29871,
29947,
29900,
13667,
29871,
29947,
29945,
13667,
29871,
29929,
29900,
29995,
470,
7621,
1135,
29871,
29929,
29945,
29995,
310,
1539,
12451,
2129,
4392,
1312,
674,
2479,
263,
4226,
413,
653,
327,
668,
29889,
13,
29961,
29900,
29900,
29953,
29896,
29962,
512,
1790,
7232,
397,
2073,
29892,
263,
13638,
310,
278,
7232,
719,
8927,
20805,
9101,
505,
385,
633,
8945,
413,
653,
327,
668,
29889,
739,
338,
640,
16305,
630,
393,
7621,
1135,
29871,
29945,
29900,
13667,
29871,
29945,
29945,
13667,
29871,
29953,
29900,
13667,
29871,
29953,
29945,
13667,
29871,
29955,
29900,
13667,
29871,
29955,
29945,
13667,
29871,
29947,
29900,
13667,
29871,
29947,
29945,
13667,
29871,
29929,
29900,
29995,
470,
7621,
1135,
29871,
29929,
29945,
29995,
310,
1539,
12451,
2129,
4392,
1312,
674,
2479,
385,
633,
8945,
413,
653,
327,
668,
29889,
512,
3058,
7232,
397,
7862,
29892,
278,
633,
8945,
413,
653,
327,
668,
338,
13602,
1156,
278,
9101,
505,
1063,
4185,
2955,
363,
7621,
1135,
29871,
29945,
29892,
29871,
29953,
29892,
29871,
29955,
29892,
29871,
29947,
29892,
29871,
29929,
29892,
29871,
29896,
29900,
29892,
29871,
29896,
29896,
29892,
29871,
29896,
29906,
29892,
29871,
29896,
29941,
29892,
29871,
29896,
29946,
29892,
29871,
29896,
29945,
29892,
470,
29871,
29906,
29900,
1209,
1179,
29889,
512,
697,
2702,
7232,
397,
2073,
29892,
278,
633,
8945,
413,
653,
327,
668,
7199,
4637,
263,
534,
275,
16103,
310,
472,
3203,
697,
1120,
359,
290,
284,
25173,
359,
608,
29892,
988,
262,
278,
1120,
359,
290,
284,
25173,
359,
608,
338,
4629,
515,
278,
2318,
19849,
310,
25173,
359,
290,
267,
29871,
29896,
29892,
29871,
29955,
29892,
29871,
29947,
29892,
29871,
29896,
29906,
29892,
29871,
29896,
29946,
29892,
322,
29871,
29896,
29955,
29889,
512,
1790,
7232,
397,
2073,
29892,
278,
633,
8945,
413,
653,
327,
668,
7199,
4637,
263,
534,
275,
16103,
310,
901,
1135,
697,
1120,
359,
290,
284,
25173,
359,
608,
29892,
988,
262,
472,
3203,
697,
310,
278,
901,
1135,
697,
1120,
359,
290,
284,
25173,
359,
290,
267,
338,
4629,
515,
278,
2318,
19849,
310,
25173,
359,
290,
267,
29871,
29896,
29892,
29871,
29955,
29892,
29871,
29947,
29892,
29871,
29896,
29906,
29892,
29871,
29896,
29946,
29892,
322,
29871,
29896,
29955,
29889,
512,
697,
7232,
397,
2073,
29892,
278,
1120,
359,
290,
284,
25173,
359,
608,
338,
25173,
359,
608,
29871,
29896,
29906,
470,
29871,
29896,
29955,
29889,
512,
1790,
7232,
397,
2073,
29892,
278,
633,
8945,
413,
653,
327,
668,
7199,
4637,
385,
5684,
7916,
25173,
359,
608,
29889,
512,
697,
7232,
397,
2073,
29892,
278,
413,
653,
327,
668,
7199,
4637,
1023,
1060,
25173,
359,
290,
267,
322,
697,
612,
25173,
359,
608,
29889,
739,
338,
884,
640,
16305,
630,
393,
1301,
2029,
800,
310,
25173,
359,
290,
267,
1122,
6403,
29892,
322,
1316,
1301,
2029,
800,
526,
427,
2388,
465,
287,
2629,
278,
1840,
376,
370,
8945,
413,
653,
327,
668,
1213,
422,
2109,
800,
310,
278,
363,
2412,
292,
25173,
359,
290,
284,
633,
8945,
1907,
322,
916,
25173,
359,
290,
284,
633,
8945,
1907,
526,
884,
427,
2388,
465,
287,
491,
278,
297,
7316,
29889,
13,
29961,
29900,
29900,
29953,
29906,
29962,
450,
5541,
2187,
322,
3519,
7199,
895,
263,
2362,
284,
15795,
18254,
374,
296,
1650,
29889,
1094,
1304,
1244,
262,
29892,
2362,
284,
15795,
18254,
374,
296,
1650,
14637,
304,
263,
29544,
310,
4497,
1372,
393,
3867,
9101,
411,
4094,
322,
3058,
21610,
297,
6388,
293,
29871,
1080,
18853,
363,
4226,
3038,
1539,
19388,
1608,
29892,
7344,
938,
336,
29899,
322,
4805,
29899,
3729,
1070,
2897,
29885,
13574,
17346,
29892,
3867,
263,
1559,
833,
29882,
2941,
10492,
408,
385,
5864,
2752,
29892,
322,
3867,
263,
6835,
292,
1788,
304,
7344,
278,
18350,
2629,
278,
4824,
29875,
5996,
282,
29950,
3464,
29889,
1222,
9422,
310,
2362,
284,
15795,
18254,
374,
296,
6851,
3160,
29892,
541,
526,
451,
9078,
304,
29892,
360,
352,
915,
26454,
29915,
29879,
3382,
2164,
382,
20860,
29915,
29879,
3436,
1974,
313,
29928,
2303,
29924,
511,
3080,
3039,
11044,
2556,
3436,
1974,
313,
2303,
29924,
511,
4886,
284,
3436,
1974,
382,
20860,
313,
29933,
2303,
511,
390,
29925,
10403,
29871,
29896,
29953,
29946,
29900,
29892,
7904,
29915,
29879,
383,
29899,
29896,
29900,
29892,
7904,
29915,
29879,
383,
29899,
29896,
29906,
29892,
19883,
29899,
8140,
3039,
11044,
2556,
3436,
1974,
313,
30110,
2303,
29924,
511,
29196,
29915,
29879,
3080,
3039,
11044,
2556,
3436,
1974,
313,
29954,
29899,
2303,
29924,
511,
322,
1317,
29883,
994,
29915,
29879,
3382,
2164,
360,
352,
915,
26454,
29915,
29879,
3436,
1974,
29892,
322,
3737,
486,
1973,
727,
974,
29889,
512,
697,
3153,
7232,
397,
2073,
29892,
278,
2362,
284,
15795,
18254,
374,
296,
1650,
338,
385,
14235,
29871,
29945,
29900,
29901,
29945,
29900,
29544,
310,
360,
2303,
29924,
322,
7904,
29915,
29879,
383,
29896,
29906,
29889,
13,
29961,
29900,
29900,
29953,
29941,
29962,
739,
338,
640,
16305,
630,
393,
278,
15259,
508,
4340,
7199,
895,
9637,
3161,
29889,
20597,
3161,
508,
367,
20848,
7825,
5584,
29892,
363,
1342,
29892,
515,
22855,
403,
305,
29889,
10050,
29899,
2576,
11407,
6455,
310,
9637,
3161,
3160,
541,
526,
451,
9078,
304,
752,
3885,
7199,
5921,
29892,
394,
9735,
398,
29892,
521,
5095,
457,
29892,
5394,
29888,
403,
29892,
13977,
29892,
13840,
29885,
1974,
29892,
274,
711,
1997,
29892,
25173,
1974,
29892,
330,
3504,
1974,
29892,
20892,
1974,
29892,
3104,
465,
1974,
29892,
15835,
398,
29892,
1374,
25715,
403,
322,
2320,
4515,
1974,
29889,
21220,
1342,
310,
752,
3885,
6943,
9637,
3161,
3160,
541,
526,
451,
9078,
304,
29892,
838,
6821,
29941,
29892,
4059,
6632,
29941,
29892,
6000,
29898,
29907,
29906,
29950,
29941,
29949,
29906,
29897,
29906,
29892,
315,
29881,
6821,
29906,
29892,
315,
29881,
6156,
29946,
29892,
3189,
6821,
29906,
29892,
6781,
6821,
29941,
29892,
6781,
29906,
29898,
6156,
29946,
29897,
29941,
29892,
10406,
6156,
29946,
29892,
6013,
2200,
7537,
10492,
29892,
1879,
29949,
29906,
29892,
12043,
29892,
476,
12432,
29892,
17705,
29892,
6062,
29891,
6448,
293,
22193,
29892,
341,
29876,
6156,
29946,
29892,
341,
29876,
6821,
29906,
29892,
4465,
29943,
29892,
4465,
29906,
25598,
29949,
29941,
29892,
4465,
24898,
29941,
29892,
405,
29950,
29946,
24898,
29941,
29892
] | 1,024 |
CommonCrawl
| -1.270831 | 0.981769 | 0.014912 | -0.033139 |
(NH4)6Mo7O24, NiSO4, RbCl, selenium, Na2SeO3, H2SeO3, selenite2Na, selenomethionone, SnCl2, ZnSO4, ZrOCl2, and mixtures and salts thereof. If selenium, selenite or selenomethionone is present, it is at a concentration of approximately 0.002 to approximately 0.02 mg/L. In addition, hydroxylapatite may also be present.
[0064] It is contemplated that amino acids can be added to the defined media. Non-limiting examples of such amino acids are Glycine, L-Alanine, L-Alanyl-L-Glutamine, L-Glutamine/Glutamax, L-Arginine hydrochloride, L-Asparagine-H2O, L-Aspartic acid, L-Cysteine hydrochloride-H2O, L-Cystine 2HCl, L-Glutamic Acid, L-Histidine hydrochloride-H2O, L-Isoleucine, L-Leucine, L-Lysine hydrochloride, L-Methionine, L-Phenylalanine, L-Proline, L-Hydroxyproline, L-Serine, L-Threonine, L-Tryptophan, L-Tyrosine disodium salt dihydrate, and L-Valine. In certain embodiments, the amino acid is L-Isoleucine, L-Phenylalanine, L-Proline, L-Hydroxyproline, L-Valine, and mixtures thereof.
[0065] It is also contemplated that the defined medium can comprise ascorbic acid. Preferably ascorbic acid is present at an initial concentration of approximately 1 mg/L to approximately 1000 mg/L, or from approximately 2 mg/L to approximately 500 mg/L, or from approximately 5 mg/L to approximately 100 mg/L, or from approximately 10 mg/L to approximately 100 mg/L or approximately at 50 mg/L.
[0066] In addition, the compositions and methods may also comprise other components such as scrum albumin, transferrin, L-glutamine, lipids, antibiotics, β-Mercaptoethanol, vitamins, minerals, ATP and similar components may be present. Examples of vitamins that may be present include, but are not limited to vitamins A, B1, B2, B3, B5, B6, B7, B9, B12, C, D1, D2, D3, D4, D5, E, tocotrienols, K1 and K2. One of skill in the art can determine the optimal concentration of minerals, vitamins, ATP, lipids, essential fatty acids, etc., for use in a given culture. The concentration of supplements may, for example, be from about 0.001 μM to about 1 mM or more. Specific examples of concentrations at which the supplements may be provided include, but are not limited to about 0.005 μM, 0.01 μM, 0.05 μM, 0.1 μM, 0.5 μM, 1.0 μM, 2.0 μM, 2.5 μM, 3.0 μM, 4.0 μM, 5.0 μM, 10 μM, 20 μM, 100 μM, etc. In one specific embodiment, the compositions and methods comprise vitamin B6 and glutamine. In another specific embodiment, the compositions and methods comprise vitamin C and an iron supplement. in another specific embodiment, the compositions and methods comprise vitamin K1 and vitamin A. In another specific embodiment, the compositions and methods comprise vitamin D3 and ATP. In another specific embodiment, the compositions and methods comprise vitamin B12 and
|
[
313,
29940,
29950,
29946,
29897,
29953,
22638,
29955,
29949,
29906,
29946,
29892,
15981,
6156,
29946,
29892,
390,
29890,
6821,
29892,
18866,
29892,
4465,
29906,
2008,
29949,
29941,
29892,
379,
29906,
2008,
29949,
29941,
29892,
269,
4498,
568,
29906,
13695,
29892,
269,
4498,
290,
621,
291,
650,
29892,
22639,
6821,
29906,
29892,
796,
29876,
6156,
29946,
29892,
796,
29878,
29949,
6821,
29906,
29892,
322,
3737,
486,
1973,
322,
4497,
1372,
727,
974,
29889,
960,
18866,
29892,
269,
4498,
568,
470,
269,
4498,
290,
621,
291,
650,
338,
2198,
29892,
372,
338,
472,
263,
26702,
310,
14235,
29871,
29900,
29889,
29900,
29900,
29906,
304,
14235,
29871,
29900,
29889,
29900,
29906,
286,
29887,
29914,
29931,
29889,
512,
6124,
29892,
17546,
3594,
433,
5031,
568,
1122,
884,
367,
2198,
29889,
13,
29961,
29900,
29900,
29953,
29946,
29962,
739,
338,
640,
16305,
630,
393,
626,
1789,
1274,
4841,
508,
367,
2715,
304,
278,
3342,
5745,
29889,
10050,
29899,
2576,
11407,
6455,
310,
1316,
626,
1789,
1274,
4841,
526,
402,
368,
29883,
457,
29892,
365,
29899,
2499,
273,
457,
29892,
365,
29899,
2499,
1384,
29880,
29899,
29931,
29899,
29954,
29880,
329,
314,
457,
29892,
365,
29899,
29954,
29880,
329,
314,
457,
29914,
29954,
29880,
329,
314,
1165,
29892,
365,
29899,
1433,
5359,
457,
17546,
305,
5095,
680,
29892,
365,
29899,
2887,
862,
22094,
29899,
29950,
29906,
29949,
29892,
365,
29899,
29909,
1028,
442,
293,
22193,
29892,
365,
29899,
29907,
858,
29872,
457,
17546,
305,
5095,
680,
29899,
29950,
29906,
29949,
29892,
365,
29899,
29907,
858,
457,
29871,
29906,
29950,
6821,
29892,
365,
29899,
29954,
29880,
329,
314,
293,
7255,
333,
29892,
365,
29899,
29950,
391,
333,
457,
17546,
305,
5095,
680,
29899,
29950,
29906,
29949,
29892,
365,
29899,
29902,
2170,
1682,
457,
29892,
365,
29899,
3226,
1682,
457,
29892,
365,
29899,
29931,
952,
457,
17546,
305,
5095,
680,
29892,
365,
29899,
29924,
621,
291,
457,
29892,
365,
29899,
29925,
3169,
2904,
284,
273,
457,
29892,
365,
29899,
1184,
1220,
29892,
365,
29899,
29950,
11279,
3594,
771,
1220,
29892,
365,
29899,
1748,
457,
29892,
365,
29899,
1349,
276,
265,
457,
29892,
365,
29899,
29911,
4641,
3021,
273,
29892,
365,
29899,
21314,
1883,
457,
766,
397,
1974,
15795,
652,
29882,
2941,
10492,
29892,
322,
365,
29899,
1440,
457,
29889,
512,
3058,
7232,
397,
7862,
29892,
278,
626,
1789,
22193,
338,
365,
29899,
29902,
2170,
1682,
457,
29892,
365,
29899,
29925,
3169,
2904,
284,
273,
457,
29892,
365,
29899,
1184,
1220,
29892,
365,
29899,
29950,
11279,
3594,
771,
1220,
29892,
365,
29899,
1440,
457,
29892,
322,
3737,
486,
1973,
727,
974,
29889,
13,
29961,
29900,
29900,
29953,
29945,
29962,
739,
338,
884,
640,
16305,
630,
393,
278,
3342,
18350,
508,
7199,
895,
408,
2616,
29890,
293,
22193,
29889,
4721,
571,
2197,
408,
2616,
29890,
293,
22193,
338,
2198,
472,
385,
2847,
26702,
310,
14235,
29871,
29896,
286,
29887,
29914,
29931,
304,
14235,
29871,
29896,
29900,
29900,
29900,
286,
29887,
29914,
29931,
29892,
470,
515,
14235,
29871,
29906,
286,
29887,
29914,
29931,
304,
14235,
29871,
29945,
29900,
29900,
286,
29887,
29914,
29931,
29892,
470,
515,
14235,
29871,
29945,
286,
29887,
29914,
29931,
304,
14235,
29871,
29896,
29900,
29900,
286,
29887,
29914,
29931,
29892,
470,
515,
14235,
29871,
29896,
29900,
286,
29887,
29914,
29931,
304,
14235,
29871,
29896,
29900,
29900,
286,
29887,
29914,
29931,
470,
14235,
472,
29871,
29945,
29900,
286,
29887,
29914,
29931,
29889,
13,
29961,
29900,
29900,
29953,
29953,
29962,
512,
6124,
29892,
278,
5541,
2187,
322,
3519,
1122,
884,
7199,
895,
916,
7117,
1316,
408,
885,
5848,
3769,
262,
29892,
6782,
17056,
29892,
365,
29899,
3820,
329,
314,
457,
29892,
17441,
4841,
29892,
3677,
747,
24414,
1199,
29892,
29871,
30187,
29899,
15836,
17885,
29877,
621,
273,
324,
29892,
13901,
314,
1144,
29892,
1375,
261,
1338,
29892,
27884,
322,
2788,
7117,
1122,
367,
2198,
29889,
1222,
9422,
310,
13901,
314,
1144,
393,
1122,
367,
2198,
3160,
29892,
541,
526,
451,
9078,
304,
13901,
314,
1144,
319,
29892,
350,
29896,
29892,
350,
29906,
29892,
350,
29941,
29892,
350,
29945,
29892,
350,
29953,
29892,
350,
29955,
29892,
350,
29929,
29892,
350,
29896,
29906,
29892,
315,
29892,
360,
29896,
29892,
360,
29906,
29892,
360,
29941,
29892,
360,
29946,
29892,
360,
29945,
29892,
382,
29892,
304,
26235,
19112,
3775,
29892,
476,
29896,
322,
476,
29906,
29889,
3118,
310,
19911,
297,
278,
1616,
508,
8161,
278,
14413,
26702,
310,
1375,
261,
1338,
29892,
13901,
314,
1144,
29892,
27884,
29892,
17441,
4841,
29892,
18853,
9950,
1017,
1274,
4841,
29892,
2992,
1696,
363,
671,
297,
263,
2183,
9257,
29889,
450,
26702,
310,
1462,
944,
29879,
1122,
29892,
363,
1342,
29892,
367,
515,
1048,
29871,
29900,
29889,
29900,
29900,
29896,
29871,
30167,
29924,
304,
1048,
29871,
29896,
286,
29924,
470,
901,
29889,
21220,
6455,
310,
14953,
800,
472,
607,
278,
1462,
944,
29879,
1122,
367,
4944,
3160,
29892,
541,
526,
451,
9078,
304,
1048,
29871,
29900,
29889,
29900,
29900,
29945,
29871,
30167,
29924,
29892,
29871,
29900,
29889,
29900,
29896,
29871,
30167,
29924,
29892,
29871,
29900,
29889,
29900,
29945,
29871,
30167,
29924,
29892,
29871,
29900,
29889,
29896,
29871,
30167,
29924,
29892,
29871,
29900,
29889,
29945,
29871,
30167,
29924,
29892,
29871,
29896,
29889,
29900,
29871,
30167,
29924,
29892,
29871,
29906,
29889,
29900,
29871,
30167,
29924,
29892,
29871,
29906,
29889,
29945,
29871,
30167,
29924,
29892,
29871,
29941,
29889,
29900,
29871,
30167,
29924,
29892,
29871,
29946,
29889,
29900,
29871,
30167,
29924,
29892,
29871,
29945,
29889,
29900,
29871,
30167,
29924,
29892,
29871,
29896,
29900,
29871,
30167,
29924,
29892,
29871,
29906,
29900,
29871,
30167,
29924,
29892,
29871,
29896,
29900,
29900,
29871,
30167,
29924,
29892,
2992,
29889,
512,
697,
2702,
7232,
397,
2073,
29892,
278,
5541,
2187,
322,
3519,
7199,
895,
13901,
9103,
350,
29953,
322,
3144,
329,
314,
457,
29889,
512,
1790,
2702,
7232,
397,
2073,
29892,
278,
5541,
2187,
322,
3519,
7199,
895,
13901,
9103,
315,
322,
385,
13977,
1462,
944,
29889,
297,
1790,
2702,
7232,
397,
2073,
29892,
278,
5541,
2187,
322,
3519,
7199,
895,
13901,
9103,
476,
29896,
322,
13901,
9103,
319,
29889,
512,
1790,
2702,
7232,
397,
2073,
29892,
278,
5541,
2187,
322,
3519,
7199,
895,
13901,
9103,
360,
29941,
322,
27884,
29889,
512,
1790,
2702,
7232,
397,
2073,
29892,
278,
5541,
2187,
322,
3519,
7199,
895,
13901,
9103,
350,
29896,
29906,
322
] | 1,024 |
CommonCrawl
| -1.556123 | 0.60022 | -0.330732 | -0.87365 |
transferrin. In another specific embodiment, the compositions and methods comprise tocotrienols and β-Mercaptoethanol. In another specific embodiment, the compositions and methods comprise glutamine and ATP. In another specific embodiment, the compositions and methods comprise an omega-3 fatty acid and glutamine. In another specific embodiment, the compositions and methods comprise an omega-6 fatty acid and vitamin. B1. In another specific embodiment, the compositions and methods comprise α-linolenic acid and B2.
[0067] The compositions of the present invention are essentially serum free. As used herein, "essentially serum free" refers to the absence of serum in the solutions of the present invention. Serum is not an essential ingredient to the compositions and methods of the present invention. Thus, the presence of serum in any of the compositions should only be attributable to impurities, e.g., from the starting materials or residual serum from the primary cell culture. For example, essentially scrum free medium or environment can contain less than 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1% serum wherein the presently improved bioactive maintenance capacity of the medium or environment is still observed. In a specific embodiment of the present invention, the essentially serum free composition does not contain serum or serum replacement, or only contains trace amounts of serum or serum replacement from the isolation of components of the serum or serum replacement that are added to the defined media.
[0068] The compositions and methods of the present invention also comprise a means for stimulating ErbB2 tyrosine kinase activity within differentiable cells. In one specific embodiment, the compositions and methods of the present invention comprise the presence of at least one ErbB3 ligand. Typically, an ErbB3 ligand will bind the ErbB3 receptor and dimerize with the ErbB2 receptor. The ErbB2 receptor is, in turn, generally responsible for intracellular tyrosine kinase activity within the differentiable cell.
[0069] As used herein, "ErbB3 ligand" refers to a ligand that binds to ErbB3, which in turn dimerizes to ErbB2, thus activating the tyrosine kinase activity of the ErbB2 portion of the ErbB2/ErbB3 heterodimeric receptor. Non-limiting examples of ErbB3 ligands include Neuregulin-1; splice variants and isoforms of Neuregulin-1, including but not limited to HRG-β, HRG-α, Neu Differentiation Factor (NDF), Acetylcholine Receptor-Inducing Activity (ARIA), Glial Growth Factor 2 (GGF2), and Sensory And Motor Neuron-Derived Factor (SMDF); Neuregulin-2; splice variants and isoforms of Neuregulin-2, including but not limited to NRG2-βl Epiregulin; and Biregulin.
[0070] In one embodiment, the means for stimulating ErbB2-directed tyrosine kinase activity comprise at least one ErbB3 ligand that is selected from the group consisting of Neuregulin-1, Heregulin-β (HRG-β), Heregulin-α (HRG-α), Neu differentiation factor (NDF), acetylcholine receptor-inducing activity (ARIA), glial growth factor 2 (GGF2), motor-neuron derived factor (SMDF), Neuregulin-2, Neuregulin-2β (NRG2-β), Epiregulin, Biregulin and variants and functional fragments thereof. In another specific embodiment, the compositions and methods of the present invention comprise more than one means for stimulating ErbB2-directed tyrosine kinase activity, such as, but not limited to, using more than one ErbB3 ligand.
[0071] In a more specific embodiment of the compositions and methods of the present invention, the ErbB3 ligand is HRG-β or a variant or functional
|
[
6782,
17056,
29889,
512,
1790,
2702,
7232,
397,
2073,
29892,
278,
5541,
2187,
322,
3519,
7199,
895,
304,
26235,
19112,
3775,
322,
29871,
30187,
29899,
15836,
17885,
29877,
621,
273,
324,
29889,
512,
1790,
2702,
7232,
397,
2073,
29892,
278,
5541,
2187,
322,
3519,
7199,
895,
3144,
329,
314,
457,
322,
27884,
29889,
512,
1790,
2702,
7232,
397,
2073,
29892,
278,
5541,
2187,
322,
3519,
7199,
895,
385,
2703,
2442,
29899,
29941,
9950,
1017,
22193,
322,
3144,
329,
314,
457,
29889,
512,
1790,
2702,
7232,
397,
2073,
29892,
278,
5541,
2187,
322,
3519,
7199,
895,
385,
2703,
2442,
29899,
29953,
9950,
1017,
22193,
322,
13901,
9103,
29889,
350,
29896,
29889,
512,
1790,
2702,
7232,
397,
2073,
29892,
278,
5541,
2187,
322,
3519,
7199,
895,
19883,
29899,
1915,
18975,
293,
22193,
322,
350,
29906,
29889,
13,
29961,
29900,
29900,
29953,
29955,
29962,
450,
5541,
2187,
310,
278,
2198,
297,
7316,
526,
13674,
724,
398,
3889,
29889,
1094,
1304,
1244,
262,
29892,
376,
404,
9247,
724,
398,
3889,
29908,
14637,
304,
278,
18070,
310,
724,
398,
297,
278,
6851,
310,
278,
2198,
297,
7316,
29889,
1816,
398,
338,
451,
385,
18853,
2348,
1127,
993,
304,
278,
5541,
2187,
322,
3519,
310,
278,
2198,
297,
7316,
29889,
6549,
29892,
278,
10122,
310,
724,
398,
297,
738,
310,
278,
5541,
2187,
881,
871,
367,
1098,
1091,
9246,
304,
2411,
332,
1907,
29892,
321,
29889,
29887,
1696,
515,
278,
6257,
17279,
470,
10995,
950,
724,
398,
515,
278,
7601,
3038,
9257,
29889,
1152,
1342,
29892,
13674,
885,
5848,
3889,
18350,
470,
5177,
508,
1712,
3109,
1135,
29871,
29896,
29900,
29892,
29871,
29929,
29892,
29871,
29947,
29892,
29871,
29955,
29892,
29871,
29953,
29892,
29871,
29945,
29892,
29871,
29946,
29892,
29871,
29941,
29892,
29871,
29906,
29892,
470,
29871,
29896,
29995,
724,
398,
988,
262,
278,
28681,
16710,
17799,
4925,
25413,
13284,
310,
278,
18350,
470,
5177,
338,
1603,
8900,
29889,
512,
263,
2702,
7232,
397,
2073,
310,
278,
2198,
297,
7316,
29892,
278,
13674,
724,
398,
3889,
15259,
947,
451,
1712,
724,
398,
470,
724,
398,
16920,
29892,
470,
871,
3743,
9637,
26999,
310,
724,
398,
470,
724,
398,
16920,
515,
278,
11695,
362,
310,
7117,
310,
278,
724,
398,
470,
724,
398,
16920,
393,
526,
2715,
304,
278,
3342,
5745,
29889,
13,
29961,
29900,
29900,
29953,
29947,
29962,
450,
5541,
2187,
322,
3519,
310,
278,
2198,
297,
7316,
884,
7199,
895,
263,
2794,
363,
20436,
18099,
1425,
29890,
29933,
29906,
7911,
1883,
457,
19015,
559,
6354,
2629,
17473,
519,
9101,
29889,
512,
697,
2702,
7232,
397,
2073,
29892,
278,
5541,
2187,
322,
3519,
310,
278,
2198,
297,
7316,
7199,
895,
278,
10122,
310,
472,
3203,
697,
1425,
29890,
29933,
29941,
14172,
392,
29889,
14213,
1711,
29892,
385,
1425,
29890,
29933,
29941,
14172,
392,
674,
7868,
278,
1425,
29890,
29933,
29941,
337,
14268,
322,
3964,
261,
675,
411,
278,
1425,
29890,
29933,
29906,
337,
14268,
29889,
450,
1425,
29890,
29933,
29906,
337,
14268,
338,
29892,
297,
2507,
29892,
6892,
14040,
363,
938,
945,
514,
1070,
7911,
1883,
457,
19015,
559,
6354,
2629,
278,
17473,
519,
3038,
29889,
13,
29961,
29900,
29900,
29953,
29929,
29962,
1094,
1304,
1244,
262,
29892,
376,
2110,
29890,
29933,
29941,
14172,
392,
29908,
14637,
304,
263,
14172,
392,
393,
7868,
29879,
304,
1425,
29890,
29933,
29941,
29892,
607,
297,
2507,
3964,
261,
7093,
304,
1425,
29890,
29933,
29906,
29892,
4550,
5039,
1218,
278,
7911,
1883,
457,
19015,
559,
6354,
310,
278,
1425,
29890,
29933,
29906,
11910,
310,
278,
1425,
29890,
29933,
29906,
29914,
2110,
29890,
29933,
29941,
25745,
397,
4193,
293,
337,
14268,
29889,
10050,
29899,
2576,
11407,
6455,
310,
1425,
29890,
29933,
29941,
14172,
4167,
3160,
2448,
545,
29887,
352,
262,
29899,
29896,
29936,
805,
5897,
29161,
322,
338,
29877,
9514,
310,
2448,
545,
29887,
352,
262,
29899,
29896,
29892,
3704,
541,
451,
9078,
304,
379,
29934,
29954,
29899,
30187,
29892,
379,
29934,
29954,
29899,
30110,
29892,
17574,
360,
8349,
7268,
362,
383,
7168,
313,
2797,
29943,
511,
7255,
300,
2904,
305,
26496,
830,
14268,
29899,
2568,
1682,
292,
13414,
313,
1718,
10764,
511,
19805,
284,
402,
798,
386,
383,
7168,
29871,
29906,
313,
26788,
29943,
29906,
511,
322,
317,
575,
706,
1126,
16843,
2448,
332,
265,
29899,
15383,
2347,
383,
7168,
313,
17061,
4037,
416,
2448,
545,
29887,
352,
262,
29899,
29906,
29936,
805,
5897,
29161,
322,
338,
29877,
9514,
310,
2448,
545,
29887,
352,
262,
29899,
29906,
29892,
3704,
541,
451,
9078,
304,
27759,
29954,
29906,
29899,
30187,
29880,
14055,
533,
29887,
352,
262,
29936,
322,
350,
533,
29887,
352,
262,
29889,
13,
29961,
29900,
29900,
29955,
29900,
29962,
512,
697,
7232,
397,
2073,
29892,
278,
2794,
363,
20436,
18099,
1425,
29890,
29933,
29906,
29899,
11851,
287,
7911,
1883,
457,
19015,
559,
6354,
7199,
895,
472,
3203,
697,
1425,
29890,
29933,
29941,
14172,
392,
393,
338,
4629,
515,
278,
2318,
19849,
310,
2448,
545,
29887,
352,
262,
29899,
29896,
29892,
2439,
387,
352,
262,
29899,
30187,
313,
20938,
29954,
29899,
30187,
511,
2439,
387,
352,
262,
29899,
30110,
313,
20938,
29954,
29899,
30110,
511,
17574,
1422,
11685,
7329,
313,
2797,
29943,
511,
1274,
300,
2904,
305,
26496,
337,
14268,
29899,
513,
1682,
292,
6354,
313,
1718,
10764,
511,
5857,
284,
14321,
7329,
29871,
29906,
313,
26788,
29943,
29906,
511,
10992,
29899,
16115,
265,
10723,
7329,
313,
17061,
4037,
511,
2448,
545,
29887,
352,
262,
29899,
29906,
29892,
2448,
545,
29887,
352,
262,
29899,
29906,
30187,
313,
16514,
29954,
29906,
29899,
30187,
511,
14055,
533,
29887,
352,
262,
29892,
350,
533,
29887,
352,
262,
322,
29161,
322,
13303,
22370,
727,
974,
29889,
512,
1790,
2702,
7232,
397,
2073,
29892,
278,
5541,
2187,
322,
3519,
310,
278,
2198,
297,
7316,
7199,
895,
901,
1135,
697,
2794,
363,
20436,
18099,
1425,
29890,
29933,
29906,
29899,
11851,
287,
7911,
1883,
457,
19015,
559,
6354,
29892,
1316,
408,
29892,
541,
451,
9078,
304,
29892,
773,
901,
1135,
697,
1425,
29890,
29933,
29941,
14172,
392,
29889,
13,
29961,
29900,
29900,
29955,
29896,
29962,
512,
263,
901,
2702,
7232,
397,
2073,
310,
278,
5541,
2187,
322,
3519,
310,
278,
2198,
297,
7316,
29892,
278,
1425,
29890,
29933,
29941,
14172,
392,
338,
379,
29934,
29954,
29899,
30187,
470,
263,
17305,
470,
13303
] | 1,024 |
CommonCrawl
| -1.300719 | 1.241326 | -0.495596 | -0.531343 |
fragment thereof. In one embodiment, the species from which the culture additive protein, polypeptide or variant or functional fragment thereof derives is the same as the species of cells that are cultured. For example, if mouse ES cells are cultured, an HRG-β with an amino acid sequence that is identical to the mus musculus HRG-β sequence can be used as an additive in culture and is considered to be "of the same species." In other embodiments, the species from which the biological additive derives is different from the cells being cultures. For example, if mouse ES cells are cultured, an HRG-β with an amino acid sequence that is identical to the human HRG-β sequence from can be used as an additive in culture and is considered to be "of different species."
[0072] As used herein, a "functional fragment" is a fragment or splice variant of a full length polypeptide that exerts a similar physiological or cellular effect as the full length polypeptide. The biological effect of the functional fragment need not be identical in scope or strength as the full-length polypeptide, so long as a similar physiological or cellular effect is seen. For example, a functional fragment of HRG-β can detectably stimulate ErbB2-directed tyrosine kinase.
[0073] As used herein, the term "variant" includes chimeric or fusion polypeptides, homologs, analogs, orthologs, and paralogs. In addition, a variant of a reference protein or polypeptide is a protein or polypeptide whose amino acid sequence is at least about 80% identical to the reference protein or polypeptide. In specific embodiments, the variant is at least about 85%, 90%, 95%, 95%, 97%, 98%, 99% or even 100% identical to the reference protein or polypeptide. As used herein, the terms "correspond(s) to" and "corresponding to," as they relate to sequence alignment, are intended to mean enumerated positions within the reference protein or polypeptide, e.g., wild-type human or mouse neuregulin-1, and those positions in the modified protein or polypeptide that align with the positions on the reference protein or polypeptide. Thus, when the amino acid sequence of a subject protein or polypeptide is aligned with the amino acid sequence of a reference protein or polypeptide, the sequence that "corresponds to" certain enumerated positions of the reference protein or polypeptide sequence are those that align with these positions of the reference sequence, but are not necessarily in these exact numerical positions of the reference sequence. Methods for aligning sequences for determining corresponding amino acids between sequences are described below.
[0074] A polypeptide having an amino acid sequence at least, for example, about 95% "identical" to a reference an amino acid sequence encoding, for example TGF-β, is understood to mean that the amino acid sequence of the polypeptide is identical to the reference sequence except that the amino acid sequence may include up to about five modifications per each 100 amino acids of the reference amino acid sequence encoding the reference TGF-β. In other words, to obtain a peptide having an amino acid sequence at least about 95% identical to a reference amino acid sequence, up to about 5% of the amino acid residues of the reference sequence may be deleted or substituted with another amino acid or a number of amino acids up to about 5% of the total amino acids in the reference sequence may be inserted into the reference sequence. These modifications of the reference sequence may occur at the N-terminus or C-terminus positions of the reference amino acid sequence or anywhere between those terminal positions, interspersed either individually among amino acids in the reference sequence or in one or more contiguous groups within the reference sequence.
[0075] As used herein, "identity" is a measure of the identity of nucleotide sequences or amino acid sequences compared to a reference nucleotide or amino acid sequence. In general, the sequences are aligned so that the highest order match is obtained. "Identity" per se has an art-recognized meaning and can be calculated using published techniques. (See, e.g., Computational Molecular Biology, Lesk, A.
|
[
9376,
727,
974,
29889,
512,
697,
7232,
397,
2073,
29892,
278,
6606,
515,
607,
278,
9257,
788,
3321,
26823,
29892,
1248,
668,
415,
680,
470,
17305,
470,
13303,
9376,
727,
974,
589,
3145,
338,
278,
1021,
408,
278,
6606,
310,
9101,
393,
526,
4185,
2955,
29889,
1152,
1342,
29892,
565,
9495,
17956,
9101,
526,
4185,
2955,
29892,
385,
379,
29934,
29954,
29899,
30187,
411,
385,
626,
1789,
22193,
5665,
393,
338,
13557,
304,
278,
2301,
2301,
1810,
375,
379,
29934,
29954,
29899,
30187,
5665,
508,
367,
1304,
408,
385,
788,
3321,
297,
9257,
322,
338,
5545,
304,
367,
376,
974,
278,
1021,
6606,
1213,
512,
916,
7232,
397,
7862,
29892,
278,
6606,
515,
607,
278,
4768,
5996,
788,
3321,
589,
3145,
338,
1422,
515,
278,
9101,
1641,
4185,
1973,
29889,
1152,
1342,
29892,
565,
9495,
17956,
9101,
526,
4185,
2955,
29892,
385,
379,
29934,
29954,
29899,
30187,
411,
385,
626,
1789,
22193,
5665,
393,
338,
13557,
304,
278,
5199,
379,
29934,
29954,
29899,
30187,
5665,
515,
508,
367,
1304,
408,
385,
788,
3321,
297,
9257,
322,
338,
5545,
304,
367,
376,
974,
1422,
6606,
1213,
13,
29961,
29900,
29900,
29955,
29906,
29962,
1094,
1304,
1244,
262,
29892,
263,
376,
2220,
284,
9376,
29908,
338,
263,
9376,
470,
805,
5897,
17305,
310,
263,
2989,
3309,
1248,
668,
415,
680,
393,
429,
814,
29879,
263,
2788,
4824,
29875,
5996,
470,
3038,
1070,
2779,
408,
278,
2989,
3309,
1248,
668,
415,
680,
29889,
450,
4768,
5996,
2779,
310,
278,
13303,
9376,
817,
451,
367,
13557,
297,
6874,
470,
9324,
408,
278,
2989,
29899,
2848,
1248,
668,
415,
680,
29892,
577,
1472,
408,
263,
2788,
4824,
29875,
5996,
470,
3038,
1070,
2779,
338,
3595,
29889,
1152,
1342,
29892,
263,
13303,
9376,
310,
379,
29934,
29954,
29899,
30187,
508,
6459,
2197,
20436,
5987,
1425,
29890,
29933,
29906,
29899,
11851,
287,
7911,
1883,
457,
19015,
559,
29889,
13,
29961,
29900,
29900,
29955,
29941,
29962,
1094,
1304,
1244,
262,
29892,
278,
1840,
376,
19365,
29908,
7805,
521,
4193,
293,
470,
21736,
1248,
668,
415,
2247,
29892,
3632,
1189,
29879,
29892,
3483,
12099,
29892,
14219,
1189,
29879,
29892,
322,
610,
284,
12099,
29889,
512,
6124,
29892,
263,
17305,
310,
263,
3407,
26823,
470,
1248,
668,
415,
680,
338,
263,
26823,
470,
1248,
668,
415,
680,
5069,
626,
1789,
22193,
5665,
338,
472,
3203,
1048,
29871,
29947,
29900,
29995,
13557,
304,
278,
3407,
26823,
470,
1248,
668,
415,
680,
29889,
512,
2702,
7232,
397,
7862,
29892,
278,
17305,
338,
472,
3203,
1048,
29871,
29947,
29945,
13667,
29871,
29929,
29900,
13667,
29871,
29929,
29945,
13667,
29871,
29929,
29945,
13667,
29871,
29929,
29955,
13667,
29871,
29929,
29947,
13667,
29871,
29929,
29929,
29995,
470,
1584,
29871,
29896,
29900,
29900,
29995,
13557,
304,
278,
3407,
26823,
470,
1248,
668,
415,
680,
29889,
1094,
1304,
1244,
262,
29892,
278,
4958,
376,
2616,
3636,
29898,
29879,
29897,
304,
29908,
322,
376,
2616,
3636,
292,
304,
1699,
408,
896,
29279,
304,
5665,
22239,
29892,
526,
9146,
304,
2099,
22447,
630,
11909,
2629,
278,
3407,
26823,
470,
1248,
668,
415,
680,
29892,
321,
29889,
29887,
1696,
8775,
29899,
1853,
5199,
470,
9495,
452,
545,
29887,
352,
262,
29899,
29896,
29892,
322,
1906,
11909,
297,
278,
9120,
26823,
470,
1248,
668,
415,
680,
393,
7595,
411,
278,
11909,
373,
278,
3407,
26823,
470,
1248,
668,
415,
680,
29889,
6549,
29892,
746,
278,
626,
1789,
22193,
5665,
310,
263,
4967,
26823,
470,
1248,
668,
415,
680,
338,
26118,
411,
278,
626,
1789,
22193,
5665,
310,
263,
3407,
26823,
470,
1248,
668,
415,
680,
29892,
278,
5665,
393,
376,
2616,
3636,
29879,
304,
29908,
3058,
22447,
630,
11909,
310,
278,
3407,
26823,
470,
1248,
668,
415,
680,
5665,
526,
1906,
393,
7595,
411,
1438,
11909,
310,
278,
3407,
5665,
29892,
541,
526,
451,
12695,
297,
1438,
2684,
16259,
11909,
310,
278,
3407,
5665,
29889,
8108,
29879,
363,
7595,
292,
15602,
363,
3683,
2827,
6590,
626,
1789,
1274,
4841,
1546,
15602,
526,
5439,
2400,
29889,
13,
29961,
29900,
29900,
29955,
29946,
29962,
319,
1248,
668,
415,
680,
2534,
385,
626,
1789,
22193,
5665,
472,
3203,
29892,
363,
1342,
29892,
1048,
29871,
29929,
29945,
29995,
376,
1693,
936,
29908,
304,
263,
3407,
385,
626,
1789,
22193,
5665,
8025,
29892,
363,
1342,
323,
29954,
29943,
29899,
30187,
29892,
338,
11098,
304,
2099,
393,
278,
626,
1789,
22193,
5665,
310,
278,
1248,
668,
415,
680,
338,
13557,
304,
278,
3407,
5665,
5174,
393,
278,
626,
1789,
22193,
5665,
1122,
3160,
701,
304,
1048,
5320,
26278,
639,
1269,
29871,
29896,
29900,
29900,
626,
1789,
1274,
4841,
310,
278,
3407,
626,
1789,
22193,
5665,
8025,
278,
3407,
323,
29954,
29943,
29899,
30187,
29889,
512,
916,
3838,
29892,
304,
4017,
263,
1236,
415,
680,
2534,
385,
626,
1789,
22193,
5665,
472,
3203,
1048,
29871,
29929,
29945,
29995,
13557,
304,
263,
3407,
626,
1789,
22193,
5665,
29892,
701,
304,
1048,
29871,
29945,
29995,
310,
278,
626,
1789,
22193,
10995,
1041,
310,
278,
3407,
5665,
1122,
367,
11132,
470,
5960,
277,
3860,
411,
1790,
626,
1789,
22193,
470,
263,
1353,
310,
626,
1789,
1274,
4841,
701,
304,
1048,
29871,
29945,
29995,
310,
278,
3001,
626,
1789,
1274,
4841,
297,
278,
3407,
5665,
1122,
367,
15478,
964,
278,
3407,
5665,
29889,
4525,
26278,
310,
278,
3407,
5665,
1122,
6403,
472,
278,
405,
29899,
357,
12254,
470,
315,
29899,
357,
12254,
11909,
310,
278,
3407,
626,
1789,
22193,
5665,
470,
12214,
1546,
1906,
8638,
11909,
29892,
1006,
1028,
414,
287,
2845,
29689,
4249,
626,
1789,
1274,
4841,
297,
278,
3407,
5665,
470,
297,
697,
470,
901,
640,
5526,
681,
6471,
2629,
278,
3407,
5665,
29889,
13,
29961,
29900,
29900,
29955,
29945,
29962,
1094,
1304,
1244,
262,
29892,
376,
22350,
29908,
338,
263,
5645,
310,
278,
10110,
310,
22699,
327,
680,
15602,
470,
626,
1789,
22193,
15602,
9401,
304,
263,
3407,
22699,
327,
680,
470,
626,
1789,
22193,
5665,
29889,
512,
2498,
29892,
278,
15602,
526,
26118,
577,
393,
278,
9939,
1797,
1993,
338,
7625,
29889,
376,
18415,
29908,
639,
409,
756,
385,
1616,
29899,
29423,
1891,
6593,
322,
508,
367,
12833,
773,
6369,
13698,
29889,
313,
13393,
29892,
321,
29889,
29887,
1696,
11796,
1288,
341,
1772,
16637,
3457,
3002,
29892,
2664,
29895,
29892,
319,
29889
] | 1,024 |
CommonCrawl
| -0.950218 | 1.589132 | -0.157462 | 0.166711 |
M., ed., Oxford University Press, New York (1988); Biocomputing: Informatics And Genome Projects, Smith, D. W., ed., Academic Press, New York (1993); Computer Analysis of Sequence Data, Part I, Griffin, A. M., and Griffin, H. G., eds., Humana Press, New Jersey (1994); von Heinje, G., Sequence Analysis In Molecular Biology, Academic Press (1987); and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York (1991)). While there exist several methods to measure identity between two polynucleotide or polypeptide sequences, the term "identity" is well known to skilled artisans (Carillo, H. & Lipton, D., Siam J Applied Math 48:1073 (1988)). Methods commonly employed to determine identity or similarity between two sequences include, but are not limited to, those disclosed in Guide to Huge Computers, Martin J. Bishop, ed., Academic Press, San Diego (1994) and Carillo, H. & Lipton, D., Siam J Applied Math 48:1073 (1988). Computer programs may also contain methods and algorithms that calculate identity and similarity. Examples of computer program methods to determine identity and similarity between two sequences include, but are not limited to, GCG program package (Devereux, J., et al., Nucleic Acids Research 12(i):387 (1984)), BLASTP, ExPASy, BLASTN, FASTA (Atschul, S. F., et al., J Molec Biol 215:403 (1990)) and FASTDB. Examples of methods to determine identity and similarity are discussed in Michaels, G. and Garian, R., Current Protocols in Protein Science, Vol I, John Wiley & Sons, Inc. (2000), which is incorporated by reference. In one embodiment of the present invention, the algorithm used to determine identity between two or more polypeptides is BLASTP.
[0076] In another embodiment of the present invention, the algorithm used to determine identity between two or more polypeptides is FASTDB, which is based upon the algorithm of Brutlag et al. (Comp. App. Biosci. 6:237-245 (1990), incorporated by reference). In a FASTDB sequence alignment, the query and subject sequences are amino sequences. The result of sequence alignment is in percent identity. Parameters that may be used in a FASTDB alignment of amino acid sequences to calculate percent identity include, but are not limited to: Matrix=PAM, k-tuple=2, Mismatch Penalty=1, Joining Penalty=20, Randomization Group Length=0, Cutoff Score=1, Gap Penalty=5, Gap Size Penalty 0.05, Window Size=500 or the length of the subject amino sequence, whichever is shorter.
[0077] If the subject sequence is shorter or longer than the query sequence because of N-terminus or C-terminus additions or deletions, not because of internal additions or deletions, a manual correction can be made, because the FASTDB program does not account for N-terminus and C-terminus truncations or additions of the subject sequence when calculating percent identity. For subject sequences truncated at the 5' or 3' ends, relative to the query sequence, the percent identity is corrected by calculating the number of bases of the query sequence that are N- and C-terminus to the reference sequence that are not matched/aligned, as a percent of the total bases of the query sequence. The results of the FASTDB sequence alignment determine matching/alignment. The alignment percentage is then subtracted from the percent identity, calculated by the above FASTDB program using the specified parameters, to arrive at a final percent identity score. This corrected score can be used for the purposes of determining how alignments "correspond" to each other, as well as percentage identity. Residues of the query (subject) sequences or the reference sequence that extend past the N- or C-termini of the reference or subject sequence, respectively, may be considered for the purposes of manually adjusting the percent identity score. That is,
|
[
341,
1696,
1226,
1696,
11045,
3014,
5254,
29892,
1570,
3088,
313,
29896,
29929,
29947,
29947,
416,
21184,
12097,
292,
29901,
512,
4830,
1199,
1126,
5739,
608,
8010,
29879,
29892,
7075,
29892,
360,
29889,
399,
1696,
1226,
1696,
16146,
293,
5254,
29892,
1570,
3088,
313,
29896,
29929,
29929,
29941,
416,
20972,
24352,
310,
922,
3910,
3630,
29892,
3455,
306,
29892,
22990,
262,
29892,
319,
29889,
341,
1696,
322,
22990,
262,
29892,
379,
29889,
402,
1696,
1226,
29879,
1696,
13863,
1648,
5254,
29892,
1570,
14500,
313,
29896,
29929,
29929,
29946,
416,
1005,
9515,
1324,
29892,
402,
1696,
922,
3910,
24352,
512,
341,
1772,
16637,
3457,
3002,
29892,
16146,
293,
5254,
313,
29896,
29929,
29947,
29955,
416,
322,
922,
3910,
24352,
1588,
4193,
29892,
402,
1091,
808,
586,
29892,
341,
29889,
322,
897,
9359,
1314,
29892,
435,
1696,
1226,
29879,
1696,
341,
10224,
880,
5254,
29892,
1570,
3088,
313,
29896,
29929,
29929,
29896,
8106,
5806,
727,
1863,
3196,
3519,
304,
5645,
10110,
1546,
1023,
1248,
948,
1682,
280,
327,
680,
470,
1248,
668,
415,
680,
15602,
29892,
278,
1840,
376,
22350,
29908,
338,
1532,
2998,
304,
2071,
24455,
1616,
275,
550,
313,
8179,
9093,
29892,
379,
29889,
669,
2718,
12533,
29892,
360,
1696,
317,
2829,
435,
2401,
2957,
5792,
29871,
29946,
29947,
29901,
29896,
29900,
29955,
29941,
313,
29896,
29929,
29947,
29947,
8106,
8108,
29879,
15574,
15723,
304,
8161,
10110,
470,
29501,
1546,
1023,
15602,
3160,
29892,
541,
526,
451,
9078,
304,
29892,
1906,
766,
15603,
297,
16886,
304,
379,
5710,
11796,
414,
29892,
6502,
435,
29889,
16880,
29892,
1226,
1696,
16146,
293,
5254,
29892,
3087,
16879,
313,
29896,
29929,
29929,
29946,
29897,
322,
1704,
9093,
29892,
379,
29889,
669,
2718,
12533,
29892,
360,
1696,
317,
2829,
435,
2401,
2957,
5792,
29871,
29946,
29947,
29901,
29896,
29900,
29955,
29941,
313,
29896,
29929,
29947,
29947,
467,
20972,
11104,
1122,
884,
1712,
3519,
322,
14009,
393,
8147,
10110,
322,
29501,
29889,
1222,
9422,
310,
6601,
1824,
3519,
304,
8161,
10110,
322,
29501,
1546,
1023,
15602,
3160,
29892,
541,
526,
451,
9078,
304,
29892,
402,
11135,
1824,
3577,
313,
29928,
1310,
29872,
1314,
29892,
435,
1696,
634,
394,
1696,
405,
1682,
280,
293,
7255,
4841,
10550,
29871,
29896,
29906,
29898,
29875,
1125,
29941,
29947,
29955,
313,
29896,
29929,
29947,
29946,
8243,
350,
4375,
1254,
29925,
29892,
1222,
29925,
3289,
29891,
29892,
350,
4375,
1254,
29940,
29892,
13515,
1254,
29909,
313,
4178,
816,
352,
29892,
317,
29889,
383,
1696,
634,
394,
1696,
435,
341,
1772,
29883,
3457,
324,
29871,
29906,
29896,
29945,
29901,
29946,
29900,
29941,
313,
29896,
29929,
29929,
29900,
876,
322,
13515,
1254,
4051,
29889,
1222,
9422,
310,
3519,
304,
8161,
10110,
322,
29501,
526,
15648,
297,
3375,
29874,
1379,
29892,
402,
29889,
322,
7455,
713,
29892,
390,
1696,
9626,
1019,
5770,
29879,
297,
14409,
262,
9327,
29892,
3684,
306,
29892,
2259,
399,
15168,
669,
317,
787,
29892,
9266,
29889,
313,
29906,
29900,
29900,
29900,
511,
607,
338,
11039,
630,
491,
3407,
29889,
512,
697,
7232,
397,
2073,
310,
278,
2198,
297,
7316,
29892,
278,
5687,
1304,
304,
8161,
10110,
1546,
1023,
470,
901,
1248,
668,
415,
2247,
338,
350,
4375,
1254,
29925,
29889,
13,
29961,
29900,
29900,
29955,
29953,
29962,
512,
1790,
7232,
397,
2073,
310,
278,
2198,
297,
7316,
29892,
278,
5687,
1304,
304,
8161,
10110,
1546,
1023,
470,
901,
1248,
668,
415,
2247,
338,
13515,
1254,
4051,
29892,
607,
338,
2729,
2501,
278,
5687,
310,
1771,
329,
3110,
634,
394,
29889,
313,
6843,
29889,
2401,
29889,
350,
2363,
455,
29889,
29871,
29953,
29901,
29906,
29941,
29955,
29899,
29906,
29946,
29945,
313,
29896,
29929,
29929,
29900,
511,
11039,
630,
491,
3407,
467,
512,
263,
13515,
1254,
4051,
5665,
22239,
29892,
278,
2346,
322,
4967,
15602,
526,
626,
1789,
15602,
29889,
450,
1121,
310,
5665,
22239,
338,
297,
10151,
10110,
29889,
12662,
2699,
393,
1122,
367,
1304,
297,
263,
13515,
1254,
4051,
22239,
310,
626,
1789,
22193,
15602,
304,
8147,
10151,
10110,
3160,
29892,
541,
526,
451,
9078,
304,
29901,
22513,
29922,
29925,
5194,
29892,
413,
29899,
23583,
29922,
29906,
29892,
341,
1608,
905,
7363,
18745,
29922,
29896,
29892,
3650,
2827,
7363,
18745,
29922,
29906,
29900,
29892,
16968,
2133,
6431,
365,
1477,
29922,
29900,
29892,
315,
329,
2696,
2522,
487,
29922,
29896,
29892,
402,
481,
7363,
18745,
29922,
29945,
29892,
402,
481,
21179,
7363,
18745,
29871,
29900,
29889,
29900,
29945,
29892,
18379,
21179,
29922,
29945,
29900,
29900,
470,
278,
3309,
310,
278,
4967,
626,
1789,
5665,
29892,
377,
4070,
369,
338,
20511,
29889,
13,
29961,
29900,
29900,
29955,
29955,
29962,
960,
278,
4967,
5665,
338,
20511,
470,
5520,
1135,
278,
2346,
5665,
1363,
310,
405,
29899,
357,
12254,
470,
315,
29899,
357,
12254,
788,
2187,
470,
7374,
1080,
29892,
451,
1363,
310,
7463,
788,
2187,
470,
7374,
1080,
29892,
263,
12219,
26385,
508,
367,
1754,
29892,
1363,
278,
13515,
1254,
4051,
1824,
947,
451,
3633,
363,
405,
29899,
357,
12254,
322,
315,
29899,
357,
12254,
21022,
800,
470,
788,
2187,
310,
278,
4967,
5665,
746,
25202,
10151,
10110,
29889,
1152,
4967,
15602,
21022,
630,
472,
278,
29871,
29945,
29915,
470,
29871,
29941,
29915,
10614,
29892,
6198,
304,
278,
2346,
5665,
29892,
278,
10151,
10110,
338,
24114,
491,
25202,
278,
1353,
310,
22561,
310,
278,
2346,
5665,
393,
526,
405,
29899,
322,
315,
29899,
357,
12254,
304,
278,
3407,
5665,
393,
526,
451,
19228,
29914,
13671,
29892,
408,
263,
10151,
310,
278,
3001,
22561,
310,
278,
2346,
5665,
29889,
450,
2582,
310,
278,
13515,
1254,
4051,
5665,
22239,
8161,
9686,
29914,
2520,
358,
29889,
450,
22239,
19649,
338,
769,
23197,
287,
515,
278,
10151,
10110,
29892,
12833,
491,
278,
2038,
13515,
1254,
4051,
1824,
773,
278,
6790,
4128,
29892,
304,
18331,
472,
263,
2186,
10151,
10110,
8158,
29889,
910,
24114,
8158,
508,
367,
1304,
363,
278,
11976,
310,
3683,
2827,
920,
7595,
1860,
376,
2616,
3636,
29908,
304,
1269,
916,
29892,
408,
1532,
408,
19649,
10110,
29889,
2538,
333,
1041,
310,
278,
2346,
313,
16009,
29897,
15602,
470,
278,
3407,
5665,
393,
10985,
4940,
278,
405,
29899,
470,
315,
29899,
18821,
29875,
310,
278,
3407,
470,
4967,
5665,
29892,
8307,
29892,
1122,
367,
5545,
363,
278,
11976,
310,
7522,
10365,
292,
278,
10151,
10110,
8158,
29889,
2193,
338,
29892
] | 1,024 |
CommonCrawl
| -0.943425 | 1.495692 | 0.502669 | 0.625861 |
residues that are not matched/aligned with the N- or C-termini of the comparison sequence may be counted when manually adjusting the percent identity score or alignment numbering.
[0078] For example, a 90 amino acid residue subject sequence is aligned with a 100 residue reference sequence to determine percent identity. The deletion occurs at the N-terminus of the subject sequence and therefore, the FASTDB alignment does not show a match/alignment of the first 10 residues at the N-terminus. The 10 unpaired residues represent 10% of the sequence (number of residues at the N- and C-termini not matched/total number of residues in the query sequence) so 10% is subtracted from the percent identity score calculated by the FASTDB program. If the remaining 90 residues were perfectly matched the final percent identity would be 90%. In another example, a 90 residue subject sequence is compared with a 100 reference sequence. This time the deletions are internal deletions so there are no residues at the N- or C-termini of the subject sequence which are not matched/aligned with the query. In this case the percent identity calculated by FASTDB is not manually corrected.
[0079] The invention also provides chimeric or fusion polypeptides. As used herein, a "chimeric polypeptide" or "fusion polypeptide" comprises at least a portion of a member of the reference polypeptide operatively linked to a second, different polypeptide. The second polypeptide has an amino acid sequence corresponding to a polypeptide which is not substantially identical to the reference polypeptide, and which is derived from the same or a different organism. With respect to the fusion polypeptide, the term "operatively linked" is intended to indicate that the reference polypeptide and the second polypeptide are fused to each other so that both sequences fulfill the proposed function attributed to the sequence used. The second polypeptide can be fused to the N-terminus or C-terminus of the reference polypeptide. For example, in one embodiment, the fusion polypeptide is a GST-IGF-1 fusion polypeptide in which an IGF-1 sequence is fused to the C-terminus of the GST sequences. Such fusion polypeptides can facilitate the purification of recombinant polypeptides. In another embodiment, the fusion polypeptide can contain a heterologous signal sequence at its N-terminus. In certain host cells (e.g., mammalian host cells), expression and/or secretion of a polypeptide can be increased through use of a heterologous signal sequence.
[0080] In addition to fragments and fusion polypeptides, the present invention includes homologs and analogs of naturally occurring polypeptides. "Homologs" are defined herein as two nucleic acids or polypeptides that have similar, or "identical," nucleotide or amino acid sequences, respectively. Homologs include allelic variants, orthologs, paralogs, agonists, and antagonists as defined hereafter. The term "homolog" further encompasses nucleic acid molecules that differ from a reference nucleotide sequence due to degeneracy of the genetic code and thus encode the same polypeptide as that encoded by the reference nucleotide sequence. As used herein, "naturally occurring" refers to a nucleic or amino acid sequence that occurs in nature.
[0081] An agonist of a polypeptide can retain substantially the same, or a subset, of the biological activities of the polypeptide. An antagonist of a polypeptide can inhibit one or more of the activities of the naturally occurring form of the polypeptide.
[0082] In another more specific embodiment of the compositions and methods of the present invention, the ErbB3 ligand is HRG-β or a variant or a functional fragment thereof. Additional, non-limiting examples of ErbB3 ligands are disclosed in U.S. Pat. No. 6,136,558, 6,387,638, and 7,063,961, which are incorporated by reference.
[0083] Heregulins are generally class
|
[
10995,
1041,
393,
526,
451,
19228,
29914,
13671,
411,
278,
405,
29899,
470,
315,
29899,
18821,
29875,
310,
278,
10230,
5665,
1122,
367,
29115,
746,
7522,
10365,
292,
278,
10151,
10110,
8158,
470,
22239,
1353,
292,
29889,
13,
29961,
29900,
29900,
29955,
29947,
29962,
1152,
1342,
29892,
263,
29871,
29929,
29900,
626,
1789,
22193,
10995,
434,
4967,
5665,
338,
26118,
411,
263,
29871,
29896,
29900,
29900,
10995,
434,
3407,
5665,
304,
8161,
10151,
10110,
29889,
450,
7374,
291,
10008,
472,
278,
405,
29899,
357,
12254,
310,
278,
4967,
5665,
322,
5480,
29892,
278,
13515,
1254,
4051,
22239,
947,
451,
1510,
263,
1993,
29914,
2520,
358,
310,
278,
937,
29871,
29896,
29900,
10995,
1041,
472,
278,
405,
29899,
357,
12254,
29889,
450,
29871,
29896,
29900,
443,
3274,
2859,
10995,
1041,
2755,
29871,
29896,
29900,
29995,
310,
278,
5665,
313,
4537,
310,
10995,
1041,
472,
278,
405,
29899,
322,
315,
29899,
18821,
29875,
451,
19228,
29914,
7827,
1353,
310,
10995,
1041,
297,
278,
2346,
5665,
29897,
577,
29871,
29896,
29900,
29995,
338,
23197,
287,
515,
278,
10151,
10110,
8158,
12833,
491,
278,
13515,
1254,
4051,
1824,
29889,
960,
278,
9886,
29871,
29929,
29900,
10995,
1041,
892,
7970,
19228,
278,
2186,
10151,
10110,
723,
367,
29871,
29929,
29900,
15543,
512,
1790,
1342,
29892,
263,
29871,
29929,
29900,
10995,
434,
4967,
5665,
338,
9401,
411,
263,
29871,
29896,
29900,
29900,
3407,
5665,
29889,
910,
931,
278,
7374,
1080,
526,
7463,
7374,
1080,
577,
727,
526,
694,
10995,
1041,
472,
278,
405,
29899,
470,
315,
29899,
18821,
29875,
310,
278,
4967,
5665,
607,
526,
451,
19228,
29914,
13671,
411,
278,
2346,
29889,
512,
445,
1206,
278,
10151,
10110,
12833,
491,
13515,
1254,
4051,
338,
451,
7522,
24114,
29889,
13,
29961,
29900,
29900,
29955,
29929,
29962,
450,
297,
7316,
884,
8128,
521,
4193,
293,
470,
21736,
1248,
668,
415,
2247,
29889,
1094,
1304,
1244,
262,
29892,
263,
376,
305,
4193,
293,
1248,
668,
415,
680,
29908,
470,
376,
29888,
3958,
1248,
668,
415,
680,
29908,
7199,
4637,
472,
3203,
263,
11910,
310,
263,
4509,
310,
278,
3407,
1248,
668,
415,
680,
1751,
6703,
9024,
304,
263,
1473,
29892,
1422,
1248,
668,
415,
680,
29889,
450,
1473,
1248,
668,
415,
680,
756,
385,
626,
1789,
22193,
5665,
6590,
304,
263,
1248,
668,
415,
680,
607,
338,
451,
23228,
368,
13557,
304,
278,
3407,
1248,
668,
415,
680,
29892,
322,
607,
338,
10723,
515,
278,
1021,
470,
263,
1422,
2894,
1608,
29889,
2973,
3390,
304,
278,
21736,
1248,
668,
415,
680,
29892,
278,
1840,
376,
3372,
6703,
9024,
29908,
338,
9146,
304,
12266,
393,
278,
3407,
1248,
668,
415,
680,
322,
278,
1473,
1248,
668,
415,
680,
526,
285,
3880,
304,
1269,
916,
577,
393,
1716,
15602,
6095,
5589,
278,
7972,
740,
29393,
304,
278,
5665,
1304,
29889,
450,
1473,
1248,
668,
415,
680,
508,
367,
285,
3880,
304,
278,
405,
29899,
357,
12254,
470,
315,
29899,
357,
12254,
310,
278,
3407,
1248,
668,
415,
680,
29889,
1152,
1342,
29892,
297,
697,
7232,
397,
2073,
29892,
278,
21736,
1248,
668,
415,
680,
338,
263,
402,
1254,
29899,
6259,
29943,
29899,
29896,
21736,
1248,
668,
415,
680,
297,
607,
385,
306,
29954,
29943,
29899,
29896,
5665,
338,
285,
3880,
304,
278,
315,
29899,
357,
12254,
310,
278,
402,
1254,
15602,
29889,
10506,
21736,
1248,
668,
415,
2247,
508,
16089,
10388,
278,
3708,
2450,
310,
27878,
2109,
424,
1248,
668,
415,
2247,
29889,
512,
1790,
7232,
397,
2073,
29892,
278,
21736,
1248,
668,
415,
680,
508,
1712,
263,
25745,
1189,
681,
7182,
5665,
472,
967,
405,
29899,
357,
12254,
29889,
512,
3058,
3495,
9101,
313,
29872,
29889,
29887,
1696,
286,
4850,
284,
713,
3495,
9101,
511,
4603,
322,
29914,
272,
7035,
291,
310,
263,
1248,
668,
415,
680,
508,
367,
11664,
1549,
671,
310,
263,
25745,
1189,
681,
7182,
5665,
29889,
13,
29961,
29900,
29900,
29947,
29900,
29962,
512,
6124,
304,
22370,
322,
21736,
1248,
668,
415,
2247,
29892,
278,
2198,
297,
7316,
7805,
3632,
1189,
29879,
322,
3483,
12099,
310,
18180,
13920,
292,
1248,
668,
415,
2247,
29889,
376,
24259,
1189,
29879,
29908,
526,
3342,
1244,
262,
408,
1023,
22699,
293,
1274,
4841,
470,
1248,
668,
415,
2247,
393,
505,
2788,
29892,
470,
376,
1693,
936,
1699,
22699,
327,
680,
470,
626,
1789,
22193,
15602,
29892,
8307,
29889,
15089,
1189,
29879,
3160,
4788,
506,
29161,
29892,
14219,
1189,
29879,
29892,
610,
284,
12099,
29892,
946,
265,
2879,
29892,
322,
3677,
12841,
2879,
408,
3342,
1244,
7045,
29889,
450,
1840,
376,
9706,
1189,
29908,
4340,
427,
2388,
465,
267,
22699,
293,
22193,
13206,
21337,
393,
1163,
515,
263,
3407,
22699,
327,
680,
5665,
2861,
304,
3587,
759,
4135,
310,
278,
2531,
7492,
775,
322,
4550,
19750,
278,
1021,
1248,
668,
415,
680,
408,
393,
18511,
491,
278,
3407,
22699,
327,
680,
5665,
29889,
1094,
1304,
1244,
262,
29892,
376,
29876,
1337,
635,
13920,
292,
29908,
14637,
304,
263,
22699,
293,
470,
626,
1789,
22193,
5665,
393,
10008,
297,
5469,
29889,
13,
29961,
29900,
29900,
29947,
29896,
29962,
530,
946,
265,
391,
310,
263,
1248,
668,
415,
680,
508,
11551,
23228,
368,
278,
1021,
29892,
470,
263,
11306,
29892,
310,
278,
4768,
5996,
14188,
310,
278,
1248,
668,
415,
680,
29889,
530,
3677,
12841,
391,
310,
263,
1248,
668,
415,
680,
508,
297,
6335,
277,
697,
470,
901,
310,
278,
14188,
310,
278,
18180,
13920,
292,
883,
310,
278,
1248,
668,
415,
680,
29889,
13,
29961,
29900,
29900,
29947,
29906,
29962,
512,
1790,
901,
2702,
7232,
397,
2073,
310,
278,
5541,
2187,
322,
3519,
310,
278,
2198,
297,
7316,
29892,
278,
1425,
29890,
29933,
29941,
14172,
392,
338,
379,
29934,
29954,
29899,
30187,
470,
263,
17305,
470,
263,
13303,
9376,
727,
974,
29889,
3462,
3245,
29892,
1661,
29899,
2576,
11407,
6455,
310,
1425,
29890,
29933,
29941,
14172,
4167,
526,
766,
15603,
297,
501,
29889,
29903,
29889,
4121,
29889,
1939,
29889,
29871,
29953,
29892,
29896,
29941,
29953,
29892,
29945,
29945,
29947,
29892,
29871,
29953,
29892,
29941,
29947,
29955,
29892,
29953,
29941,
29947,
29892,
322,
29871,
29955,
29892,
29900,
29953,
29941,
29892,
29929,
29953,
29896,
29892,
607,
526,
11039,
630,
491,
3407,
29889,
13,
29961,
29900,
29900,
29947,
29941,
29962,
2439,
387,
352,
1144,
526,
6892,
770
] | 1,024 |
CommonCrawl
| -0.859196 | 1.464545 | 0.167784 | 0.121175 |
ified into two major types, alpha and beta, based on two variant EGF-like domains that differ in their C-terminal portions. These EGF-like domains, however, are identical in the spacing of six cysteine residues contained therein. Based on an amino acid sequence comparison, Holmes et al. found that between the first and sixth cysteines in the EGF-like domain, HRGs were 45% similar to heparin-binding EGF-like growth factor (HB-EGF), 35% identical to amphiregulin (AR), 32% identical to TGF-α, and 27% identical to EGF.
[0084] The 44 kDa neu differentiation factor (NDF) is the rat equivalent of human HRG. Like the HRG polypeptides, NDF has an immunoglobulin (Ig) homology domain followed by an EGF-like domain and lacks a N-terminal signal peptide. Presently, there are at least six distinct fibroblastic pro-NDFs, classified as either alpha or beta polypeptides, based on the sequences of the EGF-like domains. Isoforms 1 to 4 are characterized on the basis of a variable stretch between the EGF-like domain and transmembrane domain. Thus it appears that different NDF isoforms are generated by alternative splicing and may perform distinct tissue-specific functions. See EP 505 148; WO 93/22424; and WO 94/28133, which are incorporated by reference.
[0085] In one embodiment of the present invention, the compositions and methods are free of exogenous insulin and insulin substitutes. The phrase "exogenous insulin or insulin substitutes" is used herein to indicate insulin or insulin substitutes that is/are not intentionally added to the compositions or methods of the present invention. Thus, in certain embodiments of the present invention, the methods and compositions are free of insulin or insulin substitutes that are intentionally supplied. The compositions or methods may, however, not necessarily be free of endogenous insulin. As used herein, "endogenous insulin" indicates that the cultured cells may be producing insulin of their own accord when cultured according to the methods of the present invention. Endogenous insulin also may be used to indicate residual impurities from the primary cell culture or impurities from the starting materials. In specific examples, the compositions and methods of the present contain less than 50, 45, 40, 35, 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 μg/ml of insulin.
[0086] As used herein, the term "insulin" refers to the protein, or variant or fragment thereof that binds to the insulin receptor in normal physiological concentrations and can induce signaling through the insulin receptor. The term "insulin" encompasses a protein having the polypeptide sequence of native human insulin, or of other mammalian insulin, or of any homologs or variants to these sequences. Additionally, the term insulin encompasses polypeptide fragments that are capable of binding to the insulin receptor to induce signaling through the insulin receptor. The term "insulin substitute" refers to any zinc containing compound that may be used in place of insulin to give substantially similar results as insulin. Examples of insulin substitutes include, but are not limited to zinc chloride, zinc nitrate, zinc bromide, and zinc sulfate.
[0087] To be clear, insulin-like growth factors are not insulin substitutes or homologs of insulin, as contemplated in the present invention. Accordingly, in another specific embodiment, the compositions and methods of the present invention comprise the use of at least one insulin-like growth factor (IGF) or a variant or a functional fragment thereof. In another embod
|
[
2164,
964,
1023,
4655,
4072,
29892,
15595,
322,
21762,
29892,
2729,
373,
1023,
17305,
382,
29954,
29943,
29899,
4561,
21904,
393,
1163,
297,
1009,
315,
29899,
8489,
979,
2011,
1080,
29889,
4525,
382,
29954,
29943,
29899,
4561,
21904,
29892,
3138,
29892,
526,
13557,
297,
278,
29250,
310,
4832,
274,
858,
29872,
457,
10995,
1041,
11122,
727,
262,
29889,
16564,
373,
385,
626,
1789,
22193,
5665,
10230,
29892,
4168,
4467,
634,
394,
29889,
1476,
393,
1546,
278,
937,
322,
25963,
274,
858,
29872,
1475,
297,
278,
382,
29954,
29943,
29899,
4561,
5354,
29892,
379,
29934,
29954,
29879,
892,
29871,
29946,
29945,
29995,
2788,
304,
540,
862,
262,
29899,
19672,
382,
29954,
29943,
29899,
4561,
14321,
7329,
313,
29950,
29933,
29899,
11787,
29943,
511,
29871,
29941,
29945,
29995,
13557,
304,
626,
561,
533,
29887,
352,
262,
313,
1718,
511,
29871,
29941,
29906,
29995,
13557,
304,
323,
29954,
29943,
29899,
30110,
29892,
322,
29871,
29906,
29955,
29995,
13557,
304,
382,
29954,
29943,
29889,
13,
29961,
29900,
29900,
29947,
29946,
29962,
450,
29871,
29946,
29946,
413,
27838,
17167,
1422,
11685,
7329,
313,
2797,
29943,
29897,
338,
278,
7548,
7126,
310,
5199,
379,
29934,
29954,
29889,
8502,
278,
379,
29934,
29954,
1248,
668,
415,
2247,
29892,
405,
4037,
756,
385,
5198,
348,
468,
2127,
352,
262,
313,
28600,
29897,
3632,
3002,
5354,
5643,
491,
385,
382,
29954,
29943,
29899,
4561,
5354,
322,
425,
4684,
263,
405,
29899,
8489,
979,
7182,
1236,
415,
680,
29889,
4360,
2705,
29892,
727,
526,
472,
3203,
4832,
8359,
18755,
307,
2204,
6288,
410,
29899,
2797,
29943,
29879,
29892,
770,
2164,
408,
2845,
15595,
470,
21762,
1248,
668,
415,
2247,
29892,
2729,
373,
278,
15602,
310,
278,
382,
29954,
29943,
29899,
4561,
21904,
29889,
306,
578,
9514,
29871,
29896,
304,
29871,
29946,
526,
2931,
1891,
373,
278,
8405,
310,
263,
2286,
16116,
1546,
278,
382,
29954,
29943,
29899,
4561,
5354,
322,
18750,
1590,
10800,
5354,
29889,
6549,
372,
5692,
393,
1422,
405,
4037,
338,
29877,
9514,
526,
5759,
491,
8671,
805,
506,
292,
322,
1122,
2189,
8359,
260,
15118,
29899,
14940,
3168,
29889,
2823,
16502,
29871,
29945,
29900,
29945,
29871,
29896,
29946,
29947,
29936,
399,
29949,
29871,
29929,
29941,
29914,
29906,
29906,
29946,
29906,
29946,
29936,
322,
399,
29949,
29871,
29929,
29946,
29914,
29906,
29947,
29896,
29941,
29941,
29892,
607,
526,
11039,
630,
491,
3407,
29889,
13,
29961,
29900,
29900,
29947,
29945,
29962,
512,
697,
7232,
397,
2073,
310,
278,
2198,
297,
7316,
29892,
278,
5541,
2187,
322,
3519,
526,
3889,
310,
429,
6352,
681,
1663,
352,
262,
322,
1663,
352,
262,
5960,
277,
2667,
29889,
450,
16549,
376,
735,
6352,
681,
1663,
352,
262,
470,
1663,
352,
262,
5960,
277,
2667,
29908,
338,
1304,
1244,
262,
304,
12266,
1663,
352,
262,
470,
1663,
352,
262,
5960,
277,
2667,
393,
338,
29914,
598,
451,
16392,
635,
2715,
304,
278,
5541,
2187,
470,
3519,
310,
278,
2198,
297,
7316,
29889,
6549,
29892,
297,
3058,
7232,
397,
7862,
310,
278,
2198,
297,
7316,
29892,
278,
3519,
322,
5541,
2187,
526,
3889,
310,
1663,
352,
262,
470,
1663,
352,
262,
5960,
277,
2667,
393,
526,
16392,
635,
19056,
29889,
450,
5541,
2187,
470,
3519,
1122,
29892,
3138,
29892,
451,
12695,
367,
3889,
310,
1095,
6352,
681,
1663,
352,
262,
29889,
1094,
1304,
1244,
262,
29892,
376,
355,
6352,
681,
1663,
352,
262,
29908,
14088,
393,
278,
4185,
2955,
9101,
1122,
367,
20811,
1663,
352,
262,
310,
1009,
1914,
15017,
746,
4185,
2955,
5034,
304,
278,
3519,
310,
278,
2198,
297,
7316,
29889,
2796,
6352,
681,
1663,
352,
262,
884,
1122,
367,
1304,
304,
12266,
10995,
950,
2411,
332,
1907,
515,
278,
7601,
3038,
9257,
470,
2411,
332,
1907,
515,
278,
6257,
17279,
29889,
512,
2702,
6455,
29892,
278,
5541,
2187,
322,
3519,
310,
278,
2198,
1712,
3109,
1135,
29871,
29945,
29900,
29892,
29871,
29946,
29945,
29892,
29871,
29946,
29900,
29892,
29871,
29941,
29945,
29892,
29871,
29941,
29900,
29892,
29871,
29906,
29945,
29892,
29871,
29906,
29900,
29892,
29871,
29896,
29945,
29892,
29871,
29896,
29900,
29892,
29871,
29929,
29892,
29871,
29947,
29892,
29871,
29955,
29892,
29871,
29953,
29892,
29871,
29945,
29892,
29871,
29946,
29892,
29871,
29941,
29892,
29871,
29906,
29892,
470,
29871,
29896,
29871,
30167,
29887,
29914,
828,
310,
1663,
352,
262,
29889,
13,
29961,
29900,
29900,
29947,
29953,
29962,
1094,
1304,
1244,
262,
29892,
278,
1840,
376,
1144,
352,
262,
29908,
14637,
304,
278,
26823,
29892,
470,
17305,
470,
9376,
727,
974,
393,
7868,
29879,
304,
278,
1663,
352,
262,
337,
14268,
297,
4226,
4824,
29875,
5996,
14953,
800,
322,
508,
9013,
346,
7182,
292,
1549,
278,
1663,
352,
262,
337,
14268,
29889,
450,
1840,
376,
1144,
352,
262,
29908,
427,
2388,
465,
267,
263,
26823,
2534,
278,
1248,
668,
415,
680,
5665,
310,
7531,
5199,
1663,
352,
262,
29892,
470,
310,
916,
286,
4850,
284,
713,
1663,
352,
262,
29892,
470,
310,
738,
3632,
1189,
29879,
470,
29161,
304,
1438,
15602,
29889,
19814,
29892,
278,
1840,
1663,
352,
262,
427,
2388,
465,
267,
1248,
668,
415,
680,
22370,
393,
526,
15390,
310,
9956,
304,
278,
1663,
352,
262,
337,
14268,
304,
9013,
346,
7182,
292,
1549,
278,
1663,
352,
262,
337,
14268,
29889,
450,
1840,
376,
1144,
352,
262,
23764,
29908,
14637,
304,
738,
503,
3742,
6943,
752,
618,
393,
1122,
367,
1304,
297,
2058,
310,
1663,
352,
262,
304,
2367,
23228,
368,
2788,
2582,
408,
1663,
352,
262,
29889,
1222,
9422,
310,
1663,
352,
262,
5960,
277,
2667,
3160,
29892,
541,
526,
451,
9078,
304,
503,
3742,
521,
5095,
680,
29892,
503,
3742,
21767,
10492,
29892,
503,
3742,
289,
456,
680,
29892,
322,
503,
3742,
5394,
29888,
403,
29889,
13,
29961,
29900,
29900,
29947,
29955,
29962,
1763,
367,
2821,
29892,
1663,
352,
262,
29899,
4561,
14321,
13879,
526,
451,
1663,
352,
262,
5960,
277,
2667,
470,
3632,
1189,
29879,
310,
1663,
352,
262,
29892,
408,
640,
16305,
630,
297,
278,
2198,
297,
7316,
29889,
7579,
368,
29892,
297,
1790,
2702,
7232,
397,
2073,
29892,
278,
5541,
2187,
322,
3519,
310,
278,
2198,
297,
7316,
7199,
895,
278,
671,
310,
472,
3203,
697,
1663,
352,
262,
29899,
4561,
14321,
7329,
313,
6259,
29943,
29897,
470,
263,
17305,
470,
263,
13303,
9376,
727,
974,
29889,
512,
1790,
7232,
397
] | 1,024 |
CommonCrawl
| -0.719268 | 1.267281 | -0.108372 | -0.060967 |
iment, the compositions and methods of the present invention are free of any exogenous insulin-like growth factors (IGFs). In specific embodiments, the compositions and methods of the present invention contain less than 200, 150, 100, 75, 50, 25, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 ng/ml of IGF-1.
[0088] As used herein, the term "activator of IGF-1R" refers to mitogens that play a pivotal role in regulating cell proliferation, differentiation, and apoptosis. The effects of an activator of IGF-1R are typically mediated through IGF-1R, although they can be mediated through other receptors. The IGF-1R is also involved in cell transformation induced by tumor virus proteins and oncogene products, and the interaction is regulated by a group of specific binding proteins (IGFBPs). In addition, a large group of IGFBP proteases hydrolyze IGFBPs, resulting in the release of bound IGFs that then resume their ability to interact with IGF-IR. For the purpose of this invention, the ligands, the receptors, the binding proteins, and the proteases are all considered to be activators of IGF-1R. In one embodiment, the activator of IGF-1R is IGF-1, or IGF-2. In a further embodiment, the activators of IGF-1R is an IGF-1 analog. Non-limiting examples of IGF-1 analogs include LongR3-IGF1, Des(1-3)IGF-1, [Arg3]IGF-1, [Ala31]IFG-1, Des(2,3)[Ala31]IGF-1, [Leu24]IGF1, Des(2,3)[Leu24]IGF-1, [Leu60]IGF-1, [Ala31][Leu60]IGF-1, [Leu24][Ala31]IGF-1, and combinations thereof. In a further embodiment, the IFG-1 analog is LongR3-IGF1, which is a recombinant analog of human insulin growth factor-1. It is contemplated that LongR3-IGF1 is initially present at a concentration of approximately 1 ng/ml to approximately 1000 ng/ml, more preferably approximately 5 ng/ml to approximately 500 ng/ml, more preferably approximately 50 ng/ml to approximately 500 ng/ml, more preferably approximately 100 ng/ml to approximately 300 ng/ml, or at a concentration of approximately 100 ng/ml.
[0089] In certain embodiments, the compositions and methods of the present invention comprise transforming growth factor beta (TGF-β) or a TGF-β family member or variants or functional fragments thereof. As used herein, the term "member of the TGF-β family" or the like refers to growth factors that are generally characterized by one of skill in the art as belonging to the TGF-β family, either due to homology with known members of the TGF-β family, or due to similarity in function with known members of the TGF-β family. In particular embodiments of the invention, if the member of the TGF-β family is present, the TGF-β family member of variant or functional fragment thereof activates SMAD 2 or 3. In certain embodiments, the member of the TGF-β family is selected from the group consisting of Nodal, Activin A, Activin B, TGF-β, bone morphogenic protein-2 (BMP2) and bone morphogenic protein-4 (BMP4). In one embodiment, the member of the TGF-β family is Activin A.
[0090] It is contemplated that if Nodal is present, it is initially present at a concentration of approximately 0.1 ng/ml to approximately 2000 ng/ml, more
|
[
2073,
29892,
278,
5541,
2187,
322,
3519,
310,
278,
2198,
297,
7316,
526,
3889,
310,
738,
429,
6352,
681,
1663,
352,
262,
29899,
4561,
14321,
13879,
313,
6259,
29943,
29879,
467,
512,
2702,
7232,
397,
7862,
29892,
278,
5541,
2187,
322,
3519,
310,
278,
2198,
297,
7316,
1712,
3109,
1135,
29871,
29906,
29900,
29900,
29892,
29871,
29896,
29945,
29900,
29892,
29871,
29896,
29900,
29900,
29892,
29871,
29955,
29945,
29892,
29871,
29945,
29900,
29892,
29871,
29906,
29945,
29892,
29871,
29906,
29900,
29892,
29871,
29896,
29945,
29892,
29871,
29896,
29900,
29892,
29871,
29929,
29892,
29871,
29947,
29892,
29871,
29955,
29892,
29871,
29953,
29892,
29871,
29945,
29892,
29871,
29946,
29892,
29871,
29941,
29892,
29871,
29906,
29892,
470,
29871,
29896,
8736,
29914,
828,
310,
306,
29954,
29943,
29899,
29896,
29889,
13,
29961,
29900,
29900,
29947,
29947,
29962,
1094,
1304,
1244,
262,
29892,
278,
1840,
376,
11236,
1061,
310,
306,
29954,
29943,
29899,
29896,
29934,
29908,
14637,
304,
1380,
468,
575,
393,
1708,
263,
282,
440,
7288,
6297,
297,
1072,
18099,
3038,
410,
29880,
9633,
362,
29892,
1422,
11685,
29892,
322,
16798,
415,
19263,
29889,
450,
9545,
310,
385,
5039,
1061,
310,
306,
29954,
29943,
29899,
29896,
29934,
526,
12234,
14457,
630,
1549,
306,
29954,
29943,
29899,
29896,
29934,
29892,
5998,
896,
508,
367,
14457,
630,
1549,
916,
337,
1547,
943,
29889,
450,
306,
29954,
29943,
29899,
29896,
29934,
338,
884,
9701,
297,
3038,
13852,
20974,
491,
21622,
272,
24424,
3279,
1144,
322,
373,
29883,
16603,
9316,
29892,
322,
278,
14881,
338,
1072,
7964,
491,
263,
2318,
310,
2702,
9956,
3279,
1144,
313,
6259,
18426,
29925,
29879,
467,
512,
6124,
29892,
263,
2919,
2318,
310,
306,
29954,
18426,
29925,
3279,
2129,
17546,
368,
911,
306,
29954,
18426,
29925,
29879,
29892,
9819,
297,
278,
6507,
310,
3216,
306,
29954,
29943,
29879,
393,
769,
620,
2017,
1009,
11509,
304,
16254,
411,
306,
29954,
29943,
29899,
8193,
29889,
1152,
278,
6437,
310,
445,
297,
7316,
29892,
278,
14172,
4167,
29892,
278,
337,
1547,
943,
29892,
278,
9956,
3279,
1144,
29892,
322,
278,
3279,
2129,
526,
599,
5545,
304,
367,
5039,
4097,
310,
306,
29954,
29943,
29899,
29896,
29934,
29889,
512,
697,
7232,
397,
2073,
29892,
278,
5039,
1061,
310,
306,
29954,
29943,
29899,
29896,
29934,
338,
306,
29954,
29943,
29899,
29896,
29892,
470,
306,
29954,
29943,
29899,
29906,
29889,
512,
263,
4340,
7232,
397,
2073,
29892,
278,
5039,
4097,
310,
306,
29954,
29943,
29899,
29896,
29934,
338,
385,
306,
29954,
29943,
29899,
29896,
15690,
29889,
10050,
29899,
2576,
11407,
6455,
310,
306,
29954,
29943,
29899,
29896,
3483,
12099,
3160,
6242,
29934,
29941,
29899,
6259,
29943,
29896,
29892,
2726,
29898,
29896,
29899,
29941,
29897,
6259,
29943,
29899,
29896,
29892,
518,
8559,
29941,
29962,
6259,
29943,
29899,
29896,
29892,
518,
29909,
433,
29941,
29896,
29962,
6545,
29954,
29899,
29896,
29892,
2726,
29898,
29906,
29892,
29941,
9601,
29909,
433,
29941,
29896,
29962,
6259,
29943,
29899,
29896,
29892,
518,
3226,
29884,
29906,
29946,
29962,
6259,
29943,
29896,
29892,
2726,
29898,
29906,
29892,
29941,
9601,
3226,
29884,
29906,
29946,
29962,
6259,
29943,
29899,
29896,
29892,
518,
3226,
29884,
29953,
29900,
29962,
6259,
29943,
29899,
29896,
29892,
518,
29909,
433,
29941,
29896,
3816,
3226,
29884,
29953,
29900,
29962,
6259,
29943,
29899,
29896,
29892,
518,
3226,
29884,
29906,
29946,
3816,
29909,
433,
29941,
29896,
29962,
6259,
29943,
29899,
29896,
29892,
322,
18240,
727,
974,
29889,
512,
263,
4340,
7232,
397,
2073,
29892,
278,
10762,
29954,
29899,
29896,
15690,
338,
6242,
29934,
29941,
29899,
6259,
29943,
29896,
29892,
607,
338,
263,
27878,
2109,
424,
15690,
310,
5199,
1663,
352,
262,
14321,
7329,
29899,
29896,
29889,
739,
338,
640,
16305,
630,
393,
6242,
29934,
29941,
29899,
6259,
29943,
29896,
338,
12919,
2198,
472,
263,
26702,
310,
14235,
29871,
29896,
8736,
29914,
828,
304,
14235,
29871,
29896,
29900,
29900,
29900,
8736,
29914,
828,
29892,
901,
5821,
2197,
14235,
29871,
29945,
8736,
29914,
828,
304,
14235,
29871,
29945,
29900,
29900,
8736,
29914,
828,
29892,
901,
5821,
2197,
14235,
29871,
29945,
29900,
8736,
29914,
828,
304,
14235,
29871,
29945,
29900,
29900,
8736,
29914,
828,
29892,
901,
5821,
2197,
14235,
29871,
29896,
29900,
29900,
8736,
29914,
828,
304,
14235,
29871,
29941,
29900,
29900,
8736,
29914,
828,
29892,
470,
472,
263,
26702,
310,
14235,
29871,
29896,
29900,
29900,
8736,
29914,
828,
29889,
13,
29961,
29900,
29900,
29947,
29929,
29962,
512,
3058,
7232,
397,
7862,
29892,
278,
5541,
2187,
322,
3519,
310,
278,
2198,
297,
7316,
7199,
895,
4327,
292,
14321,
7329,
21762,
313,
29911,
29954,
29943,
29899,
30187,
29897,
470,
263,
323,
29954,
29943,
29899,
30187,
3942,
4509,
470,
29161,
470,
13303,
22370,
727,
974,
29889,
1094,
1304,
1244,
262,
29892,
278,
1840,
376,
14242,
310,
278,
323,
29954,
29943,
29899,
30187,
3942,
29908,
470,
278,
763,
14637,
304,
14321,
13879,
393,
526,
6892,
2931,
1891,
491,
697,
310,
19911,
297,
278,
1616,
408,
23329,
304,
278,
323,
29954,
29943,
29899,
30187,
3942,
29892,
2845,
2861,
304,
3632,
3002,
411,
2998,
5144,
310,
278,
323,
29954,
29943,
29899,
30187,
3942,
29892,
470,
2861,
304,
29501,
297,
740,
411,
2998,
5144,
310,
278,
323,
29954,
29943,
29899,
30187,
3942,
29889,
512,
3153,
7232,
397,
7862,
310,
278,
297,
7316,
29892,
565,
278,
4509,
310,
278,
323,
29954,
29943,
29899,
30187,
3942,
338,
2198,
29892,
278,
323,
29954,
29943,
29899,
30187,
3942,
4509,
310,
17305,
470,
13303,
9376,
727,
974,
5039,
1078,
317,
1529,
29928,
29871,
29906,
470,
29871,
29941,
29889,
512,
3058,
7232,
397,
7862,
29892,
278,
4509,
310,
278,
323,
29954,
29943,
29899,
30187,
3942,
338,
4629,
515,
278,
2318,
19849,
310,
405,
397,
284,
29892,
21775,
262,
319,
29892,
21775,
262,
350,
29892,
323,
29954,
29943,
29899,
30187,
29892,
289,
650,
18131,
6352,
293,
26823,
29899,
29906,
313,
29933,
3580,
29906,
29897,
322,
289,
650,
18131,
6352,
293,
26823,
29899,
29946,
313,
29933,
3580,
29946,
467,
512,
697,
7232,
397,
2073,
29892,
278,
4509,
310,
278,
323,
29954,
29943,
29899,
30187,
3942,
338,
21775,
262,
319,
29889,
13,
29961,
29900,
29900,
29929,
29900,
29962,
739,
338,
640,
16305,
630,
393,
565,
405,
397,
284,
338,
2198,
29892,
372,
338,
12919,
2198,
472,
263,
26702,
310,
14235,
29871,
29900,
29889,
29896,
8736,
29914,
828,
304,
14235,
29871,
29906,
29900,
29900,
29900,
8736,
29914,
828,
29892,
901
] | 1,024 |
CommonCrawl
| -1.268114 | 1.16865 | -0.747656 | -0.727557 |
preferably approximately 1 ng/ml to approximately 1000 ng/ml, more preferably approximately 10 ng/ml to approximately 750 ng/ml, or more preferably approximately 25 ng/ml to approximately 500 ng/ml. It is contemplated that if used, Activin A is initially present at a concentration of approximately 0.01 ng/ml to approximately 1000 ng/ml, more preferably approximately 0.1 ng/ml to approximately 100 ng/ml, more preferably approximately 0.1 ng/ml to approximately 25 ng/ml, or most preferably at a concentration of approximately 10 ng/ml. It is contemplated that if present, TGF-β is initially present at a concentration of approximately 0.01 ng/ml to approximately 100 ng/ml, more preferably approximately 0.1 ng/ml to approximately 50 ng/ml, or more preferably approximately 0.1 ng/ml to approximately 20 ng/ml.
[0091] In additional embodiments of the present invention, the compositions and methods of the present invention are free of activators of FGF receptors. There are currently at least 22 known members of the family of fibroblast growth factors, with these factors binding to one of at least one of four FGF receptors. As used herein, the term "activator of an FGF receptor" refers to growth factors that are generally characterized by one of skill in the art as belonging to the FGF family, either due to homology with known members of the FGF family, or due to similarity in function with known members of the FGF family. In certain embodiments, the activator of an FGF receptor is an FGF, such as, but not limited to α-FGF and FGF2. In particular embodiments, the compositions and methods are free of exogenous FGF2. The phrase "exogenous FGF2" is used herein to indicate fibroblast growth factor 2, i.e., basic FGF, that is not intentionally added to the compositions or methods of the present invention. Thus, in certain embodiments of the present invention, the methods and compositions are free of intentionally supplied FGF2. The compositions or methods may, however, not necessarily be free of endogenous FGF2. As used herein, "endogenous FGF2" indicates that the cultured cells may be producing FGF2 of their own accord when cultured according to the methods of the present invention. "Endogenous FGF2" also may be used to indicate residual impurities from the primary cell culture or impurities from the starting materials. In specific examples, the compositions and methods of the present contain less than 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 ng/ml of FGF2.
[0092] It is contemplated, however, that the compositions and methods of the invention can include at least one activator of an FGF receptor, including any of the FGF polypeptides, functional fragments thereof or variants thereof. It is contemplated that if FGF2 is present, it is initially present at a concentration of approximately 0.1 ng/ml to approximately 100 ng/ml, more preferably approximately 0.5 ng/ml to approximately 50 ng/ml, more preferably approximately 1 ng/ml to approximately 25 ng/ml, more preferably approximately 1 ng/ml to approximately 12 ng/ml, or most preferably at a concentration of approximately 8 ng/ml. In another specific embodiment, the compositions and methods of the invention can include at least one activator of an FGF receptor, other than FGF2. For example, the compositions and methods of the present invention may comprise at least one of FGF-7, FGF-10 or FGF-22 or variants or functional fragments thereof. In specific embodiments, a combination of at least two of FGF-7, FGF-10 and FGF-22, or variants or functional fragments thereof, are present. In another embodiment, all three of FGF-7, FGF-10 and FGF-22, or variants or functional fragments
|
[
5821,
2197,
14235,
29871,
29896,
8736,
29914,
828,
304,
14235,
29871,
29896,
29900,
29900,
29900,
8736,
29914,
828,
29892,
901,
5821,
2197,
14235,
29871,
29896,
29900,
8736,
29914,
828,
304,
14235,
29871,
29955,
29945,
29900,
8736,
29914,
828,
29892,
470,
901,
5821,
2197,
14235,
29871,
29906,
29945,
8736,
29914,
828,
304,
14235,
29871,
29945,
29900,
29900,
8736,
29914,
828,
29889,
739,
338,
640,
16305,
630,
393,
565,
1304,
29892,
21775,
262,
319,
338,
12919,
2198,
472,
263,
26702,
310,
14235,
29871,
29900,
29889,
29900,
29896,
8736,
29914,
828,
304,
14235,
29871,
29896,
29900,
29900,
29900,
8736,
29914,
828,
29892,
901,
5821,
2197,
14235,
29871,
29900,
29889,
29896,
8736,
29914,
828,
304,
14235,
29871,
29896,
29900,
29900,
8736,
29914,
828,
29892,
901,
5821,
2197,
14235,
29871,
29900,
29889,
29896,
8736,
29914,
828,
304,
14235,
29871,
29906,
29945,
8736,
29914,
828,
29892,
470,
1556,
5821,
2197,
472,
263,
26702,
310,
14235,
29871,
29896,
29900,
8736,
29914,
828,
29889,
739,
338,
640,
16305,
630,
393,
565,
2198,
29892,
323,
29954,
29943,
29899,
30187,
338,
12919,
2198,
472,
263,
26702,
310,
14235,
29871,
29900,
29889,
29900,
29896,
8736,
29914,
828,
304,
14235,
29871,
29896,
29900,
29900,
8736,
29914,
828,
29892,
901,
5821,
2197,
14235,
29871,
29900,
29889,
29896,
8736,
29914,
828,
304,
14235,
29871,
29945,
29900,
8736,
29914,
828,
29892,
470,
901,
5821,
2197,
14235,
29871,
29900,
29889,
29896,
8736,
29914,
828,
304,
14235,
29871,
29906,
29900,
8736,
29914,
828,
29889,
13,
29961,
29900,
29900,
29929,
29896,
29962,
512,
5684,
7232,
397,
7862,
310,
278,
2198,
297,
7316,
29892,
278,
5541,
2187,
322,
3519,
310,
278,
2198,
297,
7316,
526,
3889,
310,
5039,
4097,
310,
383,
29954,
29943,
337,
1547,
943,
29889,
1670,
526,
5279,
472,
3203,
29871,
29906,
29906,
2998,
5144,
310,
278,
3942,
310,
18755,
307,
23190,
14321,
13879,
29892,
411,
1438,
13879,
9956,
304,
697,
310,
472,
3203,
697,
310,
3023,
383,
29954,
29943,
337,
1547,
943,
29889,
1094,
1304,
1244,
262,
29892,
278,
1840,
376,
11236,
1061,
310,
385,
383,
29954,
29943,
337,
14268,
29908,
14637,
304,
14321,
13879,
393,
526,
6892,
2931,
1891,
491,
697,
310,
19911,
297,
278,
1616,
408,
23329,
304,
278,
383,
29954,
29943,
3942,
29892,
2845,
2861,
304,
3632,
3002,
411,
2998,
5144,
310,
278,
383,
29954,
29943,
3942,
29892,
470,
2861,
304,
29501,
297,
740,
411,
2998,
5144,
310,
278,
383,
29954,
29943,
3942,
29889,
512,
3058,
7232,
397,
7862,
29892,
278,
5039,
1061,
310,
385,
383,
29954,
29943,
337,
14268,
338,
385,
383,
29954,
29943,
29892,
1316,
408,
29892,
541,
451,
9078,
304,
19883,
29899,
29943,
29954,
29943,
322,
383,
29954,
29943,
29906,
29889,
512,
3153,
7232,
397,
7862,
29892,
278,
5541,
2187,
322,
3519,
526,
3889,
310,
429,
6352,
681,
383,
29954,
29943,
29906,
29889,
450,
16549,
376,
735,
6352,
681,
383,
29954,
29943,
29906,
29908,
338,
1304,
1244,
262,
304,
12266,
18755,
307,
23190,
14321,
7329,
29871,
29906,
29892,
474,
29889,
29872,
1696,
6996,
383,
29954,
29943,
29892,
393,
338,
451,
16392,
635,
2715,
304,
278,
5541,
2187,
470,
3519,
310,
278,
2198,
297,
7316,
29889,
6549,
29892,
297,
3058,
7232,
397,
7862,
310,
278,
2198,
297,
7316,
29892,
278,
3519,
322,
5541,
2187,
526,
3889,
310,
16392,
635,
19056,
383,
29954,
29943,
29906,
29889,
450,
5541,
2187,
470,
3519,
1122,
29892,
3138,
29892,
451,
12695,
367,
3889,
310,
1095,
6352,
681,
383,
29954,
29943,
29906,
29889,
1094,
1304,
1244,
262,
29892,
376,
355,
6352,
681,
383,
29954,
29943,
29906,
29908,
14088,
393,
278,
4185,
2955,
9101,
1122,
367,
20811,
383,
29954,
29943,
29906,
310,
1009,
1914,
15017,
746,
4185,
2955,
5034,
304,
278,
3519,
310,
278,
2198,
297,
7316,
29889,
376,
5044,
6352,
681,
383,
29954,
29943,
29906,
29908,
884,
1122,
367,
1304,
304,
12266,
10995,
950,
2411,
332,
1907,
515,
278,
7601,
3038,
9257,
470,
2411,
332,
1907,
515,
278,
6257,
17279,
29889,
512,
2702,
6455,
29892,
278,
5541,
2187,
322,
3519,
310,
278,
2198,
1712,
3109,
1135,
29871,
29896,
29900,
29892,
29871,
29929,
29892,
29871,
29947,
29892,
29871,
29955,
29892,
29871,
29953,
29892,
29871,
29945,
29892,
29871,
29946,
29892,
29871,
29941,
29892,
29871,
29906,
29892,
470,
29871,
29896,
8736,
29914,
828,
310,
383,
29954,
29943,
29906,
29889,
13,
29961,
29900,
29900,
29929,
29906,
29962,
739,
338,
640,
16305,
630,
29892,
3138,
29892,
393,
278,
5541,
2187,
322,
3519,
310,
278,
297,
7316,
508,
3160,
472,
3203,
697,
5039,
1061,
310,
385,
383,
29954,
29943,
337,
14268,
29892,
3704,
738,
310,
278,
383,
29954,
29943,
1248,
668,
415,
2247,
29892,
13303,
22370,
727,
974,
470,
29161,
727,
974,
29889,
739,
338,
640,
16305,
630,
393,
565,
383,
29954,
29943,
29906,
338,
2198,
29892,
372,
338,
12919,
2198,
472,
263,
26702,
310,
14235,
29871,
29900,
29889,
29896,
8736,
29914,
828,
304,
14235,
29871,
29896,
29900,
29900,
8736,
29914,
828,
29892,
901,
5821,
2197,
14235,
29871,
29900,
29889,
29945,
8736,
29914,
828,
304,
14235,
29871,
29945,
29900,
8736,
29914,
828,
29892,
901,
5821,
2197,
14235,
29871,
29896,
8736,
29914,
828,
304,
14235,
29871,
29906,
29945,
8736,
29914,
828,
29892,
901,
5821,
2197,
14235,
29871,
29896,
8736,
29914,
828,
304,
14235,
29871,
29896,
29906,
8736,
29914,
828,
29892,
470,
1556,
5821,
2197,
472,
263,
26702,
310,
14235,
29871,
29947,
8736,
29914,
828,
29889,
512,
1790,
2702,
7232,
397,
2073,
29892,
278,
5541,
2187,
322,
3519,
310,
278,
297,
7316,
508,
3160,
472,
3203,
697,
5039,
1061,
310,
385,
383,
29954,
29943,
337,
14268,
29892,
916,
1135,
383,
29954,
29943,
29906,
29889,
1152,
1342,
29892,
278,
5541,
2187,
322,
3519,
310,
278,
2198,
297,
7316,
1122,
7199,
895,
472,
3203,
697,
310,
383,
29954,
29943,
29899,
29955,
29892,
383,
29954,
29943,
29899,
29896,
29900,
470,
383,
29954,
29943,
29899,
29906,
29906,
470,
29161,
470,
13303,
22370,
727,
974,
29889,
512,
2702,
7232,
397,
7862,
29892,
263,
10296,
310,
472,
3203,
1023,
310,
383,
29954,
29943,
29899,
29955,
29892,
383,
29954,
29943,
29899,
29896,
29900,
322,
383,
29954,
29943,
29899,
29906,
29906,
29892,
470,
29161,
470,
13303,
22370,
727,
974,
29892,
526,
2198,
29889,
512,
1790,
7232,
397,
2073,
29892,
599,
2211,
310,
383,
29954,
29943,
29899,
29955,
29892,
383,
29954,
29943,
29899,
29896,
29900,
322,
383,
29954,
29943,
29899,
29906,
29906,
29892,
470,
29161,
470,
13303,
22370
] | 1,024 |
CommonCrawl
| -1.425704 | 0.82863 | -1.358249 | -0.929937 |
thereof, are present. It is contemplated that if any of FGF-7, FGF-10 or FGF-22 or variants or functional fragments are present, each is initially present at a concentration of approximately 0.1 ng/ml to approximately 100 ng/ml, more specifically from approximately 0.5 ng/ml to approximately 50 ng/ml, more specifically from approximately 1 ng/ml to approximately 25 ng/ml, more specifically from approximately 1 ng/ml to approximately 12 ng/ml, or most specifically at a concentration of approximately 8 ng/ml.
[0093] In additional certain embodiments, the compositions and methods of the present invention comprise scrum albumin (SA). In specific embodiments, the SA is either bovine SA (BSA) or human SA (HAS). In still more specific embodiments, the concentration of the SA is more than about 0.2%, volume to volume (v/v), but less than about 10% v/v. In even more specific embodiments, the concentration of SA is more than about 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.2%, 1.4%, 1.6%, 1.8%, 2.0%, 2.2%, 2.4%, 2.6%, 2.8%, 3.0%, 3.2%, 3.4%, 3.6%, 3.8%, 4.0%, 4.2%, 4.4%, 4.6%, 4.8%, 5.0%, 5.2%, 5.4%, 5.6%, 5.8%, 6.0%, 6.2%, 6.4%, 6.6%, 6.8%, 7.0%, 7.2%, 7.4%, 7.6%, 7.8%, 8.0%, 8.2%, 8.4%, 8.6%, 8.8%, 9.0%, 9.2%, 9.4%, 9.6% and 9.8% (v/v).
[0094] In additional embodiments, the compositions and methods comprise at least one insoluble substrate. For example, the differentiable cells may be placed on a cell culture surface that comprises such compounds as, but is not limited to, polystyrene, polypropylene. The surface may, in turn, be coated with an insoluble substrate. In specific embodiments, the insoluble substrate is selected from the group consisting of a collagen, a fibronectin and fragments or variants thereof. Other examples of insoluble substrates include, but are not limited to, fibrin, elastin, fibronectins, laminins and nidogens.
[0095] Accordingly, the cell culture environments and methods of the present invention comprise plating the cells in an adherent culture. As used herein, the terms "plated" and "plating" refer to any process that allows a cell to be grown in adherent culture. As used herein, the term "adherent culture" refers to a cell culture system whereby cells are cultured on a solid surface, which may in turn be coated with an insoluble substrate that may in turn be coated with another surface coat of a substrate, such as those listed below, or any other chemical or biological material that allows the cells to proliferate or be stabilized in culture. The cells may or may not tightly adhere to the solid surface or to the substrate. The substrate for the adherent culture may comprise any one or combination of polyornithine, laminin, poly-lysine, purified collagen, gelatin, fibronectin, tenascin, vitronectin, entactin, heparin sulfate proteoglycans, poly glycolytic acid (PGA), poly lactic acid (PLA), and poly lactic-glycolic acid (PLGA). Furthermore, the substrate for the adherent culture may comprise the matrix laid down by a feeder layer, or laid down by the pluripotent human cell or cell culture. As used herein, the term "extracellular matrix" encompasses solid substr
|
[
727,
974,
29892,
526,
2198,
29889,
739,
338,
640,
16305,
630,
393,
565,
738,
310,
383,
29954,
29943,
29899,
29955,
29892,
383,
29954,
29943,
29899,
29896,
29900,
470,
383,
29954,
29943,
29899,
29906,
29906,
470,
29161,
470,
13303,
22370,
526,
2198,
29892,
1269,
338,
12919,
2198,
472,
263,
26702,
310,
14235,
29871,
29900,
29889,
29896,
8736,
29914,
828,
304,
14235,
29871,
29896,
29900,
29900,
8736,
29914,
828,
29892,
901,
10816,
515,
14235,
29871,
29900,
29889,
29945,
8736,
29914,
828,
304,
14235,
29871,
29945,
29900,
8736,
29914,
828,
29892,
901,
10816,
515,
14235,
29871,
29896,
8736,
29914,
828,
304,
14235,
29871,
29906,
29945,
8736,
29914,
828,
29892,
901,
10816,
515,
14235,
29871,
29896,
8736,
29914,
828,
304,
14235,
29871,
29896,
29906,
8736,
29914,
828,
29892,
470,
1556,
10816,
472,
263,
26702,
310,
14235,
29871,
29947,
8736,
29914,
828,
29889,
13,
29961,
29900,
29900,
29929,
29941,
29962,
512,
5684,
3058,
7232,
397,
7862,
29892,
278,
5541,
2187,
322,
3519,
310,
278,
2198,
297,
7316,
7199,
895,
885,
5848,
3769,
262,
313,
8132,
467,
512,
2702,
7232,
397,
7862,
29892,
278,
16698,
338,
2845,
289,
586,
457,
16698,
313,
29933,
8132,
29897,
470,
5199,
16698,
313,
29950,
3289,
467,
512,
1603,
901,
2702,
7232,
397,
7862,
29892,
278,
26702,
310,
278,
16698,
338,
901,
1135,
1048,
29871,
29900,
29889,
29906,
13667,
7977,
304,
7977,
313,
29894,
29914,
29894,
511,
541,
3109,
1135,
1048,
29871,
29896,
29900,
29995,
325,
29914,
29894,
29889,
512,
1584,
901,
2702,
7232,
397,
7862,
29892,
278,
26702,
310,
16698,
338,
901,
1135,
1048,
29871,
29900,
29889,
29941,
13667,
29871,
29900,
29889,
29946,
13667,
29871,
29900,
29889,
29945,
13667,
29871,
29900,
29889,
29953,
13667,
29871,
29900,
29889,
29955,
13667,
29871,
29900,
29889,
29947,
13667,
29871,
29900,
29889,
29929,
13667,
29871,
29896,
29889,
29900,
13667,
29871,
29896,
29889,
29906,
13667,
29871,
29896,
29889,
29946,
13667,
29871,
29896,
29889,
29953,
13667,
29871,
29896,
29889,
29947,
13667,
29871,
29906,
29889,
29900,
13667,
29871,
29906,
29889,
29906,
13667,
29871,
29906,
29889,
29946,
13667,
29871,
29906,
29889,
29953,
13667,
29871,
29906,
29889,
29947,
13667,
29871,
29941,
29889,
29900,
13667,
29871,
29941,
29889,
29906,
13667,
29871,
29941,
29889,
29946,
13667,
29871,
29941,
29889,
29953,
13667,
29871,
29941,
29889,
29947,
13667,
29871,
29946,
29889,
29900,
13667,
29871,
29946,
29889,
29906,
13667,
29871,
29946,
29889,
29946,
13667,
29871,
29946,
29889,
29953,
13667,
29871,
29946,
29889,
29947,
13667,
29871,
29945,
29889,
29900,
13667,
29871,
29945,
29889,
29906,
13667,
29871,
29945,
29889,
29946,
13667,
29871,
29945,
29889,
29953,
13667,
29871,
29945,
29889,
29947,
13667,
29871,
29953,
29889,
29900,
13667,
29871,
29953,
29889,
29906,
13667,
29871,
29953,
29889,
29946,
13667,
29871,
29953,
29889,
29953,
13667,
29871,
29953,
29889,
29947,
13667,
29871,
29955,
29889,
29900,
13667,
29871,
29955,
29889,
29906,
13667,
29871,
29955,
29889,
29946,
13667,
29871,
29955,
29889,
29953,
13667,
29871,
29955,
29889,
29947,
13667,
29871,
29947,
29889,
29900,
13667,
29871,
29947,
29889,
29906,
13667,
29871,
29947,
29889,
29946,
13667,
29871,
29947,
29889,
29953,
13667,
29871,
29947,
29889,
29947,
13667,
29871,
29929,
29889,
29900,
13667,
29871,
29929,
29889,
29906,
13667,
29871,
29929,
29889,
29946,
13667,
29871,
29929,
29889,
29953,
29995,
322,
29871,
29929,
29889,
29947,
29995,
313,
29894,
29914,
29894,
467,
13,
29961,
29900,
29900,
29929,
29946,
29962,
512,
5684,
7232,
397,
7862,
29892,
278,
5541,
2187,
322,
3519,
7199,
895,
472,
3203,
697,
1663,
324,
431,
280,
25148,
403,
29889,
1152,
1342,
29892,
278,
17473,
519,
9101,
1122,
367,
7180,
373,
263,
3038,
9257,
7101,
393,
7199,
4637,
1316,
752,
3885,
408,
29892,
541,
338,
451,
9078,
304,
29892,
1248,
858,
29891,
28593,
29892,
15680,
771,
2272,
29880,
1600,
29889,
450,
7101,
1122,
29892,
297,
2507,
29892,
367,
1302,
630,
411,
385,
1663,
324,
431,
280,
25148,
403,
29889,
512,
2702,
7232,
397,
7862,
29892,
278,
1663,
324,
431,
280,
25148,
403,
338,
4629,
515,
278,
2318,
19849,
310,
263,
784,
11541,
29892,
263,
18755,
1617,
522,
262,
322,
22370,
470,
29161,
727,
974,
29889,
5901,
6455,
310,
1663,
324,
431,
280,
25148,
1078,
3160,
29892,
541,
526,
451,
9078,
304,
29892,
285,
4626,
262,
29892,
560,
579,
262,
29892,
18755,
1617,
522,
1144,
29892,
301,
9103,
1144,
322,
302,
333,
468,
575,
29889,
13,
29961,
29900,
29900,
29929,
29945,
29962,
7579,
368,
29892,
278,
3038,
9257,
23136,
322,
3519,
310,
278,
2198,
297,
7316,
7199,
895,
715,
1218,
278,
9101,
297,
385,
594,
2276,
296,
9257,
29889,
1094,
1304,
1244,
262,
29892,
278,
4958,
376,
572,
630,
29908,
322,
376,
572,
1218,
29908,
2737,
304,
738,
1889,
393,
6511,
263,
3038,
304,
367,
21633,
297,
594,
2276,
296,
9257,
29889,
1094,
1304,
1244,
262,
29892,
278,
1840,
376,
328,
2276,
296,
9257,
29908,
14637,
304,
263,
3038,
9257,
1788,
988,
1609,
9101,
526,
4185,
2955,
373,
263,
7773,
7101,
29892,
607,
1122,
297,
2507,
367,
1302,
630,
411,
385,
1663,
324,
431,
280,
25148,
403,
393,
1122,
297,
2507,
367,
1302,
630,
411,
1790,
7101,
24296,
310,
263,
25148,
403,
29892,
1316,
408,
1906,
9904,
2400,
29892,
470,
738,
916,
22233,
470,
4768,
5996,
5518,
393,
6511,
278,
9101,
304,
410,
29880,
9633,
403,
470,
367,
16160,
1891,
297,
9257,
29889,
450,
9101,
1122,
470,
1122,
451,
19932,
368,
594,
4150,
304,
278,
7773,
7101,
470,
304,
278,
25148,
403,
29889,
450,
25148,
403,
363,
278,
594,
2276,
296,
9257,
1122,
7199,
895,
738,
697,
470,
10296,
310,
15680,
1398,
389,
457,
29892,
301,
9103,
262,
29892,
15680,
29899,
368,
29879,
457,
29892,
3708,
2164,
784,
11541,
29892,
9127,
21203,
29892,
18755,
1617,
522,
262,
29892,
3006,
6151,
262,
29892,
13901,
1617,
522,
262,
29892,
875,
627,
262,
29892,
540,
862,
262,
5394,
29888,
403,
3279,
468,
368,
29883,
550,
29892,
15680,
330,
368,
1054,
3637,
293,
22193,
313,
29925,
12739,
511,
15680,
425,
20009,
22193,
313,
29925,
4375,
511,
322,
15680,
425,
20009,
29899,
16808,
1054,
293,
22193,
313,
7390,
12739,
467,
16478,
29892,
278,
25148,
403,
363,
278,
594,
2276,
296,
9257,
1122,
7199,
895,
278,
4636,
12341,
1623,
491,
263,
1238,
2447,
7546,
29892,
470,
12341,
1623,
491,
278,
715,
332,
666,
327,
296,
5199,
3038,
470,
3038,
9257,
29889,
1094,
1304,
1244,
262,
29892,
278,
1840,
376,
1062,
945,
514,
1070,
4636,
29908,
427,
2388,
465,
267,
7773,
25148
] | 1,024 |
CommonCrawl
| -1.412118 | 0.571669 | -0.878562 | -0.996976 |
ates such as but not limited to those described above, as well as the matrix laid down by a feeder cell layer or by the pluripotent human cell or cell culture. In one embodiment, the cells are plated on MATRIGEL®-coated plates. In another embodiment, the cells are plated on fibronectin-coated plates. In certain embodiments, if the cells are plated on fibronectin, the plates are prepared by coating with 10 μg/ml human plasma fibronectin (Invitrogen, #33016-015), diluted in tissue grade water, for 2-3 hours at room temperature. In another embodiment, serum can be placed in the medium for up to 24 hours to allow cells to plate to the plastic. If using serum to promote the attachment of the cells, the media is then removed and the compositions, which are essentially serum-free, are added to the plated cells.
[0096] The compositions and methods of the present invention contemplate that the differentiable cells are cultured in conditions that are essentially free of a feeder cell or feeder layer. As used herein, a "feeder cell" is a cell that grows in vitro, that is co-cultured with a target cell and stabilizes the target cell in its current state of differentiation. As used herein, a "feeder cell layer" can be used interchangeably with the term "feeder cell." As used herein, the term "essentially free of a feeder cell" refers to tissue culture conditions that do not contain feeder cells, or that contain a de minimus number of feeder cells. By "de minims", it is meant that number of feeder cells that are carried over to the instant culture conditions from previous culture conditions where the differentiable cells may have been cultured on feeder cells. In one embodiment of the above method, conditioned medium is obtained from a feeder cell that stabilizes the target cell in its current state of differentiation. In another embodiment, the defined medium is a non-conditioned medium, which is a medium that is not obtained from a feeder cell.
[0097] As used herein, the term "stabilize," when used in reference to the differentiation state of a cell or culture of cells, indicates that the cells will continue to proliferate over multiple passages in culture, and preferably indefinitely in culture, where most, if not all, of the cells in the culture are of the same differentiation state. In addition, when the stabilized cells divide, the division typically yield cells of the same cell type or yield cells of the same differentiation state. A stabilized cell or cell population in general, does not further differentiate or de-differentiate if the cell culture conditions are not altered, and the eels continue to be passaged and are not overgrown. In one embodiment, the cell that is stabilized is capable of proliferation in the stable state indefinitely, or for at least more than 2 passages. In a more specific embodiment, the cells are stable for more than 3 passages, 4 passages, 5 passages, 6 passages, 7 passages, 8 passages, 9 passages, more than 10 passages, more than 15 passages, more than 20 passages, more than 25 passages, or more than 30 passages. In one embodiment, the cell is stable for greater than approximately I month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, or 11 months of continuous passaging. In another embodiment, the cell is stable for greater than approximately 1 year of continuous passaging. In one embodiment, stem cells are maintained in culture in a pluripotent state by routine passage in the defined medium until it is desired that they be differentiated. As used herein, the term "proliferate" refers to an increase in the number cells in a cell culture.
[0098] In certain embodiments, the compositions and methods comprise an inactivator of BMP signaling. As used herein, an "inactivator of BMP signaling" refers to an agent that antagonizes the activity of one or more BMP proteins or any of their upstream or downstream signaling components through any of its possible signaling pathways. The
|
[
1078,
1316,
408,
541,
451,
9078,
304,
1906,
5439,
2038,
29892,
408,
1532,
408,
278,
4636,
12341,
1623,
491,
263,
1238,
2447,
3038,
7546,
470,
491,
278,
715,
332,
666,
327,
296,
5199,
3038,
470,
3038,
9257,
29889,
512,
697,
7232,
397,
2073,
29892,
278,
9101,
526,
715,
630,
373,
341,
1299,
3960,
1692,
29931,
30342,
29899,
1111,
630,
715,
1078,
29889,
512,
1790,
7232,
397,
2073,
29892,
278,
9101,
526,
715,
630,
373,
18755,
1617,
522,
262,
29899,
1111,
630,
715,
1078,
29889,
512,
3058,
7232,
397,
7862,
29892,
565,
278,
9101,
526,
715,
630,
373,
18755,
1617,
522,
262,
29892,
278,
715,
1078,
526,
13240,
491,
1302,
1218,
411,
29871,
29896,
29900,
29871,
30167,
29887,
29914,
828,
5199,
715,
25392,
18755,
1617,
522,
262,
313,
12165,
277,
307,
1885,
29892,
396,
29941,
29941,
29900,
29896,
29953,
29899,
29900,
29896,
29945,
511,
21749,
3860,
297,
260,
15118,
19468,
4094,
29892,
363,
29871,
29906,
29899,
29941,
6199,
472,
5716,
10430,
29889,
512,
1790,
7232,
397,
2073,
29892,
724,
398,
508,
367,
7180,
297,
278,
18350,
363,
701,
304,
29871,
29906,
29946,
6199,
304,
2758,
9101,
304,
15284,
304,
278,
715,
6288,
29889,
960,
773,
724,
398,
304,
27391,
278,
26305,
310,
278,
9101,
29892,
278,
5745,
338,
769,
6206,
322,
278,
5541,
2187,
29892,
607,
526,
13674,
724,
398,
29899,
9021,
29892,
526,
2715,
304,
278,
715,
630,
9101,
29889,
13,
29961,
29900,
29900,
29929,
29953,
29962,
450,
5541,
2187,
322,
3519,
310,
278,
2198,
297,
7316,
640,
331,
2341,
393,
278,
17473,
519,
9101,
526,
4185,
2955,
297,
5855,
393,
526,
13674,
3889,
310,
263,
1238,
2447,
3038,
470,
1238,
2447,
7546,
29889,
1094,
1304,
1244,
262,
29892,
263,
376,
1725,
2447,
3038,
29908,
338,
263,
3038,
393,
25088,
297,
13901,
307,
29892,
393,
338,
1302,
29899,
29883,
499,
2955,
411,
263,
3646,
3038,
322,
16160,
7093,
278,
3646,
3038,
297,
967,
1857,
2106,
310,
1422,
11685,
29889,
1094,
1304,
1244,
262,
29892,
263,
376,
1725,
2447,
3038,
7546,
29908,
508,
367,
1304,
1006,
3167,
2197,
411,
278,
1840,
376,
1725,
2447,
3038,
1213,
1094,
1304,
1244,
262,
29892,
278,
1840,
376,
404,
9247,
3889,
310,
263,
1238,
2447,
3038,
29908,
14637,
304,
260,
15118,
9257,
5855,
393,
437,
451,
1712,
1238,
2447,
9101,
29892,
470,
393,
1712,
263,
316,
6260,
375,
1353,
310,
1238,
2447,
9101,
29889,
2648,
376,
311,
6260,
29879,
613,
372,
338,
6839,
393,
1353,
310,
1238,
2447,
9101,
393,
526,
8988,
975,
304,
278,
14426,
9257,
5855,
515,
3517,
9257,
5855,
988,
278,
17473,
519,
9101,
1122,
505,
1063,
4185,
2955,
373,
1238,
2447,
9101,
29889,
512,
697,
7232,
397,
2073,
310,
278,
2038,
1158,
29892,
4195,
287,
18350,
338,
7625,
515,
263,
1238,
2447,
3038,
393,
16160,
7093,
278,
3646,
3038,
297,
967,
1857,
2106,
310,
1422,
11685,
29889,
512,
1790,
7232,
397,
2073,
29892,
278,
3342,
18350,
338,
263,
1661,
29899,
16122,
287,
18350,
29892,
607,
338,
263,
18350,
393,
338,
451,
7625,
515,
263,
1238,
2447,
3038,
29889,
13,
29961,
29900,
29900,
29929,
29955,
29962,
1094,
1304,
1244,
262,
29892,
278,
1840,
376,
303,
4427,
675,
1699,
746,
1304,
297,
3407,
304,
278,
1422,
11685,
2106,
310,
263,
3038,
470,
9257,
310,
9101,
29892,
14088,
393,
278,
9101,
674,
6773,
304,
410,
29880,
9633,
403,
975,
2999,
1209,
1179,
297,
9257,
29892,
322,
5821,
2197,
297,
1753,
18639,
297,
9257,
29892,
988,
1556,
29892,
565,
451,
599,
29892,
310,
278,
9101,
297,
278,
9257,
526,
310,
278,
1021,
1422,
11685,
2106,
29889,
512,
6124,
29892,
746,
278,
16160,
1891,
9101,
16429,
29892,
278,
8542,
12234,
7709,
9101,
310,
278,
1021,
3038,
1134,
470,
7709,
9101,
310,
278,
1021,
1422,
11685,
2106,
29889,
319,
16160,
1891,
3038,
470,
3038,
4665,
297,
2498,
29892,
947,
451,
4340,
17473,
403,
470,
316,
29899,
29881,
8349,
7268,
403,
565,
278,
3038,
9257,
5855,
526,
451,
10551,
287,
29892,
322,
278,
321,
1379,
6773,
304,
367,
1209,
4063,
322,
526,
451,
975,
29887,
4708,
29889,
512,
697,
7232,
397,
2073,
29892,
278,
3038,
393,
338,
16160,
1891,
338,
15390,
310,
410,
29880,
9633,
362,
297,
278,
13714,
2106,
297,
1753,
18639,
29892,
470,
363,
472,
3203,
901,
1135,
29871,
29906,
1209,
1179,
29889,
512,
263,
901,
2702,
7232,
397,
2073,
29892,
278,
9101,
526,
13714,
363,
901,
1135,
29871,
29941,
1209,
1179,
29892,
29871,
29946,
1209,
1179,
29892,
29871,
29945,
1209,
1179,
29892,
29871,
29953,
1209,
1179,
29892,
29871,
29955,
1209,
1179,
29892,
29871,
29947,
1209,
1179,
29892,
29871,
29929,
1209,
1179,
29892,
901,
1135,
29871,
29896,
29900,
1209,
1179,
29892,
901,
1135,
29871,
29896,
29945,
1209,
1179,
29892,
901,
1135,
29871,
29906,
29900,
1209,
1179,
29892,
901,
1135,
29871,
29906,
29945,
1209,
1179,
29892,
470,
901,
1135,
29871,
29941,
29900,
1209,
1179,
29889,
512,
697,
7232,
397,
2073,
29892,
278,
3038,
338,
13714,
363,
7621,
1135,
14235,
306,
4098,
29892,
29871,
29906,
7378,
29892,
29871,
29941,
7378,
29892,
29871,
29946,
7378,
29892,
29871,
29945,
7378,
29892,
29871,
29953,
7378,
29892,
29871,
29955,
7378,
29892,
29871,
29947,
7378,
29892,
29871,
29929,
7378,
29892,
29871,
29896,
29900,
7378,
29892,
470,
29871,
29896,
29896,
7378,
310,
9126,
1209,
6751,
29889,
512,
1790,
7232,
397,
2073,
29892,
278,
3038,
338,
13714,
363,
7621,
1135,
14235,
29871,
29896,
1629,
310,
9126,
1209,
6751,
29889,
512,
697,
7232,
397,
2073,
29892,
20805,
9101,
526,
19949,
297,
9257,
297,
263,
715,
332,
666,
327,
296,
2106,
491,
26529,
13382,
297,
278,
3342,
18350,
2745,
372,
338,
7429,
393,
896,
367,
17473,
630,
29889,
1094,
1304,
1244,
262,
29892,
278,
1840,
376,
771,
29880,
9633,
403,
29908,
14637,
304,
385,
7910,
297,
278,
1353,
9101,
297,
263,
3038,
9257,
29889,
13,
29961,
29900,
29900,
29929,
29947,
29962,
512,
3058,
7232,
397,
7862,
29892,
278,
5541,
2187,
322,
3519,
7199,
895,
385,
297,
11236,
1061,
310,
350,
3580,
7182,
292,
29889,
1094,
1304,
1244,
262,
29892,
385,
376,
262,
11236,
1061,
310,
350,
3580,
7182,
292,
29908,
14637,
304,
385,
10823,
393,
3677,
12841,
7093,
278,
6354,
310,
697,
470,
901,
350,
3580,
3279,
1144,
470,
738,
310,
1009,
701,
5461,
470,
1623,
5461,
7182,
292,
7117,
1549,
738,
310,
967,
1950,
7182,
292,
2224,
1994,
29889,
450
] | 1,024 |
CommonCrawl
| -0.89316 | 1.127121 | -0.61182 | 0.142678 |
compound(s) used to inactivate BMP signaling can be any compound known in the art, or later discovered. Non-limiting examples of inactivators of BMP signaling include dominant-negative, truncated BMP receptor, soluble BMP receptors, BMP receptor-Fe chimeras, noggin, follistatin, chordin, gremlin, cerberus/DAN family proteins, ventropin, high dose activin, and amnionless.
[0099] In certain embodiments, the compositions and methods can comprise at least one hormone, cytokine, adipokine, growth hormone or variant or functional fragment thereof. It is currently contemplated that in certain embodiments, the growth hormone present in the defined medium will be of the same species as the differentiable cells that are cultured with the defined media. Thus, for example, if a human cell is cultured, the growth hormone is human growth hormone. The use of growth hormone that is from a species different than the cultured cells is also contemplated. Preferably the hormone, cytokine, adipokine and/or growth hormone is present at an initial concentration of approximately 0.001 ng/ml to approximately 1000 ng/ml, more preferably approximately 0.001 ng/ml to approximately 250 ng/ml, or more preferably approximately 0.01 ng/ml to approximately 150 ng/ml.
[0100] Examples of cytokines and adipokines that may be included in the compositions and methods of the present invention include, but are not limited to, the four α-helix bundle family of cytokines, the interleukin (IL-1) family of cytokines, the IL-17 family of cytokines and the chemokine family of cytokines. Of course, the invention contemplates members and subclasses of each of these families of cytokines, such as, but not limited to, the CC chemokines, the CXC chemokines, the C chemokines and the CX3C chemokines, interferons, interleukins, lymphotoxins, c-kit ligand, granulocyte-macrophage colony-stimulating factor, (GM-CSF), monocyte-macrophage colony-stimulating factor (M-CSF), granulocyte colony-stimulating factor (G-CSF), leptin, adiponectin, resistin, plasminogen activator inhibitor-1 (PAI-1), tumor necrosis factor-alpha (TNFα), tumor necrosis factor-beta (TNFβ), leukemia inhibitory factor, visfatin, retinol binding protein 4 (RBP4), erythropoietin (EPO), thrombopoietin (THPO). Of course, one of skill in the art will understand that the invention contemplates variants or functional fragments of the above-listed factors.
[0101] The present invention relates to methods of culturing differentiable cells, with the methods comprising plating differentiable cells on a cell culture surface, providing a basal salt nutrient solution to the cells and providing a means for stimulating ErbB2-directed tyrosine kinase activity in the cells.
[0102] In one embodiment; differentiable cells are contacted with the at least one of the compositions of the invention in the absence of serum or serum replacement, and in the absence of a feeder cell layer, such that the cells are maintained in an undifferentiated state for at least one month. Pluripotency can be determined through characterization of the cells with respect to surface markers, transcriptional markers, karyotype, and ability to differentiate to cells of the three germ layers. These characteristics are well known to those of ordinary skill in the art.
[0103] In another embodiment, the differentiable cells are cultured in suspension, using the cell media of the present invention. The term "suspension" as used in the context of cell culturing is used as it is in the art. Namely, cell culture suspensions are cell culture environments where the cells do not adhere to a surface. One of skill in the art will be familiar with suspension culture techniques, including, but not limited to
|
[
752,
618,
29898,
29879,
29897,
1304,
304,
297,
11236,
403,
350,
3580,
7182,
292,
508,
367,
738,
752,
618,
2998,
297,
278,
1616,
29892,
470,
2678,
10943,
29889,
10050,
29899,
2576,
11407,
6455,
310,
297,
11236,
4097,
310,
350,
3580,
7182,
292,
3160,
28526,
29899,
22198,
29892,
21022,
630,
350,
3580,
337,
14268,
29892,
899,
431,
280,
350,
3580,
337,
1547,
943,
29892,
350,
3580,
337,
14268,
29899,
8263,
521,
4193,
294,
29892,
11313,
5359,
29892,
900,
1761,
21203,
29892,
521,
4194,
29892,
1395,
828,
262,
29892,
5147,
495,
375,
29914,
29928,
2190,
3942,
3279,
1144,
29892,
9712,
1336,
262,
29892,
1880,
437,
344,
5039,
262,
29892,
322,
626,
29876,
291,
2222,
29889,
13,
29961,
29900,
29900,
29929,
29929,
29962,
512,
3058,
7232,
397,
7862,
29892,
278,
5541,
2187,
322,
3519,
508,
7199,
895,
472,
3203,
697,
298,
555,
650,
29892,
5094,
17082,
457,
29892,
594,
666,
554,
457,
29892,
14321,
298,
555,
650,
470,
17305,
470,
13303,
9376,
727,
974,
29889,
739,
338,
5279,
640,
16305,
630,
393,
297,
3058,
7232,
397,
7862,
29892,
278,
14321,
298,
555,
650,
2198,
297,
278,
3342,
18350,
674,
367,
310,
278,
1021,
6606,
408,
278,
17473,
519,
9101,
393,
526,
4185,
2955,
411,
278,
3342,
5745,
29889,
6549,
29892,
363,
1342,
29892,
565,
263,
5199,
3038,
338,
4185,
2955,
29892,
278,
14321,
298,
555,
650,
338,
5199,
14321,
298,
555,
650,
29889,
450,
671,
310,
14321,
298,
555,
650,
393,
338,
515,
263,
6606,
1422,
1135,
278,
4185,
2955,
9101,
338,
884,
640,
16305,
630,
29889,
4721,
571,
2197,
278,
298,
555,
650,
29892,
5094,
17082,
457,
29892,
594,
666,
554,
457,
322,
29914,
272,
14321,
298,
555,
650,
338,
2198,
472,
385,
2847,
26702,
310,
14235,
29871,
29900,
29889,
29900,
29900,
29896,
8736,
29914,
828,
304,
14235,
29871,
29896,
29900,
29900,
29900,
8736,
29914,
828,
29892,
901,
5821,
2197,
14235,
29871,
29900,
29889,
29900,
29900,
29896,
8736,
29914,
828,
304,
14235,
29871,
29906,
29945,
29900,
8736,
29914,
828,
29892,
470,
901,
5821,
2197,
14235,
29871,
29900,
29889,
29900,
29896,
8736,
29914,
828,
304,
14235,
29871,
29896,
29945,
29900,
8736,
29914,
828,
29889,
13,
29961,
29900,
29896,
29900,
29900,
29962,
1222,
9422,
310,
5094,
17082,
1475,
322,
594,
666,
554,
1475,
393,
1122,
367,
5134,
297,
278,
5541,
2187,
322,
3519,
310,
278,
2198,
297,
7316,
3160,
29892,
541,
526,
451,
9078,
304,
29892,
278,
3023,
19883,
29899,
3952,
861,
11846,
3942,
310,
5094,
17082,
1475,
29892,
278,
1006,
280,
2679,
262,
313,
6227,
29899,
29896,
29897,
3942,
310,
5094,
17082,
1475,
29892,
278,
17674,
29899,
29896,
29955,
3942,
310,
5094,
17082,
1475,
322,
278,
8950,
554,
457,
3942,
310,
5094,
17082,
1475,
29889,
4587,
3236,
29892,
278,
297,
7316,
640,
331,
9884,
5144,
322,
1014,
13203,
310,
1269,
310,
1438,
13175,
310,
5094,
17082,
1475,
29892,
1316,
408,
29892,
541,
451,
9078,
304,
29892,
278,
19178,
8950,
554,
1475,
29892,
278,
315,
29990,
29907,
8950,
554,
1475,
29892,
278,
315,
8950,
554,
1475,
322,
278,
315,
29990,
29941,
29907,
8950,
554,
1475,
29892,
1006,
571,
787,
29892,
1006,
280,
2679,
1144,
29892,
301,
962,
561,
327,
2251,
1144,
29892,
274,
29899,
7354,
14172,
392,
29892,
3803,
352,
22502,
371,
29899,
8628,
19783,
482,
784,
2592,
29899,
303,
326,
18099,
7329,
29892,
313,
21576,
29899,
9295,
29943,
511,
1601,
22502,
371,
29899,
8628,
19783,
482,
784,
2592,
29899,
303,
326,
18099,
7329,
313,
29924,
29899,
9295,
29943,
511,
3803,
352,
22502,
371,
784,
2592,
29899,
303,
326,
18099,
7329,
313,
29954,
29899,
9295,
29943,
511,
454,
415,
262,
29892,
594,
666,
265,
522,
262,
29892,
9241,
262,
29892,
715,
294,
1195,
6352,
5039,
1061,
297,
6335,
2105,
29899,
29896,
313,
7228,
29902,
29899,
29896,
511,
21622,
272,
452,
29883,
1883,
275,
7329,
29899,
2312,
313,
29911,
22498,
30110,
511,
21622,
272,
452,
29883,
1883,
275,
7329,
29899,
3571,
313,
29911,
22498,
30187,
511,
454,
2679,
29747,
297,
6335,
5047,
7329,
29892,
1998,
29888,
21203,
29892,
3240,
262,
324,
9956,
26823,
29871,
29946,
313,
29934,
29933,
29925,
29946,
511,
604,
1541,
307,
1129,
2035,
262,
313,
29923,
13152,
511,
266,
456,
29890,
12861,
2035,
262,
313,
29911,
3954,
29949,
467,
4587,
3236,
29892,
697,
310,
19911,
297,
278,
1616,
674,
2274,
393,
278,
297,
7316,
640,
331,
9884,
29161,
470,
13303,
22370,
310,
278,
2038,
29899,
1761,
287,
13879,
29889,
13,
29961,
29900,
29896,
29900,
29896,
29962,
450,
2198,
297,
7316,
1104,
1078,
304,
3519,
310,
4185,
3864,
17473,
519,
9101,
29892,
411,
278,
3519,
7199,
5921,
715,
1218,
17473,
519,
9101,
373,
263,
3038,
9257,
7101,
29892,
13138,
263,
2362,
284,
15795,
18254,
374,
296,
1650,
304,
278,
9101,
322,
13138,
263,
2794,
363,
20436,
18099,
1425,
29890,
29933,
29906,
29899,
11851,
287,
7911,
1883,
457,
19015,
559,
6354,
297,
278,
9101,
29889,
13,
29961,
29900,
29896,
29900,
29906,
29962,
512,
697,
7232,
397,
2073,
29936,
17473,
519,
9101,
526,
6958,
287,
411,
278,
472,
3203,
697,
310,
278,
5541,
2187,
310,
278,
297,
7316,
297,
278,
18070,
310,
724,
398,
470,
724,
398,
16920,
29892,
322,
297,
278,
18070,
310,
263,
1238,
2447,
3038,
7546,
29892,
1316,
393,
278,
9101,
526,
19949,
297,
385,
563,
8349,
7268,
630,
2106,
363,
472,
3203,
697,
4098,
29889,
1858,
332,
666,
327,
3819,
508,
367,
10087,
1549,
2931,
2133,
310,
278,
9101,
411,
3390,
304,
7101,
29320,
29892,
1301,
3395,
284,
29320,
29892,
413,
653,
327,
668,
29892,
322,
11509,
304,
17473,
403,
304,
9101,
310,
278,
2211,
22593,
15359,
29889,
4525,
21862,
526,
1532,
2998,
304,
1906,
310,
15311,
19911,
297,
278,
1616,
29889,
13,
29961,
29900,
29896,
29900,
29941,
29962,
512,
1790,
7232,
397,
2073,
29892,
278,
17473,
519,
9101,
526,
4185,
2955,
297,
8872,
2673,
29892,
773,
278,
3038,
5745,
310,
278,
2198,
297,
7316,
29889,
450,
1840,
376,
29879,
375,
29886,
2673,
29908,
408,
1304,
297,
278,
3030,
310,
3038,
4185,
3864,
338,
1304,
408,
372,
338,
297,
278,
1616,
29889,
13041,
873,
29892,
3038,
9257,
8872,
5580,
526,
3038,
9257,
23136,
988,
278,
9101,
437,
451,
594,
4150,
304,
263,
7101,
29889,
3118,
310,
19911,
297,
278,
1616,
674,
367,
9985,
411,
8872,
2673,
9257,
13698,
29892,
3704,
29892,
541,
451,
9078,
304
] | 1,024 |
CommonCrawl
| -0.982823 | 1.464545 | -0.031264 | 0.006072 |
, the use of equipment such as flow hoods, incubators and/or equipment used to keep the cells in constant motion, e.g., rotator platforms, shakers, etc, if necessary. As used herein, cells are "in motion" if they are moving, or if their immediate environment is moving relative to the cells. If the cells are kept "in motion", the motion will, in one embodiment, be a "gentle motion" that is designed to avoid or prevent exposing the cells to sheer stress.
[0104] In one particular embodiment, the differentiable cells are expanded in a suspension culture, using the cell media of the present invention. In another particular embodiment, the differentiable cells expand in suspension, but do not differentiate. The term "expand" in the context of cell culture is used as it is the art, Namely, expand is used to indicate cellular proliferation to increase the number of viable cells. In a specific embodiment, the cells are expanded in a culture suspension by culturing for more than about one day, i.e., about 24 hours. In a more specific embodiment, the cells are expanded in a suspension culture by culturing for at least 2, 3, 4, 5, 6, 7 days, or at least 2, 3, 4, 5, 6, 7, 8 weeks.
[0105] In general, the cell medium compositions of the present invention are refreshed at least once every day, but the medium can be changed more often or less often, depending of the specific needs and circumstances of the suspension culture. For example, the medium may be refreshed about every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 hours, or any fraction thereof. In additional examples, the medium may be refreshed less often such as, but not limited to, every 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9 or every 2 or more days, or any time frame in between.
[0106] In general, the cells that are cultured in suspension in the medium compositions of the present invention are "split" every week or so, but the cells can be split more often or less often, depending of the specific needs and circumstances of the suspension culture. For example, the cells may be split every 1, 2, 3, 4, 5, 6 ,7 ,8, 9, 10, 11, 12, 13, 14 or more days, or any time frame in between. As used herein, the term "split" in the context of cell culture is used as it is in the art. Namely, cell culture splitting, or passaging, is the collection of cells from a previous culture and subsequent transfer of small number of collected (harvested) cells into a new cell culture vessel. In general, passaging cells allows the cells to continue to grow in a healthy cell culture environment. One of skill in the art will be familiar with the process and methods of cell culture passaging, which may, but not necessarily, involve the use enzymatic or non-enzymatic methods that may be used to disaggregate cells that have clumped together during their growth expansion.
[0107] In some instances, a degree of cell death may occur in the cultured (suspended and adherent) cells immediately after passaging. In one embodiment, the differentiable cells can "recover" from passaging, by delaying the refreshing of the cell medium for more than 24 hours. Thereafter, the cell medium may be changed more frequently. In another embodiment, the cell culture medium can further comprise an inhibitor of cell death. For example, Wartanabe et al., recently disclosed the use of a Rho-associated kinase inhibitor, Y27632, to protect human ES cells after dissociation. See Watanbe, K., et al., Nat. Biotechnol., 25(6):681-686 (2007), which is incorpor
|
[
29892,
278,
671,
310,
21083,
1316,
408,
4972,
298,
2092,
29879,
29892,
5528,
431,
4097,
322,
29914,
272,
21083,
1304,
304,
3013,
278,
9101,
297,
4868,
10884,
29892,
321,
29889,
29887,
1696,
5731,
1061,
21796,
29892,
528,
21079,
29892,
2992,
29892,
565,
5181,
29889,
1094,
1304,
1244,
262,
29892,
9101,
526,
376,
262,
10884,
29908,
565,
896,
526,
8401,
29892,
470,
565,
1009,
16800,
5177,
338,
8401,
6198,
304,
278,
9101,
29889,
960,
278,
9101,
526,
8126,
376,
262,
10884,
613,
278,
10884,
674,
29892,
297,
697,
7232,
397,
2073,
29892,
367,
263,
376,
5362,
280,
10884,
29908,
393,
338,
8688,
304,
4772,
470,
5557,
14060,
292,
278,
9101,
304,
1183,
261,
22884,
29889,
13,
29961,
29900,
29896,
29900,
29946,
29962,
512,
697,
3153,
7232,
397,
2073,
29892,
278,
17473,
519,
9101,
526,
17832,
297,
263,
8872,
2673,
9257,
29892,
773,
278,
3038,
5745,
310,
278,
2198,
297,
7316,
29889,
512,
1790,
3153,
7232,
397,
2073,
29892,
278,
17473,
519,
9101,
7985,
297,
8872,
2673,
29892,
541,
437,
451,
17473,
403,
29889,
450,
1840,
376,
18837,
29908,
297,
278,
3030,
310,
3038,
9257,
338,
1304,
408,
372,
338,
278,
1616,
29892,
13041,
873,
29892,
7985,
338,
1304,
304,
12266,
3038,
1070,
410,
29880,
9633,
362,
304,
7910,
278,
1353,
310,
3516,
519,
9101,
29889,
512,
263,
2702,
7232,
397,
2073,
29892,
278,
9101,
526,
17832,
297,
263,
9257,
8872,
2673,
491,
4185,
3864,
363,
901,
1135,
1048,
697,
2462,
29892,
474,
29889,
29872,
1696,
1048,
29871,
29906,
29946,
6199,
29889,
512,
263,
901,
2702,
7232,
397,
2073,
29892,
278,
9101,
526,
17832,
297,
263,
8872,
2673,
9257,
491,
4185,
3864,
363,
472,
3203,
29871,
29906,
29892,
29871,
29941,
29892,
29871,
29946,
29892,
29871,
29945,
29892,
29871,
29953,
29892,
29871,
29955,
3841,
29892,
470,
472,
3203,
29871,
29906,
29892,
29871,
29941,
29892,
29871,
29946,
29892,
29871,
29945,
29892,
29871,
29953,
29892,
29871,
29955,
29892,
29871,
29947,
11405,
29889,
13,
29961,
29900,
29896,
29900,
29945,
29962,
512,
2498,
29892,
278,
3038,
18350,
5541,
2187,
310,
278,
2198,
297,
7316,
526,
11086,
287,
472,
3203,
2748,
1432,
2462,
29892,
541,
278,
18350,
508,
367,
3939,
901,
4049,
470,
3109,
4049,
29892,
8679,
310,
278,
2702,
4225,
322,
14209,
310,
278,
8872,
2673,
9257,
29889,
1152,
1342,
29892,
278,
18350,
1122,
367,
11086,
287,
1048,
1432,
29871,
29896,
29892,
29871,
29906,
29892,
29871,
29941,
29892,
29871,
29946,
29892,
29871,
29945,
29892,
29871,
29953,
29892,
29871,
29955,
29892,
29871,
29947,
29892,
29871,
29929,
29892,
29871,
29896,
29900,
29892,
29871,
29896,
29896,
29892,
29871,
29896,
29906,
29892,
29871,
29896,
29941,
29892,
29871,
29896,
29946,
29892,
29871,
29896,
29945,
29892,
29871,
29896,
29953,
29892,
29871,
29896,
29955,
29892,
29871,
29896,
29947,
29892,
29871,
29896,
29929,
29892,
29871,
29906,
29900,
29892,
29871,
29906,
29896,
29892,
29871,
29906,
29906,
29892,
29871,
29906,
29941,
470,
29871,
29906,
29946,
6199,
29892,
470,
738,
15958,
727,
974,
29889,
512,
5684,
6455,
29892,
278,
18350,
1122,
367,
11086,
287,
3109,
4049,
1316,
408,
29892,
541,
451,
9078,
304,
29892,
1432,
29871,
29896,
29889,
29896,
29892,
29871,
29896,
29889,
29906,
29892,
29871,
29896,
29889,
29941,
29892,
29871,
29896,
29889,
29946,
29892,
29871,
29896,
29889,
29945,
29892,
29871,
29896,
29889,
29953,
29892,
29871,
29896,
29889,
29955,
29892,
29871,
29896,
29889,
29947,
29892,
29871,
29896,
29889,
29929,
470,
1432,
29871,
29906,
470,
901,
3841,
29892,
470,
738,
931,
3515,
297,
1546,
29889,
13,
29961,
29900,
29896,
29900,
29953,
29962,
512,
2498,
29892,
278,
9101,
393,
526,
4185,
2955,
297,
8872,
2673,
297,
278,
18350,
5541,
2187,
310,
278,
2198,
297,
7316,
526,
376,
5451,
29908,
1432,
4723,
470,
577,
29892,
541,
278,
9101,
508,
367,
6219,
901,
4049,
470,
3109,
4049,
29892,
8679,
310,
278,
2702,
4225,
322,
14209,
310,
278,
8872,
2673,
9257,
29889,
1152,
1342,
29892,
278,
9101,
1122,
367,
6219,
1432,
29871,
29896,
29892,
29871,
29906,
29892,
29871,
29941,
29892,
29871,
29946,
29892,
29871,
29945,
29892,
29871,
29953,
1919,
29955,
1919,
29947,
29892,
29871,
29929,
29892,
29871,
29896,
29900,
29892,
29871,
29896,
29896,
29892,
29871,
29896,
29906,
29892,
29871,
29896,
29941,
29892,
29871,
29896,
29946,
470,
901,
3841,
29892,
470,
738,
931,
3515,
297,
1546,
29889,
1094,
1304,
1244,
262,
29892,
278,
1840,
376,
5451,
29908,
297,
278,
3030,
310,
3038,
9257,
338,
1304,
408,
372,
338,
297,
278,
1616,
29889,
13041,
873,
29892,
3038,
9257,
24368,
29892,
470,
1209,
6751,
29892,
338,
278,
4333,
310,
9101,
515,
263,
3517,
9257,
322,
15352,
6782,
310,
2319,
1353,
310,
16531,
313,
8222,
29894,
2868,
29897,
9101,
964,
263,
716,
3038,
9257,
21239,
29889,
512,
2498,
29892,
1209,
6751,
9101,
6511,
278,
9101,
304,
6773,
304,
6548,
297,
263,
9045,
29891,
3038,
9257,
5177,
29889,
3118,
310,
19911,
297,
278,
1616,
674,
367,
9985,
411,
278,
1889,
322,
3519,
310,
3038,
9257,
1209,
6751,
29892,
607,
1122,
29892,
541,
451,
12695,
29892,
25135,
278,
671,
427,
14022,
2454,
470,
1661,
29899,
4666,
962,
2454,
3519,
393,
1122,
367,
1304,
304,
22941,
7642,
403,
9101,
393,
505,
1067,
3427,
287,
4208,
2645,
1009,
14321,
13184,
29889,
13,
29961,
29900,
29896,
29900,
29955,
29962,
512,
777,
8871,
29892,
263,
7426,
310,
3038,
4892,
1122,
6403,
297,
278,
4185,
2955,
313,
29879,
375,
29886,
2760,
322,
594,
2276,
296,
29897,
9101,
7389,
1156,
1209,
6751,
29889,
512,
697,
7232,
397,
2073,
29892,
278,
17473,
519,
9101,
508,
376,
3757,
957,
29908,
515,
1209,
6751,
29892,
491,
9055,
292,
278,
2143,
690,
2790,
310,
278,
3038,
18350,
363,
901,
1135,
29871,
29906,
29946,
6199,
29889,
1670,
7045,
29892,
278,
3038,
18350,
1122,
367,
3939,
901,
13672,
29889,
512,
1790,
7232,
397,
2073,
29892,
278,
3038,
9257,
18350,
508,
4340,
7199,
895,
385,
297,
6335,
2105,
310,
3038,
4892,
29889,
1152,
1342,
29892,
399,
442,
273,
4302,
634,
394,
1696,
10325,
766,
15603,
278,
671,
310,
263,
390,
1251,
29899,
21264,
630,
19015,
559,
297,
6335,
2105,
29892,
612,
29906,
29955,
29953,
29941,
29906,
29892,
304,
12566,
5199,
17956,
9101,
1156,
766,
2839,
362,
29889,
2823,
14269,
273,
915,
29892,
476,
1696,
634,
394,
1696,
19259,
29889,
3457,
866,
3049,
324,
1696,
29871,
29906,
29945,
29898,
29953,
1125,
29953,
29947,
29896,
29899,
29953,
29947,
29953,
313,
29906,
29900,
29900,
29955,
511,
607,
338,
11039
] | 1,024 |
CommonCrawl
| -1.086071 | 0.50678 | -0.854353 | -0.035037 |
ated by reference. In additional embodiments, the cell culture medium may comprise caspase inhibitors, growth factors or other trophic factors to prevent or attenuate cell death immediately after passaging. Specific examples of compounds that may be used include, but are not limited to, HA 1077, Dihydrochloride, Hydroxyfasudil, Rho Kinase Inhibitor, Rho-Kinase Inhibitor II, Rho Kinase Inhibitor III, Kinase Inhibitor IV and Y27632 all of which are commercially available. In still another embodiment, the compounds or factors used to prevent or attenuate cell death during or immediately after cell passaging may be removed from the cell culture medium after the cells have recovered from the passaging process.
[0108] In additional embodiments, the compositions and methods the present invention may also comprise the presence or use of surfactants. In one particular embodiment, the compositions and methods comprise at least one surfactant in the context of a suspension culture. Surfactants are well-known in the art and, generally speaking, are amphiphilic in nature. In specific embodiments, the present invention comprises the use of at least one surfactant that is either anionic, cationic, non-ionic or zwitterionic. The concentration of the surfactant used in the compositions and methods of the present invention is a matter of routine screening and optimization. For example, Owen et al., reported the use of surfactants in cell culture techniques for HeLa cells and human amniotic cells. See Owen et al., J. Cell. Sci., 32:363-376 (1978), which is incorporated by reference. Examples of surfactants that may be used include, but are not limited to, Sodium dodecyl sulfate (SDS), ammonium lauryl sulfate, and other alkyl sulfate salts, Sodium laureth sulfate (SLES), Alkyl benzene sulfonate, Soaps, or fatty acid salts, Cetyl trimethylammonium bromide (CTAB) (hexadecyl trimethyl ammonium bromide), and other alkyltrimethylammonium salts, Cetylpyridinium chloride (CPC), Polyethoxylated tallow amine (POEA), Benzalkonium chloride (BAC), Benzethonium chloride (BZT), Dodecyl betaine, Dodecyl dimethylamine oxide, Cocamidopropyl betaine, Coco ampho glycinate, Alkyl poly(ethylene oxide), Copolymers of poly(ethylene oxide) and poly(propylene oxide) such as Pluronic F68, Alkyl polyglucosides, such as, but not limited to, Octyl glucoside, Decyl maltoside, Fatty alcohols, Cetyl alcohol, Oleyl alcohol, Cocamide MEA, cocamidc DEA and cocamidc TEA.
[0109] It is contemplated that the differentiable cells can be passaged using enzymatic, non-enzymatic, or manual dissociation methods prior to and/or after contact with the defined medium of the invention. Non-limiting examples of enzymatic dissociation methods include the use of proteases such as trypsin, collagenase, dispase, and ACCUTASE®. In one embodiment, ACCUTASE® is used to passage the contacted cells. When enzymatic passaging methods are used, the resultant culture can comprise a mixture of singlets, doublets, triplets, and clumps of cells that vary in size depending on the enzyme used. A non-limiting example of a non-enzymatic dissociation method is a cell dispersal buffer. Manual passaging techniques have been well described in the art, such as in Schulz et al., 2004 Stem Cells, 22(7):1218-38. The choice of passaging method is influenced by the choice of extracellular matrix, if one is present, and is easily determined by one of ordinary skill in the art.
[0110] In one specific embodiment, methods of culturing differentiable cells comprise providing
|
[
630,
491,
3407,
29889,
512,
5684,
7232,
397,
7862,
29892,
278,
3038,
9257,
18350,
1122,
7199,
895,
3209,
29886,
559,
297,
6335,
17259,
29892,
14321,
13879,
470,
916,
3147,
561,
293,
13879,
304,
5557,
470,
472,
841,
27240,
3038,
4892,
7389,
1156,
1209,
6751,
29889,
21220,
6455,
310,
752,
3885,
393,
1122,
367,
1304,
3160,
29892,
541,
526,
451,
9078,
304,
29892,
379,
29909,
29871,
29896,
29900,
29955,
29955,
29892,
4671,
29882,
11279,
305,
5095,
680,
29892,
379,
11279,
3594,
13629,
566,
309,
29892,
390,
1251,
20311,
559,
512,
6335,
2105,
29892,
390,
1251,
29899,
29968,
262,
559,
512,
6335,
2105,
1944,
29892,
390,
1251,
20311,
559,
512,
6335,
2105,
4786,
29892,
20311,
559,
512,
6335,
2105,
6599,
322,
612,
29906,
29955,
29953,
29941,
29906,
599,
310,
607,
526,
7825,
5584,
3625,
29889,
512,
1603,
1790,
7232,
397,
2073,
29892,
278,
752,
3885,
470,
13879,
1304,
304,
5557,
470,
472,
841,
27240,
3038,
4892,
2645,
470,
7389,
1156,
3038,
1209,
6751,
1122,
367,
6206,
515,
278,
3038,
9257,
18350,
1156,
278,
9101,
505,
24776,
515,
278,
1209,
6751,
1889,
29889,
13,
29961,
29900,
29896,
29900,
29947,
29962,
512,
5684,
7232,
397,
7862,
29892,
278,
5541,
2187,
322,
3519,
278,
2198,
297,
7316,
1122,
884,
7199,
895,
278,
10122,
470,
671,
310,
1190,
17028,
1934,
29889,
512,
697,
3153,
7232,
397,
2073,
29892,
278,
5541,
2187,
322,
3519,
7199,
895,
472,
3203,
697,
1190,
17028,
424,
297,
278,
3030,
310,
263,
8872,
2673,
9257,
29889,
6298,
17028,
1934,
526,
1532,
29899,
5203,
297,
278,
1616,
322,
29892,
6892,
13590,
29892,
526,
626,
2876,
561,
309,
293,
297,
5469,
29889,
512,
2702,
7232,
397,
7862,
29892,
278,
2198,
297,
7316,
7199,
4637,
278,
671,
310,
472,
3203,
697,
1190,
17028,
424,
393,
338,
2845,
385,
26629,
29892,
274,
362,
293,
29892,
1661,
29899,
26629,
470,
5263,
5171,
26629,
29889,
450,
26702,
310,
278,
1190,
17028,
424,
1304,
297,
278,
5541,
2187,
322,
3519,
310,
278,
2198,
297,
7316,
338,
263,
4383,
310,
26529,
4315,
292,
322,
13883,
29889,
1152,
1342,
29892,
438,
15556,
634,
394,
1696,
8967,
278,
671,
310,
1190,
17028,
1934,
297,
3038,
9257,
13698,
363,
940,
5661,
9101,
322,
5199,
626,
1240,
13574,
9101,
29889,
2823,
438,
15556,
634,
394,
1696,
435,
29889,
19413,
29889,
5636,
1696,
29871,
29941,
29906,
29901,
29941,
29953,
29941,
29899,
29941,
29955,
29953,
313,
29896,
29929,
29955,
29947,
511,
607,
338,
11039,
630,
491,
3407,
29889,
1222,
9422,
310,
1190,
17028,
1934,
393,
1122,
367,
1304,
3160,
29892,
541,
526,
451,
9078,
304,
29892,
317,
397,
1974,
270,
356,
1270,
29880,
5394,
29888,
403,
313,
7230,
29903,
511,
626,
3712,
1974,
425,
2857,
29880,
5394,
29888,
403,
29892,
322,
916,
29827,
2904,
5394,
29888,
403,
4497,
1372,
29892,
317,
397,
1974,
425,
545,
386,
5394,
29888,
403,
313,
29903,
17101,
511,
838,
29895,
2904,
3856,
29920,
1600,
5394,
18753,
403,
29892,
1105,
2547,
29892,
470,
9950,
1017,
22193,
4497,
1372,
29892,
15018,
2904,
17151,
621,
2904,
314,
3712,
1974,
289,
456,
680,
313,
1783,
2882,
29897,
313,
20970,
1943,
1270,
29880,
17151,
621,
2904,
626,
3712,
1974,
289,
456,
680,
511,
322,
916,
29827,
2904,
15450,
621,
2904,
314,
3712,
1974,
4497,
1372,
29892,
15018,
2904,
2272,
2429,
262,
1974,
521,
5095,
680,
313,
6271,
29907,
511,
21755,
621,
2251,
2904,
630,
260,
9536,
626,
457,
313,
13152,
29923,
29909,
511,
4111,
29920,
2235,
265,
1974,
521,
5095,
680,
313,
29933,
2477,
511,
4111,
29920,
621,
265,
1974,
521,
5095,
680,
313,
29933,
29999,
29911,
511,
360,
356,
1270,
29880,
1010,
9874,
29892,
360,
356,
1270,
29880,
3964,
621,
2904,
314,
457,
19100,
680,
29892,
315,
542,
314,
333,
459,
1336,
2904,
1010,
9874,
29892,
315,
6235,
626,
561,
29877,
330,
368,
16381,
403,
29892,
838,
29895,
2904,
15680,
29898,
621,
2904,
1600,
19100,
680,
511,
10061,
324,
962,
414,
310,
15680,
29898,
621,
2904,
1600,
19100,
680,
29897,
322,
15680,
29898,
771,
2272,
29880,
1600,
19100,
680,
29897,
1316,
408,
1858,
332,
8927,
383,
29953,
29947,
29892,
838,
29895,
2904,
15680,
3820,
1682,
359,
2247,
29892,
1316,
408,
29892,
541,
451,
9078,
304,
29892,
4756,
2904,
3144,
1682,
359,
680,
29892,
3826,
2904,
286,
1997,
359,
680,
29892,
383,
271,
1017,
27231,
29882,
3775,
29892,
15018,
2904,
27231,
5391,
29892,
438,
2330,
29880,
27231,
5391,
29892,
315,
542,
314,
680,
22986,
29909,
29892,
274,
542,
314,
333,
29883,
5012,
29909,
322,
274,
542,
314,
333,
29883,
17067,
29909,
29889,
13,
29961,
29900,
29896,
29900,
29929,
29962,
739,
338,
640,
16305,
630,
393,
278,
17473,
519,
9101,
508,
367,
1209,
4063,
773,
427,
14022,
2454,
29892,
1661,
29899,
4666,
962,
2454,
29892,
470,
12219,
766,
2839,
362,
3519,
7536,
304,
322,
29914,
272,
1156,
6958,
411,
278,
3342,
18350,
310,
278,
297,
7316,
29889,
10050,
29899,
2576,
11407,
6455,
310,
427,
14022,
2454,
766,
2839,
362,
3519,
3160,
278,
671,
310,
3279,
2129,
1316,
408,
1018,
567,
262,
29892,
784,
11541,
559,
29892,
12272,
559,
29892,
322,
319,
4174,
2692,
8127,
30342,
29889,
512,
697,
7232,
397,
2073,
29892,
319,
4174,
2692,
8127,
30342,
338,
1304,
304,
13382,
278,
6958,
287,
9101,
29889,
1932,
427,
14022,
2454,
1209,
6751,
3519,
526,
1304,
29892,
278,
1121,
424,
9257,
508,
7199,
895,
263,
29544,
310,
1809,
10376,
29892,
3765,
1372,
29892,
21954,
1372,
29892,
322,
1067,
17204,
310,
9101,
393,
13100,
297,
2159,
8679,
373,
278,
427,
14022,
29872,
1304,
29889,
319,
1661,
29899,
2576,
11407,
1342,
310,
263,
1661,
29899,
4666,
962,
2454,
766,
2839,
362,
1158,
338,
263,
3038,
29106,
284,
6835,
29889,
2315,
950,
1209,
6751,
13698,
505,
1063,
1532,
5439,
297,
278,
1616,
29892,
1316,
408,
297,
12988,
29920,
634,
394,
1696,
29871,
29906,
29900,
29900,
29946,
624,
331,
315,
10071,
29892,
29871,
29906,
29906,
29898,
29955,
1125,
29896,
29906,
29896,
29947,
29899,
29941,
29947,
29889,
450,
7348,
310,
1209,
6751,
1158,
338,
28482,
491,
278,
7348,
310,
1294,
945,
514,
1070,
4636,
29892,
565,
697,
338,
2198,
29892,
322,
338,
5948,
10087,
491,
697,
310,
15311,
19911,
297,
278,
1616,
29889,
13,
29961,
29900,
29896,
29896,
29900,
29962,
512,
697,
2702,
7232,
397,
2073,
29892,
3519,
310,
4185,
3864,
17473,
519,
9101,
7199,
895,
13138
] | 1,024 |
CommonCrawl
| -1.064335 | 1.028489 | -0.538746 | -0.497824 |
a dissociation solution to a layer of differentiable cells that are contained in a culture chamber prior to dissociation, where the dissociation breaks apart the layer of cells into single cells. After dissociation, the single cells are placed into a new tissue culture chamber with a stem cell culture solution, wherein the stem cell culture solution comprises a basal salt nutrient solution and an ErbB3 ligand. Once cultured, the single stem cells are placed in conditions that permit growth and division of the single cells. In another specific embodiment, the methods of culturing differentiable cells comprise providing a dissociation solution to an aggregation differentiable cells that are contained in a culture chamber prior, where the dissociation breaks apart the aggregates of cells into single cells or smaller aggregates of cells.
[0111] The disaggregation solution used in the methods of the present invention can be any disaggregation solution capable of breaking apart or disaggregating the cells into single cells, without causing extensive toxicity to the cells. Examples of disaggregation solutions include, but are not limited to, trypsin, ACCUTASE®, 0.25% Trypsin/EDTA, TrypLE, or VERSENE® (EDTA) and trypsin. The methods of the present invention need not result in every cell of the confluent layer or suspension being disaggregated into single cells, provided that at least a few single cells are disaggregated and capable of being re-cultured.
[0112] Either at the beginning of culture, or after passaging, the differentiable cells can be seeded at any density, including a single cell in a culture chamber. The cell density of the seeded cells may be adjusted depending on a variety of factors, including but not limited to the use of adherent or suspension cultures, the specific recipe of the cell culture media used, the growth conditions and the contemplated use of the cultured cells. Examples of cell culture densities include, but are not limited to, 0.01×105 cells/ml, 0.05×105 cells/ml, 0.1×105 cells/ml, 0.5×105 cells/ml, 1.0×105 cells/ml, 1.2×105 cells/ml, 1.4×105 cells/ml, 1.6×105 cells/ml, 1.8×105 cells/ml, 2.0×105 cells/ml, 3.0×105 cells/ml, 4.0×105 cells/ml, 5.0×105 cells/ml, 6.0×105 cells/ml, 7.0×105 cells/ml, 8.0×105 cells/ml, 9.0×105 cells/ml, or 10.0×105 cells/ml, or more or any value in between.
[0113] Differentiable cells may also be utilized to screen for molecules or factors that influence their plasticity or other characteristics. For example, differentiable cells could be used to identify agents that induce apoptosis, differentiation or proliferation, as well as similar effects in differentiated lineages that have been generated from the differentiable cells.
[0114] Because the compositions and methods of the present invention allow for single cell passaging, differentiable cells have been successfully cultured in high-throughput settings, such as, but not limited to, 96-well plates and 384-well plates. FIG. 16 shows the morphology and alkaline phosphatase staining of BG02 cells that were cultured in DC-HAIF in both a 96-well and 384-well plate, using the methods described herein. Briefly, hESCs cells that were split, using ACCUTASE®, and plated in 96-well and 384-well plates and cultured showed a similar plating efficiency as what is observed using other culture dishes. In addition, the cells formed colonies, and were expanded successfully over 5 days in the smaller environments. These smaller cultures remained morphologically undifferentiated, and stained uniformly positive for alkaline phosphatase, a marker of undifferentiated cells. Furthermore, hESCs could also be grown in 96-well culture devices (not shown) that provide real-
|
[
263,
766,
2839,
362,
1650,
304,
263,
7546,
310,
17473,
519,
9101,
393,
526,
11122,
297,
263,
9257,
24171,
7536,
304,
766,
2839,
362,
29892,
988,
278,
766,
2839,
362,
16706,
12435,
278,
7546,
310,
9101,
964,
2323,
9101,
29889,
2860,
766,
2839,
362,
29892,
278,
2323,
9101,
526,
7180,
964,
263,
716,
260,
15118,
9257,
24171,
411,
263,
20805,
3038,
9257,
1650,
29892,
988,
262,
278,
20805,
3038,
9257,
1650,
7199,
4637,
263,
2362,
284,
15795,
18254,
374,
296,
1650,
322,
385,
1425,
29890,
29933,
29941,
14172,
392,
29889,
9038,
4185,
2955,
29892,
278,
2323,
20805,
9101,
526,
7180,
297,
5855,
393,
14257,
14321,
322,
8542,
310,
278,
2323,
9101,
29889,
512,
1790,
2702,
7232,
397,
2073,
29892,
278,
3519,
310,
4185,
3864,
17473,
519,
9101,
7199,
895,
13138,
263,
766,
2839,
362,
1650,
304,
385,
11404,
362,
17473,
519,
9101,
393,
526,
11122,
297,
263,
9257,
24171,
7536,
29892,
988,
278,
766,
2839,
362,
16706,
12435,
278,
11404,
1078,
310,
9101,
964,
2323,
9101,
470,
7968,
11404,
1078,
310,
9101,
29889,
13,
29961,
29900,
29896,
29896,
29896,
29962,
450,
22941,
7642,
362,
1650,
1304,
297,
278,
3519,
310,
278,
2198,
297,
7316,
508,
367,
738,
22941,
7642,
362,
1650,
15390,
310,
16679,
12435,
470,
22941,
7642,
1218,
278,
9101,
964,
2323,
9101,
29892,
1728,
10805,
20607,
304,
29916,
24798,
304,
278,
9101,
29889,
1222,
9422,
310,
22941,
7642,
362,
6851,
3160,
29892,
541,
526,
451,
9078,
304,
29892,
1018,
567,
262,
29892,
319,
4174,
2692,
8127,
30342,
29892,
29871,
29900,
29889,
29906,
29945,
29995,
3967,
567,
262,
29914,
3352,
6040,
29892,
3967,
29886,
1307,
29892,
470,
478,
23598,
1430,
29923,
30342,
313,
3352,
6040,
29897,
322,
1018,
567,
262,
29889,
450,
3519,
310,
278,
2198,
297,
7316,
817,
451,
1121,
297,
1432,
3038,
310,
278,
18669,
8122,
7546,
470,
8872,
2673,
1641,
22941,
7642,
630,
964,
2323,
9101,
29892,
4944,
393,
472,
3203,
263,
2846,
2323,
9101,
526,
22941,
7642,
630,
322,
15390,
310,
1641,
337,
29899,
29883,
499,
2955,
29889,
13,
29961,
29900,
29896,
29896,
29906,
29962,
20370,
472,
278,
6763,
310,
9257,
29892,
470,
1156,
1209,
6751,
29892,
278,
17473,
519,
9101,
508,
367,
16717,
287,
472,
738,
9027,
29892,
3704,
263,
2323,
3038,
297,
263,
9257,
24171,
29889,
450,
3038,
9027,
310,
278,
16717,
287,
9101,
1122,
367,
10365,
287,
8679,
373,
263,
12875,
310,
13879,
29892,
3704,
541,
451,
9078,
304,
278,
671,
310,
594,
2276,
296,
470,
8872,
2673,
4185,
1973,
29892,
278,
2702,
9522,
412,
310,
278,
3038,
9257,
5745,
1304,
29892,
278,
14321,
5855,
322,
278,
640,
16305,
630,
671,
310,
278,
4185,
2955,
9101,
29889,
1222,
9422,
310,
3038,
9257,
6245,
1907,
3160,
29892,
541,
526,
451,
9078,
304,
29892,
29871,
29900,
29889,
29900,
29896,
30122,
29896,
29900,
29945,
9101,
29914,
828,
29892,
29871,
29900,
29889,
29900,
29945,
30122,
29896,
29900,
29945,
9101,
29914,
828,
29892,
29871,
29900,
29889,
29896,
30122,
29896,
29900,
29945,
9101,
29914,
828,
29892,
29871,
29900,
29889,
29945,
30122,
29896,
29900,
29945,
9101,
29914,
828,
29892,
29871,
29896,
29889,
29900,
30122,
29896,
29900,
29945,
9101,
29914,
828,
29892,
29871,
29896,
29889,
29906,
30122,
29896,
29900,
29945,
9101,
29914,
828,
29892,
29871,
29896,
29889,
29946,
30122,
29896,
29900,
29945,
9101,
29914,
828,
29892,
29871,
29896,
29889,
29953,
30122,
29896,
29900,
29945,
9101,
29914,
828,
29892,
29871,
29896,
29889,
29947,
30122,
29896,
29900,
29945,
9101,
29914,
828,
29892,
29871,
29906,
29889,
29900,
30122,
29896,
29900,
29945,
9101,
29914,
828,
29892,
29871,
29941,
29889,
29900,
30122,
29896,
29900,
29945,
9101,
29914,
828,
29892,
29871,
29946,
29889,
29900,
30122,
29896,
29900,
29945,
9101,
29914,
828,
29892,
29871,
29945,
29889,
29900,
30122,
29896,
29900,
29945,
9101,
29914,
828,
29892,
29871,
29953,
29889,
29900,
30122,
29896,
29900,
29945,
9101,
29914,
828,
29892,
29871,
29955,
29889,
29900,
30122,
29896,
29900,
29945,
9101,
29914,
828,
29892,
29871,
29947,
29889,
29900,
30122,
29896,
29900,
29945,
9101,
29914,
828,
29892,
29871,
29929,
29889,
29900,
30122,
29896,
29900,
29945,
9101,
29914,
828,
29892,
470,
29871,
29896,
29900,
29889,
29900,
30122,
29896,
29900,
29945,
9101,
29914,
828,
29892,
470,
901,
470,
738,
995,
297,
1546,
29889,
13,
29961,
29900,
29896,
29896,
29941,
29962,
360,
8349,
7268,
519,
9101,
1122,
884,
367,
3667,
1891,
304,
4315,
363,
13206,
21337,
470,
13879,
393,
9949,
1009,
715,
6288,
537,
470,
916,
21862,
29889,
1152,
1342,
29892,
17473,
519,
9101,
1033,
367,
1304,
304,
12439,
19518,
393,
9013,
346,
16798,
415,
19263,
29892,
1422,
11685,
470,
410,
29880,
9633,
362,
29892,
408,
1532,
408,
2788,
9545,
297,
17473,
630,
1196,
1179,
393,
505,
1063,
5759,
515,
278,
17473,
519,
9101,
29889,
13,
29961,
29900,
29896,
29896,
29946,
29962,
7311,
278,
5541,
2187,
322,
3519,
310,
278,
2198,
297,
7316,
2758,
363,
2323,
3038,
1209,
6751,
29892,
17473,
519,
9101,
505,
1063,
8472,
4185,
2955,
297,
1880,
29899,
20678,
649,
6055,
29892,
1316,
408,
29892,
541,
451,
9078,
304,
29892,
29871,
29929,
29953,
29899,
5872,
715,
1078,
322,
29871,
29941,
29947,
29946,
29899,
5872,
715,
1078,
29889,
383,
6259,
29889,
29871,
29896,
29953,
3697,
278,
18131,
3002,
322,
394,
11311,
457,
1374,
25715,
271,
559,
380,
17225,
310,
350,
29954,
29900,
29906,
9101,
393,
892,
4185,
2955,
297,
13681,
29899,
15715,
6545,
297,
1716,
263,
29871,
29929,
29953,
29899,
5872,
322,
29871,
29941,
29947,
29946,
29899,
5872,
15284,
29892,
773,
278,
3519,
5439,
1244,
262,
29889,
1771,
2575,
368,
29892,
298,
2890,
29907,
29879,
9101,
393,
892,
6219,
29892,
773,
319,
4174,
2692,
8127,
30342,
29892,
322,
715,
630,
297,
29871,
29929,
29953,
29899,
5872,
322,
29871,
29941,
29947,
29946,
29899,
5872,
715,
1078,
322,
4185,
2955,
10018,
263,
2788,
715,
1218,
19201,
408,
825,
338,
8900,
773,
916,
9257,
270,
17006,
29889,
512,
6124,
29892,
278,
9101,
8429,
8104,
583,
29892,
322,
892,
17832,
8472,
975,
29871,
29945,
3841,
297,
278,
7968,
23136,
29889,
4525,
7968,
4185,
1973,
9488,
18131,
1189,
1711,
563,
8349,
7268,
630,
29892,
322,
380,
7114,
26018,
6374,
363,
394,
11311,
457,
1374,
25715,
271,
559,
29892,
263,
17456,
310,
563,
8349,
7268,
630,
9101,
29889,
16478,
29892,
298,
2890,
29907,
29879,
1033,
884,
367,
21633,
297,
29871,
29929,
29953,
29899,
5872,
9257,
9224,
313,
1333,
4318,
29897,
393,
3867,
1855,
29899
] | 1,024 |
CommonCrawl
| -0.84697 | 1.062231 | -0.194671 | 0.252723 |
time measurements of impedance, which can be used to measure cell proliferation and viability using the RT-CES® methods from ACEA Biosciences, Inc. (www.aceabio.com). Such an approach would enable a label-free identification and quantitation of subtle or immediate effects on differentiable cells, as well as measurements of proliferation, apoptosis and changes to morphology, in real time.
[0115] The compositions and methods of the invention may contain virtually any combination of the components set out above or described elsewhere herein, provided the compositions and methods comprise a basal salt nutrient solution and a means for stimulating ErbB2 directed tyrosine kinase activity. As one skilled in the art would recognize, the components of the compositions and methods of the invention will vary according to the protocol design. Accordingly, one embodiment of the present invention relates to culturing differentiable cells in 96-well plates and/or 384-well plates. Indeed, using the methods and compositions of the present invention, the cell culture chamber, i.e., the culture dish, is no longer limited to specific dimensions. Thus, the methods of the present invention is not limited to specific culture chamber dimensions.
[0116] The compositions and methods described herein have several useful features. For example, the compositions and methods described herein are useful for modeling the early stages of human development. Furthermore, the compositions and methods described herein can also serve for therapeutic intervention in disease states, such as neurodegenerative disorders, diabetes mellitus or renal failure, such as by the development of pure tissue or cell type.
[0117] The cell types that differentiate from differentiable cells have several uses in various fields of research and development including but not limited to drug discovery, drug development and testing, toxicology, production of cells for therapeutic purposes as well as basic science research. These cell types express molecules that are of interest in a wide range of research fields. These include the molecules known to be required for the functioning of the various cell types as described in standard reference texts. These molecules include, but are not limited to, cytokines, growth factors, cytokine receptors, extracellular matrix, transcription factors, secreted polypeptides and other molecules, and growth factor receptors.
[0118] It is contemplated that the differentiable cells of the invention can be differentiated through contact with a cell differentiation environment. As used herein, the term "cell differentiation environment" refers to a cell culture condition wherein the differentiable cells are induced to differentiate, or are induced to become a human cell culture enriched in differentiated cells. Preferably, the differentiated cell lineage induced by the growth factor will be homogeneous in nature. The term "homogeneous," refers to a population that contains more than approximately 50%, 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the desired cell lineage.
[0119] A cell differentiating medium or environment may be utilized to partially, terminally, or reversibly differentiate the differentiable cells of the present invention. In accordance with the invention the medium of the cell differentiation environment may contain a variety of components including, for example, KODMEM medium (Knockout Dulbecco's Modified Eagle's Medium), DMEM, Ham's F12 medium, FBS (fetal bovine serum), FGF2 (fibroblast growth factor 2), KSR or hLIF (human leukemia inhibitory factor). The cell differentiation environment can also contain supplements such as L-Glutamine, NEAA (non-essential amino acids), P/S (penicillin/streptomycin), N2, B27 and β-mercaptoethanol (β-ME). It is contemplated that additional factors may be added to the cell differentiation environment, including, but not limited to, fibronectin, laminin, heparin, heparin sulfate, retinoic acid, members of the epidermal growth factor family (EG
|
[
2230,
20398,
310,
25275,
749,
29892,
607,
508,
367,
1304,
304,
5645,
3038,
410,
29880,
9633,
362,
322,
3516,
3097,
773,
278,
390,
29911,
29899,
27266,
30342,
3519,
515,
319,
4741,
29909,
350,
2363,
455,
2063,
29892,
9266,
29889,
313,
1636,
29889,
815,
370,
601,
29889,
510,
467,
10506,
385,
2948,
723,
9025,
263,
3858,
29899,
9021,
29769,
322,
4323,
7018,
310,
12059,
280,
470,
16800,
9545,
373,
17473,
519,
9101,
29892,
408,
1532,
408,
20398,
310,
410,
29880,
9633,
362,
29892,
16798,
415,
19263,
322,
3620,
304,
18131,
3002,
29892,
297,
1855,
931,
29889,
13,
29961,
29900,
29896,
29896,
29945,
29962,
450,
5541,
2187,
322,
3519,
310,
278,
297,
7316,
1122,
1712,
4610,
1474,
738,
10296,
310,
278,
7117,
731,
714,
2038,
470,
5439,
17551,
1244,
262,
29892,
4944,
278,
5541,
2187,
322,
3519,
7199,
895,
263,
2362,
284,
15795,
18254,
374,
296,
1650,
322,
263,
2794,
363,
20436,
18099,
1425,
29890,
29933,
29906,
10624,
7911,
1883,
457,
19015,
559,
6354,
29889,
1094,
697,
2071,
24455,
297,
278,
1616,
723,
18720,
29892,
278,
7117,
310,
278,
5541,
2187,
322,
3519,
310,
278,
297,
7316,
674,
13100,
5034,
304,
278,
9608,
2874,
29889,
7579,
368,
29892,
697,
7232,
397,
2073,
310,
278,
2198,
297,
7316,
1104,
1078,
304,
4185,
3864,
17473,
519,
9101,
297,
29871,
29929,
29953,
29899,
5872,
715,
1078,
322,
29914,
272,
29871,
29941,
29947,
29946,
29899,
5872,
715,
1078,
29889,
14598,
29892,
773,
278,
3519,
322,
5541,
2187,
310,
278,
2198,
297,
7316,
29892,
278,
3038,
9257,
24171,
29892,
474,
29889,
29872,
1696,
278,
9257,
270,
728,
29892,
338,
694,
5520,
9078,
304,
2702,
13391,
29889,
6549,
29892,
278,
3519,
310,
278,
2198,
297,
7316,
338,
451,
9078,
304,
2702,
9257,
24171,
13391,
29889,
13,
29961,
29900,
29896,
29896,
29953,
29962,
450,
5541,
2187,
322,
3519,
5439,
1244,
262,
505,
3196,
5407,
5680,
29889,
1152,
1342,
29892,
278,
5541,
2187,
322,
3519,
5439,
1244,
262,
526,
5407,
363,
1904,
292,
278,
4688,
22950,
310,
5199,
5849,
29889,
16478,
29892,
278,
5541,
2187,
322,
3519,
5439,
1244,
262,
508,
884,
9080,
363,
266,
1572,
412,
329,
293,
1006,
7316,
297,
17135,
5922,
29892,
1316,
408,
452,
2192,
311,
4738,
1230,
766,
20488,
29892,
652,
370,
10778,
286,
514,
277,
375,
470,
4325,
284,
10672,
29892,
1316,
408,
491,
278,
5849,
310,
8296,
260,
15118,
470,
3038,
1134,
29889,
13,
29961,
29900,
29896,
29896,
29955,
29962,
450,
3038,
4072,
393,
17473,
403,
515,
17473,
519,
9101,
505,
3196,
3913,
297,
5164,
4235,
310,
5925,
322,
5849,
3704,
541,
451,
9078,
304,
15721,
20699,
29892,
15721,
5849,
322,
6724,
29892,
304,
27375,
3002,
29892,
5802,
310,
9101,
363,
266,
1572,
412,
329,
293,
11976,
408,
1532,
408,
6996,
10466,
5925,
29889,
4525,
3038,
4072,
4653,
13206,
21337,
393,
526,
310,
4066,
297,
263,
9377,
3464,
310,
5925,
4235,
29889,
4525,
3160,
278,
13206,
21337,
2998,
304,
367,
3734,
363,
278,
740,
292,
310,
278,
5164,
3038,
4072,
408,
5439,
297,
3918,
3407,
26442,
29889,
4525,
13206,
21337,
3160,
29892,
541,
526,
451,
9078,
304,
29892,
5094,
17082,
1475,
29892,
14321,
13879,
29892,
5094,
17082,
457,
337,
1547,
943,
29892,
1294,
945,
514,
1070,
4636,
29892,
1301,
3395,
13879,
29892,
7035,
287,
1248,
668,
415,
2247,
322,
916,
13206,
21337,
29892,
322,
14321,
7329,
337,
1547,
943,
29889,
13,
29961,
29900,
29896,
29896,
29947,
29962,
739,
338,
640,
16305,
630,
393,
278,
17473,
519,
9101,
310,
278,
297,
7316,
508,
367,
17473,
630,
1549,
6958,
411,
263,
3038,
1422,
11685,
5177,
29889,
1094,
1304,
1244,
262,
29892,
278,
1840,
376,
3729,
1422,
11685,
5177,
29908,
14637,
304,
263,
3038,
9257,
4195,
988,
262,
278,
17473,
519,
9101,
526,
20974,
304,
17473,
403,
29892,
470,
526,
20974,
304,
4953,
263,
5199,
3038,
9257,
427,
4018,
287,
297,
17473,
630,
9101,
29889,
4721,
571,
2197,
29892,
278,
17473,
630,
3038,
1196,
482,
20974,
491,
278,
14321,
7329,
674,
367,
3632,
23724,
297,
5469,
29889,
450,
1840,
376,
9706,
23724,
1699,
14637,
304,
263,
4665,
393,
3743,
901,
1135,
14235,
29871,
29945,
29900,
13667,
29871,
29953,
29900,
13667,
29871,
29955,
29900,
13667,
29871,
29947,
29900,
13667,
29871,
29947,
29945,
13667,
29871,
29947,
29953,
13667,
29871,
29947,
29955,
13667,
29871,
29947,
29947,
13667,
29871,
29947,
29929,
13667,
29871,
29929,
29900,
13667,
29871,
29929,
29896,
13667,
29871,
29929,
29906,
13667,
29871,
29929,
29941,
13667,
29871,
29929,
29946,
13667,
29871,
29929,
29945,
13667,
29871,
29929,
29953,
13667,
29871,
29929,
29955,
13667,
29871,
29929,
29947,
13667,
470,
29871,
29929,
29929,
29995,
310,
278,
7429,
3038,
1196,
482,
29889,
13,
29961,
29900,
29896,
29896,
29929,
29962,
319,
3038,
17473,
1218,
18350,
470,
5177,
1122,
367,
3667,
1891,
304,
22039,
29892,
6624,
635,
29892,
470,
18764,
14981,
17473,
403,
278,
17473,
519,
9101,
310,
278,
2198,
297,
7316,
29889,
512,
15017,
749,
411,
278,
297,
7316,
278,
18350,
310,
278,
3038,
1422,
11685,
5177,
1122,
1712,
263,
12875,
310,
7117,
3704,
29892,
363,
1342,
29892,
476,
13668,
2303,
29924,
18350,
313,
29968,
1217,
384,
449,
360,
352,
915,
26454,
29915,
29879,
3382,
2164,
382,
20860,
29915,
29879,
3436,
1974,
511,
360,
2303,
29924,
29892,
7904,
29915,
29879,
383,
29896,
29906,
18350,
29892,
383,
9851,
313,
29888,
26310,
289,
586,
457,
724,
398,
511,
383,
29954,
29943,
29906,
313,
29888,
747,
307,
23190,
14321,
7329,
29871,
29906,
511,
476,
14098,
470,
298,
5265,
29943,
313,
26029,
454,
2679,
29747,
297,
6335,
5047,
7329,
467,
450,
3038,
1422,
11685,
5177,
508,
884,
1712,
1462,
944,
29879,
1316,
408,
365,
29899,
29954,
29880,
329,
314,
457,
29892,
14693,
6344,
313,
5464,
29899,
404,
2556,
626,
1789,
1274,
4841,
511,
349,
29914,
29903,
313,
2238,
293,
453,
262,
29914,
13045,
415,
290,
11078,
262,
511,
405,
29906,
29892,
350,
29906,
29955,
322,
29871,
30187,
29899,
1050,
17885,
29877,
621,
273,
324,
313,
30187,
29899,
2303,
467,
739,
338,
640,
16305,
630,
393,
5684,
13879,
1122,
367,
2715,
304,
278,
3038,
1422,
11685,
5177,
29892,
3704,
29892,
541,
451,
9078,
304,
29892,
18755,
1617,
522,
262,
29892,
301,
9103,
262,
29892,
540,
862,
262,
29892,
540,
862,
262,
5394,
29888,
403,
29892,
3240,
1789,
293,
22193,
29892,
5144,
310,
278,
321,
5935,
837,
284,
14321,
7329,
3942,
313,
11787
] | 1,024 |
CommonCrawl
| -0.283179 | 1.417825 | 0.323178 | 0.767528 |
Fs), members of the fibroblast growth factor family (FGFs) including FGF2 and/or FGF8, members of the platelet derived growth factor family (PDGFs), transforming growth factor (TGF)/bone morphogenetic protein (BMP)/growth and differentiation factor (GDF) factor family antagonists including but not limited to noggin, follistatin, chordin, gremlin, cerberus/DAN family proteins, ventropin, high dose activin, and amnionless or variants or functional fragments thereof. TGF/BMP/GDF antagonists could also be added in the form of TGF/BMP/GDF receptor-Fe chimeras. Other factors that may be added include molecules that can activate or inactivate signaling through Notch receptor family, including but not limited to proteins of the Delta-like and Jagged families as well as inhibitors of Notch processing or cleavage, or variants or functional fragments thereof. Other growth factors may include members of the insulin like growth factor family (IGF), insulin, the wingless related (WNT) factor family, and the hedgehog factor family or variants or functional fragments thereof. Additional factors may be added to promote mesendoderm stem/progenitor, endoderm stem/progenitor, mesoderm stem/progenitor, or definitive endoderm stem/progenitor proliferation and survival as well as survival and differentiation of derivatives of these progenitors.
[0120] The compositions described herein are useful for the screening of test compounds to determine whether a test compound modulates pluripotency, proliferation, and/or differentiation of differentiable cells. Pluripotency, proliferation and/or differentiation of differentiable cells can be readily ascertained by one of ordinary skill in the art. Non-limiting methods include examining cell morphology, the expression of various markers, teratoma formation, cell counts and measurements of impedance.
[0121] The progression of the differentiable cells to the desired cell lineage, or its maintenance in an undifferentiated state can be monitored by quantitating expression of marker genes characteristic of the desired cell lineage as well as the lack of expression of marker genes characteristic of differentiable cell types. One method of quantitating gene expression of such marker genes is through the use of quantitative PCR (Q-PCR). Methods of performing Q-PCR are well known in the art. Other methods that are known in the art can also be used to quantitate marker gene expression. Marker gene expression can be detected by using antibodies specific for the marker gene of interest.
[0122] In certain embodiments, the screening method encompasses methods of identifying a compound capable of modulating pluripotency, proliferation and/or differentiation of a differentiable cell, comprising (a) providing a differentiable cell; (b) culturing the cell in a composition comprising a basal salt nutrient solution and an ErbB3 ligand, wherein the composition is essentially serum free; (c) contacting the cell with a test compound; and determining whether an increase or decrease in pluripotency, proliferation and/or differentiation occurs in the cell contacted with the compound, said increase being an indication that the compound modulates pluripotency, proliferation and/or differentiation. In certain embodiments, the ErbB3 ligand is HRG-β. In other embodiments, the ErbB3 ligand can be substituted with a test compound, to determine the effects of the test compound. For example, the effects on pluripotency, proliferation and/or differentiation that occurs with the test compound can be compared to the effects on pluripotency, proliferation and/or differentiation that occurs with the ErbB3 ligand to determine the effects of the test compound on the differentiable cells. It is contemplated that any of the compositions described herein can be used in the screening methods of the present invention.
[0123] In yet another embodiment, the cells can be cultured in the absence of an ErbB3 ligand (ErbB2-directed tyrosine kinase activity) to determine the effects of the absence of an ErbB3 lig
|
[
29943,
29879,
511,
5144,
310,
278,
18755,
307,
23190,
14321,
7329,
3942,
313,
29943,
29954,
29943,
29879,
29897,
3704,
383,
29954,
29943,
29906,
322,
29914,
272,
383,
29954,
29943,
29947,
29892,
5144,
310,
278,
15284,
1026,
10723,
14321,
7329,
3942,
313,
25014,
29954,
29943,
29879,
511,
4327,
292,
14321,
7329,
313,
29911,
29954,
29943,
6802,
15933,
18131,
6352,
7492,
26823,
313,
29933,
3580,
6802,
29887,
798,
386,
322,
1422,
11685,
7329,
313,
29954,
4037,
29897,
7329,
3942,
3677,
12841,
2879,
3704,
541,
451,
9078,
304,
11313,
5359,
29892,
900,
1761,
21203,
29892,
521,
4194,
29892,
1395,
828,
262,
29892,
5147,
495,
375,
29914,
29928,
2190,
3942,
3279,
1144,
29892,
9712,
1336,
262,
29892,
1880,
437,
344,
5039,
262,
29892,
322,
626,
29876,
291,
2222,
470,
29161,
470,
13303,
22370,
727,
974,
29889,
323,
29954,
29943,
29914,
29933,
3580,
29914,
29954,
4037,
3677,
12841,
2879,
1033,
884,
367,
2715,
297,
278,
883,
310,
323,
29954,
29943,
29914,
29933,
3580,
29914,
29954,
4037,
337,
14268,
29899,
8263,
521,
4193,
294,
29889,
5901,
13879,
393,
1122,
367,
2715,
3160,
13206,
21337,
393,
508,
5039,
403,
470,
297,
11236,
403,
7182,
292,
1549,
2216,
305,
337,
14268,
3942,
29892,
3704,
541,
451,
9078,
304,
3279,
1144,
310,
278,
360,
2554,
29899,
4561,
322,
24423,
3192,
13175,
408,
1532,
408,
297,
6335,
17259,
310,
2216,
305,
9068,
470,
4531,
485,
482,
29892,
470,
29161,
470,
13303,
22370,
727,
974,
29889,
5901,
14321,
13879,
1122,
3160,
5144,
310,
278,
1663,
352,
262,
763,
14321,
7329,
3942,
313,
6259,
29943,
511,
1663,
352,
262,
29892,
278,
21612,
2222,
4475,
313,
16048,
29911,
29897,
7329,
3942,
29892,
322,
278,
298,
12864,
29882,
468,
7329,
3942,
470,
29161,
470,
13303,
22370,
727,
974,
29889,
3462,
3245,
13879,
1122,
367,
2715,
304,
27391,
4883,
355,
397,
837,
20805,
29914,
771,
1885,
2105,
29892,
1095,
397,
837,
20805,
29914,
771,
1885,
2105,
29892,
4883,
397,
837,
20805,
29914,
771,
1885,
2105,
29892,
470,
8422,
573,
1095,
397,
837,
20805,
29914,
771,
1885,
2105,
410,
29880,
9633,
362,
322,
10503,
2561,
408,
1532,
408,
10503,
2561,
322,
1422,
11685,
310,
25748,
310,
1438,
410,
1885,
17259,
29889,
13,
29961,
29900,
29896,
29906,
29900,
29962,
450,
5541,
2187,
5439,
1244,
262,
526,
5407,
363,
278,
4315,
292,
310,
1243,
752,
3885,
304,
8161,
3692,
263,
1243,
752,
618,
878,
352,
1078,
715,
332,
666,
327,
3819,
29892,
410,
29880,
9633,
362,
29892,
322,
29914,
272,
1422,
11685,
310,
17473,
519,
9101,
29889,
1858,
332,
666,
327,
3819,
29892,
410,
29880,
9633,
362,
322,
29914,
272,
1422,
11685,
310,
17473,
519,
9101,
508,
367,
28520,
408,
6327,
7114,
491,
697,
310,
15311,
19911,
297,
278,
1616,
29889,
10050,
29899,
2576,
11407,
3519,
3160,
4392,
2827,
3038,
18131,
3002,
29892,
278,
4603,
310,
5164,
29320,
29892,
1935,
271,
4125,
12409,
29892,
3038,
18139,
322,
20398,
310,
25275,
749,
29889,
13,
29961,
29900,
29896,
29906,
29896,
29962,
450,
410,
11476,
310,
278,
17473,
519,
9101,
304,
278,
7429,
3038,
1196,
482,
29892,
470,
967,
25413,
297,
385,
563,
8349,
7268,
630,
2106,
508,
367,
11819,
287,
491,
4323,
277,
1218,
4603,
310,
17456,
2531,
267,
17443,
310,
278,
7429,
3038,
1196,
482,
408,
1532,
408,
278,
10225,
310,
4603,
310,
17456,
2531,
267,
17443,
310,
17473,
519,
3038,
4072,
29889,
3118,
1158,
310,
4323,
277,
1218,
18530,
4603,
310,
1316,
17456,
2531,
267,
338,
1549,
278,
671,
310,
4323,
23378,
9609,
29934,
313,
29984,
29899,
9026,
29934,
467,
8108,
29879,
310,
15859,
660,
29899,
9026,
29934,
526,
1532,
2998,
297,
278,
1616,
29889,
5901,
3519,
393,
526,
2998,
297,
278,
1616,
508,
884,
367,
1304,
304,
4323,
10388,
17456,
18530,
4603,
29889,
4485,
261,
18530,
4603,
508,
367,
17809,
491,
773,
3677,
747,
397,
583,
2702,
363,
278,
17456,
18530,
310,
4066,
29889,
13,
29961,
29900,
29896,
29906,
29906,
29962,
512,
3058,
7232,
397,
7862,
29892,
278,
4315,
292,
1158,
427,
2388,
465,
267,
3519,
310,
2893,
9215,
263,
752,
618,
15390,
310,
878,
18099,
715,
332,
666,
327,
3819,
29892,
410,
29880,
9633,
362,
322,
29914,
272,
1422,
11685,
310,
263,
17473,
519,
3038,
29892,
7199,
5921,
313,
29874,
29897,
13138,
263,
17473,
519,
3038,
29936,
313,
29890,
29897,
4185,
3864,
278,
3038,
297,
263,
15259,
7199,
5921,
263,
2362,
284,
15795,
18254,
374,
296,
1650,
322,
385,
1425,
29890,
29933,
29941,
14172,
392,
29892,
988,
262,
278,
15259,
338,
13674,
724,
398,
3889,
29936,
313,
29883,
29897,
6958,
292,
278,
3038,
411,
263,
1243,
752,
618,
29936,
322,
3683,
2827,
3692,
385,
7910,
470,
23806,
297,
715,
332,
666,
327,
3819,
29892,
410,
29880,
9633,
362,
322,
29914,
272,
1422,
11685,
10008,
297,
278,
3038,
6958,
287,
411,
278,
752,
618,
29892,
1497,
7910,
1641,
385,
4221,
362,
393,
278,
752,
618,
878,
352,
1078,
715,
332,
666,
327,
3819,
29892,
410,
29880,
9633,
362,
322,
29914,
272,
1422,
11685,
29889,
512,
3058,
7232,
397,
7862,
29892,
278,
1425,
29890,
29933,
29941,
14172,
392,
338,
379,
29934,
29954,
29899,
30187,
29889,
512,
916,
7232,
397,
7862,
29892,
278,
1425,
29890,
29933,
29941,
14172,
392,
508,
367,
5960,
277,
3860,
411,
263,
1243,
752,
618,
29892,
304,
8161,
278,
9545,
310,
278,
1243,
752,
618,
29889,
1152,
1342,
29892,
278,
9545,
373,
715,
332,
666,
327,
3819,
29892,
410,
29880,
9633,
362,
322,
29914,
272,
1422,
11685,
393,
10008,
411,
278,
1243,
752,
618,
508,
367,
9401,
304,
278,
9545,
373,
715,
332,
666,
327,
3819,
29892,
410,
29880,
9633,
362,
322,
29914,
272,
1422,
11685,
393,
10008,
411,
278,
1425,
29890,
29933,
29941,
14172,
392,
304,
8161,
278,
9545,
310,
278,
1243,
752,
618,
373,
278,
17473,
519,
9101,
29889,
739,
338,
640,
16305,
630,
393,
738,
310,
278,
5541,
2187,
5439,
1244,
262,
508,
367,
1304,
297,
278,
4315,
292,
3519,
310,
278,
2198,
297,
7316,
29889,
13,
29961,
29900,
29896,
29906,
29941,
29962,
512,
3447,
1790,
7232,
397,
2073,
29892,
278,
9101,
508,
367,
4185,
2955,
297,
278,
18070,
310,
385,
1425,
29890,
29933,
29941,
14172,
392,
313,
2110,
29890,
29933,
29906,
29899,
11851,
287,
7911,
1883,
457,
19015,
559,
6354,
29897,
304,
8161,
278,
9545,
310,
278,
18070,
310,
385,
1425,
29890,
29933,
29941,
14172
] | 1,024 |
CommonCrawl
| -0.765458 | 1.708528 | 0.127437 | 0.509493 |
and (ErbB2-directed tyrosine kinase activity) on the cells.
[0124] Using the methods described herein, compositions comprising the desired cell lineage that are substantially free of other cell types can be produced. Alternatively, compositions comprising mixtures of the differentiable cells and the desired cell lineage can also be produced.
[0125] In some embodiments of the present invention, cells of the desired cell lineage can be isolated by using an affinity tag that is specific for such cells. One example of an affinity tag specific for a target cell is an antibody that is specific to a marker polypeptide that is present on the cell surface of the target cell but which is not substantially present on other cell types that would be found in a cell culture produced by the methods described herein.
[0126] The present invention also relates to kits, wherein the kit comprises a basal salt nutrient solution and at least one compound capable of stimulating ErbB2-directed tyrosine kinase activity. In one embodiment, the kits comprise at least one ErbB3 ligand, as described herein. In another embodiment, the kits comprise more than one ErbB3 ligand. In another embodiment, the kits comprise at least one of TGF-β or a TGF-β family member or a variant or functional fragment thereof as described herein. In yet another embodiment, the kits comprise More than one of TGF-β or a TGF-β family member or a variant or functional fragment thereof. In still another embodiment, the kits comprise at least one fibroblast growth factor or variant or functional fragment thereof. In another embodiment, the kits comprise more than one fibroblast growth factor or variant or functional fragment thereof In a specific embodiment, the kits comprise at least one of FGF-7, FGF-10, FGF-22 or variants or functional fragments thereof. In another embodiment, the kits comprise serum albumin. In still another embodiment, the kits comprise serum and/or at least one insoluble substrate as described herein and/or at least one disaggregation solution.
[0127] The kits of the invention may contain virtually any combination of the components set out above or described elsewhere herein. As one skilled in the art would recognize, the components supplied with kits of the invention will vary with the intended use for the kits. Thus, kits may be designed to perform various functions set out in this application and the components of such kits will vary accordingly.
[0128] Throughout this application, various publications are referenced. The disclosures of all of these publications and those references cited within those publications in their entireties are hereby incorporated by reference into this application in their entirety in order to more fully describe the state of the art to which this invention pertains.
[0129] The human embryonic stem cell line BG01v (BresaGen, Inc., Athens, Ga.) was used in some of the experiments described herein. The BG01v hESC line is a karyotypically variant cell line, which exhibits stable karyotype containing specific trisomies (karyotype: 49, XXY,+12,+17). Parent cultures were maintained as described previously (Schulz et al., 2003, BMC Neurosci., 4:27; Schulz et al., 2004, Stem Cells, 22(7):1218-38; Rosler et al., 2004, Dev. Dynamics, 229:259-274; Brimble et al., 2004 Stem Cells Dev., 13:585-596). Briefly, the cells were grown in dishes coated with MATRIGEL® or fibronectin, in conditioned media from mouse embryonic fibroblasts (MEFs) (MEF-CM) comprising DMEM:F12 with 20% KSR, 8 ng/ml FGF2, 2 mM L-Glutamine, 1× non-essential amino acids, 0.5 U/ml penicillin, 0
|
[
392,
313,
2110,
29890,
29933,
29906,
29899,
11851,
287,
7911,
1883,
457,
19015,
559,
6354,
29897,
373,
278,
9101,
29889,
13,
29961,
29900,
29896,
29906,
29946,
29962,
5293,
278,
3519,
5439,
1244,
262,
29892,
5541,
2187,
7199,
5921,
278,
7429,
3038,
1196,
482,
393,
526,
23228,
368,
3889,
310,
916,
3038,
4072,
508,
367,
7371,
29889,
20360,
29892,
5541,
2187,
7199,
5921,
3737,
486,
1973,
310,
278,
17473,
519,
9101,
322,
278,
7429,
3038,
1196,
482,
508,
884,
367,
7371,
29889,
13,
29961,
29900,
29896,
29906,
29945,
29962,
512,
777,
7232,
397,
7862,
310,
278,
2198,
297,
7316,
29892,
9101,
310,
278,
7429,
3038,
1196,
482,
508,
367,
23968,
491,
773,
385,
2756,
13593,
4055,
393,
338,
2702,
363,
1316,
9101,
29889,
3118,
1342,
310,
385,
2756,
13593,
4055,
2702,
363,
263,
3646,
3038,
338,
385,
3677,
747,
1486,
393,
338,
2702,
304,
263,
17456,
1248,
668,
415,
680,
393,
338,
2198,
373,
278,
3038,
7101,
310,
278,
3646,
3038,
541,
607,
338,
451,
23228,
368,
2198,
373,
916,
3038,
4072,
393,
723,
367,
1476,
297,
263,
3038,
9257,
7371,
491,
278,
3519,
5439,
1244,
262,
29889,
13,
29961,
29900,
29896,
29906,
29953,
29962,
450,
2198,
297,
7316,
884,
1104,
1078,
304,
413,
1169,
29892,
988,
262,
278,
413,
277,
7199,
4637,
263,
2362,
284,
15795,
18254,
374,
296,
1650,
322,
472,
3203,
697,
752,
618,
15390,
310,
20436,
18099,
1425,
29890,
29933,
29906,
29899,
11851,
287,
7911,
1883,
457,
19015,
559,
6354,
29889,
512,
697,
7232,
397,
2073,
29892,
278,
413,
1169,
7199,
895,
472,
3203,
697,
1425,
29890,
29933,
29941,
14172,
392,
29892,
408,
5439,
1244,
262,
29889,
512,
1790,
7232,
397,
2073,
29892,
278,
413,
1169,
7199,
895,
901,
1135,
697,
1425,
29890,
29933,
29941,
14172,
392,
29889,
512,
1790,
7232,
397,
2073,
29892,
278,
413,
1169,
7199,
895,
472,
3203,
697,
310,
323,
29954,
29943,
29899,
30187,
470,
263,
323,
29954,
29943,
29899,
30187,
3942,
4509,
470,
263,
17305,
470,
13303,
9376,
727,
974,
408,
5439,
1244,
262,
29889,
512,
3447,
1790,
7232,
397,
2073,
29892,
278,
413,
1169,
7199,
895,
5853,
1135,
697,
310,
323,
29954,
29943,
29899,
30187,
470,
263,
323,
29954,
29943,
29899,
30187,
3942,
4509,
470,
263,
17305,
470,
13303,
9376,
727,
974,
29889,
512,
1603,
1790,
7232,
397,
2073,
29892,
278,
413,
1169,
7199,
895,
472,
3203,
697,
18755,
307,
23190,
14321,
7329,
470,
17305,
470,
13303,
9376,
727,
974,
29889,
512,
1790,
7232,
397,
2073,
29892,
278,
413,
1169,
7199,
895,
901,
1135,
697,
18755,
307,
23190,
14321,
7329,
470,
17305,
470,
13303,
9376,
727,
974,
512,
263,
2702,
7232,
397,
2073,
29892,
278,
413,
1169,
7199,
895,
472,
3203,
697,
310,
383,
29954,
29943,
29899,
29955,
29892,
383,
29954,
29943,
29899,
29896,
29900,
29892,
383,
29954,
29943,
29899,
29906,
29906,
470,
29161,
470,
13303,
22370,
727,
974,
29889,
512,
1790,
7232,
397,
2073,
29892,
278,
413,
1169,
7199,
895,
724,
398,
3769,
262,
29889,
512,
1603,
1790,
7232,
397,
2073,
29892,
278,
413,
1169,
7199,
895,
724,
398,
322,
29914,
272,
472,
3203,
697,
1663,
324,
431,
280,
25148,
403,
408,
5439,
1244,
262,
322,
29914,
272,
472,
3203,
697,
22941,
7642,
362,
1650,
29889,
13,
29961,
29900,
29896,
29906,
29955,
29962,
450,
413,
1169,
310,
278,
297,
7316,
1122,
1712,
4610,
1474,
738,
10296,
310,
278,
7117,
731,
714,
2038,
470,
5439,
17551,
1244,
262,
29889,
1094,
697,
2071,
24455,
297,
278,
1616,
723,
18720,
29892,
278,
7117,
19056,
411,
413,
1169,
310,
278,
297,
7316,
674,
13100,
411,
278,
9146,
671,
363,
278,
413,
1169,
29889,
6549,
29892,
413,
1169,
1122,
367,
8688,
304,
2189,
5164,
3168,
731,
714,
297,
445,
2280,
322,
278,
7117,
310,
1316,
413,
1169,
674,
13100,
16205,
29889,
13,
29961,
29900,
29896,
29906,
29947,
29962,
17044,
449,
445,
2280,
29892,
5164,
25964,
526,
16180,
29889,
450,
766,
11291,
1973,
310,
599,
310,
1438,
25964,
322,
1906,
9282,
274,
1573,
2629,
1906,
25964,
297,
1009,
4152,
2938,
526,
1244,
1609,
11039,
630,
491,
3407,
964,
445,
2280,
297,
1009,
4152,
1017,
297,
1797,
304,
901,
8072,
8453,
278,
2106,
310,
278,
1616,
304,
607,
445,
297,
7316,
639,
2408,
29879,
29889,
13,
29961,
29900,
29896,
29906,
29929,
29962,
450,
5199,
7232,
719,
8927,
20805,
3038,
1196,
350,
29954,
29900,
29896,
29894,
313,
29933,
690,
29874,
15462,
29892,
9266,
1696,
9193,
575,
29892,
10415,
1846,
471,
1304,
297,
777,
310,
278,
15729,
5439,
1244,
262,
29889,
450,
350,
29954,
29900,
29896,
29894,
298,
2890,
29907,
1196,
338,
263,
413,
653,
327,
1478,
1711,
17305,
3038,
1196,
29892,
607,
10371,
1169,
13714,
413,
653,
327,
668,
6943,
2702,
534,
275,
290,
583,
313,
29895,
653,
327,
668,
29901,
29871,
29946,
29929,
29892,
6193,
29979,
29892,
29974,
29896,
29906,
29892,
29974,
29896,
29955,
467,
22280,
4185,
1973,
892,
19949,
408,
5439,
9251,
313,
4504,
352,
29920,
634,
394,
1696,
29871,
29906,
29900,
29900,
29941,
29892,
350,
12513,
17574,
1883,
455,
1696,
29871,
29946,
29901,
29906,
29955,
29936,
12988,
29920,
634,
394,
1696,
29871,
29906,
29900,
29900,
29946,
29892,
624,
331,
315,
10071,
29892,
29871,
29906,
29906,
29898,
29955,
1125,
29896,
29906,
29896,
29947,
29899,
29941,
29947,
29936,
5678,
1358,
634,
394,
1696,
29871,
29906,
29900,
29900,
29946,
29892,
9481,
29889,
22554,
1199,
29892,
29871,
29906,
29906,
29929,
29901,
29906,
29945,
29929,
29899,
29906,
29955,
29946,
29936,
1771,
326,
569,
634,
394,
1696,
29871,
29906,
29900,
29900,
29946,
624,
331,
315,
10071,
9481,
1696,
29871,
29896,
29941,
29901,
29945,
29947,
29945,
29899,
29945,
29929,
29953,
467,
1771,
2575,
368,
29892,
278,
9101,
892,
21633,
297,
270,
17006,
1302,
630,
411,
341,
1299,
3960,
1692,
29931,
30342,
470,
18755,
1617,
522,
262,
29892,
297,
4195,
287,
5745,
515,
9495,
7232,
719,
8927,
18755,
307,
2204,
19416,
313,
2303,
29943,
29879,
29897,
313,
2303,
29943,
29899,
24494,
29897,
7199,
5921,
360,
2303,
29924,
29901,
29943,
29896,
29906,
411,
29871,
29906,
29900,
29995,
476,
14098,
29892,
29871,
29947,
8736,
29914,
828,
383,
29954,
29943,
29906,
29892,
29871,
29906,
286,
29924,
365,
29899,
29954,
29880,
329,
314,
457,
29892,
29871,
29896,
30122,
1661,
29899,
404,
2556,
626,
1789,
1274,
4841,
29892,
29871,
29900,
29889,
29945,
501,
29914,
828,
6584,
293,
453,
262,
29892,
29871,
29900
] | 1,024 |
CommonCrawl
| -1.159432 | 0.846799 | -0.353596 | -0.223503 |
.5 U/ml streptomycin, 0.1 mM β-mercaptoethanol (Sigma, St. Louis, Mo., USA), with collagenase passaging.
[0130] The defined culture (DC) media tested herein comprised DMEM/F12, 2 mM Glutamax, 1× non-essential amino acids, 0.5 U/ml penicillin, 0.5 U/ml streptomycin, 10 μg/ml transferrin (all from Invitrogen, Carlsbad, Calif., USA) 0.1 mM β-mercaptoethanol (Sigma), 0.2% fatty acid-free Cohn's fraction V BSA (Serologicals), 1× Trace Element mixes A, B and C (Cellgro) and 50 μg/ml Ascorbic Acid (Sigma). Variable levels of recombinant growth factors were used, including FGF2 (Sigma), LongR3-IGF1 (JRH Biosciences), Heregulin-β EGF domain (HRGβ, Peprotech), TGFβ (R&D systems), nodal (R&D systems), LIF (R&D systems), EGF (R&D systems), TGFα (R&D systems), HRGα (R&D systems), BMP4 (R&D systems), and Activin A (R&D Systems). LongR3-IGF1 is a modified version of IGF1 that has reduced affinity for IGF1 binding proteins, some of which are expressed in hESCs. DC-HAIF is the defined culture media as above, containing 10 ng/ml HRG-β, 10 ng/ml Activin A, 200 ng/ml LR-IGF1 and 8 ng/ml FGF2. DC-HAI is defined culture media as above containing 10 ng/ml HRG-β, 10 ng/ml Activin A, and 200 ng/ml LR-IGF1. In both DC-HAIF and DC-HAI, the LR-IGF1 component can, of course be replaced with IFG1.
[0131] MATRIGEL® coated dishes were prepared by diluting Growth Factor Reduced BD MATRIGEL® matrix (BD Biosciences, Franklin Lakes, N.J., USA) to a final concentration range of about 1:30 to about 1:1000 in cold DMEM/F-12. In one embodiment, the concentration of MATRIGEL® is about 1:200. 1 ml/35 mm dish was used to coat dishes for 1-2 hours at room temperature or at least overnight at 4° C. Plates were stored up to one week at 4° C. MATRIGEL® solution was removed immediately before use.
[0132] For the tested conditions, parent cultures were plated into 6-well dishes for comparison of multiple conditions. Cultures were typically plated directly into the test conditions. The cultures were assessed every day and graded based on morphological criteria 4 to 5 days after plating. The grading scale of 1 to 5 involved examining the whole culture and assessing overall proportion of undifferentiated colonies, their relative size, and proportion of colonies or parts of colonies exhibiting obvious differentiation. Grade 5 indicates "ideal" cultures, with large undifferentiated colonies and negligible differentiation. Grade 4 indicates a very good culture, but with some obvious differentiation. Grade 3 indicates an acceptable culture, but with around half the colonies exhibiting obvious differentiation. Grade 2 cultures are predominantly differentiated, with occasional putative undifferentiated cells. Grade 1 cultures contain differentiated colonies or the cultures did not adhere or did not survive. Cultures that exhibited good expansion of undifferentiated cells were passaged to assess longer-term culture in these conditions.
Expression of ErbB1-3, Nrg1 and ADAM19 in BG01v Cells
[0133] Real time RT-PCR was used to demonstrate expression of ErbB1-3, Neuregulin and ADAM-19
|
[
29889,
29945,
501,
29914,
828,
12243,
415,
290,
11078,
262,
29892,
29871,
29900,
29889,
29896,
286,
29924,
29871,
30187,
29899,
1050,
17885,
29877,
621,
273,
324,
313,
10142,
29892,
624,
29889,
5899,
29892,
4546,
1696,
8278,
511,
411,
784,
11541,
559,
1209,
6751,
29889,
13,
29961,
29900,
29896,
29941,
29900,
29962,
450,
3342,
9257,
313,
12696,
29897,
5745,
9528,
1244,
262,
7199,
3368,
360,
2303,
29924,
29914,
29943,
29896,
29906,
29892,
29871,
29906,
286,
29924,
8467,
329,
314,
1165,
29892,
29871,
29896,
30122,
1661,
29899,
404,
2556,
626,
1789,
1274,
4841,
29892,
29871,
29900,
29889,
29945,
501,
29914,
828,
6584,
293,
453,
262,
29892,
29871,
29900,
29889,
29945,
501,
29914,
828,
12243,
415,
290,
11078,
262,
29892,
29871,
29896,
29900,
29871,
30167,
29887,
29914,
828,
6782,
17056,
313,
497,
515,
15518,
277,
307,
1885,
29892,
1704,
3137,
12313,
29892,
3037,
361,
1696,
8278,
29897,
29871,
29900,
29889,
29896,
286,
29924,
29871,
30187,
29899,
1050,
17885,
29877,
621,
273,
324,
313,
10142,
511,
29871,
29900,
29889,
29906,
29995,
9950,
1017,
22193,
29899,
9021,
315,
6547,
29915,
29879,
15958,
478,
350,
8132,
313,
1748,
5996,
29879,
511,
29871,
29896,
30122,
20597,
10619,
6837,
267,
319,
29892,
350,
322,
315,
313,
4617,
17170,
29897,
322,
29871,
29945,
29900,
29871,
30167,
29887,
29914,
828,
1094,
2616,
29890,
293,
7255,
333,
313,
10142,
467,
28736,
11174,
310,
27878,
2109,
424,
14321,
13879,
892,
1304,
29892,
3704,
383,
29954,
29943,
29906,
313,
10142,
511,
6242,
29934,
29941,
29899,
6259,
29943,
29896,
313,
29967,
29934,
29950,
350,
2363,
455,
2063,
511,
2439,
387,
352,
262,
29899,
30187,
382,
29954,
29943,
5354,
313,
20938,
29954,
30187,
29892,
3938,
771,
11345,
511,
323,
29954,
29943,
30187,
313,
29934,
29987,
29928,
6757,
511,
18778,
284,
313,
29934,
29987,
29928,
6757,
511,
365,
6545,
313,
29934,
29987,
29928,
6757,
511,
382,
29954,
29943,
313,
29934,
29987,
29928,
6757,
511,
323,
29954,
29943,
30110,
313,
29934,
29987,
29928,
6757,
511,
379,
29934,
29954,
30110,
313,
29934,
29987,
29928,
6757,
511,
350,
3580,
29946,
313,
29934,
29987,
29928,
6757,
511,
322,
21775,
262,
319,
313,
29934,
29987,
29928,
23985,
467,
6242,
29934,
29941,
29899,
6259,
29943,
29896,
338,
263,
9120,
1873,
310,
306,
29954,
29943,
29896,
393,
756,
12212,
2756,
13593,
363,
306,
29954,
29943,
29896,
9956,
3279,
1144,
29892,
777,
310,
607,
526,
13384,
297,
298,
2890,
29907,
29879,
29889,
13681,
29899,
15715,
6545,
338,
278,
3342,
9257,
5745,
408,
2038,
29892,
6943,
29871,
29896,
29900,
8736,
29914,
828,
379,
29934,
29954,
29899,
30187,
29892,
29871,
29896,
29900,
8736,
29914,
828,
21775,
262,
319,
29892,
29871,
29906,
29900,
29900,
8736,
29914,
828,
365,
29934,
29899,
6259,
29943,
29896,
322,
29871,
29947,
8736,
29914,
828,
383,
29954,
29943,
29906,
29889,
13681,
29899,
15715,
29902,
338,
3342,
9257,
5745,
408,
2038,
6943,
29871,
29896,
29900,
8736,
29914,
828,
379,
29934,
29954,
29899,
30187,
29892,
29871,
29896,
29900,
8736,
29914,
828,
21775,
262,
319,
29892,
322,
29871,
29906,
29900,
29900,
8736,
29914,
828,
365,
29934,
29899,
6259,
29943,
29896,
29889,
512,
1716,
13681,
29899,
15715,
6545,
322,
13681,
29899,
15715,
29902,
29892,
278,
365,
29934,
29899,
6259,
29943,
29896,
4163,
508,
29892,
310,
3236,
367,
8611,
411,
10762,
29954,
29896,
29889,
13,
29961,
29900,
29896,
29941,
29896,
29962,
341,
1299,
3960,
1692,
29931,
30342,
1302,
630,
270,
17006,
892,
13240,
491,
21749,
17068,
402,
798,
386,
383,
7168,
4367,
29884,
1133,
350,
29928,
341,
1299,
3960,
1692,
29931,
30342,
4636,
313,
29121,
350,
2363,
455,
2063,
29892,
21504,
365,
6926,
29892,
405,
29889,
29967,
1696,
8278,
29897,
304,
263,
2186,
26702,
3464,
310,
1048,
29871,
29896,
29901,
29941,
29900,
304,
1048,
29871,
29896,
29901,
29896,
29900,
29900,
29900,
297,
11220,
360,
2303,
29924,
29914,
29943,
29899,
29896,
29906,
29889,
512,
697,
7232,
397,
2073,
29892,
278,
26702,
310,
341,
1299,
3960,
1692,
29931,
30342,
338,
1048,
29871,
29896,
29901,
29906,
29900,
29900,
29889,
29871,
29896,
286,
29880,
29914,
29941,
29945,
5654,
270,
728,
471,
1304,
304,
24296,
270,
17006,
363,
29871,
29896,
29899,
29906,
6199,
472,
5716,
10430,
470,
472,
3203,
975,
11147,
472,
29871,
29946,
30073,
315,
29889,
1858,
1078,
892,
6087,
701,
304,
697,
4723,
472,
29871,
29946,
30073,
315,
29889,
341,
1299,
3960,
1692,
29931,
30342,
1650,
471,
6206,
7389,
1434,
671,
29889,
13,
29961,
29900,
29896,
29941,
29906,
29962,
1152,
278,
9528,
5855,
29892,
3847,
4185,
1973,
892,
715,
630,
964,
29871,
29953,
29899,
5872,
270,
17006,
363,
10230,
310,
2999,
5855,
29889,
7965,
1973,
892,
12234,
715,
630,
4153,
964,
278,
1243,
5855,
29889,
450,
4185,
1973,
892,
1223,
11517,
1432,
2462,
322,
4656,
287,
2729,
373,
18131,
5996,
16614,
29871,
29946,
304,
29871,
29945,
3841,
1156,
715,
1218,
29889,
450,
4656,
292,
6287,
310,
29871,
29896,
304,
29871,
29945,
9701,
4392,
2827,
278,
3353,
9257,
322,
24809,
292,
12463,
18618,
310,
563,
8349,
7268,
630,
8104,
583,
29892,
1009,
6198,
2159,
29892,
322,
18618,
310,
8104,
583,
470,
5633,
310,
8104,
583,
10371,
11407,
6924,
1422,
11685,
29889,
4989,
311,
29871,
29945,
14088,
376,
680,
284,
29908,
4185,
1973,
29892,
411,
2919,
563,
8349,
7268,
630,
8104,
583,
322,
3480,
3473,
1821,
1422,
11685,
29889,
4989,
311,
29871,
29946,
14088,
263,
1407,
1781,
9257,
29892,
541,
411,
777,
6924,
1422,
11685,
29889,
4989,
311,
29871,
29941,
14088,
385,
22691,
9257,
29892,
541,
411,
2820,
4203,
278,
8104,
583,
10371,
11407,
6924,
1422,
11685,
29889,
4989,
311,
29871,
29906,
4185,
1973,
526,
758,
24130,
10835,
17473,
630,
29892,
411,
14882,
1848,
1925,
1230,
563,
8349,
7268,
630,
9101,
29889,
4989,
311,
29871,
29896,
4185,
1973,
1712,
17473,
630,
8104,
583,
470,
278,
4185,
1973,
1258,
451,
594,
4150,
470,
1258,
451,
10503,
573,
29889,
7965,
1973,
393,
10371,
1573,
1781,
13184,
310,
563,
8349,
7268,
630,
9101,
892,
1209,
4063,
304,
24809,
5520,
29899,
8489,
9257,
297,
1438,
5855,
29889,
13,
10960,
310,
1425,
29890,
29933,
29896,
29899,
29941,
29892,
405,
11007,
29896,
322,
11033,
5194,
29896,
29929,
297,
350,
29954,
29900,
29896,
29894,
315,
10071,
13,
29961,
29900,
29896,
29941,
29941,
29962,
8195,
931,
390,
29911,
29899,
9026,
29934,
471,
1304,
304,
22222,
4603,
310,
1425,
29890,
29933,
29896,
29899,
29941,
29892,
2448,
545,
29887,
352,
262,
322,
11033,
5194,
29899,
29896,
29929
] | 1,024 |
CommonCrawl
| -1.126827 | 0.841608 | -0.237484 | -0.121681 |
in BG01v cells (FIG. 1). BG01v cells cultured in DC media as described above, containing 100 ng/ml LongR3-IGF1 (LR-IGF1), 8 ng/ml FGF2 and 1 ng/ml. Activin A were harvested and RNA was prepared using the RNeasy mini kit (Qiagen) according to the manufacturer's instructions. First strand cDNA was prepared using the iScript kit (Biorad) and real time PCR was carried out using a MJ Research Opticon thermal cycler.
[0134] TaqMan assays on demand (Applied Biosystems) for ADAM19 (Hs00224960_ml), EGFR (Hs00193306_ml), ErbB2 (Hs00170433_ml), ErbB3 (Hs00176538_ml), NRG1 (Hs00247620_ml), OCT4 (Hs00742896_sl) and control GAPDH were used with TaqMan universal PCR (Applied Biosystems). The real time amplification plots are shown in FIG. 1, demonstrating expression of these transcripts in undifferentiated BG01v cells.
Inhibition of ErbB2 Slows Proliferation of BG01v Cells
[0135] The EGF domain family of ligands bind to the ErbB family of receptor tyrosine kinases. To examine the effect of known inhibitors of ErbB tyrosine kinases in hESCs, BG01v cells were plated in 6 well trays on MATRIGEL® diluted at 1:1000, in defined culture medium (DC) containing 100 ng/ml LongR3-IGF1, 8 ng/ml FGF2 and 1 ng/ml Activin A. On the next day, DMSO (carrier control), 50 nM-20 μM AG1478 (an ErbB1 inhibitor), or 100 nM-20 μM AG879 (an ErbB2 inhibitor) was added with fresh medium. The cells were cultured for an additional 5 days, with daily media changes. The cultures were then fixed and stained for alkaline phosphatase activity.
[0136] Subconfluent colonies of AP+ BG01v cells observed (FIGS. 2A, and B) in control and AG1478 cultured cells, indicating that neither DMSO nor AG1478 (50 nM-20 μM) had an apparent affect on cell proliferation. AG879, however, substantially inhibited cell growth at 5 μM (FIG. 2C) and caused cell death at 20 μM (not shown). The cultures grown in AG879 did not appear to differentiate and appeared to maintain a pluripotent morphology and alkaline phosphatase activity, indicating that AG879 appeared to inhibit proliferation without inducing differentiation, suggesting that BG01v cells are reliant on ErbB2 signaling for cell survival. Conversely, BG01v cells grown in similar conditions as above do not appear to be reliant on ErbB1 signal for proliferation.
BG01v Cells are Maintained in Defined Media Containing Heregulin
[0137] Expression of ErbB2 and ErbB3 and the inhibition of proliferation with AG879 suggested that BG01v cells have active endogenous ErbB signaling and that they may also respond to exogenous HRG-β. BG01v cells were grown in DC medium containing 10 ng/ml HRG-β, 200 ng/ml LongR3-IGF1, 8 ng/ml FGF2 and 10 ng/ml Activin A, on MATRIGEL® diluted 1:1000 (FIGS. 3A and B). These cells were grown for 4 passages, or >20 days, exhibited undifferentiated morph
|
[
297,
350,
29954,
29900,
29896,
29894,
9101,
313,
18667,
29889,
29871,
29896,
467,
350,
29954,
29900,
29896,
29894,
9101,
4185,
2955,
297,
13681,
5745,
408,
5439,
2038,
29892,
6943,
29871,
29896,
29900,
29900,
8736,
29914,
828,
6242,
29934,
29941,
29899,
6259,
29943,
29896,
313,
29519,
29899,
6259,
29943,
29896,
511,
29871,
29947,
8736,
29914,
828,
383,
29954,
29943,
29906,
322,
29871,
29896,
8736,
29914,
828,
29889,
21775,
262,
319,
892,
4023,
29894,
2868,
322,
390,
3521,
471,
13240,
773,
278,
390,
8139,
8995,
20629,
413,
277,
313,
29984,
29875,
5370,
29897,
5034,
304,
278,
12012,
9945,
29915,
29879,
11994,
29889,
3824,
851,
392,
274,
29928,
3521,
471,
13240,
773,
278,
474,
4081,
413,
277,
313,
29933,
1611,
328,
29897,
322,
1855,
931,
9609,
29934,
471,
8988,
714,
773,
263,
341,
29967,
10550,
20693,
4144,
26963,
5094,
12121,
29889,
13,
29961,
29900,
29896,
29941,
29946,
29962,
10523,
29939,
2517,
1223,
1036,
373,
9667,
313,
2052,
2957,
350,
2363,
973,
29879,
29897,
363,
11033,
5194,
29896,
29929,
313,
29950,
29879,
29900,
29900,
29906,
29906,
29946,
29929,
29953,
29900,
29918,
828,
511,
382,
29954,
15860,
313,
29950,
29879,
29900,
29900,
29896,
29929,
29941,
29941,
29900,
29953,
29918,
828,
511,
1425,
29890,
29933,
29906,
313,
29950,
29879,
29900,
29900,
29896,
29955,
29900,
29946,
29941,
29941,
29918,
828,
511,
1425,
29890,
29933,
29941,
313,
29950,
29879,
29900,
29900,
29896,
29955,
29953,
29945,
29941,
29947,
29918,
828,
511,
27759,
29954,
29896,
313,
29950,
29879,
29900,
29900,
29906,
29946,
29955,
29953,
29906,
29900,
29918,
828,
511,
438,
1783,
29946,
313,
29950,
29879,
29900,
29900,
29955,
29946,
29906,
29947,
29929,
29953,
29918,
2536,
29897,
322,
2761,
402,
3301,
29928,
29950,
892,
1304,
411,
10523,
29939,
2517,
15968,
9609,
29934,
313,
2052,
2957,
350,
2363,
973,
29879,
467,
450,
1855,
931,
20563,
2450,
24580,
526,
4318,
297,
383,
6259,
29889,
29871,
29896,
29892,
9004,
1218,
4603,
310,
1438,
1301,
924,
29879,
297,
563,
8349,
7268,
630,
350,
29954,
29900,
29896,
29894,
9101,
29889,
13,
797,
6335,
654,
310,
1425,
29890,
29933,
29906,
317,
677,
29879,
1019,
29880,
9633,
362,
310,
350,
29954,
29900,
29896,
29894,
315,
10071,
13,
29961,
29900,
29896,
29941,
29945,
29962,
450,
382,
29954,
29943,
5354,
3942,
310,
14172,
4167,
7868,
304,
278,
1425,
29890,
29933,
3942,
310,
337,
14268,
7911,
1883,
457,
19015,
2129,
29889,
1763,
25917,
278,
2779,
310,
2998,
297,
6335,
17259,
310,
1425,
29890,
29933,
7911,
1883,
457,
19015,
2129,
297,
298,
2890,
29907,
29879,
29892,
350,
29954,
29900,
29896,
29894,
9101,
892,
715,
630,
297,
29871,
29953,
1532,
260,
764,
29879,
373,
341,
1299,
3960,
1692,
29931,
30342,
21749,
3860,
472,
29871,
29896,
29901,
29896,
29900,
29900,
29900,
29892,
297,
3342,
9257,
18350,
313,
12696,
29897,
6943,
29871,
29896,
29900,
29900,
8736,
29914,
828,
6242,
29934,
29941,
29899,
6259,
29943,
29896,
29892,
29871,
29947,
8736,
29914,
828,
383,
29954,
29943,
29906,
322,
29871,
29896,
8736,
29914,
828,
21775,
262,
319,
29889,
1551,
278,
2446,
2462,
29892,
360,
4345,
29949,
313,
4287,
4336,
2761,
511,
29871,
29945,
29900,
302,
29924,
29899,
29906,
29900,
29871,
30167,
29924,
16369,
29896,
29946,
29955,
29947,
313,
273,
1425,
29890,
29933,
29896,
297,
6335,
2105,
511,
470,
29871,
29896,
29900,
29900,
302,
29924,
29899,
29906,
29900,
29871,
30167,
29924,
16369,
29947,
29955,
29929,
313,
273,
1425,
29890,
29933,
29906,
297,
6335,
2105,
29897,
471,
2715,
411,
10849,
18350,
29889,
450,
9101,
892,
4185,
2955,
363,
385,
5684,
29871,
29945,
3841,
29892,
411,
14218,
5745,
3620,
29889,
450,
4185,
1973,
892,
769,
4343,
322,
380,
7114,
363,
394,
11311,
457,
1374,
25715,
271,
559,
6354,
29889,
13,
29961,
29900,
29896,
29941,
29953,
29962,
3323,
535,
1579,
8122,
8104,
583,
310,
12279,
29974,
350,
29954,
29900,
29896,
29894,
9101,
8900,
313,
3738,
10749,
29889,
29871,
29906,
29909,
29892,
322,
350,
29897,
297,
2761,
322,
16369,
29896,
29946,
29955,
29947,
4185,
2955,
9101,
29892,
23941,
393,
9561,
360,
4345,
29949,
3643,
16369,
29896,
29946,
29955,
29947,
313,
29945,
29900,
302,
29924,
29899,
29906,
29900,
29871,
30167,
29924,
29897,
750,
385,
20295,
6602,
373,
3038,
410,
29880,
9633,
362,
29889,
16369,
29947,
29955,
29929,
29892,
3138,
29892,
23228,
368,
297,
6335,
1573,
3038,
14321,
472,
29871,
29945,
29871,
30167,
29924,
313,
18667,
29889,
29871,
29906,
29907,
29897,
322,
8581,
3038,
4892,
472,
29871,
29906,
29900,
29871,
30167,
29924,
313,
1333,
4318,
467,
450,
4185,
1973,
21633,
297,
16369,
29947,
29955,
29929,
1258,
451,
2615,
304,
17473,
403,
322,
7470,
304,
7344,
263,
715,
332,
666,
327,
296,
18131,
3002,
322,
394,
11311,
457,
1374,
25715,
271,
559,
6354,
29892,
23941,
393,
16369,
29947,
29955,
29929,
7470,
304,
297,
6335,
277,
410,
29880,
9633,
362,
1728,
9013,
3277,
1422,
11685,
29892,
26233,
393,
350,
29954,
29900,
29896,
29894,
9101,
526,
12536,
424,
373,
1425,
29890,
29933,
29906,
7182,
292,
363,
3038,
10503,
2561,
29889,
1281,
874,
873,
29892,
350,
29954,
29900,
29896,
29894,
9101,
21633,
297,
2788,
5855,
408,
2038,
437,
451,
2615,
304,
367,
12536,
424,
373,
1425,
29890,
29933,
29896,
7182,
363,
410,
29880,
9633,
362,
29889,
13,
29933,
29954,
29900,
29896,
29894,
315,
10071,
526,
341,
2365,
7114,
297,
5282,
1312,
8213,
2866,
17225,
2439,
387,
352,
262,
13,
29961,
29900,
29896,
29941,
29955,
29962,
21444,
310,
1425,
29890,
29933,
29906,
322,
1425,
29890,
29933,
29941,
322,
278,
297,
6335,
654,
310,
410,
29880,
9633,
362,
411,
16369,
29947,
29955,
29929,
7829,
393,
350,
29954,
29900,
29896,
29894,
9101,
505,
6136,
1095,
6352,
681,
1425,
29890,
29933,
7182,
292,
322,
393,
896,
1122,
884,
10049,
304,
429,
6352,
681,
379,
29934,
29954,
29899,
30187,
29889,
350,
29954,
29900,
29896,
29894,
9101,
892,
21633,
297,
13681,
18350,
6943,
29871,
29896,
29900,
8736,
29914,
828,
379,
29934,
29954,
29899,
30187,
29892,
29871,
29906,
29900,
29900,
8736,
29914,
828,
6242,
29934,
29941,
29899,
6259,
29943,
29896,
29892,
29871,
29947,
8736,
29914,
828,
383,
29954,
29943,
29906,
322,
29871,
29896,
29900,
8736,
29914,
828,
21775,
262,
319,
29892,
373,
341,
1299,
3960,
1692,
29931,
30342,
21749,
3860,
29871,
29896,
29901,
29896,
29900,
29900,
29900,
313,
3738,
10749,
29889,
29871,
29941,
29909,
322,
350,
467,
4525,
9101,
892,
21633,
363,
29871,
29946,
1209,
1179,
29892,
470,
1405,
29906,
29900,
3841,
29892,
10371,
1573,
563,
8349,
7268,
630,
18131
] | 1,024 |
CommonCrawl
| -0.864631 | 1.220561 | 0.008187 | 0.317231 |
ology and did not show elevated spontaneous differentiation.
[0138] Furthermore, BG01v cells were also maintained for 2 passages, or >13 days, in DC medium comprising 10 ng/ml HRGβ, 200 ng/ml LongR3-IGF1, and 10 ng/ml Activin A. These cultures proliferated normally and exhibited very low spontaneous differentiation, demonstrating that BG01v cells could be maintained in defined conditions with HRGβ in the absence of FGF2.
The Role of ErbB2-Directed Tyrosine Kinase in ES Cells
[0139] RT-PCR demonstrated that mESCs express ADAM19, Neuregulin1 (Nrg1), and ErbB1-4 (FIG. 4A). In mESCs, ErbB2 and 3 appeared to be expressed at higher levels than ErbB1, with low levels of ErbB4 being detected. These data suggest that endogenous HRG-β could be involved in driving mESC self-renewal.
[0140] The expression of the ErbB receptor transcripts in mouse embryonic fibroblasts (MEFs) was also examined (FIG. 4B). MEFs are a heterogenous population of cells derived from E12.5-13.5 viscera that have been used historically to maintain mouse and human EC cells and ES cells. Expression of Nrg1 and Adam19 in this population suggests that the HRG-β ectodomain is also present in MEF-conditioned media and may exert significant effects upon pluripotency.
[0141] AG1478 and AG879 were used to examine the role of HRG/ErbB signaling in mouse ES cells. R1 mouse ES cells were maintained in standard conditions in DMEM, 10% FBS, 10% KSR, 0.5 U/ml penicillin, 0.5 U/ml streptomycin, 1×NEAA, 1 mM sodium pyruvate, 1000 U/ml LIF (ESGRO), 0.1 mM β-ME, and were passaged with 0.5% trypsin/EDTA. 2×105 cells/well were plated in 6 well trays on MATRIGEL® diluted at 1:1000. The day after plating, DMSO (carrier control), 1-50 μM AG1478, or 1-50 μM AG879 was added with fresh medium. The cells were cultured an additional 8 days, with daily media changes. The cultures were then fixed and stained for alkaline phosphatase activity.
[0142] DMSO and 1-50 μM AG1478 had no apparent affect on cell proliferation, with subconfluent colonies of alkaline phosphatase positive mESCs observed (FIGS. 5A-C). However, AG879 substantially inhibited cell growth at 50 μM (compare FIGS. 5D and 5F) and may have slowed proliferation at 20 μM (FIG. 5E). mESCs grown in AG879 did not appear to differentiate and maintained a pluripotent morphology, and alkaline phosphatase activity.
[0143] The results indicate that AG879 appeared to inhibit proliferation, without inducing differentiation, of mESCs, suggesting that mESCs require ErbB2 signaling for proliferation. Conversely, mESCs do not appear to be reliant on an ErbB1 signal for proliferation. The concentration of AG879 required to inhibit proliferation was ˜10× higher for mESCs than that for BG01v cells grown in defined conditions, indicating that either the serum used in the mESC conditions may have interfered with the activity of the drug, that AG879 has a lower affinity for the mouse ErbB2 tyrosine kinase than for human ErbB2 tyrosine kinase, or that ErbB2 may
|
[
3002,
322,
1258,
451,
1510,
11858,
630,
805,
609,
23584,
1422,
11685,
29889,
13,
29961,
29900,
29896,
29941,
29947,
29962,
16478,
29892,
350,
29954,
29900,
29896,
29894,
9101,
892,
884,
19949,
363,
29871,
29906,
1209,
1179,
29892,
470,
1405,
29896,
29941,
3841,
29892,
297,
13681,
18350,
7199,
5921,
29871,
29896,
29900,
8736,
29914,
828,
379,
29934,
29954,
30187,
29892,
29871,
29906,
29900,
29900,
8736,
29914,
828,
6242,
29934,
29941,
29899,
6259,
29943,
29896,
29892,
322,
29871,
29896,
29900,
8736,
29914,
828,
21775,
262,
319,
29889,
4525,
4185,
1973,
410,
29880,
9633,
630,
12891,
322,
10371,
1573,
1407,
4482,
805,
609,
23584,
1422,
11685,
29892,
9004,
1218,
393,
350,
29954,
29900,
29896,
29894,
9101,
1033,
367,
19949,
297,
3342,
5855,
411,
379,
29934,
29954,
30187,
297,
278,
18070,
310,
383,
29954,
29943,
29906,
29889,
13,
1576,
1528,
280,
310,
1425,
29890,
29933,
29906,
29899,
17392,
287,
9656,
1883,
457,
20311,
559,
297,
17956,
315,
10071,
13,
29961,
29900,
29896,
29941,
29929,
29962,
390,
29911,
29899,
9026,
29934,
28585,
393,
286,
2890,
29907,
29879,
4653,
11033,
5194,
29896,
29929,
29892,
2448,
545,
29887,
352,
262,
29896,
313,
29940,
11007,
29896,
511,
322,
1425,
29890,
29933,
29896,
29899,
29946,
313,
18667,
29889,
29871,
29946,
29909,
467,
512,
286,
2890,
29907,
29879,
29892,
1425,
29890,
29933,
29906,
322,
29871,
29941,
7470,
304,
367,
13384,
472,
6133,
11174,
1135,
1425,
29890,
29933,
29896,
29892,
411,
4482,
11174,
310,
1425,
29890,
29933,
29946,
1641,
17809,
29889,
4525,
848,
4368,
393,
1095,
6352,
681,
379,
29934,
29954,
29899,
30187,
1033,
367,
9701,
297,
19500,
286,
2890,
29907,
1583,
29899,
1267,
809,
284,
29889,
13,
29961,
29900,
29896,
29946,
29900,
29962,
450,
4603,
310,
278,
1425,
29890,
29933,
337,
14268,
1301,
924,
29879,
297,
9495,
7232,
719,
8927,
18755,
307,
2204,
19416,
313,
2303,
29943,
29879,
29897,
471,
884,
4392,
1312,
313,
18667,
29889,
29871,
29946,
29933,
467,
22986,
29943,
29879,
526,
263,
25745,
6352,
681,
4665,
310,
9101,
10723,
515,
382,
29896,
29906,
29889,
29945,
29899,
29896,
29941,
29889,
29945,
1998,
23322,
393,
505,
1063,
1304,
3603,
1711,
304,
7344,
9495,
322,
5199,
17522,
9101,
322,
17956,
9101,
29889,
21444,
310,
405,
11007,
29896,
322,
11783,
29896,
29929,
297,
445,
4665,
14661,
393,
278,
379,
29934,
29954,
29899,
30187,
321,
312,
397,
290,
475,
338,
884,
2198,
297,
22986,
29943,
29899,
16122,
287,
5745,
322,
1122,
429,
814,
7282,
9545,
2501,
715,
332,
666,
327,
3819,
29889,
13,
29961,
29900,
29896,
29946,
29896,
29962,
16369,
29896,
29946,
29955,
29947,
322,
16369,
29947,
29955,
29929,
892,
1304,
304,
25917,
278,
6297,
310,
379,
29934,
29954,
29914,
2110,
29890,
29933,
7182,
292,
297,
9495,
17956,
9101,
29889,
390,
29896,
9495,
17956,
9101,
892,
19949,
297,
3918,
5855,
297,
360,
2303,
29924,
29892,
29871,
29896,
29900,
29995,
383,
9851,
29892,
29871,
29896,
29900,
29995,
476,
14098,
29892,
29871,
29900,
29889,
29945,
501,
29914,
828,
6584,
293,
453,
262,
29892,
29871,
29900,
29889,
29945,
501,
29914,
828,
12243,
415,
290,
11078,
262,
29892,
29871,
29896,
30122,
8186,
6344,
29892,
29871,
29896,
286,
29924,
20892,
1974,
11451,
582,
29894,
403,
29892,
29871,
29896,
29900,
29900,
29900,
501,
29914,
828,
365,
6545,
313,
2890,
29954,
1672,
511,
29871,
29900,
29889,
29896,
286,
29924,
29871,
30187,
29899,
2303,
29892,
322,
892,
1209,
4063,
411,
29871,
29900,
29889,
29945,
29995,
1018,
567,
262,
29914,
3352,
6040,
29889,
29871,
29906,
30122,
29896,
29900,
29945,
9101,
29914,
5872,
892,
715,
630,
297,
29871,
29953,
1532,
260,
764,
29879,
373,
341,
1299,
3960,
1692,
29931,
30342,
21749,
3860,
472,
29871,
29896,
29901,
29896,
29900,
29900,
29900,
29889,
450,
2462,
1156,
715,
1218,
29892,
360,
4345,
29949,
313,
4287,
4336,
2761,
511,
29871,
29896,
29899,
29945,
29900,
29871,
30167,
29924,
16369,
29896,
29946,
29955,
29947,
29892,
470,
29871,
29896,
29899,
29945,
29900,
29871,
30167,
29924,
16369,
29947,
29955,
29929,
471,
2715,
411,
10849,
18350,
29889,
450,
9101,
892,
4185,
2955,
385,
5684,
29871,
29947,
3841,
29892,
411,
14218,
5745,
3620,
29889,
450,
4185,
1973,
892,
769,
4343,
322,
380,
7114,
363,
394,
11311,
457,
1374,
25715,
271,
559,
6354,
29889,
13,
29961,
29900,
29896,
29946,
29906,
29962,
360,
4345,
29949,
322,
29871,
29896,
29899,
29945,
29900,
29871,
30167,
29924,
16369,
29896,
29946,
29955,
29947,
750,
694,
20295,
6602,
373,
3038,
410,
29880,
9633,
362,
29892,
411,
1014,
535,
1579,
8122,
8104,
583,
310,
394,
11311,
457,
1374,
25715,
271,
559,
6374,
286,
2890,
29907,
29879,
8900,
313,
3738,
10749,
29889,
29871,
29945,
29909,
29899,
29907,
467,
2398,
29892,
16369,
29947,
29955,
29929,
23228,
368,
297,
6335,
1573,
3038,
14321,
472,
29871,
29945,
29900,
29871,
30167,
29924,
313,
18307,
383,
6259,
29903,
29889,
29871,
29945,
29928,
322,
29871,
29945,
29943,
29897,
322,
1122,
505,
5232,
287,
410,
29880,
9633,
362,
472,
29871,
29906,
29900,
29871,
30167,
29924,
313,
18667,
29889,
29871,
29945,
29923,
467,
286,
2890,
29907,
29879,
21633,
297,
16369,
29947,
29955,
29929,
1258,
451,
2615,
304,
17473,
403,
322,
19949,
263,
715,
332,
666,
327,
296,
18131,
3002,
29892,
322,
394,
11311,
457,
1374,
25715,
271,
559,
6354,
29889,
13,
29961,
29900,
29896,
29946,
29941,
29962,
450,
2582,
12266,
393,
16369,
29947,
29955,
29929,
7470,
304,
297,
6335,
277,
410,
29880,
9633,
362,
29892,
1728,
9013,
3277,
1422,
11685,
29892,
310,
286,
2890,
29907,
29879,
29892,
26233,
393,
286,
2890,
29907,
29879,
1996,
1425,
29890,
29933,
29906,
7182,
292,
363,
410,
29880,
9633,
362,
29889,
1281,
874,
873,
29892,
286,
2890,
29907,
29879,
437,
451,
2615,
304,
367,
12536,
424,
373,
385,
1425,
29890,
29933,
29896,
7182,
363,
410,
29880,
9633,
362,
29889,
450,
26702,
310,
16369,
29947,
29955,
29929,
3734,
304,
297,
6335,
277,
410,
29880,
9633,
362,
471,
29871,
31793,
29896,
29900,
30122,
6133,
363,
286,
2890,
29907,
29879,
1135,
393,
363,
350,
29954,
29900,
29896,
29894,
9101,
21633,
297,
3342,
5855,
29892,
23941,
393,
2845,
278,
724,
398,
1304,
297,
278,
286,
2890,
29907,
5855,
1122,
505,
1006,
571,
287,
411,
278,
6354,
310,
278,
15721,
29892,
393,
16369,
29947,
29955,
29929,
756,
263,
5224,
2756,
13593,
363,
278,
9495,
1425,
29890,
29933,
29906,
7911,
1883,
457,
19015,
559,
1135,
363,
5199,
1425,
29890,
29933,
29906,
7911,
1883,
457,
19015,
559,
29892,
470,
393,
1425,
29890,
29933,
29906,
1122
] | 1,024 |
CommonCrawl
| -0.774968 | 1.288046 | 0.295103 | 0.404508 |
play slightly different roles with the different species of ES cells.
[0144] Another highly selective inhibitor of the ErbB2 tyrosine kinase, tyrphostin AG825 (Murillo, et al. 2001 Cancer Res 61, 7408-7412), was used to investigate the role of ErbB2 in human ESCs. AG825 significantly inhibited proliferation of hESCs growing in conditioned medium (CM) (FIG. 6A). AG825 inhibited proliferation without widespread cell death, and viable hESCs could be maintained for >5 days (not shown). Western blotting showed that AG825 inhibited autophosphorylation of ErbB2 at tyrosine-1248 in starved/heregulin (HRG) pulsed hESCs growing in DC-HAIF (FIG. 6B). Thus, disruption of ErbB2 signaling severely inhibited hESC proliferation. To establish hESCs in defined growth conditions, cultures could be passaged directly from CM conditions into DC-HAIF and exhibited minimal spontaneous differentiation (FIG. 6C). Colony and cell-counting assays confirmed that LongR3-IGF1 and HRG played the major roles in self-renewal and proliferation in the context of one of the embodiments of the present invention (FIGS. 6D, 6E). Phosphorylation of IGF1R, IR, FGF2α, ErbB2, and ErbB3 was also observed in both steady-state DC-HAIF cultures, and in starved cultures that were pulsed with DC-HAIF (FIG. 6F).
Culture of Mouse ES Cells in Defined Conditions
[0145] To further examine the role of HRG/ErbB2 signaling in mouse ES cells, the proliferation of R1 ES cells was examined in DC medium using a combination of growth factors. 1×105 cells/well were plated in 6-well trays, coated with 0.2% gelatin, in DC containing combinations of 10 ng/ml HRG-β, 100 ng/ml LongR3-IGF1, 1 ng/ml Activin A, 1000 U/ml mouse LIF or 10 ng/ml BMP4 (Table 1, below). Proliferation was observed over 8 days.
[0146] Viable colonies only grew in conditions containing at least LIF/HRG-β or LIF/BMP4 (Table 1). No additional obvious benefit was observed when LongR3-IGF1 or Activin were added to these combinations. Normal proliferation was observed in a control parental culture, and no viable colonies were observed in defined media without any growth factors.
TABLE-US-00001 TABLE 1 HRG IGF Activin LIF BMP4 Growth + No + + Yes + + No + + + Yes + + + No + + + + Yes + + No + + + Yes + + Yes + + + Yes
[0147] A quantitative assay was performed by plating 2×105 cells/well in 6-well trays on 1:1000 MATRIGEL®, in selected combinations of 10 or 50 ng/ml HRG-β, 10 ng/ml EGF, 1000 U/ml LIF or 10 ng/ml BMP4. The cultures were grown for 8 days, fixed, and the number of alkaline phosphatase colonies was counted (FIG. 7A). No colonies were observed in defined conditions without growth factors, and <45 colonies were observed with HRG-β, HRG-β/EGF and HRG-β/BMP combinations. While 1358 colonies were observed in LIF alone, 4114 and 3734 colonies were observed in the 10 ng/ml HRG-β/LIF and 50 ng/ml HRG-β/LIF combinations, respectively. This indicated that in defined conditions, LIF alone provided a substantial pluripotency signal, and HRG
|
[
1708,
10029,
1422,
16178,
411,
278,
1422,
6606,
310,
17956,
9101,
29889,
13,
29961,
29900,
29896,
29946,
29946,
29962,
7280,
10712,
1831,
573,
297,
6335,
2105,
310,
278,
1425,
29890,
29933,
29906,
7911,
1883,
457,
19015,
559,
29892,
260,
4316,
561,
520,
262,
16369,
29947,
29906,
29945,
313,
29924,
332,
9093,
29892,
634,
394,
29889,
29871,
29906,
29900,
29900,
29896,
1815,
2265,
2538,
29871,
29953,
29896,
29892,
29871,
29955,
29946,
29900,
29947,
29899,
29955,
29946,
29896,
29906,
511,
471,
1304,
304,
23033,
278,
6297,
310,
1425,
29890,
29933,
29906,
297,
5199,
382,
7187,
29879,
29889,
16369,
29947,
29906,
29945,
16951,
297,
6335,
1573,
410,
29880,
9633,
362,
310,
298,
2890,
29907,
29879,
15678,
297,
4195,
287,
18350,
313,
24494,
29897,
313,
18667,
29889,
29871,
29953,
29909,
467,
16369,
29947,
29906,
29945,
297,
6335,
1573,
410,
29880,
9633,
362,
1728,
281,
2247,
29886,
949,
3038,
4892,
29892,
322,
3516,
519,
298,
2890,
29907,
29879,
1033,
367,
19949,
363,
1405,
29945,
3841,
313,
1333,
4318,
467,
10504,
1999,
327,
1259,
10018,
393,
16369,
29947,
29906,
29945,
297,
6335,
1573,
1120,
3021,
25715,
706,
18411,
310,
1425,
29890,
29933,
29906,
472,
7911,
1883,
457,
29899,
29896,
29906,
29946,
29947,
297,
5810,
1490,
29914,
2276,
387,
352,
262,
313,
20938,
29954,
29897,
282,
7273,
287,
298,
2890,
29907,
29879,
15678,
297,
13681,
29899,
15715,
6545,
313,
18667,
29889,
29871,
29953,
29933,
467,
6549,
29892,
766,
18953,
310,
1425,
29890,
29933,
29906,
7182,
292,
2775,
873,
297,
6335,
1573,
298,
2890,
29907,
410,
29880,
9633,
362,
29889,
1763,
10127,
298,
2890,
29907,
29879,
297,
3342,
14321,
5855,
29892,
4185,
1973,
1033,
367,
1209,
4063,
4153,
515,
315,
29924,
5855,
964,
13681,
29899,
15715,
6545,
322,
10371,
1573,
13114,
805,
609,
23584,
1422,
11685,
313,
18667,
29889,
29871,
29953,
29907,
467,
1530,
2592,
322,
3038,
29899,
2798,
292,
1223,
1036,
16725,
393,
6242,
29934,
29941,
29899,
6259,
29943,
29896,
322,
379,
29934,
29954,
5318,
278,
4655,
16178,
297,
1583,
29899,
1267,
809,
284,
322,
410,
29880,
9633,
362,
297,
278,
3030,
310,
697,
310,
278,
7232,
397,
7862,
310,
278,
2198,
297,
7316,
313,
3738,
10749,
29889,
29871,
29953,
29928,
29892,
29871,
29953,
29923,
467,
1963,
25715,
706,
18411,
310,
306,
29954,
29943,
29896,
29934,
29892,
23292,
29892,
383,
29954,
29943,
29906,
30110,
29892,
1425,
29890,
29933,
29906,
29892,
322,
1425,
29890,
29933,
29941,
471,
884,
8900,
297,
1716,
27357,
29899,
3859,
13681,
29899,
15715,
6545,
4185,
1973,
29892,
322,
297,
5810,
1490,
4185,
1973,
393,
892,
282,
7273,
287,
411,
13681,
29899,
15715,
6545,
313,
18667,
29889,
29871,
29953,
29943,
467,
13,
27926,
310,
25992,
17956,
315,
10071,
297,
5282,
1312,
11790,
2187,
13,
29961,
29900,
29896,
29946,
29945,
29962,
1763,
4340,
25917,
278,
6297,
310,
379,
29934,
29954,
29914,
2110,
29890,
29933,
29906,
7182,
292,
297,
9495,
17956,
9101,
29892,
278,
410,
29880,
9633,
362,
310,
390,
29896,
17956,
9101,
471,
4392,
1312,
297,
13681,
18350,
773,
263,
10296,
310,
14321,
13879,
29889,
29871,
29896,
30122,
29896,
29900,
29945,
9101,
29914,
5872,
892,
715,
630,
297,
29871,
29953,
29899,
5872,
260,
764,
29879,
29892,
1302,
630,
411,
29871,
29900,
29889,
29906,
29995,
9127,
21203,
29892,
297,
13681,
6943,
18240,
310,
29871,
29896,
29900,
8736,
29914,
828,
379,
29934,
29954,
29899,
30187,
29892,
29871,
29896,
29900,
29900,
8736,
29914,
828,
6242,
29934,
29941,
29899,
6259,
29943,
29896,
29892,
29871,
29896,
8736,
29914,
828,
21775,
262,
319,
29892,
29871,
29896,
29900,
29900,
29900,
501,
29914,
828,
9495,
365,
6545,
470,
29871,
29896,
29900,
8736,
29914,
828,
350,
3580,
29946,
313,
3562,
29871,
29896,
29892,
2400,
467,
1019,
29880,
9633,
362,
471,
8900,
975,
29871,
29947,
3841,
29889,
13,
29961,
29900,
29896,
29946,
29953,
29962,
10630,
519,
8104,
583,
871,
13631,
297,
5855,
6943,
472,
3203,
365,
6545,
29914,
20938,
29954,
29899,
30187,
470,
365,
6545,
29914,
29933,
3580,
29946,
313,
3562,
29871,
29896,
467,
1939,
5684,
6924,
14169,
471,
8900,
746,
6242,
29934,
29941,
29899,
6259,
29943,
29896,
470,
21775,
262,
892,
2715,
304,
1438,
18240,
29889,
21981,
410,
29880,
9633,
362,
471,
8900,
297,
263,
2761,
3847,
284,
9257,
29892,
322,
694,
3516,
519,
8104,
583,
892,
8900,
297,
3342,
5745,
1728,
738,
14321,
13879,
29889,
13,
21009,
29899,
3308,
29899,
29900,
29900,
29900,
29900,
29896,
10911,
29871,
29896,
379,
29934,
29954,
306,
29954,
29943,
21775,
262,
365,
6545,
350,
3580,
29946,
402,
798,
386,
718,
1939,
718,
718,
3869,
718,
718,
1939,
718,
718,
718,
3869,
718,
718,
718,
1939,
718,
718,
718,
718,
3869,
718,
718,
1939,
718,
718,
718,
3869,
718,
718,
3869,
718,
718,
718,
3869,
13,
29961,
29900,
29896,
29946,
29955,
29962,
319,
4323,
23378,
1223,
388,
471,
8560,
491,
715,
1218,
29871,
29906,
30122,
29896,
29900,
29945,
9101,
29914,
5872,
297,
29871,
29953,
29899,
5872,
260,
764,
29879,
373,
29871,
29896,
29901,
29896,
29900,
29900,
29900,
341,
1299,
3960,
1692,
29931,
30342,
29892,
297,
4629,
18240,
310,
29871,
29896,
29900,
470,
29871,
29945,
29900,
8736,
29914,
828,
379,
29934,
29954,
29899,
30187,
29892,
29871,
29896,
29900,
8736,
29914,
828,
382,
29954,
29943,
29892,
29871,
29896,
29900,
29900,
29900,
501,
29914,
828,
365,
6545,
470,
29871,
29896,
29900,
8736,
29914,
828,
350,
3580,
29946,
29889,
450,
4185,
1973,
892,
21633,
363,
29871,
29947,
3841,
29892,
4343,
29892,
322,
278,
1353,
310,
394,
11311,
457,
1374,
25715,
271,
559,
8104,
583,
471,
29115,
313,
18667,
29889,
29871,
29955,
29909,
467,
1939,
8104,
583,
892,
8900,
297,
3342,
5855,
1728,
14321,
13879,
29892,
322,
529,
29946,
29945,
8104,
583,
892,
8900,
411,
379,
29934,
29954,
29899,
30187,
29892,
379,
29934,
29954,
29899,
30187,
29914,
11787,
29943,
322,
379,
29934,
29954,
29899,
30187,
29914,
29933,
3580,
18240,
29889,
5806,
29871,
29896,
29941,
29945,
29947,
8104,
583,
892,
8900,
297,
365,
6545,
7432,
29892,
29871,
29946,
29896,
29896,
29946,
322,
29871,
29941,
29955,
29941,
29946,
8104,
583,
892,
8900,
297,
278,
29871,
29896,
29900,
8736,
29914,
828,
379,
29934,
29954,
29899,
30187,
29914,
5265,
29943,
322,
29871,
29945,
29900,
8736,
29914,
828,
379,
29934,
29954,
29899,
30187,
29914,
5265,
29943,
18240,
29892,
8307,
29889,
910,
18694,
393,
297,
3342,
5855,
29892,
365,
6545,
7432,
4944,
263,
23228,
715,
332,
666,
327,
3819,
7182,
29892,
322,
379,
29934,
29954
] | 1,024 |
CommonCrawl
| -0.803497 | 1.038871 | 0.274481 | 0.350118 |
-β exhibited a large synergistic effect with LIF, more than doubling the number of Proliferating mESC colonies in this assay. Low magnification images of this assay also indicate this synergistic proliferative effect (FIGS. 7B-G).
Characterization of Pluripotency of Human Embryonic Stem Cells (hESCs) Maintained in DC-HAIF
[0148] Multiple approaches were used to confirm the maintenance of plasticity of hESCs in DC-HAIF. BG02 cells cultured in DC-HAIF for 6 months (25 passages) maintained the potential to form complex teratomas (FIG. 8A) and representatives of the three germ layers in vitro (FIG. 8B). Transcriptional analyses were used to compare global expression in hESCs cells (Liu et al 2006, BMC Dev Biol 6, 20) maintained in CM and DC-HAIF. Greater than 11,600 transcripts were detected in BG02 cells grown in DC-HAIF for 10 and 32 passages, and BG02 cells grown in CM for 64 passages. There were about 10364 transcripts common to all populations (FIG. 8C), including known hESC markers such as CD9, DNMT3, NANOG, OCT4, TERT and UTF1 (not shown). High correlation coefficients were observed in comparisons of CM and DC-HAIF cultures (R2select=0.928), as well as in early and late passage cells (R2select=0.959) (FIG. 8D). Hierarchical clustering analysis demonstrated that BG02 cells maintained in DC-HAIF grouped tightly and retained a close similarity to BG02 and BG03 cells maintained in CM (FIG. 8E). These data are consistent with previous analyses showing that undifferentiated hESCs clustered tightly compared to embryoid bodies or fibroblasts (Liu et al 2006, BMC Dev Biol 6, 20). Thus, cells maintained in the compositions of the present invention are able to maintain key markers of pluirpotency. Accordingly, the compositions of the present invention can be used as a simple medium for supporting self-renewal of differentiable cells.
Maintenance of Human Embryonic Stem Cells (hESCs) on Humanized Extracellular Matrices (ECMs) in DC-HAIF
[0149] To investigate the role of ErbB2 signaling and develop a defined media for hESCs, DC-HAIF cultures were initially expanded on culture dished coated with growth factor-reduced MATRIGEL® 1:30, but could also be maintained successfully long-term on this substrate diluted 1:200 (FIG. 9A), or 1:1000. BG02 and CyT49 hESCs could also be maintained for >5 passages on tissue culture dishes coated with human serum (FIG. 9B); human fibronectin (FIG. 9C); or VITROGRO® (FIG. 9D), which is a proprietary humanized ECM.
Single Cell Passaging of Human Embryonic Stem Cells (hESCs)
[0150] Multiple research groups have demonstrated that certain triplodies, notably of hChr12 and 17, are accumulated in hESCs under certain sub-optimal culture conditions (Baker et al., 2007 Nat. Biotech.25(2):207-215). The appearance of triploidies seems to be most directly related to poor cell survival when cultures are split to single cells at passaging, providing a presumed strong selective growth advantage for cells harboring these aneuploidies. Conversely, hESCs growing in one embodiment of the present invention, DC-HAIF, maintained high viability at plating after being split to single cells (FIG. 10A-D). BG01 and BG02 cells maintained a normal karyotype (FIG. 10E) after being passaged with ACCUTASE® for >18 and 19 passages respectively. The maintenance of
|
[
29899,
30187,
10371,
1573,
263,
2919,
5222,
15064,
4695,
2779,
411,
365,
6545,
29892,
901,
1135,
4662,
21435,
278,
1353,
310,
1019,
29880,
9633,
1218,
286,
2890,
29907,
8104,
583,
297,
445,
1223,
388,
29889,
17511,
9119,
2450,
4558,
310,
445,
1223,
388,
884,
12266,
445,
5222,
15064,
4695,
410,
29880,
9633,
1230,
2779,
313,
3738,
10749,
29889,
29871,
29955,
29933,
29899,
29954,
467,
13,
20755,
2133,
310,
1858,
332,
666,
327,
3819,
310,
12968,
28735,
719,
8927,
624,
331,
315,
10071,
313,
29882,
2890,
29907,
29879,
29897,
341,
2365,
7114,
297,
13681,
29899,
15715,
6545,
13,
29961,
29900,
29896,
29946,
29947,
29962,
26905,
13501,
892,
1304,
304,
9659,
278,
25413,
310,
715,
6288,
537,
310,
298,
2890,
29907,
29879,
297,
13681,
29899,
15715,
6545,
29889,
350,
29954,
29900,
29906,
9101,
4185,
2955,
297,
13681,
29899,
15715,
6545,
363,
29871,
29953,
7378,
313,
29906,
29945,
1209,
1179,
29897,
19949,
278,
7037,
304,
883,
4280,
1935,
8678,
294,
313,
18667,
29889,
29871,
29947,
29909,
29897,
322,
2755,
5056,
310,
278,
2211,
22593,
15359,
297,
13901,
307,
313,
18667,
29889,
29871,
29947,
29933,
467,
4103,
3395,
284,
3483,
952,
267,
892,
1304,
304,
7252,
5534,
4603,
297,
298,
2890,
29907,
29879,
9101,
313,
29931,
5871,
634,
394,
29871,
29906,
29900,
29900,
29953,
29892,
350,
12513,
9481,
3457,
324,
29871,
29953,
29892,
29871,
29906,
29900,
29897,
19949,
297,
315,
29924,
322,
13681,
29899,
15715,
6545,
29889,
4122,
1008,
1135,
29871,
29896,
29896,
29892,
29953,
29900,
29900,
1301,
924,
29879,
892,
17809,
297,
350,
29954,
29900,
29906,
9101,
21633,
297,
13681,
29899,
15715,
6545,
363,
29871,
29896,
29900,
322,
29871,
29941,
29906,
1209,
1179,
29892,
322,
350,
29954,
29900,
29906,
9101,
21633,
297,
315,
29924,
363,
29871,
29953,
29946,
1209,
1179,
29889,
1670,
892,
1048,
29871,
29896,
29900,
29941,
29953,
29946,
1301,
924,
29879,
3619,
304,
599,
23093,
313,
18667,
29889,
29871,
29947,
29907,
511,
3704,
2998,
298,
2890,
29907,
29320,
1316,
408,
7307,
29929,
29892,
360,
29940,
11490,
29941,
29892,
405,
2190,
29949,
29954,
29892,
438,
1783,
29946,
29892,
323,
20161,
322,
18351,
29896,
313,
1333,
4318,
467,
5057,
19869,
16127,
892,
8900,
297,
5734,
14125,
310,
315,
29924,
322,
13681,
29899,
15715,
6545,
4185,
1973,
313,
29934,
29906,
2622,
29922,
29900,
29889,
29929,
29906,
29947,
511,
408,
1532,
408,
297,
4688,
322,
5683,
13382,
9101,
313,
29934,
29906,
2622,
29922,
29900,
29889,
29929,
29945,
29929,
29897,
313,
18667,
29889,
29871,
29947,
29928,
467,
12433,
1279,
936,
16993,
3241,
7418,
28585,
393,
350,
29954,
29900,
29906,
9101,
19949,
297,
13681,
29899,
15715,
6545,
27831,
19932,
368,
322,
26060,
263,
3802,
29501,
304,
350,
29954,
29900,
29906,
322,
350,
29954,
29900,
29941,
9101,
19949,
297,
315,
29924,
313,
18667,
29889,
29871,
29947,
29923,
467,
4525,
848,
526,
13747,
411,
3517,
3483,
952,
267,
6445,
393,
563,
8349,
7268,
630,
298,
2890,
29907,
29879,
9867,
287,
19932,
368,
9401,
304,
7232,
719,
3398,
17873,
470,
18755,
307,
2204,
19416,
313,
29931,
5871,
634,
394,
29871,
29906,
29900,
29900,
29953,
29892,
350,
12513,
9481,
3457,
324,
29871,
29953,
29892,
29871,
29906,
29900,
467,
6549,
29892,
9101,
19949,
297,
278,
5541,
2187,
310,
278,
2198,
297,
7316,
526,
2221,
304,
7344,
1820,
29320,
310,
715,
29884,
381,
17765,
3819,
29889,
7579,
368,
29892,
278,
5541,
2187,
310,
278,
2198,
297,
7316,
508,
367,
1304,
408,
263,
2560,
18350,
363,
20382,
1583,
29899,
1267,
809,
284,
310,
17473,
519,
9101,
29889,
13,
6330,
841,
749,
310,
12968,
28735,
719,
8927,
624,
331,
315,
10071,
313,
29882,
2890,
29907,
29879,
29897,
373,
12968,
1891,
7338,
945,
514,
1070,
5345,
11669,
313,
11206,
29924,
29879,
29897,
297,
13681,
29899,
15715,
6545,
13,
29961,
29900,
29896,
29946,
29929,
29962,
1763,
23033,
278,
6297,
310,
1425,
29890,
29933,
29906,
7182,
292,
322,
2693,
263,
3342,
5745,
363,
298,
2890,
29907,
29879,
29892,
13681,
29899,
15715,
6545,
4185,
1973,
892,
12919,
17832,
373,
9257,
270,
3276,
1302,
630,
411,
14321,
7329,
29899,
9313,
1133,
341,
1299,
3960,
1692,
29931,
30342,
29871,
29896,
29901,
29941,
29900,
29892,
541,
1033,
884,
367,
19949,
8472,
1472,
29899,
8489,
373,
445,
25148,
403,
21749,
3860,
29871,
29896,
29901,
29906,
29900,
29900,
313,
18667,
29889,
29871,
29929,
29909,
511,
470,
29871,
29896,
29901,
29896,
29900,
29900,
29900,
29889,
350,
29954,
29900,
29906,
322,
8045,
29911,
29946,
29929,
298,
2890,
29907,
29879,
1033,
884,
367,
19949,
363,
1405,
29945,
1209,
1179,
373,
260,
15118,
9257,
270,
17006,
1302,
630,
411,
5199,
724,
398,
313,
18667,
29889,
29871,
29929,
29933,
416,
5199,
18755,
1617,
522,
262,
313,
18667,
29889,
29871,
29929,
29907,
416,
470,
478,
1806,
1672,
29954,
1672,
30342,
313,
18667,
29889,
29871,
29929,
29928,
511,
607,
338,
263,
24440,
653,
5199,
1891,
17522,
29924,
29889,
13,
15771,
19413,
6978,
6751,
310,
12968,
28735,
719,
8927,
624,
331,
315,
10071,
313,
29882,
2890,
29907,
29879,
29897,
13,
29961,
29900,
29896,
29945,
29900,
29962,
26905,
5925,
6471,
505,
28585,
393,
3058,
3367,
572,
397,
583,
29892,
451,
2197,
310,
298,
11626,
29896,
29906,
322,
29871,
29896,
29955,
29892,
526,
18414,
7964,
297,
298,
2890,
29907,
29879,
1090,
3058,
1014,
29899,
3670,
3039,
9257,
5855,
313,
29933,
5790,
634,
394,
1696,
29871,
29906,
29900,
29900,
29955,
19259,
29889,
3457,
866,
305,
29889,
29906,
29945,
29898,
29906,
1125,
29906,
29900,
29955,
29899,
29906,
29896,
29945,
467,
450,
10097,
310,
17487,
417,
333,
583,
2444,
304,
367,
1556,
4153,
4475,
304,
6460,
3038,
10503,
2561,
746,
4185,
1973,
526,
6219,
304,
2323,
9101,
472,
1209,
6751,
29892,
13138,
263,
2225,
21571,
4549,
1831,
573,
14321,
10631,
363,
9101,
4023,
4089,
292,
1438,
385,
29872,
786,
417,
333,
583,
29889,
1281,
874,
873,
29892,
298,
2890,
29907,
29879,
15678,
297,
697,
7232,
397,
2073,
310,
278,
2198,
297,
7316,
29892,
13681,
29899,
15715,
6545,
29892,
19949,
1880,
3516,
3097,
472,
715,
1218,
1156,
1641,
6219,
304,
2323,
9101,
313,
18667,
29889,
29871,
29896,
29900,
29909,
29899,
29928,
467,
350,
29954,
29900,
29896,
322,
350,
29954,
29900,
29906,
9101,
19949,
263,
4226,
413,
653,
327,
668,
313,
18667,
29889,
29871,
29896,
29900,
29923,
29897,
1156,
1641,
1209,
4063,
411,
319,
4174,
2692,
8127,
30342,
363,
1405,
29896,
29947,
322,
29871,
29896,
29929,
1209,
1179,
8307,
29889,
450,
25413,
310
] | 1,024 |
CommonCrawl
| -0.108481 | 1.495692 | 0.976528 | 1.083747 |
normal karyotype in cells demonstrated that disaggregation of hESC cultures to single cells did not inherently lead to the accumulation of these trisomics in hESCs maintained in DC-HAIF. BG01 and BG02 cultures were also passaged by disaggregation to single cells with multiple passaging agents (FIG. 11). Cultures were split with ACCUTASE®, 0.25% Trypsin/EDTA, TrypLE, or VERSENE® (EDTA) for 5 passages and karyotyped. The data demonstrate that culturing and passaging hESCs in the compositions of the present invention maintained a normal karyotype in at least two human embryonic cell lines, using a variety of cell disaggregation reagents.
[0151] Large-scale expansion of undifferentiated hESCs is also possible, using the compositions of the present invention. A starting confluent culture of BG02 cells in a 60 mm plate was expanded in DC-HAIF through 4 passages to generate >1.12×1010 cells in 20 days in a single experiment. The cultures remained undifferentiated, as demonstrated by >85% of the cells in the batch maintaining expression of markers of pluirpotency such as OCT4, CD9, SSEA-4, TRA-1-81 when examined by flow cytometry (FIG. 12A). Expression of other markers of pluripotency was also observed by RT-PCR analysis, while markers of differentiated lineages α-fetoprotein, MSX1 and HAND1 were not detected (FIG. 12B). Fluorescence in situ hybridization analysis demonstrated that the cells cultured and passaged in DC-HAIF maintained expected copy numbers for hChr12 (98% 2-copy), hChr17 (98% 2-copy), hChrX (95% 1-copy) and hChrY (98% 1-copy) (FIG. 12C). Karyotyping analysis also demonstrated that a normal euploid chromosome content and banding profile was maintained in these cells
Insulin and IGF1 Exert Different Effects on hESCs when Applied at Physiological Concentrations
[0152] Essentially all of the reported culture conditions for hESCs to date include supraphysiological levels of insulin, which can stimulate both IR and IGF1R. To distinguish the activities that insulin and insulin-substitutes exert, compared to IGF1, hESCs are cultured in defined media conditions in physiological levels of these growth factors. The concentrations of insulin and IGF1 are titrated from about 0.2 to about 200 ng/ml and cell proliferation is monitored by counting cells after 5 days. Cultures that expand successfully are serially passaged 5 times. Physiological levels of IGF1 support the expansion of hESC cultures, whereas physiological levels of insulin do not, indicating that the activity of insulin or insulin-substitutes cannot replace IGF1, and that IGF1 and insulin (or insulin substitutes) represent separate classes of biological activities with regard to action on hESCs.
Methods for Screening the Effects of Supplements
[0153] To initially examine the effects of Vitamin B12 and Vitamin B6 on the growth or differentiation hESCs growing at an intermediate density, BG02 cells are split using ACCUTASE® and 1×105 cells/well are plated in 6-well trays in defined culture (DC) media. The DC media contains 10 ng/ml HRG-β, 200 ng/ml LongR3-IGF1, and 10 ng/ml FGF10. Vitamin B6 (0.5 μM) and/or Vitamin B12 (0.5 μM) are added to experimental wells. Cell numbers in each condition are counted after 7 days. Cell counting and colony counting of both experimental and control wells will provide insight on the effects of Vitamin B6 and Vitamin B12 on cell growth.
[0154] In addition, markers of differentiation
|
[
4226,
413,
653,
327,
668,
297,
9101,
28585,
393,
22941,
7642,
362,
310,
298,
2890,
29907,
4185,
1973,
304,
2323,
9101,
1258,
451,
7846,
2705,
3275,
304,
278,
18414,
2785,
310,
1438,
534,
275,
290,
1199,
297,
298,
2890,
29907,
29879,
19949,
297,
13681,
29899,
15715,
6545,
29889,
350,
29954,
29900,
29896,
322,
350,
29954,
29900,
29906,
4185,
1973,
892,
884,
1209,
4063,
491,
22941,
7642,
362,
304,
2323,
9101,
411,
2999,
1209,
6751,
19518,
313,
18667,
29889,
29871,
29896,
29896,
467,
7965,
1973,
892,
6219,
411,
319,
4174,
2692,
8127,
30342,
29892,
29871,
29900,
29889,
29906,
29945,
29995,
3967,
567,
262,
29914,
3352,
6040,
29892,
3967,
29886,
1307,
29892,
470,
478,
23598,
1430,
29923,
30342,
313,
3352,
6040,
29897,
363,
29871,
29945,
1209,
1179,
322,
413,
653,
327,
1478,
287,
29889,
450,
848,
22222,
393,
4185,
3864,
322,
1209,
6751,
298,
2890,
29907,
29879,
297,
278,
5541,
2187,
310,
278,
2198,
297,
7316,
19949,
263,
4226,
413,
653,
327,
668,
297,
472,
3203,
1023,
5199,
7232,
719,
8927,
3038,
3454,
29892,
773,
263,
12875,
310,
3038,
22941,
7642,
362,
337,
351,
1237,
29889,
13,
29961,
29900,
29896,
29945,
29896,
29962,
8218,
479,
29899,
7052,
13184,
310,
563,
8349,
7268,
630,
298,
2890,
29907,
29879,
338,
884,
1950,
29892,
773,
278,
5541,
2187,
310,
278,
2198,
297,
7316,
29889,
319,
6257,
18669,
8122,
9257,
310,
350,
29954,
29900,
29906,
9101,
297,
263,
29871,
29953,
29900,
5654,
15284,
471,
17832,
297,
13681,
29899,
15715,
6545,
1549,
29871,
29946,
1209,
1179,
304,
5706,
1405,
29896,
29889,
29896,
29906,
30122,
29896,
29900,
29896,
29900,
9101,
297,
29871,
29906,
29900,
3841,
297,
263,
2323,
7639,
29889,
450,
4185,
1973,
9488,
563,
8349,
7268,
630,
29892,
408,
28585,
491,
1405,
29947,
29945,
29995,
310,
278,
9101,
297,
278,
9853,
7344,
292,
4603,
310,
29320,
310,
715,
29884,
381,
17765,
3819,
1316,
408,
438,
1783,
29946,
29892,
7307,
29929,
29892,
317,
1660,
29909,
29899,
29946,
29892,
323,
4717,
29899,
29896,
29899,
29947,
29896,
746,
4392,
1312,
491,
4972,
274,
3637,
7843,
313,
18667,
29889,
29871,
29896,
29906,
29909,
467,
21444,
310,
916,
29320,
310,
715,
332,
666,
327,
3819,
471,
884,
8900,
491,
390,
29911,
29899,
9026,
29934,
7418,
29892,
1550,
29320,
310,
17473,
630,
1196,
1179,
19883,
29899,
29888,
300,
459,
4859,
262,
29892,
10888,
29990,
29896,
322,
379,
9468,
29896,
892,
451,
17809,
313,
18667,
29889,
29871,
29896,
29906,
29933,
467,
2379,
29884,
2361,
29883,
663,
297,
2990,
7498,
19515,
2133,
7418,
28585,
393,
278,
9101,
4185,
2955,
322,
1209,
4063,
297,
13681,
29899,
15715,
6545,
19949,
3806,
3509,
3694,
363,
298,
11626,
29896,
29906,
313,
29929,
29947,
29995,
29871,
29906,
29899,
8552,
511,
298,
11626,
29896,
29955,
313,
29929,
29947,
29995,
29871,
29906,
29899,
8552,
511,
298,
11626,
29990,
313,
29929,
29945,
29995,
29871,
29896,
29899,
8552,
29897,
322,
298,
11626,
29979,
313,
29929,
29947,
29995,
29871,
29896,
29899,
8552,
29897,
313,
18667,
29889,
29871,
29896,
29906,
29907,
467,
476,
653,
327,
1478,
292,
7418,
884,
28585,
393,
263,
4226,
321,
786,
417,
333,
25173,
359,
608,
2793,
322,
3719,
292,
8722,
471,
19949,
297,
1438,
9101,
13,
797,
29879,
352,
262,
322,
306,
29954,
29943,
29896,
1222,
814,
360,
15622,
26475,
29879,
373,
298,
2890,
29907,
29879,
746,
2401,
2957,
472,
11661,
29875,
5996,
1281,
1760,
29878,
800,
13,
29961,
29900,
29896,
29945,
29906,
29962,
11044,
9247,
599,
310,
278,
8967,
9257,
5855,
363,
298,
2890,
29907,
29879,
304,
2635,
3160,
13159,
1140,
952,
29875,
5996,
11174,
310,
1663,
352,
262,
29892,
607,
508,
20436,
5987,
1716,
23292,
322,
306,
29954,
29943,
29896,
29934,
29889,
1763,
20820,
278,
14188,
393,
1663,
352,
262,
322,
1663,
352,
262,
29899,
22492,
277,
2667,
429,
814,
29892,
9401,
304,
306,
29954,
29943,
29896,
29892,
298,
2890,
29907,
29879,
526,
4185,
2955,
297,
3342,
5745,
5855,
297,
4824,
29875,
5996,
11174,
310,
1438,
14321,
13879,
29889,
450,
14953,
800,
310,
1663,
352,
262,
322,
306,
29954,
29943,
29896,
526,
4329,
29878,
630,
515,
1048,
29871,
29900,
29889,
29906,
304,
1048,
29871,
29906,
29900,
29900,
8736,
29914,
828,
322,
3038,
410,
29880,
9633,
362,
338,
11819,
287,
491,
21248,
9101,
1156,
29871,
29945,
3841,
29889,
7965,
1973,
393,
7985,
8472,
526,
7797,
368,
1209,
4063,
29871,
29945,
3064,
29889,
11661,
29875,
5996,
11174,
310,
306,
29954,
29943,
29896,
2304,
278,
13184,
310,
298,
2890,
29907,
4185,
1973,
29892,
13452,
4824,
29875,
5996,
11174,
310,
1663,
352,
262,
437,
451,
29892,
23941,
393,
278,
6354,
310,
1663,
352,
262,
470,
1663,
352,
262,
29899,
22492,
277,
2667,
2609,
5191,
306,
29954,
29943,
29896,
29892,
322,
393,
306,
29954,
29943,
29896,
322,
1663,
352,
262,
313,
272,
1663,
352,
262,
5960,
277,
2667,
29897,
2755,
5004,
4413,
310,
4768,
5996,
14188,
411,
4880,
304,
3158,
373,
298,
2890,
29907,
29879,
29889,
13,
26112,
363,
22666,
292,
278,
26475,
29879,
310,
9179,
944,
29879,
13,
29961,
29900,
29896,
29945,
29941,
29962,
1763,
12919,
25917,
278,
9545,
310,
22491,
9103,
350,
29896,
29906,
322,
22491,
9103,
350,
29953,
373,
278,
14321,
470,
1422,
11685,
298,
2890,
29907,
29879,
15678,
472,
385,
19697,
9027,
29892,
350,
29954,
29900,
29906,
9101,
526,
6219,
773,
319,
4174,
2692,
8127,
30342,
322,
29871,
29896,
30122,
29896,
29900,
29945,
9101,
29914,
5872,
526,
715,
630,
297,
29871,
29953,
29899,
5872,
260,
764,
29879,
297,
3342,
9257,
313,
12696,
29897,
5745,
29889,
450,
13681,
5745,
3743,
29871,
29896,
29900,
8736,
29914,
828,
379,
29934,
29954,
29899,
30187,
29892,
29871,
29906,
29900,
29900,
8736,
29914,
828,
6242,
29934,
29941,
29899,
6259,
29943,
29896,
29892,
322,
29871,
29896,
29900,
8736,
29914,
828,
383,
29954,
29943,
29896,
29900,
29889,
22491,
9103,
350,
29953,
313,
29900,
29889,
29945,
29871,
30167,
29924,
29897,
322,
29914,
272,
22491,
9103,
350,
29896,
29906,
313,
29900,
29889,
29945,
29871,
30167,
29924,
29897,
526,
2715,
304,
17986,
1532,
29879,
29889,
19413,
3694,
297,
1269,
4195,
526,
29115,
1156,
29871,
29955,
3841,
29889,
19413,
21248,
322,
784,
2592,
21248,
310,
1716,
17986,
322,
2761,
1532,
29879,
674,
3867,
25483,
373,
278,
9545,
310,
22491,
9103,
350,
29953,
322,
22491,
9103,
350,
29896,
29906,
373,
3038,
14321,
29889,
13,
29961,
29900,
29896,
29945,
29946,
29962,
512,
6124,
29892,
29320,
310,
1422,
11685
] | 1,024 |
CommonCrawl
| -0.279783 | 1.329575 | 0.797205 | 1.286127 |
, such as OCT4 can be assayed in the experimental well to determine the effects of the additives and supplements to the differentiation state of the differentiable cells.
Culturing of hESCs in the Absence of FGF2
[0155] BG02 cells were maintained long term in DC-HAI, for 20 passages (FIG. 13A), and BG01 cells were also serially passaged in DC-HAI, both in the absence of FGF2. The cultures did not deteriorate or exhibit overt differentiation, and exhibited normal expansion of undifferentiated colonies throughout the culture period. The maintenance of a normal male karyotype in a BG02 culture was demonstrated after 6 passages in DC-HAI (FIG. 13B, 20/20 normal metaphase spreads).
[0156] Transcriptional analyses were used to compare global expression in hESCs cells maintained in DC-HAIF and DC-HAI. Total cellular RNA was isolated from hESCs using Trizol (Invitrogen) and was treated with DNase I (Invitrogen) according to the manufacturer's suggested protocol. Sample amplification was performed with 100 ng of total RNA using the Illumina RNA Amplification kit and labeling was achieved by incorporation of biotin-16-UTP (Perkin Elmer Life and Analytical Sciences) at a ratio of 1:1 with unlabeled UTP. Labeled, amplified material (700 ng per array) was hybridized to Illumina Sentrix Human-6 Expression Beadchips containing 47,296 transcript probes according to the manufacturer's instructions (Illumina, Inc.). Arrays were scanned with an Illumina Bead Array Reader confocal scanner and primary data processing, background subtraction, and data analysis were performed using Illumina BeadStudio software according to the manufacturer's instructions. A minimum detection confidence score of 0.99 (a computed cutoff indicating the target sequence signal was distinguishable from the negative controls) was used to discriminate the presence or absence of transcript expression. Data analysis was performed using parallel approaches described for other hESC samples (Liu et al 2006, BMC Dev Biol 6:20). Hierarchical clustering was performed as described previously (Liu et al 2005, BMC Dev Biol 6:20), and was based on average linkage and Euclidean distances as the similarity metric using differentially expressed genes identified by ANOVA (p<0.05). Detailed descriptions of the sensitivity and quality control tests used in array manufacture and algorithms used in the Bead studio software are available from Illumina, Inc (San Diego, Calif.). The majority of transcripts detected were expressed in both DC-HAIF and DC-HAI BG02 cultures, including known hESC markers such as CD9, DNMT3, NANOG, OCT4, TERT and UTF1 (not shown). High correlation coefficients were observed in comparisons of DC-HAIF and DC-HAI cultures (R2 select=0.961) (FIG. 14). Hierarchical clustering analysis demonstrated that BG02 cells maintained in DC-HAI grouped tightly and retained a close similarity to cells maintained in DC-HAIF, as well as BG02 and other hESC lines in multiple culture formats (FIG. 15). These data are consistent with previous analyses showing that undifferentiated hESCs clustered tightly compared to embryoid bodies or fibroblasts (Liu et al 2006, BMC Dev Biol 6:20).
[0157] Furthermore, BG02 cells maintained in DC-HAI differentiated to representatives of mesoderm, endoderm and ectoderm in complex teratomas formed in SCID-beige mice (not shown), formally demonstrating the maintenance of pluripotency in cultures grown in the absence of exogenous FGF2.
[0158] To examine if exogenous FGF2 was required in the context of single cell passaging, BG01 cells were passaged with ACCUTASE® and grown in defined conditions containing only 10 ng/ml HRG-β and 200 ng/ml Long
|
[
29892,
1316,
408,
438,
1783,
29946,
508,
367,
1223,
388,
287,
297,
278,
17986,
1532,
304,
8161,
278,
9545,
310,
278,
788,
277,
3145,
322,
1462,
944,
29879,
304,
278,
1422,
11685,
2106,
310,
278,
17473,
519,
9101,
29889,
13,
29907,
499,
3864,
310,
298,
2890,
29907,
29879,
297,
278,
24650,
663,
310,
383,
29954,
29943,
29906,
13,
29961,
29900,
29896,
29945,
29945,
29962,
350,
29954,
29900,
29906,
9101,
892,
19949,
1472,
1840,
297,
13681,
29899,
15715,
29902,
29892,
363,
29871,
29906,
29900,
1209,
1179,
313,
18667,
29889,
29871,
29896,
29941,
29909,
511,
322,
350,
29954,
29900,
29896,
9101,
892,
884,
7797,
368,
1209,
4063,
297,
13681,
29899,
15715,
29902,
29892,
1716,
297,
278,
18070,
310,
383,
29954,
29943,
29906,
29889,
450,
4185,
1973,
1258,
451,
270,
1308,
1611,
403,
470,
10371,
277,
975,
29873,
1422,
11685,
29892,
322,
10371,
1573,
4226,
13184,
310,
563,
8349,
7268,
630,
8104,
583,
10106,
278,
9257,
3785,
29889,
450,
25413,
310,
263,
4226,
14263,
413,
653,
327,
668,
297,
263,
350,
29954,
29900,
29906,
9257,
471,
28585,
1156,
29871,
29953,
1209,
1179,
297,
13681,
29899,
15715,
29902,
313,
18667,
29889,
29871,
29896,
29941,
29933,
29892,
29871,
29906,
29900,
29914,
29906,
29900,
4226,
1539,
12451,
559,
9677,
29879,
467,
13,
29961,
29900,
29896,
29945,
29953,
29962,
4103,
3395,
284,
3483,
952,
267,
892,
1304,
304,
7252,
5534,
4603,
297,
298,
2890,
29907,
29879,
9101,
19949,
297,
13681,
29899,
15715,
6545,
322,
13681,
29899,
15715,
29902,
29889,
14990,
3038,
1070,
390,
3521,
471,
23968,
515,
298,
2890,
29907,
29879,
773,
323,
7485,
324,
313,
12165,
277,
307,
1885,
29897,
322,
471,
14914,
411,
360,
29940,
559,
306,
313,
12165,
277,
307,
1885,
29897,
5034,
304,
278,
12012,
9945,
29915,
29879,
7829,
9608,
29889,
21029,
20563,
2450,
471,
8560,
411,
29871,
29896,
29900,
29900,
8736,
310,
3001,
390,
3521,
773,
278,
8408,
398,
1099,
390,
3521,
1913,
572,
2450,
413,
277,
322,
3858,
292,
471,
14363,
491,
11039,
362,
310,
4768,
327,
262,
29899,
29896,
29953,
29899,
2692,
29925,
313,
5894,
9089,
1260,
1050,
4634,
322,
11597,
3637,
936,
17253,
29897,
472,
263,
11959,
310,
29871,
29896,
29901,
29896,
411,
443,
29880,
24025,
501,
3557,
29889,
365,
24025,
29892,
20563,
2164,
5518,
313,
29955,
29900,
29900,
8736,
639,
1409,
29897,
471,
7498,
19515,
1891,
304,
8408,
398,
1099,
28048,
2126,
12968,
29899,
29953,
21444,
350,
1479,
4161,
567,
6943,
29871,
29946,
29955,
29892,
29906,
29929,
29953,
1301,
924,
2070,
267,
5034,
304,
278,
12012,
9945,
29915,
29879,
11994,
313,
10002,
398,
1099,
29892,
9266,
6250,
4398,
29879,
892,
885,
11310,
411,
385,
8408,
398,
1099,
350,
1479,
4398,
830,
1664,
1970,
18642,
885,
7310,
322,
7601,
848,
9068,
29892,
3239,
1014,
3018,
428,
29892,
322,
848,
7418,
892,
8560,
773,
8408,
398,
1099,
350,
1479,
28867,
7047,
5034,
304,
278,
12012,
9945,
29915,
29879,
11994,
29889,
319,
9212,
15326,
16420,
8158,
310,
29871,
29900,
29889,
29929,
29929,
313,
29874,
15712,
5700,
2696,
23941,
278,
3646,
5665,
7182,
471,
20820,
519,
515,
278,
8178,
11761,
29897,
471,
1304,
304,
2313,
5632,
16976,
278,
10122,
470,
18070,
310,
1301,
924,
4603,
29889,
3630,
7418,
471,
8560,
773,
8943,
13501,
5439,
363,
916,
298,
2890,
29907,
11916,
313,
29931,
5871,
634,
394,
29871,
29906,
29900,
29900,
29953,
29892,
350,
12513,
9481,
3457,
324,
29871,
29953,
29901,
29906,
29900,
467,
12433,
1279,
936,
16993,
3241,
471,
8560,
408,
5439,
9251,
313,
29931,
5871,
634,
394,
29871,
29906,
29900,
29900,
29945,
29892,
350,
12513,
9481,
3457,
324,
29871,
29953,
29901,
29906,
29900,
511,
322,
471,
2729,
373,
6588,
1544,
482,
322,
382,
27511,
24610,
408,
278,
29501,
12714,
773,
16712,
368,
13384,
2531,
267,
15659,
491,
319,
6632,
20449,
313,
29886,
29966,
29900,
29889,
29900,
29945,
467,
360,
11881,
2342,
1980,
310,
278,
4771,
24858,
322,
11029,
2761,
6987,
1304,
297,
1409,
12012,
545,
322,
14009,
1304,
297,
278,
350,
1479,
8693,
7047,
526,
3625,
515,
8408,
398,
1099,
29892,
9266,
313,
22509,
16879,
29892,
3037,
361,
6250,
450,
13638,
310,
1301,
924,
29879,
17809,
892,
13384,
297,
1716,
13681,
29899,
15715,
6545,
322,
13681,
29899,
15715,
29902,
350,
29954,
29900,
29906,
4185,
1973,
29892,
3704,
2998,
298,
2890,
29907,
29320,
1316,
408,
7307,
29929,
29892,
360,
29940,
11490,
29941,
29892,
405,
2190,
29949,
29954,
29892,
438,
1783,
29946,
29892,
323,
20161,
322,
18351,
29896,
313,
1333,
4318,
467,
5057,
19869,
16127,
892,
8900,
297,
5734,
14125,
310,
13681,
29899,
15715,
6545,
322,
13681,
29899,
15715,
29902,
4185,
1973,
313,
29934,
29906,
1831,
29922,
29900,
29889,
29929,
29953,
29896,
29897,
313,
18667,
29889,
29871,
29896,
29946,
467,
12433,
1279,
936,
16993,
3241,
7418,
28585,
393,
350,
29954,
29900,
29906,
9101,
19949,
297,
13681,
29899,
15715,
29902,
27831,
19932,
368,
322,
26060,
263,
3802,
29501,
304,
9101,
19949,
297,
13681,
29899,
15715,
6545,
29892,
408,
1532,
408,
350,
29954,
29900,
29906,
322,
916,
298,
2890,
29907,
3454,
297,
2999,
9257,
21971,
313,
18667,
29889,
29871,
29896,
29945,
467,
4525,
848,
526,
13747,
411,
3517,
3483,
952,
267,
6445,
393,
563,
8349,
7268,
630,
298,
2890,
29907,
29879,
9867,
287,
19932,
368,
9401,
304,
7232,
719,
3398,
17873,
470,
18755,
307,
2204,
19416,
313,
29931,
5871,
634,
394,
29871,
29906,
29900,
29900,
29953,
29892,
350,
12513,
9481,
3457,
324,
29871,
29953,
29901,
29906,
29900,
467,
13,
29961,
29900,
29896,
29945,
29955,
29962,
16478,
29892,
350,
29954,
29900,
29906,
9101,
19949,
297,
13681,
29899,
15715,
29902,
17473,
630,
304,
2755,
5056,
310,
4883,
397,
837,
29892,
1095,
397,
837,
322,
321,
312,
397,
837,
297,
4280,
1935,
8678,
294,
8429,
297,
12314,
1367,
29899,
915,
2231,
286,
625,
313,
1333,
4318,
511,
28269,
9004,
1218,
278,
25413,
310,
715,
332,
666,
327,
3819,
297,
4185,
1973,
21633,
297,
278,
18070,
310,
429,
6352,
681,
383,
29954,
29943,
29906,
29889,
13,
29961,
29900,
29896,
29945,
29947,
29962,
1763,
25917,
565,
429,
6352,
681,
383,
29954,
29943,
29906,
471,
3734,
297,
278,
3030,
310,
2323,
3038,
1209,
6751,
29892,
350,
29954,
29900,
29896,
9101,
892,
1209,
4063,
411,
319,
4174,
2692,
8127,
30342,
322,
21633,
297,
3342,
5855,
6943,
871,
29871,
29896,
29900,
8736,
29914,
828,
379,
29934,
29954,
29899,
30187,
322,
29871,
29906,
29900,
29900,
8736,
29914,
828,
6242
] | 1,024 |
CommonCrawl
| -0.3382 | 1.495692 | 0.637609 | 0.992676 |
R3-IGF1 (DC-HI). These DC-HI cultures were maintained for 10 passages, and did not exhibit overt differentiation or a slowing of proliferation.
[0159] These studies clearly demonstrated that the provision of exogenous FGF2 is not required when hESCs are maintained in defined media minimally containing heregulin and IGF1. Furthermore cultures absent FGF2 retained key properties of pluripotency, including transcriptional profile and differentiation to mesoderm, endoderm and ectoderm in vivo.
Suspension Cultures
[0160] Starting cultures of BG02 cells were maintained in DC-HAIF medium on dishes coated with 1:200 matrigel, as described herein and were split by passaging with ACCUTASE®. To initiate suspension culture, BG02 cells were disaggregated with ACCUTASE® and placed in low attachment 6-well trays at a density of 1.6, 3, or 6×105 cells/ml (0.5, 1, or 2×106 cells in 3 ml volumes) in DC-HAIF medium. The trays were placed on a rotating platform at 80-100 rpm in a humidified incubator with 5% CO2. Under these conditions hESCs coalesced into small spheres of morphologically viable cells within 24 hours.
[0161] The medium in the wells was changed on the second day, and every day thereafter. Suspension aggregates continued to proliferate, growing larger over time without obvious signs of differentiation (FIG. 17). Some of the spheres continued to aggregate over the course of the culture, as some aggregates became much larger than the majority. In addition, non-spherical aggregates could be observed in the process of merging during the first few days of the culture. To limit this continued aggregation, 38 μg/ml DNascI was included in some suspension cultures for the first 24 hours. This approach appeared to conducive to the initial aggregation, with relatively larger, but fewer, aggregates formed in the presence of DNaseI. It is not clear, however, if the DNaseI treatment reduced the subsequent merging of spheres and exposure to DNaseI consistently made these aggregates harder to break up when splitting.
[0162] Suspension cultures were disaggregated with ACCUTASE® approximately every 7 days and new spheres were established. While the densities varied in different experiments, spheres established within this range of densities (1.6-6×105 cells/ml) could be maintained in culture for more than 12 passages, or >80 days, without morphological signs of differentiation. FISH analyses of serially passaged suspension hESCs were also performed to assess the chromosome number for common aneuplodies. BG02 cells that had been grown in suspension for 6 passages exhibited normal counts for hChr 12 (96% two copy, n=788), hChr 17 (97% two copy, n=587), hChr X (97% one copy, n=724) and hChr Y (98% one copy, n=689).
Expansion of Differentiable Cells in Suspension Culture
[0163] Unlike embryoid body culture in the presence of serum or inducers of differentiation, suspension aggregates of hESCs in DC-HAIF did not appear to differentiate. Obvious visceral endoderm was not observed, neither was the formation of structures resembling proamniotic cavities, both classic signs of embryoid body differentiation. To examine the lack of differentiation more closely, cultures were plated back into adherent conditions on MATRIGEL® diluted 1:200 and cultured in DC-HAIF. These cultures were also primarily undifferentiated, and did not exhibit obvious morphological signs of increased differentiation such as the presence of larger, flattened cells, or structured regions.
[0164] Cell counting was used to assess the relative growth rates of cells in suspension compared to adherent culture. In this experiment, an adherent culture of BG02 cells was passaged with
|
[
29934,
29941,
29899,
6259,
29943,
29896,
313,
12696,
29899,
17628,
467,
4525,
13681,
29899,
17628,
4185,
1973,
892,
19949,
363,
29871,
29896,
29900,
1209,
1179,
29892,
322,
1258,
451,
10371,
277,
975,
29873,
1422,
11685,
470,
263,
5232,
292,
310,
410,
29880,
9633,
362,
29889,
13,
29961,
29900,
29896,
29945,
29929,
29962,
4525,
11898,
9436,
28585,
393,
278,
25161,
310,
429,
6352,
681,
383,
29954,
29943,
29906,
338,
451,
3734,
746,
298,
2890,
29907,
29879,
526,
19949,
297,
3342,
5745,
6260,
635,
6943,
902,
387,
352,
262,
322,
306,
29954,
29943,
29896,
29889,
16478,
4185,
1973,
29207,
383,
29954,
29943,
29906,
26060,
1820,
4426,
310,
715,
332,
666,
327,
3819,
29892,
3704,
1301,
3395,
284,
8722,
322,
1422,
11685,
304,
4883,
397,
837,
29892,
1095,
397,
837,
322,
321,
312,
397,
837,
297,
325,
4243,
29889,
13,
29903,
375,
29886,
2673,
7965,
1973,
13,
29961,
29900,
29896,
29953,
29900,
29962,
23748,
4185,
1973,
310,
350,
29954,
29900,
29906,
9101,
892,
19949,
297,
13681,
29899,
15715,
6545,
18350,
373,
270,
17006,
1302,
630,
411,
29871,
29896,
29901,
29906,
29900,
29900,
1775,
8966,
295,
29892,
408,
5439,
1244,
262,
322,
892,
6219,
491,
1209,
6751,
411,
319,
4174,
2692,
8127,
30342,
29889,
1763,
14511,
403,
8872,
2673,
9257,
29892,
350,
29954,
29900,
29906,
9101,
892,
22941,
7642,
630,
411,
319,
4174,
2692,
8127,
30342,
322,
7180,
297,
4482,
26305,
29871,
29953,
29899,
5872,
260,
764,
29879,
472,
263,
9027,
310,
29871,
29896,
29889,
29953,
29892,
29871,
29941,
29892,
470,
29871,
29953,
30122,
29896,
29900,
29945,
9101,
29914,
828,
313,
29900,
29889,
29945,
29892,
29871,
29896,
29892,
470,
29871,
29906,
30122,
29896,
29900,
29953,
9101,
297,
29871,
29941,
286,
29880,
18167,
29897,
297,
13681,
29899,
15715,
6545,
18350,
29889,
450,
260,
764,
29879,
892,
7180,
373,
263,
5731,
1218,
7481,
472,
29871,
29947,
29900,
29899,
29896,
29900,
29900,
364,
3358,
297,
263,
3165,
333,
2164,
5528,
431,
1061,
411,
29871,
29945,
29995,
4810,
29906,
29889,
7634,
1438,
5855,
298,
2890,
29907,
29879,
1302,
2122,
1133,
964,
2319,
26903,
267,
310,
18131,
1189,
1711,
3516,
519,
9101,
2629,
29871,
29906,
29946,
6199,
29889,
13,
29961,
29900,
29896,
29953,
29896,
29962,
450,
18350,
297,
278,
1532,
29879,
471,
3939,
373,
278,
1473,
2462,
29892,
322,
1432,
2462,
727,
7045,
29889,
9511,
29886,
2673,
11404,
1078,
7572,
304,
410,
29880,
9633,
403,
29892,
15678,
7200,
975,
931,
1728,
6924,
18906,
310,
1422,
11685,
313,
18667,
29889,
29871,
29896,
29955,
467,
3834,
310,
278,
26903,
267,
7572,
304,
20431,
975,
278,
3236,
310,
278,
9257,
29892,
408,
777,
11404,
1078,
3897,
1568,
7200,
1135,
278,
13638,
29889,
512,
6124,
29892,
1661,
29899,
29879,
8096,
936,
11404,
1078,
1033,
367,
8900,
297,
278,
1889,
310,
2778,
3460,
2645,
278,
937,
2846,
3841,
310,
278,
9257,
29889,
1763,
4046,
445,
7572,
11404,
362,
29892,
29871,
29941,
29947,
29871,
30167,
29887,
29914,
828,
360,
29940,
6151,
29902,
471,
5134,
297,
777,
8872,
2673,
4185,
1973,
363,
278,
937,
29871,
29906,
29946,
6199,
29889,
910,
2948,
7470,
304,
13417,
455,
345,
304,
278,
2847,
11404,
362,
29892,
411,
13774,
7200,
29892,
541,
28145,
29892,
11404,
1078,
8429,
297,
278,
10122,
310,
360,
29940,
559,
29902,
29889,
739,
338,
451,
2821,
29892,
3138,
29892,
565,
278,
360,
29940,
559,
29902,
14502,
12212,
278,
15352,
2778,
3460,
310,
26903,
267,
322,
14060,
545,
304,
360,
29940,
559,
29902,
5718,
2705,
1754,
1438,
11404,
1078,
22622,
304,
2867,
701,
746,
24368,
29889,
13,
29961,
29900,
29896,
29953,
29906,
29962,
9511,
29886,
2673,
4185,
1973,
892,
22941,
7642,
630,
411,
319,
4174,
2692,
8127,
30342,
14235,
1432,
29871,
29955,
3841,
322,
716,
26903,
267,
892,
7841,
29889,
5806,
278,
6245,
1907,
23821,
297,
1422,
15729,
29892,
26903,
267,
7841,
2629,
445,
3464,
310,
6245,
1907,
313,
29896,
29889,
29953,
29899,
29953,
30122,
29896,
29900,
29945,
9101,
29914,
828,
29897,
1033,
367,
19949,
297,
9257,
363,
901,
1135,
29871,
29896,
29906,
1209,
1179,
29892,
470,
1405,
29947,
29900,
3841,
29892,
1728,
18131,
5996,
18906,
310,
1422,
11685,
29889,
383,
3235,
29950,
3483,
952,
267,
310,
7797,
368,
1209,
4063,
8872,
2673,
298,
2890,
29907,
29879,
892,
884,
8560,
304,
24809,
278,
25173,
359,
608,
1353,
363,
3619,
385,
12932,
572,
397,
583,
29889,
350,
29954,
29900,
29906,
9101,
393,
750,
1063,
21633,
297,
8872,
2673,
363,
29871,
29953,
1209,
1179,
10371,
1573,
4226,
18139,
363,
298,
11626,
29871,
29896,
29906,
313,
29929,
29953,
29995,
1023,
3509,
29892,
302,
29922,
29955,
29947,
29947,
511,
298,
11626,
29871,
29896,
29955,
313,
29929,
29955,
29995,
1023,
3509,
29892,
302,
29922,
29945,
29947,
29955,
511,
298,
11626,
1060,
313,
29929,
29955,
29995,
697,
3509,
29892,
302,
29922,
29955,
29906,
29946,
29897,
322,
298,
11626,
612,
313,
29929,
29947,
29995,
697,
3509,
29892,
302,
29922,
29953,
29947,
29929,
467,
13,
9544,
9454,
310,
360,
8349,
7268,
519,
315,
10071,
297,
9511,
29886,
2673,
14062,
13,
29961,
29900,
29896,
29953,
29941,
29962,
853,
4561,
7232,
719,
3398,
3573,
9257,
297,
278,
10122,
310,
724,
398,
470,
9013,
22543,
310,
1422,
11685,
29892,
8872,
2673,
11404,
1078,
310,
298,
2890,
29907,
29879,
297,
13681,
29899,
15715,
6545,
1258,
451,
2615,
304,
17473,
403,
29889,
4250,
2366,
1998,
2265,
284,
1095,
397,
837,
471,
451,
8900,
29892,
9561,
471,
278,
12409,
310,
12286,
620,
1590,
1847,
410,
314,
1240,
13574,
18346,
1907,
29892,
1716,
22037,
18906,
310,
7232,
719,
3398,
3573,
1422,
11685,
29889,
1763,
25917,
278,
10225,
310,
1422,
11685,
901,
16467,
29892,
4185,
1973,
892,
715,
630,
1250,
964,
594,
2276,
296,
5855,
373,
341,
1299,
3960,
1692,
29931,
30342,
21749,
3860,
29871,
29896,
29901,
29906,
29900,
29900,
322,
4185,
2955,
297,
13681,
29899,
15715,
6545,
29889,
4525,
4185,
1973,
892,
884,
19434,
563,
8349,
7268,
630,
29892,
322,
1258,
451,
10371,
277,
6924,
18131,
5996,
18906,
310,
11664,
1422,
11685,
1316,
408,
278,
10122,
310,
7200,
29892,
1652,
8606,
287,
9101,
29892,
470,
2281,
2955,
12786,
29889,
13,
29961,
29900,
29896,
29953,
29946,
29962,
19413,
21248,
471,
1304,
304,
24809,
278,
6198,
14321,
19257,
310,
9101,
297,
8872,
2673,
9401,
304,
594,
2276,
296,
9257,
29889,
512,
445,
7639,
29892,
385,
594,
2276,
296,
9257,
310,
350,
29954,
29900,
29906,
9101,
471,
1209,
4063,
411
] | 1,024 |
CommonCrawl
| -0.376239 | 1.199797 | 0.492358 | 0.911723 |
ACCUTASE®, and about 1×106 cells were placed in parallel suspension or adherent culture wells. Individual wells were counted on days 1-6 and plotted on a log scale (FIG. 18). While a higher initial proportion of hESCs were viable after 24 hours in adherent culture (˜90% vs ˜14%), growth rates were comparable thereafter. This indicated that hESCs could proliferate just as rapidly in suspension culture as in traditional adherent culture. Cell counts performed during passaging allows one to gauge the amount of expansion possible in this simple suspension system. In several cultures seeded with 5×105 cells, approximately 107 cells, or more, were generated after 7 days. The expansion after 7 days in suspension culture equated to about a 20-fold or more expansion, with the largest expansion observed being ˜24× the input cell number.
Characteristics of Differentiable Cells Expanded in Suspension Culture
[0165] Quantitative RT-PCR (qPCR) was used to compare gene expression in hESCs grown in suspension and adherent culture in DC-HAIF. Comparable levels of OCT4, a marker of pluripotent cells, were observed in both culture formats, confirming that cultures maintained in suspension were primarily undifferentiated. SOX17, a marker of definitive endoderm, was not expressed in either population of hESCs. The qPCR analysis also examined the potential of suspension hESCs to differentiate to definitive endoderm, as aggregates in suspension. Adherent and suspension hESCs were differentiated using parallel conditions. hESC cultures were treated with RPMI containing 2% BSA, 100 ng/ml Activin A, 8 ng/ml FGF2 and 25 ng/ml Wnt3A for 24 hours, followed by 2 days in the same medium without Wnt3A. The expression of OCT4 was downregulated, and expression of SOX17 upregulated similarly in both definitive endoderm samples compared to undifferentiated hESCs. This differentiation analysis confirmed that hESCs cultured in suspension in DC-HAIF maintained their differentiation potential, as evidenced by the likely formation of definitive endoderm.
Addition of an Apoptosis Inhibitor in Suspension Culture
[0166] To attenuate the loss of cells after initial passaging in suspension, an inhibitor of apoptosis was added to the medium. Cells were passaged as in Example 12, except that Y-27632, an inhibitor of p160-Rho-associated coiled-coil kinase (ROCK), was added to the medium.
[0167] Suspension aggregates of BG02 cells were formed by seeding 2×106 single cells in 6-well dishes in 3 ml DC-HAIF medium, at 100 rpm on a rotating platform in an incubator (Table 2, Experiment A). 10 μM Y27632 ROCK inhibitor was added to test wells for the course of the experiment and the cultures observed daily and counted after 24 hours (day 1) and after 4 or 5 days. As shown in FIG. 20, addition of Y27632 had a profound effect on the initial aggregation phase of suspension culture. Compared to cells aggregated in medium without inhibitor, much larger aggregates were formed in the presence of Y27632 (FIG. 20). Cell counting confirmed that more viable cells were present in the presence of inhibitor (Table 2, Experiment A). This difference in cell number persisted throughout the course of the culture period, with more cells also observed on day 4, compared to cultures without inhibitor. As with previous suspension culture experiments, cells exposed to Y27632 could also be serially passaged, and maintained in an undifferentiated state (not shown). When the aggregates were split again, almost twice as many cells were observed with Y27632 treatment (Table 2 Experiment A). RT-PCR analysis demonstrated that BG02 cells grown in suspension culture in the presence of Y27632 remained undifferentiated (
|
[
319,
4174,
2692,
8127,
30342,
29892,
322,
1048,
29871,
29896,
30122,
29896,
29900,
29953,
9101,
892,
7180,
297,
8943,
8872,
2673,
470,
594,
2276,
296,
9257,
1532,
29879,
29889,
1894,
23352,
1532,
29879,
892,
29115,
373,
3841,
29871,
29896,
29899,
29953,
322,
715,
15048,
373,
263,
1480,
6287,
313,
18667,
29889,
29871,
29896,
29947,
467,
5806,
263,
6133,
2847,
18618,
310,
298,
2890,
29907,
29879,
892,
3516,
519,
1156,
29871,
29906,
29946,
6199,
297,
594,
2276,
296,
9257,
313,
31793,
29929,
29900,
29995,
7186,
29871,
31793,
29896,
29946,
29995,
511,
14321,
19257,
892,
5734,
519,
727,
7045,
29889,
910,
18694,
393,
298,
2890,
29907,
29879,
1033,
410,
29880,
9633,
403,
925,
408,
19328,
297,
8872,
2673,
9257,
408,
297,
13807,
594,
2276,
296,
9257,
29889,
19413,
18139,
8560,
2645,
1209,
6751,
6511,
697,
304,
22931,
278,
5253,
310,
13184,
1950,
297,
445,
2560,
8872,
2673,
1788,
29889,
512,
3196,
4185,
1973,
16717,
287,
411,
29871,
29945,
30122,
29896,
29900,
29945,
9101,
29892,
14235,
29871,
29896,
29900,
29955,
9101,
29892,
470,
901,
29892,
892,
5759,
1156,
29871,
29955,
3841,
29889,
450,
13184,
1156,
29871,
29955,
3841,
297,
8872,
2673,
9257,
1592,
630,
304,
1048,
263,
29871,
29906,
29900,
29899,
8771,
470,
901,
13184,
29892,
411,
278,
10150,
13184,
8900,
1641,
29871,
31793,
29906,
29946,
30122,
278,
1881,
3038,
1353,
29889,
13,
20755,
6765,
310,
360,
8349,
7268,
519,
315,
10071,
12027,
392,
287,
297,
9511,
29886,
2673,
14062,
13,
29961,
29900,
29896,
29953,
29945,
29962,
22746,
23378,
390,
29911,
29899,
9026,
29934,
313,
29939,
9026,
29934,
29897,
471,
1304,
304,
7252,
18530,
4603,
297,
298,
2890,
29907,
29879,
21633,
297,
8872,
2673,
322,
594,
2276,
296,
9257,
297,
13681,
29899,
15715,
6545,
29889,
28663,
519,
11174,
310,
438,
1783,
29946,
29892,
263,
17456,
310,
715,
332,
666,
327,
296,
9101,
29892,
892,
8900,
297,
1716,
9257,
21971,
29892,
9659,
292,
393,
4185,
1973,
19949,
297,
8872,
2673,
892,
19434,
563,
8349,
7268,
630,
29889,
7791,
29990,
29896,
29955,
29892,
263,
17456,
310,
8422,
573,
1095,
397,
837,
29892,
471,
451,
13384,
297,
2845,
4665,
310,
298,
2890,
29907,
29879,
29889,
450,
3855,
9026,
29934,
7418,
884,
4392,
1312,
278,
7037,
310,
8872,
2673,
298,
2890,
29907,
29879,
304,
17473,
403,
304,
8422,
573,
1095,
397,
837,
29892,
408,
11404,
1078,
297,
8872,
2673,
29889,
2087,
2276,
296,
322,
8872,
2673,
298,
2890,
29907,
29879,
892,
17473,
630,
773,
8943,
5855,
29889,
298,
2890,
29907,
4185,
1973,
892,
14914,
411,
390,
29925,
10403,
6943,
29871,
29906,
29995,
350,
8132,
29892,
29871,
29896,
29900,
29900,
8736,
29914,
828,
21775,
262,
319,
29892,
29871,
29947,
8736,
29914,
828,
383,
29954,
29943,
29906,
322,
29871,
29906,
29945,
8736,
29914,
828,
399,
593,
29941,
29909,
363,
29871,
29906,
29946,
6199,
29892,
5643,
491,
29871,
29906,
3841,
297,
278,
1021,
18350,
1728,
399,
593,
29941,
29909,
29889,
450,
4603,
310,
438,
1783,
29946,
471,
1623,
1727,
7964,
29892,
322,
4603,
310,
7791,
29990,
29896,
29955,
701,
1727,
7964,
22829,
297,
1716,
8422,
573,
1095,
397,
837,
11916,
9401,
304,
563,
8349,
7268,
630,
298,
2890,
29907,
29879,
29889,
910,
1422,
11685,
7418,
16725,
393,
298,
2890,
29907,
29879,
4185,
2955,
297,
8872,
2673,
297,
13681,
29899,
15715,
6545,
19949,
1009,
1422,
11685,
7037,
29892,
408,
3415,
3615,
1133,
491,
278,
5517,
12409,
310,
8422,
573,
1095,
397,
837,
29889,
13,
2528,
654,
310,
385,
319,
1129,
415,
19263,
512,
6335,
2105,
297,
9511,
29886,
2673,
14062,
13,
29961,
29900,
29896,
29953,
29953,
29962,
1763,
472,
841,
27240,
278,
6410,
310,
9101,
1156,
2847,
1209,
6751,
297,
8872,
2673,
29892,
385,
297,
6335,
2105,
310,
16798,
415,
19263,
471,
2715,
304,
278,
18350,
29889,
315,
10071,
892,
1209,
4063,
408,
297,
8741,
29871,
29896,
29906,
29892,
5174,
393,
612,
29899,
29906,
29955,
29953,
29941,
29906,
29892,
385,
297,
6335,
2105,
310,
282,
29896,
29953,
29900,
29899,
29934,
1251,
29899,
21264,
630,
1302,
2356,
29899,
1111,
309,
19015,
559,
313,
1672,
7077,
511,
471,
2715,
304,
278,
18350,
29889,
13,
29961,
29900,
29896,
29953,
29955,
29962,
9511,
29886,
2673,
11404,
1078,
310,
350,
29954,
29900,
29906,
9101,
892,
8429,
491,
16717,
292,
29871,
29906,
30122,
29896,
29900,
29953,
2323,
9101,
297,
29871,
29953,
29899,
5872,
270,
17006,
297,
29871,
29941,
286,
29880,
13681,
29899,
15715,
6545,
18350,
29892,
472,
29871,
29896,
29900,
29900,
364,
3358,
373,
263,
5731,
1218,
7481,
297,
385,
5528,
431,
1061,
313,
3562,
29871,
29906,
29892,
1222,
15362,
319,
467,
29871,
29896,
29900,
29871,
30167,
29924,
612,
29906,
29955,
29953,
29941,
29906,
16641,
7077,
297,
6335,
2105,
471,
2715,
304,
1243,
1532,
29879,
363,
278,
3236,
310,
278,
7639,
322,
278,
4185,
1973,
8900,
14218,
322,
29115,
1156,
29871,
29906,
29946,
6199,
313,
3250,
29871,
29896,
29897,
322,
1156,
29871,
29946,
470,
29871,
29945,
3841,
29889,
1094,
4318,
297,
383,
6259,
29889,
29871,
29906,
29900,
29892,
6124,
310,
612,
29906,
29955,
29953,
29941,
29906,
750,
263,
2600,
618,
2779,
373,
278,
2847,
11404,
362,
8576,
310,
8872,
2673,
9257,
29889,
3831,
1965,
304,
9101,
11404,
630,
297,
18350,
1728,
297,
6335,
2105,
29892,
1568,
7200,
11404,
1078,
892,
8429,
297,
278,
10122,
310,
612,
29906,
29955,
29953,
29941,
29906,
313,
18667,
29889,
29871,
29906,
29900,
467,
19413,
21248,
16725,
393,
901,
3516,
519,
9101,
892,
2198,
297,
278,
10122,
310,
297,
6335,
2105,
313,
3562,
29871,
29906,
29892,
1222,
15362,
319,
467,
910,
4328,
297,
3038,
1353,
3736,
12652,
10106,
278,
3236,
310,
278,
9257,
3785,
29892,
411,
901,
9101,
884,
8900,
373,
2462,
29871,
29946,
29892,
9401,
304,
4185,
1973,
1728,
297,
6335,
2105,
29889,
1094,
411,
3517,
8872,
2673,
9257,
15729,
29892,
9101,
19884,
304,
612,
29906,
29955,
29953,
29941,
29906,
1033,
884,
367,
7797,
368,
1209,
4063,
29892,
322,
19949,
297,
385,
563,
8349,
7268,
630,
2106,
313,
1333,
4318,
467,
1932,
278,
11404,
1078,
892,
6219,
1449,
29892,
4359,
8951,
408,
1784,
9101,
892,
8900,
411,
612,
29906,
29955,
29953,
29941,
29906,
14502,
313,
3562,
29871,
29906,
1222,
15362,
319,
467,
390,
29911,
29899,
9026,
29934,
7418,
28585,
393,
350,
29954,
29900,
29906,
9101,
21633,
297,
8872,
2673,
9257,
297,
278,
10122,
310,
612,
29906,
29955,
29953,
29941,
29906,
9488,
563,
8349,
7268,
630,
313
] | 1,024 |
CommonCrawl
| -0.278594 | 1.220561 | 0.689612 | 1.129282 |
FIG. 21).
[0168] As previous experiments had shown that growth rates of cells in suspension and adherent culture were similar after the initial 24 hours, an experiment was performed where Y27632 was removed after this initial period (Table 2, Experiment B). Consistent with these previous observations, Y27632 enhanced initial survival and aggregation of hESCs after initial passage, but removing the inhibitor after 24 hours did not negatively impact the number and viability count of cells analyzed on day 5. 1.4×107 (+Y27632) and 1.8×107 (+/-Y27632) viable cells were generated when inhibitor was present compared to 3.9×106 cells in untreated cultures. This analysis confirmed that Y27632 had the largest impact during the first 24 hours of suspension hESC culture.
[0169] Because of the enhanced survival and aggregation observed in the presence of Y27632, an experiment was performed to examine if it was possible to reduce the number of cells used to seed suspension cultures (Table 2, Experiment C). Previous experiments had indicated that seeding ES cells at a low density of about 5×105 cells per 3 ml DC-HAIF, or less, did not work well. To determine if addition of a ROCK inhibitor would allow cell seeding at lower densities, a range of cell concentrations (from about 2×106 total cells down to about 1×105 total cells was used to seed suspension cultures in 6-well trays cells in 3 ml DC-HAIF. 10 μM Y27632 was added to all conditions, and the cell number and viability assessed on day 5. Successful aggregation and expansion was observed even at low seeding densities. An approxiate 13 fold expansion of viable cells was observed even cultures that were only seeded with 1×105 cells. Inhibition of ROCK with Y27632 therefore facilitated initial survival of hESCs at much lower densities in this suspension system.
TABLE-US-00002 TABLE 2 Suspension Cultures with and without an Apoptosis Inhibitor Cell counts: total (viable, %) Expt. Treatment Seeding p0, day 1 p0, day 4 p1, day 4 A HAIF 2 × 106 1.9 × 106 (3.5 × 105, 19%) 1.8 × 106 (1.3 × 106, 75%) 2.5 × 106 (2.2 × 106, 88%) +Y27632 2 × 106 1.6 × 106 (1.2 × 106, 74%) 7.8 × 106 (7.1 × 106, 91%) 4.6 × 106 (4.2 × 106, 91%) p0, day 5 B HAIF 2 × 106 2.9 × 106 (5.5 × 105, 26%) .sup. 4.8 × 106 (3.9 × 106, 81.3%) +Y27632 2 × 106 1.9 × 106 (1.4 × 106, 73%) 1.5 × 107 (1.4 × 107, 92%) +/-Y27632 2 × 106 N/A 1.9 × 107 (1.8 × 107, 96%) C +Y27632 2 × 106 1.9 × 106 (1.6 × 106, 84%) 1.4 × 107 (1.2 × 107, 90%) 1 × 106 8.7 × 105 (6.6 × 105, 76%) 8.6 × 106 (7.8 × 106, 91%)
|
[
18667,
29889,
29871,
29906,
29896,
467,
13,
29961,
29900,
29896,
29953,
29947,
29962,
1094,
3517,
15729,
750,
4318,
393,
14321,
19257,
310,
9101,
297,
8872,
2673,
322,
594,
2276,
296,
9257,
892,
2788,
1156,
278,
2847,
29871,
29906,
29946,
6199,
29892,
385,
7639,
471,
8560,
988,
612,
29906,
29955,
29953,
29941,
29906,
471,
6206,
1156,
445,
2847,
3785,
313,
3562,
29871,
29906,
29892,
1222,
15362,
350,
467,
2138,
9696,
411,
1438,
3517,
13917,
29892,
612,
29906,
29955,
29953,
29941,
29906,
427,
29308,
2847,
10503,
2561,
322,
11404,
362,
310,
298,
2890,
29907,
29879,
1156,
2847,
13382,
29892,
541,
11077,
278,
297,
6335,
2105,
1156,
29871,
29906,
29946,
6199,
1258,
451,
3480,
6703,
10879,
278,
1353,
322,
3516,
3097,
2302,
310,
9101,
29537,
287,
373,
2462,
29871,
29945,
29889,
29871,
29896,
29889,
29946,
30122,
29896,
29900,
29955,
20532,
29979,
29906,
29955,
29953,
29941,
29906,
29897,
322,
29871,
29896,
29889,
29947,
30122,
29896,
29900,
29955,
20532,
24028,
29979,
29906,
29955,
29953,
29941,
29906,
29897,
3516,
519,
9101,
892,
5759,
746,
297,
6335,
2105,
471,
2198,
9401,
304,
29871,
29941,
29889,
29929,
30122,
29896,
29900,
29953,
9101,
297,
443,
2484,
630,
4185,
1973,
29889,
910,
7418,
16725,
393,
612,
29906,
29955,
29953,
29941,
29906,
750,
278,
10150,
10879,
2645,
278,
937,
29871,
29906,
29946,
6199,
310,
8872,
2673,
298,
2890,
29907,
9257,
29889,
13,
29961,
29900,
29896,
29953,
29929,
29962,
7311,
310,
278,
427,
29308,
10503,
2561,
322,
11404,
362,
8900,
297,
278,
10122,
310,
612,
29906,
29955,
29953,
29941,
29906,
29892,
385,
7639,
471,
8560,
304,
25917,
565,
372,
471,
1950,
304,
10032,
278,
1353,
310,
9101,
1304,
304,
16717,
8872,
2673,
4185,
1973,
313,
3562,
29871,
29906,
29892,
1222,
15362,
315,
467,
4721,
2366,
15729,
750,
18694,
393,
16717,
292,
17956,
9101,
472,
263,
4482,
9027,
310,
1048,
29871,
29945,
30122,
29896,
29900,
29945,
9101,
639,
29871,
29941,
286,
29880,
13681,
29899,
15715,
6545,
29892,
470,
3109,
29892,
1258,
451,
664,
1532,
29889,
1763,
8161,
565,
6124,
310,
263,
16641,
7077,
297,
6335,
2105,
723,
2758,
3038,
16717,
292,
472,
5224,
6245,
1907,
29892,
263,
3464,
310,
3038,
14953,
800,
313,
3166,
1048,
29871,
29906,
30122,
29896,
29900,
29953,
3001,
9101,
1623,
304,
1048,
29871,
29896,
30122,
29896,
29900,
29945,
3001,
9101,
471,
1304,
304,
16717,
8872,
2673,
4185,
1973,
297,
29871,
29953,
29899,
5872,
260,
764,
29879,
9101,
297,
29871,
29941,
286,
29880,
13681,
29899,
15715,
6545,
29889,
29871,
29896,
29900,
29871,
30167,
29924,
612,
29906,
29955,
29953,
29941,
29906,
471,
2715,
304,
599,
5855,
29892,
322,
278,
3038,
1353,
322,
3516,
3097,
1223,
11517,
373,
2462,
29871,
29945,
29889,
21397,
1319,
11404,
362,
322,
13184,
471,
8900,
1584,
472,
4482,
16717,
292,
6245,
1907,
29889,
530,
2134,
5389,
403,
29871,
29896,
29941,
900,
29881,
13184,
310,
3516,
519,
9101,
471,
8900,
1584,
4185,
1973,
393,
892,
871,
16717,
287,
411,
29871,
29896,
30122,
29896,
29900,
29945,
9101,
29889,
512,
6335,
654,
310,
16641,
7077,
411,
612,
29906,
29955,
29953,
29941,
29906,
5480,
16089,
22731,
2847,
10503,
2561,
310,
298,
2890,
29907,
29879,
472,
1568,
5224,
6245,
1907,
297,
445,
8872,
2673,
1788,
29889,
13,
21009,
29899,
3308,
29899,
29900,
29900,
29900,
29900,
29906,
10911,
29871,
29906,
9511,
29886,
2673,
7965,
1973,
411,
322,
1728,
385,
319,
1129,
415,
19263,
512,
6335,
2105,
19413,
18139,
29901,
3001,
313,
1403,
519,
29892,
28233,
1222,
415,
29889,
6479,
271,
358,
922,
21219,
282,
29900,
29892,
2462,
29871,
29896,
282,
29900,
29892,
2462,
29871,
29946,
282,
29896,
29892,
2462,
29871,
29946,
319,
379,
29909,
6545,
29871,
29906,
13105,
29871,
29896,
29900,
29953,
29871,
29896,
29889,
29929,
13105,
29871,
29896,
29900,
29953,
313,
29941,
29889,
29945,
13105,
29871,
29896,
29900,
29945,
29892,
29871,
29896,
29929,
10997,
29871,
29896,
29889,
29947,
13105,
29871,
29896,
29900,
29953,
313,
29896,
29889,
29941,
13105,
29871,
29896,
29900,
29953,
29892,
29871,
29955,
29945,
10997,
29871,
29906,
29889,
29945,
13105,
29871,
29896,
29900,
29953,
313,
29906,
29889,
29906,
13105,
29871,
29896,
29900,
29953,
29892,
29871,
29947,
29947,
10997,
718,
29979,
29906,
29955,
29953,
29941,
29906,
29871,
29906,
13105,
29871,
29896,
29900,
29953,
29871,
29896,
29889,
29953,
13105,
29871,
29896,
29900,
29953,
313,
29896,
29889,
29906,
13105,
29871,
29896,
29900,
29953,
29892,
29871,
29955,
29946,
10997,
29871,
29955,
29889,
29947,
13105,
29871,
29896,
29900,
29953,
313,
29955,
29889,
29896,
13105,
29871,
29896,
29900,
29953,
29892,
29871,
29929,
29896,
10997,
29871,
29946,
29889,
29953,
13105,
29871,
29896,
29900,
29953,
313,
29946,
29889,
29906,
13105,
29871,
29896,
29900,
29953,
29892,
29871,
29929,
29896,
10997,
282,
29900,
29892,
2462,
29871,
29945,
350,
379,
29909,
6545,
29871,
29906,
13105,
29871,
29896,
29900,
29953,
29871,
29906,
29889,
29929,
13105,
29871,
29896,
29900,
29953,
313,
29945,
29889,
29945,
13105,
29871,
29896,
29900,
29945,
29892,
29871,
29906,
29953,
10997,
869,
12587,
29889,
29871,
29946,
29889,
29947,
13105,
29871,
29896,
29900,
29953,
313,
29941,
29889,
29929,
13105,
29871,
29896,
29900,
29953,
29892,
29871,
29947,
29896,
29889,
29941,
10997,
718,
29979,
29906,
29955,
29953,
29941,
29906,
29871,
29906,
13105,
29871,
29896,
29900,
29953,
29871,
29896,
29889,
29929,
13105,
29871,
29896,
29900,
29953,
313,
29896,
29889,
29946,
13105,
29871,
29896,
29900,
29953,
29892,
29871,
29955,
29941,
10997,
29871,
29896,
29889,
29945,
13105,
29871,
29896,
29900,
29955,
313,
29896,
29889,
29946,
13105,
29871,
29896,
29900,
29955,
29892,
29871,
29929,
29906,
10997,
718,
24028,
29979,
29906,
29955,
29953,
29941,
29906,
29871,
29906,
13105,
29871,
29896,
29900,
29953,
405,
29914,
29909,
29871,
29896,
29889,
29929,
13105,
29871,
29896,
29900,
29955,
313,
29896,
29889,
29947,
13105,
29871,
29896,
29900,
29955,
29892,
29871,
29929,
29953,
10997,
315,
718,
29979,
29906,
29955,
29953,
29941,
29906,
29871,
29906,
13105,
29871,
29896,
29900,
29953,
29871,
29896,
29889,
29929,
13105,
29871,
29896,
29900,
29953,
313,
29896,
29889,
29953,
13105,
29871,
29896,
29900,
29953,
29892,
29871,
29947,
29946,
10997,
29871,
29896,
29889,
29946,
13105,
29871,
29896,
29900,
29955,
313,
29896,
29889,
29906,
13105,
29871,
29896,
29900,
29955,
29892,
29871,
29929,
29900,
10997,
29871,
29896,
13105,
29871,
29896,
29900,
29953,
29871,
29947,
29889,
29955,
13105,
29871,
29896,
29900,
29945,
313,
29953,
29889,
29953,
13105,
29871,
29896,
29900,
29945,
29892,
29871,
29955,
29953,
10997,
29871,
29947,
29889,
29953,
13105,
29871,
29896,
29900,
29953,
313,
29955,
29889,
29947,
13105,
29871,
29896,
29900,
29953,
29892,
29871,
29929,
29896,
10997
] | 1,024 |
CommonCrawl
| -1.226 | 0.247223 | -0.323559 | -0.07488 |
You can find me at www.MelindaMaxfield.com
I am also on Facebook and Twitter.
Melinda Jane Maxfield ("Melinda") operates www.blog.spacefairy.com and may operate other websites. It is Melinda's policy to respect your privacy regarding any information we may collect while operating our websites.
Like most website operators, Melinda collects non-personally-identifying information of the sort that web browsers and servers typically make available, such as the browser type, language preference, referring site, and the date and time of each visitor request. Melinda's purpose in collecting non-personally identifying information is to better understand how Melinda's visitors use its website. From time to time, Melinda may release non-personally-identifying information in the aggregate, e.g., by publishing a report on trends in the usage of its website.
Melinda also collects potentially personally-identifying information like Internet Protocol (IP) addresses for logged in users and for users leaving comments on www.blog.spacefairy.com blogs/sites. Melinda only discloses logged in user and commenter IP addresses under the same circumstances that it uses and discloses personally-identifying information as described below, except that commenter IP addresses and email addresses are visible and disclosed to the administrators of the blog/site where the comment was left.
Certain visitors to Melinda's websites choose to interact with Melinda in ways that require Melinda to gather personally-identifying information. The amount and type of information that Melinda gathers depends on the nature of the interaction. For example, we ask visitors who sign up at www.blog.spacefairy.com to provide a username and email address. Those who engage in transactions with Melinda are asked to provide additional information, including as necessary the personal and financial information required to process those transactions. In each case, Melinda collects such information only insofar as is necessary or appropriate to fulfill the purpose of the visitor's interaction with Melinda. Melinda does not disclose personally-identifying information other than as described below. And visitors can always refuse to supply personally-identifying information, with the caveat that it may prevent them from engaging in certain website-related activities.
Melinda may collect statistics about the behavior of visitors to its websites. Melinda may display this information publicly or provide it to others. However, Melinda does not disclose personally-identifying information other than as described below.
Melinda discloses potentially personally-identifying and personally-identifying information only to those of its employees, contractors and affiliated organizations that (i) need to know that information in order to process it on Melinda's behalf or to provide services available at Melinda's websites, and (ii) that have agreed not to disclose it to others. Some of those employees, contractors and affiliated organizations may be located outside of your home country; by using Melinda's websites, you consent to the transfer of such information to them. Melinda will not rent or sell potentially personally-identifying and personally-identifying information to anyone. Other than to its employees, contractors and affiliated organizations, as described above, Melinda discloses potentially personally-identifying and personally-identifying information only in response to a subpoena, court order or other governmental request, or when Melinda believes in good faith that disclosure is reasonably necessary to protect the property or rights of Melinda, third parties or the public at large. If you are a registered user of an Melinda website and have supplied your email address, Melinda may occasionally send you an email to tell you about new features, solicit your feedback, or just keep you up to date with what's going on with Melinda and our products. If you send us a request (for example via email or via one of our feedback mechanisms), we reserve the right to publish it in order to help us clarify or respond to your request or to help us support other users. Melinda takes all measures reasonably necessary to protect against the unauthorized access, use, alteration or destruction of potentially personally-identifying and personally-identifying information.
A cookie is a string of information that a website stores on a visitor's computer, and that the visitor's browser provides to the website each time the visitor returns. Melinda uses cookies to help Melinda identify and track visitors, their usage of Melinda website, and their website access preferences. Melinda visitors who do not wish to have cookies placed on their computers should set their browsers to refuse cookies before using Melinda's websites, with the drawback that certain features of Melinda's websites may not function properly without the aid of cookies.
|
[
887,
508,
1284,
592,
472,
7821,
29889,
29924,
295,
11054,
7976,
2671,
29889,
510,
13,
29902,
626,
884,
373,
13327,
322,
20147,
29889,
13,
29924,
295,
11054,
10447,
5918,
2671,
4852,
29924,
295,
11054,
1159,
1751,
1078,
7821,
29889,
7312,
29889,
3493,
29888,
1466,
29891,
29889,
510,
322,
1122,
21994,
916,
28007,
29889,
739,
338,
6286,
11054,
29915,
29879,
8898,
304,
3390,
596,
5999,
4135,
11211,
738,
2472,
591,
1122,
6314,
1550,
13598,
1749,
28007,
29889,
13,
27552,
1556,
4700,
12768,
29892,
6286,
11054,
6314,
29879,
1661,
29899,
10532,
635,
29899,
1693,
9215,
2472,
310,
278,
2656,
393,
1856,
14376,
322,
12424,
12234,
1207,
3625,
29892,
1316,
408,
278,
4714,
1134,
29892,
4086,
24583,
29892,
16811,
3268,
29892,
322,
278,
2635,
322,
931,
310,
1269,
27682,
2009,
29889,
6286,
11054,
29915,
29879,
6437,
297,
6314,
292,
1661,
29899,
10532,
635,
2893,
9215,
2472,
338,
304,
2253,
2274,
920,
6286,
11054,
29915,
29879,
26824,
671,
967,
4700,
29889,
3645,
931,
304,
931,
29892,
6286,
11054,
1122,
6507,
1661,
29899,
10532,
635,
29899,
1693,
9215,
2472,
297,
278,
20431,
29892,
321,
29889,
29887,
1696,
491,
27256,
263,
3461,
373,
534,
1975,
297,
278,
8744,
310,
967,
4700,
29889,
13,
29924,
295,
11054,
884,
6314,
29879,
19998,
22345,
29899,
1693,
9215,
2472,
763,
4685,
1019,
5770,
313,
5690,
29897,
14157,
363,
13817,
297,
4160,
322,
363,
4160,
10124,
6589,
373,
7821,
29889,
7312,
29889,
3493,
29888,
1466,
29891,
29889,
510,
12618,
29879,
29914,
16315,
29889,
6286,
11054,
871,
766,
11291,
267,
13817,
297,
1404,
322,
3440,
261,
5641,
14157,
1090,
278,
1021,
14209,
393,
372,
3913,
322,
766,
11291,
267,
22345,
29899,
1693,
9215,
2472,
408,
5439,
2400,
29892,
5174,
393,
3440,
261,
5641,
14157,
322,
4876,
14157,
526,
7962,
322,
766,
15603,
304,
278,
6343,
4097,
310,
278,
12618,
29914,
2746,
988,
278,
3440,
471,
2175,
29889,
13,
29907,
13946,
26824,
304,
6286,
11054,
29915,
29879,
28007,
6755,
304,
16254,
411,
6286,
11054,
297,
5837,
393,
1996,
6286,
11054,
304,
11705,
22345,
29899,
1693,
9215,
2472,
29889,
450,
5253,
322,
1134,
310,
2472,
393,
6286,
11054,
330,
19467,
7111,
373,
278,
5469,
310,
278,
14881,
29889,
1152,
1342,
29892,
591,
2244,
26824,
1058,
1804,
701,
472,
7821,
29889,
7312,
29889,
3493,
29888,
1466,
29891,
29889,
510,
304,
3867,
263,
8952,
322,
4876,
3211,
29889,
16025,
1058,
3033,
482,
297,
22160,
411,
6286,
11054,
526,
4433,
304,
3867,
5684,
2472,
29892,
3704,
408,
5181,
278,
7333,
322,
18161,
2472,
3734,
304,
1889,
1906,
22160,
29889,
512,
1269,
1206,
29892,
6286,
11054,
6314,
29879,
1316,
2472,
871,
297,
578,
15641,
408,
338,
5181,
470,
8210,
304,
6095,
5589,
278,
6437,
310,
278,
27682,
29915,
29879,
14881,
411,
6286,
11054,
29889,
6286,
11054,
947,
451,
2313,
2226,
22345,
29899,
1693,
9215,
2472,
916,
1135,
408,
5439,
2400,
29889,
1126,
26824,
508,
2337,
26506,
304,
11421,
22345,
29899,
1693,
9215,
2472,
29892,
411,
278,
24230,
271,
393,
372,
1122,
5557,
963,
515,
3033,
6751,
297,
3058,
4700,
29899,
12817,
14188,
29889,
13,
29924,
295,
11054,
1122,
6314,
13964,
1048,
278,
6030,
310,
26824,
304,
967,
28007,
29889,
6286,
11054,
1122,
2479,
445,
2472,
970,
368,
470,
3867,
372,
304,
4045,
29889,
2398,
29892,
6286,
11054,
947,
451,
2313,
2226,
22345,
29899,
1693,
9215,
2472,
916,
1135,
408,
5439,
2400,
29889,
13,
29924,
295,
11054,
766,
11291,
267,
19998,
22345,
29899,
1693,
9215,
322,
22345,
29899,
1693,
9215,
2472,
871,
304,
1906,
310,
967,
22873,
29892,
8078,
943,
322,
23736,
630,
25700,
393,
313,
29875,
29897,
817,
304,
1073,
393,
2472,
297,
1797,
304,
1889,
372,
373,
6286,
11054,
29915,
29879,
2306,
3131,
470,
304,
3867,
5786,
3625,
472,
6286,
11054,
29915,
29879,
28007,
29892,
322,
313,
2236,
29897,
393,
505,
15502,
451,
304,
2313,
2226,
372,
304,
4045,
29889,
3834,
310,
1906,
22873,
29892,
8078,
943,
322,
23736,
630,
25700,
1122,
367,
5982,
5377,
310,
596,
3271,
4234,
29936,
491,
773,
6286,
11054,
29915,
29879,
28007,
29892,
366,
20218,
304,
278,
6782,
310,
1316,
2472,
304,
963,
29889,
6286,
11054,
674,
451,
23990,
470,
19417,
19998,
22345,
29899,
1693,
9215,
322,
22345,
29899,
1693,
9215,
2472,
304,
5019,
29889,
5901,
1135,
304,
967,
22873,
29892,
8078,
943,
322,
23736,
630,
25700,
29892,
408,
5439,
2038,
29892,
6286,
11054,
766,
11291,
267,
19998,
22345,
29899,
1693,
9215,
322,
22345,
29899,
1693,
9215,
2472,
871,
297,
2933,
304,
263,
1014,
1129,
2386,
29892,
8973,
1797,
470,
916,
5874,
284,
2009,
29892,
470,
746,
6286,
11054,
1339,
17180,
297,
1781,
10847,
393,
766,
25071,
338,
2769,
2197,
5181,
304,
12566,
278,
2875,
470,
10462,
310,
6286,
11054,
29892,
4654,
13973,
470,
278,
970,
472,
2919,
29889,
960,
366,
526,
263,
15443,
1404,
310,
385,
6286,
11054,
4700,
322,
505,
19056,
596,
4876,
3211,
29892,
6286,
11054,
1122,
23025,
3638,
366,
385,
4876,
304,
2649,
366,
1048,
716,
5680,
29892,
26978,
277,
596,
16705,
29892,
470,
925,
3013,
366,
701,
304,
2635,
411,
825,
29915,
29879,
2675,
373,
411,
6286,
11054,
322,
1749,
9316,
29889,
960,
366,
3638,
502,
263,
2009,
313,
1454,
1342,
3025,
4876,
470,
3025,
697,
310,
1749,
16705,
7208,
12903,
511,
591,
23986,
278,
1492,
304,
9805,
372,
297,
1797,
304,
1371,
502,
15544,
470,
10049,
304,
596,
2009,
470,
304,
1371,
502,
2304,
916,
4160,
29889,
6286,
11054,
4893,
599,
15366,
2769,
2197,
5181,
304,
12566,
2750,
278,
1185,
329,
2015,
1891,
2130,
29892,
671,
29892,
10551,
362,
470,
22104,
310,
19998,
22345,
29899,
1693,
9215,
322,
22345,
29899,
1693,
9215,
2472,
29889,
13,
29909,
15327,
338,
263,
1347,
310,
2472,
393,
263,
4700,
14422,
373,
263,
27682,
29915,
29879,
6601,
29892,
322,
393,
278,
27682,
29915,
29879,
4714,
8128,
304,
278,
4700,
1269,
931,
278,
27682,
3639,
29889,
6286,
11054,
3913,
21046,
304,
1371,
6286,
11054,
12439,
322,
5702,
26824,
29892,
1009,
8744,
310,
6286,
11054,
4700,
29892,
322,
1009,
4700,
2130,
5821,
2063,
29889,
6286,
11054,
26824,
1058,
437,
451,
6398,
304,
505,
21046,
7180,
373,
1009,
23226,
881,
731,
1009,
14376,
304,
26506,
21046,
1434,
773,
6286,
11054,
29915,
29879,
28007,
29892,
411,
278,
4216,
1627,
393,
3058,
5680,
310,
6286,
11054,
29915,
29879,
28007,
1122,
451,
740,
6284,
1728,
278,
16226,
310,
21046,
29889,
13
] | 1,024 |
CommonCrawl
| -1.126827 | -1.381497 | -1.967945 | -1.327108 |
HomeFind a JobSummer 2023 - Data Analytics Intern
Summer 2023 - Data Analytics Intern
Smithfield, VA / Lisle, IL
Job/Internship Description
ID 2022-26026
Job Locations US-VA-Smithfield | US-IL-Lisle
Category Intern/Trainee
Type Intern
FLSA Status Non-Exempt
Shift Corporate - Day Shift
US-VA-Smithfield
US-IL-Lisle
The Smithfield internship program offers students a paid opportunity to work in an environment that fosters real-life experiences, explores career options and applies your education. You will become a part of key management meetings and participate in meaningful projects. Our goal is to provide a working environment that not only encourages creativity, but offers equality, diversity, mutual respect and productivity. Our summer internships are typically 10 weeks.
The Data Analytics Summer Intern is responsible for assisting with various Data Analytics projects, and initiatives. The projects will include solving business problems using Python, SQL, and Power BI.
Core Responsibilities
Assist Data Analytics team by executing standard reports and, running / maintaining ad hoc queries.
Participate in the development of reports, metrics, models, scorecards, and dashboards by gathering data and transforming it into stories and insights that drive business recommendations and decision-making.
Build reports and dashboards in Microsoft Power BI, Analysis for Office, Bex, and other reporting tools
Participate in hosted training sessions on how to use the various tools and reports created by the department
Communicate effectively with report stakeholders to ensure strong understanding of the methodology behind all calculations
Perform ad hoc analysis using data from various systems and explain results of analysis to management
Provide comprehensive analytical and fact-based reporting to support and/or identify opportunities to impact operational results, and support strategic initiatives.
To perform this job successfully, an individual must be able to perform each essential duty satisfactorily. The requirements listed below are representative of the knowledge, skill, and/or ability required. Reasonable accommodations may be made to enable individuals to perform the essential functions.
Completion of sophomore year at an accredited four year university.
Pursuing a bachelor's degree in Computer Science, Information Technology, Business or Engineering.
Must be in good academic standing.
Knowledge of basic Information Technology principals and techniques.
Proficient use of computers.
Ability to express ideas clearly and concisely, both orally and in writing.
Proactive thinker and self-motivated.
Strong time, project, and organizational management skills.
The ability to work independently.
Ability to work well with others in fast paced, dynamic environment.
Ability to be respectful, approachable and team oriented while building strong working relationships and a positive work environment.
Careers and Benefits
To learn more about Smithfield's benefits, visit smithfieldfoods.com/careerbenefits.
More than 63,000 employees globally drive our success. We strive to create a fair, ethical and rewarding work environment.
Working at Smithfield isn't just a job - it's the foundation for a lifelong career with training designed to help you advance professionally.
Our people matter. That's why we offer excellent, comprehensive benefits packages to our full-time employees.
Sustainability is ingrained in our culture and guides how we operate. We believe in innovating for the future.
About Smithfield Foods
Headquartered in Smithfield, Va. since 1936, Smithfield Foods, Inc. is an American food company with agricultural roots and a global reach. With more than 60,000 jobs globally, we are dedicated to producing "Good food. Responsibly" and serve as one of the world's leading vertically integrated protein companies. We have pioneered sustainability standards for more than two decades, including our industry-leading commitments to become carbon negative in our U.S. company-owned operations and reduce GHG emissions 30 percent across our entire U.S. value chain by 2030. We believe in the power of protein to end food insecurity and have donated hundreds of millions of food servings to our communities. Smithfield boasts a portfolio of high-quality iconic brands, such as Smithfield, Eckrich and Nathan's Famous, among many
|
[
8778,
12542,
263,
17163,
11139,
1050,
29871,
29906,
29900,
29906,
29941,
448,
3630,
11597,
22026,
2422,
13,
11139,
1050,
29871,
29906,
29900,
29906,
29941,
448,
3630,
11597,
22026,
2422,
13,
29209,
2671,
29892,
478,
29909,
847,
15285,
280,
29892,
17674,
13,
11947,
29914,
17579,
3527,
12953,
13,
1367,
29871,
29906,
29900,
29906,
29906,
29899,
29906,
29953,
29900,
29906,
29953,
13,
11947,
5976,
800,
3148,
29899,
20449,
29899,
29209,
2671,
891,
3148,
29899,
6227,
29899,
29931,
275,
280,
13,
10900,
2422,
29914,
5323,
457,
29872,
13,
1542,
2422,
13,
10536,
8132,
16034,
10050,
29899,
1252,
3456,
13,
29657,
12767,
403,
448,
8373,
1383,
2027,
13,
3308,
29899,
20449,
29899,
29209,
2671,
13,
3308,
29899,
6227,
29899,
29931,
275,
280,
13,
1576,
7075,
2671,
2836,
3527,
1824,
16688,
8041,
263,
12530,
15130,
304,
664,
297,
385,
5177,
393,
9926,
414,
1855,
29899,
19264,
27482,
29892,
3902,
2361,
6413,
3987,
322,
16058,
596,
9793,
29889,
887,
674,
4953,
263,
760,
310,
1820,
10643,
5870,
886,
322,
5221,
403,
297,
6593,
1319,
9279,
29889,
8680,
7306,
338,
304,
3867,
263,
1985,
5177,
393,
451,
871,
18443,
267,
907,
28157,
29892,
541,
16688,
17193,
29892,
6894,
537,
29892,
5478,
950,
3390,
322,
3234,
2068,
29889,
8680,
11801,
2836,
9981,
526,
12234,
29871,
29896,
29900,
11405,
29889,
13,
1576,
3630,
11597,
22026,
13329,
2422,
338,
14040,
363,
6985,
292,
411,
5164,
3630,
11597,
22026,
9279,
29892,
322,
14511,
5056,
29889,
450,
9279,
674,
3160,
17069,
5381,
4828,
773,
5132,
29892,
3758,
29892,
322,
9206,
350,
29902,
29889,
13,
9203,
2538,
29886,
787,
747,
9770,
13,
7900,
391,
3630,
11597,
22026,
3815,
491,
14012,
3918,
13676,
322,
29892,
2734,
847,
7344,
292,
594,
298,
542,
9365,
29889,
13,
7439,
12654,
403,
297,
278,
5849,
310,
13676,
29892,
21556,
29892,
4733,
29892,
8158,
28160,
29892,
322,
12569,
24691,
491,
11705,
292,
848,
322,
4327,
292,
372,
964,
15874,
322,
1663,
5861,
393,
7899,
5381,
6907,
800,
322,
10608,
29899,
28990,
29889,
13,
8893,
13676,
322,
12569,
24691,
297,
7783,
9206,
350,
29902,
29892,
24352,
363,
11367,
29892,
350,
735,
29892,
322,
916,
23415,
8492,
13,
7439,
12654,
403,
297,
17791,
6694,
21396,
373,
920,
304,
671,
278,
5164,
8492,
322,
13676,
2825,
491,
278,
14311,
13,
5261,
2523,
403,
17583,
411,
3461,
380,
1296,
8948,
414,
304,
9801,
4549,
8004,
310,
278,
1158,
3002,
5742,
599,
17203,
13,
5894,
689,
594,
298,
542,
7418,
773,
848,
515,
5164,
6757,
322,
5649,
2582,
310,
7418,
304,
10643,
13,
1184,
29894,
680,
15171,
6270,
16114,
936,
322,
2114,
29899,
6707,
23415,
304,
2304,
322,
29914,
272,
12439,
28602,
1907,
304,
10879,
1751,
1288,
2582,
29892,
322,
2304,
16650,
293,
14511,
5056,
29889,
13,
1762,
2189,
445,
4982,
8472,
29892,
385,
5375,
1818,
367,
2221,
304,
2189,
1269,
18853,
13360,
5119,
7168,
2354,
29889,
450,
11780,
9904,
2400,
526,
21097,
310,
278,
7134,
29892,
19911,
29892,
322,
29914,
272,
11509,
3734,
29889,
830,
1658,
519,
24803,
800,
1122,
367,
1754,
304,
9025,
15724,
304,
2189,
278,
18853,
3168,
29889,
13,
28958,
310,
269,
3021,
290,
487,
1629,
472,
385,
1035,
1127,
1573,
3023,
1629,
16372,
29889,
13,
29925,
1295,
26420,
263,
289,
496,
12779,
29915,
29879,
7426,
297,
20972,
9327,
29892,
10343,
17968,
29892,
15197,
470,
22557,
29889,
13,
29924,
504,
367,
297,
1781,
21567,
13407,
29889,
13,
29968,
3707,
5485,
310,
6996,
10343,
17968,
3420,
1338,
322,
13698,
29889,
13,
1184,
9639,
993,
671,
310,
23226,
29889,
13,
4920,
1793,
304,
4653,
7014,
9436,
322,
3022,
275,
873,
29892,
1716,
470,
635,
322,
297,
5007,
29889,
13,
1184,
4925,
1348,
261,
322,
1583,
29899,
14817,
440,
630,
29889,
13,
5015,
549,
931,
29892,
2060,
29892,
322,
8674,
1288,
10643,
25078,
29889,
13,
1576,
11509,
304,
664,
25499,
29889,
13,
4920,
1793,
304,
664,
1532,
411,
4045,
297,
5172,
22906,
287,
29892,
7343,
5177,
29889,
13,
4920,
1793,
304,
367,
3390,
1319,
29892,
2948,
519,
322,
3815,
7769,
287,
1550,
5214,
4549,
1985,
21702,
322,
263,
6374,
664,
5177,
29889,
13,
29907,
598,
414,
322,
4111,
1389,
1169,
13,
1762,
5110,
901,
1048,
7075,
2671,
29915,
29879,
23633,
29892,
6493,
1560,
389,
2671,
1181,
19653,
29889,
510,
29914,
18020,
261,
1785,
1389,
1169,
29889,
13,
20761,
1135,
29871,
29953,
29941,
29892,
29900,
29900,
29900,
22873,
13149,
635,
7899,
1749,
2551,
29889,
1334,
380,
4401,
304,
1653,
263,
6534,
29892,
11314,
936,
322,
20751,
292,
664,
5177,
29889,
13,
5531,
292,
472,
7075,
2671,
3508,
29915,
29873,
925,
263,
4982,
448,
372,
29915,
29879,
278,
22778,
363,
263,
11747,
295,
549,
6413,
411,
6694,
8688,
304,
1371,
366,
6564,
6351,
635,
29889,
13,
29949,
332,
2305,
4383,
29889,
2193,
29915,
29879,
2020,
591,
5957,
15129,
29892,
15171,
6270,
23633,
9741,
304,
1749,
2989,
29899,
2230,
22873,
29889,
13,
29903,
504,
475,
3097,
338,
2348,
22042,
297,
1749,
9257,
322,
1410,
2247,
920,
591,
21994,
29889,
1334,
4658,
297,
24233,
1218,
363,
278,
5434,
29889,
13,
28173,
7075,
2671,
25453,
29879,
13,
5494,
339,
4254,
287,
297,
7075,
2671,
29892,
12549,
29889,
1951,
29871,
29896,
29929,
29941,
29953,
29892,
7075,
2671,
25453,
29879,
29892,
9266,
29889,
338,
385,
3082,
9687,
5001,
411,
18032,
3631,
16778,
322,
263,
5534,
6159,
29889,
2973,
901,
1135,
29871,
29953,
29900,
29892,
29900,
29900,
29900,
17643,
13149,
635,
29892,
591,
526,
16955,
304,
20811,
376,
18420,
9687,
29889,
2538,
29886,
787,
14981,
29908,
322,
9080,
408,
697,
310,
278,
3186,
29915,
29879,
8236,
4837,
1711,
23387,
26823,
14582,
29889,
1334,
505,
29323,
14561,
15075,
475,
3097,
20801,
363,
901,
1135,
1023,
1602,
3076,
29892,
3704,
1749,
13661,
29899,
25369,
9063,
1860,
304,
4953,
22004,
8178,
297,
1749,
501,
29889,
29903,
29889,
5001,
29899,
26689,
6931,
322,
10032,
402,
29950,
29954,
953,
6847,
29871,
29941,
29900,
10151,
4822,
1749,
4152,
501,
29889,
29903,
29889,
995,
9704,
491,
29871,
29906,
29900,
29941,
29900,
29889,
1334,
4658,
297,
278,
3081,
310,
26823,
304,
1095,
9687,
297,
8926,
322,
505,
1016,
630,
21006,
310,
14746,
310,
9687,
3348,
886,
304,
1749,
23507,
29889,
7075,
2671,
1045,
19416,
263,
2011,
25648,
310,
1880,
29899,
29567,
9849,
293,
1506,
4167,
29892,
1316,
408,
7075,
2671,
29892,
382,
384,
4018,
322,
27650,
29915,
29879,
27458,
681,
29892,
4249,
1784
] | 1,024 |
CommonCrawl
| -0.23631 | -0.632676 | -0.183015 | 0.856069 |
On 7 & 8 March 2018 more than 200 international visitors from the mechanical, plant, and electrical engineering industries informed themselves about trends and developments such as Industry 4.0 and cost optimisiation with Strategic Parts Management, boosting market potential with Electronic Product Catalogs and Building Information Modeling (BIM).
The opening keynote presentation of the 19th Industry Forum is available upon request.
Andreas Brandauer, born in 1965, has been with Siemens Industry Software GmbH for more than 12 years. As Senior PreSales Consultant, he has experience in the areas of mechatronic development in mechanical engineering, virtual commissioning and early real-time simulation.
Dipl. Ing. Peter Zander is in charge of organization, care and advancement of CAD-and PDM systems onsite KUKA Systems Assembly & Test. He is responsible for the unification of CAD, PDM, ERP and standard parts management.
Marcel Nagel has been at SCHUNK GmbH & Co. KG since finishing his Bachelors in mechanical engineering in 2002. Throughout his career at SCHUNK, he has been focused in the areas of development automation along with product and portfolio management. Since 2013 he has been the Head of Central Product and Portfolio Management.
Tobias Wild is working as a Development Engineer for mechanical components at the Swiss company Stadler Rail. In the year 2017 Tobias Wild was responsible for the integration of CADENAS PARTsolutions at Stadler.
A Mechanical Engineer with over 20 years' experience in industrial lifting & positioning solutions for client projects worldwide. Specialist linear motion and power transmission skills gained in a variety of roles at Power Jacks including engineering, production, sales and marketing. A flair for product marketing sees Bruce Hamper Head the Marketing Team that has seen Power Jacks enter and expand in new markets globally including offices in Germany and China.
Steve started his career as a Mechanical Design Engineer in 1983 designing automation systems. While in this area Steve developed several software methods that would increase productivity for Mechanical Engineers. In 1994 he joined PHD, Inc. serving various roles in Marketing, Finance and Information Technology where he is responsible for the creation and distribution of engineering software including Customer CAD, sizing software, animations, and web sites including all other Information Technology responsibilities for PHD, Inc and its subsidiaries.
Catriona manages WARR Hyperloop's inventory and leads the levitation and contactless propulsion teams. She is a Master student in engineering physics at TU Munich and is developing the next generation of particle detectors for the Large Hadron Collider at CERN. Working both in the massive international collaboration that is CERN and in the small, dynamic Hyperloop team gives insights into how organizations of different sizes manage design and development processes.
Since July 2016, Anthea Waidele has been the Marketing Assistant for the Afag Group. Her focus is on online marketing and the production of print documents. Among other things, she is responsible for maintaining the Afag website and social media channels.
Jakob Prechtl is an Industrial Master in Metal and Mechanical Engineering. After working for several years in the machine tool industry, he set up his own business in 1990 with an engineering office in the area of development and design and gradually expanded it. Since 2009 he is Partner and Managing Director of Prechtl Engineering GmbH with main focus on machine tools, special equipment, automotive, wind energy and medical technology. The resulting self made product "Portable surgical microscope for developing countries" was spun off in 2016 into a separate company "Prechtl TechSolutions", whose sole owner is Jakob Prechtl.
Sergio Salsedo has many years' experience in Software Development in high tech environments. He has also been responsible for PLM consulting services in his career. Mr. Salsedo is not only the Co-founder of Focus PLM, an Aras Gold Partner, but also the companies CEO and provides CAD connectors amongst other integrated solutions.
Luca Borghi, CEO of CADENAS Italiana S.r.l., has been a member of CADENAS since 2009. Since then he has held various positions, including technical manager for the pre- and post-sales activities PARTsolutions/PARTwarehouse for Italy and Spain
|
[
1551,
29871,
29955,
669,
29871,
29947,
4779,
29871,
29906,
29900,
29896,
29947,
901,
1135,
29871,
29906,
29900,
29900,
6121,
26824,
515,
278,
28310,
29892,
8024,
29892,
322,
3546,
16888,
21639,
6397,
2722,
23388,
6053,
1048,
534,
1975,
322,
2693,
1860,
1316,
408,
12157,
719,
29871,
29946,
29889,
29900,
322,
3438,
5994,
10770,
362,
411,
3767,
1845,
293,
3455,
29879,
15057,
29892,
14505,
292,
9999,
7037,
411,
28251,
293,
10969,
5725,
29879,
322,
17166,
10343,
8125,
292,
313,
29933,
7833,
467,
13,
1576,
8718,
1589,
948,
866,
24329,
310,
278,
29871,
29896,
29929,
386,
12157,
719,
29474,
338,
3625,
2501,
2009,
29889,
13,
2855,
12588,
18007,
7547,
29892,
6345,
297,
29871,
29896,
29929,
29953,
29945,
29892,
756,
1063,
411,
317,
3768,
575,
12157,
719,
18540,
18156,
363,
901,
1135,
29871,
29896,
29906,
2440,
29889,
1094,
24260,
4721,
29903,
2122,
2138,
499,
424,
29892,
540,
756,
7271,
297,
278,
10161,
310,
592,
13496,
1617,
293,
5849,
297,
28310,
21639,
29892,
6901,
12969,
292,
322,
4688,
1855,
29899,
2230,
17402,
29889,
13,
12130,
572,
29889,
22607,
29889,
5310,
796,
3825,
338,
297,
8323,
310,
13013,
29892,
2562,
322,
3061,
27967,
310,
315,
3035,
29899,
392,
349,
23560,
6757,
373,
2746,
476,
19960,
29909,
23985,
13266,
669,
4321,
29889,
940,
338,
14040,
363,
278,
443,
2450,
310,
315,
3035,
29892,
349,
23560,
29892,
8982,
29925,
322,
3918,
5633,
10643,
29889,
13,
7083,
2242,
14461,
295,
756,
1063,
472,
317,
3210,
3904,
29968,
18156,
669,
3189,
29889,
476,
29954,
1951,
28321,
670,
13620,
295,
943,
297,
28310,
21639,
297,
29871,
29906,
29900,
29900,
29906,
29889,
17044,
449,
670,
6413,
472,
317,
3210,
3904,
29968,
29892,
540,
756,
1063,
21309,
297,
278,
10161,
310,
5849,
3345,
362,
3412,
411,
3234,
322,
2011,
25648,
10643,
29889,
4001,
29871,
29906,
29900,
29896,
29941,
540,
756,
1063,
278,
12252,
310,
8068,
10969,
322,
3371,
25648,
15057,
29889,
13,
29911,
711,
3173,
11821,
338,
1985,
408,
263,
14650,
10863,
261,
363,
28310,
7117,
472,
278,
26182,
5001,
10347,
1358,
9620,
29889,
512,
278,
1629,
29871,
29906,
29900,
29896,
29955,
22354,
3173,
11821,
471,
14040,
363,
278,
13465,
310,
315,
3035,
1430,
3289,
349,
8322,
2929,
17925,
472,
10347,
1358,
29889,
13,
29909,
27439,
936,
10863,
261,
411,
975,
29871,
29906,
29900,
2440,
29915,
7271,
297,
18408,
11747,
1259,
669,
2602,
292,
6851,
363,
3132,
9279,
3186,
8157,
29889,
12630,
391,
5608,
10884,
322,
3081,
22713,
25078,
17515,
297,
263,
12875,
310,
16178,
472,
9206,
5457,
29879,
3704,
21639,
29892,
5802,
29892,
16538,
322,
9999,
292,
29889,
319,
17422,
381,
363,
3234,
9999,
292,
18553,
18885,
7904,
546,
12252,
278,
4485,
15133,
8583,
393,
756,
3595,
9206,
5457,
29879,
3896,
322,
7985,
297,
716,
2791,
1691,
13149,
635,
3704,
25222,
297,
9556,
322,
7551,
29889,
13,
7789,
345,
4687,
670,
6413,
408,
263,
27439,
936,
12037,
10863,
261,
297,
29871,
29896,
29929,
29947,
29941,
2874,
292,
3345,
362,
6757,
29889,
5806,
297,
445,
4038,
13981,
8906,
3196,
7047,
3519,
393,
723,
7910,
3234,
2068,
363,
27439,
936,
10863,
414,
29889,
512,
29871,
29896,
29929,
29929,
29946,
540,
8772,
349,
26124,
29892,
9266,
29889,
16330,
5164,
16178,
297,
4485,
15133,
29892,
4231,
749,
322,
10343,
17968,
988,
540,
338,
14040,
363,
278,
11265,
322,
4978,
310,
21639,
7047,
3704,
21886,
315,
3035,
29892,
269,
5281,
7047,
29892,
3778,
800,
29892,
322,
1856,
11840,
3704,
599,
916,
10343,
17968,
5544,
747,
9770,
363,
349,
26124,
29892,
9266,
322,
967,
11684,
8819,
4314,
29889,
13,
29907,
8141,
16017,
767,
1179,
399,
1718,
29934,
26078,
7888,
29915,
29879,
11817,
706,
322,
11981,
278,
14453,
7018,
322,
6958,
2222,
3107,
25381,
10907,
29889,
2296,
338,
263,
9082,
8368,
297,
21639,
17558,
472,
323,
29965,
13564,
436,
322,
338,
14338,
278,
2446,
12623,
310,
16445,
6459,
943,
363,
278,
8218,
479,
14302,
1617,
13435,
1241,
472,
315,
1001,
29940,
29889,
24176,
1716,
297,
278,
20364,
6121,
24771,
393,
338,
315,
1001,
29940,
322,
297,
278,
2319,
29892,
7343,
26078,
7888,
3815,
4076,
1663,
5861,
964,
920,
25700,
310,
1422,
15786,
10933,
2874,
322,
5849,
10174,
29889,
13,
23036,
5468,
29871,
29906,
29900,
29896,
29953,
29892,
530,
1552,
29874,
22552,
680,
280,
756,
1063,
278,
4485,
15133,
4007,
22137,
363,
278,
10393,
351,
6431,
29889,
2439,
8569,
338,
373,
7395,
9999,
292,
322,
278,
5802,
310,
1596,
10701,
29889,
17302,
916,
2712,
29892,
1183,
338,
14040,
363,
7344,
292,
278,
10393,
351,
4700,
322,
5264,
5745,
18196,
29889,
13,
29967,
557,
711,
4721,
2570,
29880,
338,
385,
12157,
9315,
9082,
297,
24992,
322,
27439,
936,
22557,
29889,
2860,
1985,
363,
3196,
2440,
297,
278,
4933,
5780,
13661,
29892,
540,
731,
701,
670,
1914,
5381,
297,
29871,
29896,
29929,
29929,
29900,
411,
385,
21639,
8034,
297,
278,
4038,
310,
5849,
322,
2874,
322,
22020,
17832,
372,
29889,
4001,
29871,
29906,
29900,
29900,
29929,
540,
338,
3455,
1089,
322,
2315,
6751,
15944,
310,
4721,
2570,
29880,
22557,
18156,
411,
1667,
8569,
373,
4933,
8492,
29892,
4266,
21083,
29892,
3345,
327,
573,
29892,
8805,
5864,
322,
16083,
15483,
29889,
450,
9819,
1583,
1754,
3234,
376,
2290,
519,
25300,
936,
20710,
1883,
4338,
363,
14338,
10916,
29908,
471,
805,
348,
1283,
297,
29871,
29906,
29900,
29896,
29953,
964,
263,
5004,
5001,
376,
6572,
2570,
29880,
1920,
305,
13296,
17925,
613,
5069,
14419,
12271,
338,
27850,
4721,
2570,
29880,
29889,
13,
1748,
5346,
317,
1338,
23162,
756,
1784,
2440,
29915,
7271,
297,
18540,
14650,
297,
1880,
734,
305,
23136,
29889,
940,
756,
884,
1063,
14040,
363,
16507,
29924,
8799,
292,
5786,
297,
670,
6413,
29889,
3237,
29889,
317,
1338,
23162,
338,
451,
871,
278,
3189,
29899,
11940,
261,
310,
383,
5421,
16507,
29924,
29892,
385,
826,
294,
6650,
3455,
1089,
29892,
541,
884,
278,
14582,
14645,
29949,
322,
8128,
315,
3035,
4511,
943,
22611,
916,
23387,
6851,
29889,
13,
29931,
18393,
20918,
2918,
29892,
14645,
29949,
310,
315,
3035,
1430,
3289,
28732,
317,
29889,
29878,
29889,
29880,
1696,
756,
1063,
263,
4509,
310,
315,
3035,
1430,
3289,
1951,
29871,
29906,
29900,
29900,
29929,
29889,
4001,
769,
540,
756,
4934,
5164,
11909,
29892,
3704,
16905,
8455,
363,
278,
758,
29899,
322,
1400,
29899,
29879,
2122,
14188,
349,
8322,
2929,
17925,
29914,
26092,
2519,
8697,
363,
12730,
322,
13616
] | 1,024 |
C4
| 0.193665 | 0.364023 | 0.626849 | 0.451308 |
April 2007 Monthly Archive
Does Media Reporting Of Suicides and Homicides Promote Copycats?
I won't give a detailed answer to this question here (it seems to be no), but there is an article making the reddit rounds now that I need to kill before it becomes another meme (like that other badly reported story about psychiatry.)
The article is from BMJ 2002, called Influences of the Media On Suicide, and it puts its conclusion right at the top:
Reporting and portrayal of suicidal behaviour in the media may have potentially negative influences and facilitate suicidal acts by people exposed to such stimuli. Recent systematic reviews by others and ourselves (unpublished) have found overwhelming evidence for such effects.1 (emphasis mine)
And it offers about 8 references in support. And so now every nut with a microphone can proclaim it loudly: it's the media's fault.
We may want to take a pause and examine these 8 references: none of them offer anything close to "overwhelming evidence." For example:
Reference 1-- the one directly cited for the above statements-- is indicative of the type of "overwhelming evidence" that exists. The study finds that media reporting of suicide is extensive and detailed, but not that there is a clear link to future suicides.
In the summary, the authors use phrases like, "dearth of literature," "evidence is less reliable," "few studies permitting/demonstrating [the link]," "does not demonstrate consistency," "many studies fail to demonstrate" over 11 times in the 3 pages describing the studies.
Despite this, they are sure the link exists-- but they don't actually show the link, they infer a link. The authors repeat phrases, "it is fair to conclude that the evidence suggests an association [exists]" "tends to suggest," "probably reasonable to regard the association is causal" 13 times in two pages. Under these criteria, it's reasonable to assume the Matrix is real.
Reference 3 (not even linked correctly) is a letter to the editor, describing two cases, where the method of suicide was affected by internet, but not the decision to commit suicide. And the methods were rather weak: one guy took two pills of castor oil, and the other woman tried to drink water. No, I'm not kidding.
Reference 5 is frequently cited in support of media's impact. It supposedly says that a TV show with a Tylenol OD caused more Tylenol ODs: 20% of these suicidal viewers said it influenced their decision to attempt suicide in the first week post broadcast. Maybe-- that 20% is really 6 people. And most had attempted Tylenol OD in the past. Oh, and the authors note that while 17% of the suicidal viewers' choice of Tylenol was influenced by the show, some of them chose not to use it because of the show.
Reference 12 is probably the most cited reference in this field. In 1978 Vienna built a subway, which soon became a popular method of suicide. So the government established guidelines for reporting-- specifically, that the method not be mentioned-- and subways suicides decreased by 80%. Fantastic. Overall suicide rates didn't change, though. Too bad.
So much for the "overwhelming evidence" for a soon to be media soundbite.
The article doesn't make a good case for media influencing the decision to kill yourself, though I'll admit that it may influence the method. And that's where it gets tricky.
It's important to make a distinction between copycat suicides and copycat homicides: more poeple die in the latter, and, let's postulate, they didn't want to die. That has to be part of the calculus in media reporting. Copying suicide by water (instead of pills) is different than copying a 30 person massacre (instead of killing, say, one person.)
But you have to weigh this against the societal costs. The solution offered in these articles is to restrict media reporting. I think we can agree that the media are neither liberal nor conservative, but sensationalists, their bias is titillation. But to allow anyone, especially government, to affect the content of reporting-- literally, the information we are allowed to have-- seems exactly the wrong solution to a problem which may not actually exist. (e.
|
[
3786,
29871,
29906,
29900,
29900,
29955,
23471,
368,
9000,
13,
25125,
8213,
13969,
292,
4587,
2166,
293,
2247,
322,
15089,
293,
2247,
9705,
866,
14187,
29883,
1446,
29973,
13,
29902,
2113,
29915,
29873,
2367,
263,
13173,
1234,
304,
445,
1139,
1244,
313,
277,
2444,
304,
367,
694,
511,
541,
727,
338,
385,
4274,
3907,
278,
337,
1289,
277,
364,
3885,
1286,
393,
306,
817,
304,
12088,
1434,
372,
7415,
1790,
286,
2004,
313,
4561,
393,
916,
28042,
8967,
5828,
1048,
11643,
7163,
719,
1846,
13,
1576,
4274,
338,
515,
350,
29924,
29967,
29871,
29906,
29900,
29900,
29906,
29892,
2000,
512,
25044,
2063,
310,
278,
8213,
1551,
2166,
293,
680,
29892,
322,
372,
15223,
967,
15997,
1492,
472,
278,
2246,
29901,
13,
13020,
292,
322,
2011,
764,
284,
310,
23921,
23670,
10468,
297,
278,
5745,
1122,
505,
19998,
8178,
7112,
2063,
322,
16089,
10388,
23921,
23670,
14741,
491,
2305,
19884,
304,
1316,
20436,
14549,
29889,
3599,
296,
1788,
2454,
21804,
491,
4045,
322,
20278,
313,
348,
5467,
3726,
29897,
505,
1476,
975,
1332,
295,
4056,
10757,
363,
1316,
9545,
29889,
29896,
313,
7278,
25101,
7903,
29897,
13,
2855,
372,
16688,
1048,
29871,
29947,
9282,
297,
2304,
29889,
1126,
577,
1286,
1432,
18254,
411,
263,
9200,
6710,
508,
9580,
8342,
372,
22526,
368,
29901,
372,
29915,
29879,
278,
5745,
29915,
29879,
12570,
29889,
13,
4806,
1122,
864,
304,
2125,
263,
19957,
322,
25917,
1438,
29871,
29947,
9282,
29901,
5642,
310,
963,
5957,
3099,
3802,
304,
376,
957,
1332,
295,
4056,
10757,
1213,
1152,
1342,
29901,
13,
7422,
29871,
29896,
489,
278,
697,
4153,
274,
1573,
363,
278,
2038,
9506,
489,
338,
4221,
1230,
310,
278,
1134,
310,
376,
957,
1332,
295,
4056,
10757,
29908,
393,
4864,
29889,
450,
6559,
14061,
393,
5745,
23415,
310,
23921,
680,
338,
20607,
322,
13173,
29892,
541,
451,
393,
727,
338,
263,
2821,
1544,
304,
5434,
23921,
2247,
29889,
13,
797,
278,
15837,
29892,
278,
15717,
671,
12216,
2129,
763,
29892,
376,
311,
28696,
310,
12845,
1699,
376,
5750,
5084,
338,
3109,
23279,
1699,
376,
29888,
809,
11898,
3635,
5367,
29914,
2310,
265,
710,
1218,
518,
1552,
1544,
1402,
29908,
376,
13221,
451,
22222,
5718,
3819,
1699,
376,
13011,
11898,
4418,
304,
22222,
29908,
975,
29871,
29896,
29896,
3064,
297,
278,
29871,
29941,
6515,
20766,
278,
11898,
29889,
13,
4002,
29886,
568,
445,
29892,
896,
526,
1854,
278,
1544,
4864,
489,
541,
896,
1016,
29915,
29873,
2869,
1510,
278,
1544,
29892,
896,
10115,
263,
1544,
29889,
450,
15717,
12312,
12216,
2129,
29892,
376,
277,
338,
6534,
304,
17668,
393,
278,
10757,
14661,
385,
15477,
518,
9933,
18017,
376,
29873,
1975,
304,
4368,
1699,
376,
771,
14815,
15590,
304,
4880,
278,
15477,
338,
3269,
284,
29908,
29871,
29896,
29941,
3064,
297,
1023,
6515,
29889,
7634,
1438,
16614,
29892,
372,
29915,
29879,
15590,
304,
5251,
278,
22513,
338,
1855,
29889,
13,
7422,
29871,
29941,
313,
1333,
1584,
9024,
5149,
29897,
338,
263,
5497,
304,
278,
6920,
29892,
20766,
1023,
4251,
29892,
988,
278,
1158,
310,
23921,
680,
471,
15201,
491,
8986,
29892,
541,
451,
278,
10608,
304,
9063,
23921,
680,
29889,
1126,
278,
3519,
892,
3265,
8062,
29901,
697,
1410,
29891,
3614,
1023,
282,
6090,
310,
4320,
272,
17182,
29892,
322,
278,
916,
6114,
1898,
304,
13748,
4094,
29889,
1939,
29892,
306,
29915,
29885,
451,
413,
2205,
292,
29889,
13,
7422,
29871,
29945,
338,
13672,
274,
1573,
297,
2304,
310,
5745,
29915,
29879,
10879,
29889,
739,
7424,
368,
4083,
393,
263,
5648,
1510,
411,
263,
9656,
2435,
324,
438,
29928,
8581,
901,
9656,
2435,
324,
438,
29928,
29879,
29901,
29871,
29906,
29900,
29995,
310,
1438,
23921,
23670,
1776,
414,
1497,
372,
28482,
1009,
10608,
304,
4218,
23921,
680,
297,
278,
937,
4723,
1400,
12672,
29889,
7198,
489,
393,
29871,
29906,
29900,
29995,
338,
2289,
29871,
29953,
2305,
29889,
1126,
1556,
750,
16388,
9656,
2435,
324,
438,
29928,
297,
278,
4940,
29889,
6439,
29892,
322,
278,
15717,
4443,
393,
1550,
29871,
29896,
29955,
29995,
310,
278,
23921,
23670,
1776,
414,
29915,
7348,
310,
9656,
2435,
324,
471,
28482,
491,
278,
1510,
29892,
777,
310,
963,
12784,
451,
304,
671,
372,
1363,
310,
278,
1510,
29889,
13,
7422,
29871,
29896,
29906,
338,
3117,
278,
1556,
274,
1573,
3407,
297,
445,
1746,
29889,
512,
29871,
29896,
29929,
29955,
29947,
25309,
4240,
263,
1014,
1582,
29892,
607,
4720,
3897,
263,
5972,
1158,
310,
23921,
680,
29889,
1105,
278,
5874,
7841,
1410,
10652,
1475,
363,
23415,
489,
10816,
29892,
393,
278,
1158,
451,
367,
5276,
489,
322,
1014,
1994,
23921,
2247,
9263,
1463,
491,
29871,
29947,
29900,
15543,
18089,
6288,
29889,
6811,
497,
23921,
680,
19257,
3282,
29915,
29873,
1735,
29892,
2466,
29889,
1763,
29877,
4319,
29889,
13,
6295,
1568,
363,
278,
376,
957,
1332,
295,
4056,
10757,
29908,
363,
263,
4720,
304,
367,
5745,
6047,
29890,
568,
29889,
13,
1576,
4274,
1838,
29915,
29873,
1207,
263,
1781,
1206,
363,
5745,
13787,
3277,
278,
10608,
304,
12088,
7535,
29892,
2466,
306,
29915,
645,
20000,
393,
372,
1122,
9949,
278,
1158,
29889,
1126,
393,
29915,
29879,
988,
372,
4947,
28722,
29889,
13,
3112,
29915,
29879,
4100,
304,
1207,
263,
21578,
1546,
3509,
4117,
23921,
2247,
322,
3509,
4117,
3632,
293,
2247,
29901,
901,
772,
29872,
552,
762,
297,
278,
7480,
29892,
322,
29892,
1235,
29915,
29879,
1400,
5987,
29892,
896,
3282,
29915,
29873,
864,
304,
762,
29889,
2193,
756,
304,
367,
760,
310,
278,
24282,
297,
5745,
23415,
29889,
14187,
292,
23921,
680,
491,
4094,
313,
2611,
1479,
310,
282,
6090,
29897,
338,
1422,
1135,
17596,
263,
29871,
29941,
29900,
2022,
4158,
21160,
313,
2611,
1479,
310,
23393,
29892,
1827,
29892,
697,
2022,
1846,
13,
6246,
366,
505,
304,
591,
1141,
445,
2750,
278,
5374,
26310,
21544,
29889,
450,
1650,
12520,
297,
1438,
7456,
338,
304,
9250,
5745,
23415,
29889,
306,
1348,
591,
508,
8661,
393,
278,
5745,
526,
9561,
26054,
3643,
8976,
1230,
29892,
541,
4771,
1288,
2879,
29892,
1009,
24003,
338,
4329,
453,
362,
29889,
1205,
304,
2758,
5019,
29892,
7148,
5874,
29892,
304,
6602,
278,
2793,
310,
23415,
489,
22830,
29892,
278,
2472,
591,
526,
6068,
304,
505,
489,
2444,
3721,
278,
2743,
1650,
304,
263,
1108,
607,
1122,
451,
2869,
1863,
29889,
313,
29872,
29889
] | 1,024 |
CommonCrawl
| -0.179889 | 0.208289 | 0.664507 | 0.192009 |
g. I know it seems prurient, but I actually want to know all the details of David Kelly's suicide.)
Not to mention that if you say the media are partly responsible, then you're saying that you're less responsible.
(More on copycat suicides here, and on university suicides/copycats here and here.)
A Quick Word on Porn's Effect On Your Penis
A reader commented that I was minimizing porn's negative effect on women, that ubiquitous internet porn has damaged womens' psyches irreparably. That it makes women have to conform to some impossible standard.
Nay.
Porn is not the problem. I'm not saying it's a tremendous boon to society, but you can't blame porn for failing relationships, the pressure on women to attain impossible standards of appearance and performance; and male disinterest in normal sexual relationships.
Certainly porn puts pressure on women, but the effect is not directly from porn, or even from men. Here's an example that the reader offered: porn forces the women to shave. Not exactly-- they want to shave. Why they want to is a cultural discussion, but it isn't because men are explicitly commanding them to do so.
Certainly, porn has affected men. Ok, women want to shave; why do men suddenly want to, also? And, I'd expect that a frequent porn user (whatever that exactly is) might have some difficulty with arousal in normal (or repeat) circumstances.
But there's a greater problem that can't be blamed on porn. Every comic since Marx (Groucho, not the other nut) has joked about how men want sex and women don't. But in the past three or four years, I've heard comics make the opposite jokes: women want it, men could just as easily pass it up. Men are disinterested in sex with their established partners. As comic Mark Maron put it, "[I prefer masturbation because sex] takes up too much energy and it involves other people." Men always are ready for new women, but what happens to sex with your partner over time? Sure, ordinarily it may decline a little, but this is different: this is male disinterest, "lack of energy," lack of motivation to keep a connection with one's partner alive. The penis may still go up-- but everything else is gone.
Let's face it, porn may make women feel inadequate, but how the hell adequate can a woman feel if her boyfriend/husband would rather watch TV than have sex? "But I'm tired." How tired could you possibly be?
So there are two parts to the problem. The easy, and smaller, part is media/porn objectification of women, and its effects on women and men. But the second, more crucial part is male "impotence" (metaphorical) and apathy. Let me be clear about this: porn might magnify this effect; but it doesn't cause it.
I know no girl in the world is going to believe this, but it's true: if you ask the average guy over 30 if they'd rather be with a girl they have been with many times before or masturbate, they'll pick masturbate. You know why? Because their soul bailed out when they were 15-- because they are narcissists. What in life is worth aspiring to? You don't feel a part of anything bigger, everything seems distant, unreal. Everyone is waiting for something to happen, for their life to "start"-- they're 40 and they're still waiting. (As Mike Birbiglia joked, "I'm not going to get married until I'm absolutely certain nothing else good can happen in my life.") Concepts like loyalty don't even get a token nod, because today they seem outright preposterous.
And men have a distorted view of what it means to be loved. They want to be loved not for who they are, but who they think they are. "I'm an actor." "I'm a major force in WoW." "I'm a fiscal conservative but a social liberal." What he wants is his girlfriend to say, "I love him because he is such an intellectual, he knows so much about politics." What he doesn't want is her to say, "I love him because he's good to me."
"Sure it'd be better to be with a
|
[
29887,
29889,
306,
1073,
372,
2444,
544,
332,
993,
29892,
541,
306,
2869,
864,
304,
1073,
599,
278,
4902,
310,
4699,
21872,
29915,
29879,
23921,
680,
1846,
13,
3664,
304,
3585,
393,
565,
366,
1827,
278,
5745,
526,
22669,
14040,
29892,
769,
366,
29915,
276,
5934,
393,
366,
29915,
276,
3109,
14040,
29889,
13,
29898,
20761,
373,
3509,
4117,
23921,
2247,
1244,
29892,
322,
373,
16372,
23921,
2247,
29914,
8552,
29883,
1446,
1244,
322,
1244,
1846,
13,
29909,
26141,
10803,
373,
349,
1398,
29915,
29879,
26475,
1551,
3575,
7363,
275,
13,
29909,
9591,
19952,
393,
306,
471,
6260,
5281,
1277,
29876,
29915,
29879,
8178,
2779,
373,
5866,
29892,
393,
318,
5365,
28358,
681,
8986,
1277,
29876,
756,
5625,
4063,
281,
290,
575,
29915,
11643,
267,
19950,
862,
2197,
29889,
2193,
372,
3732,
5866,
505,
304,
14670,
304,
777,
9301,
3918,
29889,
13,
29940,
388,
29889,
13,
29925,
1398,
338,
451,
278,
1108,
29889,
306,
29915,
29885,
451,
5934,
372,
29915,
29879,
263,
14586,
355,
681,
1045,
265,
304,
12459,
29892,
541,
366,
508,
29915,
29873,
1999,
420,
1277,
29876,
363,
17581,
21702,
29892,
278,
12959,
373,
5866,
304,
1098,
475,
9301,
20801,
310,
10097,
322,
4180,
29936,
322,
14263,
766,
1639,
342,
297,
4226,
18287,
21702,
29889,
13,
29907,
13946,
368,
1277,
29876,
15223,
12959,
373,
5866,
29892,
541,
278,
2779,
338,
451,
4153,
515,
1277,
29876,
29892,
470,
1584,
515,
1757,
29889,
2266,
29915,
29879,
385,
1342,
393,
278,
9591,
12520,
29901,
1277,
29876,
8249,
278,
5866,
304,
528,
1351,
29889,
2216,
3721,
489,
896,
864,
304,
528,
1351,
29889,
3750,
896,
864,
304,
338,
263,
16375,
10679,
29892,
541,
372,
3508,
29915,
29873,
1363,
1757,
526,
9479,
1899,
292,
963,
304,
437,
577,
29889,
13,
29907,
13946,
368,
29892,
1277,
29876,
756,
15201,
1757,
29889,
3674,
29892,
5866,
864,
304,
528,
1351,
29936,
2020,
437,
1757,
11584,
864,
304,
29892,
884,
29973,
1126,
29892,
306,
29915,
29881,
2149,
393,
263,
17091,
1277,
29876,
1404,
313,
1332,
5564,
393,
3721,
338,
29897,
1795,
505,
777,
14656,
411,
564,
681,
284,
297,
4226,
313,
272,
12312,
29897,
14209,
29889,
13,
6246,
727,
29915,
29879,
263,
7621,
1108,
393,
508,
29915,
29873,
367,
1999,
2795,
373,
1277,
29876,
29889,
7569,
419,
293,
1951,
28579,
313,
3338,
283,
1859,
29892,
451,
278,
916,
18254,
29897,
756,
432,
12504,
1048,
920,
1757,
864,
7916,
322,
5866,
1016,
29915,
29873,
29889,
1205,
297,
278,
4940,
2211,
470,
3023,
2440,
29892,
306,
29915,
345,
6091,
419,
1199,
1207,
278,
11564,
432,
23195,
29901,
5866,
864,
372,
29892,
1757,
1033,
925,
408,
5948,
1209,
372,
701,
29889,
7567,
526,
766,
1639,
2868,
297,
7916,
411,
1009,
7841,
22056,
29889,
1094,
419,
293,
4485,
1085,
265,
1925,
372,
29892,
14704,
29902,
5821,
286,
579,
9265,
362,
1363,
7916,
29962,
4893,
701,
2086,
1568,
5864,
322,
372,
20789,
916,
2305,
1213,
7567,
2337,
526,
7960,
363,
716,
5866,
29892,
541,
825,
5930,
304,
7916,
411,
596,
18096,
975,
931,
29973,
18585,
29892,
22420,
6275,
372,
1122,
4845,
457,
263,
2217,
29892,
541,
445,
338,
1422,
29901,
445,
338,
14263,
766,
1639,
342,
29892,
376,
2364,
310,
5864,
1699,
10225,
310,
17385,
362,
304,
3013,
263,
3957,
411,
697,
29915,
29879,
18096,
18758,
29889,
450,
6584,
275,
1122,
1603,
748,
701,
489,
541,
4129,
1683,
338,
7695,
29889,
13,
12024,
29915,
29879,
3700,
372,
29892,
1277,
29876,
1122,
1207,
5866,
4459,
297,
1943,
339,
403,
29892,
541,
920,
278,
23927,
19967,
339,
403,
508,
263,
6114,
4459,
565,
902,
8023,
18326,
29914,
14116,
4980,
723,
3265,
6505,
5648,
1135,
505,
7916,
29973,
376,
6246,
306,
29915,
29885,
23407,
1213,
1128,
23407,
1033,
366,
10075,
367,
29973,
13,
6295,
727,
526,
1023,
5633,
304,
278,
1108,
29889,
450,
4780,
29892,
322,
7968,
29892,
760,
338,
5745,
29914,
29886,
1398,
1203,
2450,
310,
5866,
29892,
322,
967,
9545,
373,
5866,
322,
1757,
29889,
1205,
278,
1473,
29892,
901,
7618,
1455,
760,
338,
14263,
376,
6574,
327,
663,
29908,
313,
2527,
481,
2015,
936,
29897,
322,
263,
2084,
29891,
29889,
2803,
592,
367,
2821,
1048,
445,
29901,
1277,
29876,
1795,
9119,
1598,
445,
2779,
29936,
541,
372,
1838,
29915,
29873,
4556,
372,
29889,
13,
29902,
1073,
694,
7826,
297,
278,
3186,
338,
2675,
304,
4658,
445,
29892,
541,
372,
29915,
29879,
1565,
29901,
565,
366,
2244,
278,
6588,
1410,
29891,
975,
29871,
29941,
29900,
565,
896,
29915,
29881,
3265,
367,
411,
263,
7826,
896,
505,
1063,
411,
1784,
3064,
1434,
470,
286,
579,
9265,
403,
29892,
896,
29915,
645,
5839,
286,
579,
9265,
403,
29889,
887,
1073,
2020,
29973,
7311,
1009,
10752,
9922,
2356,
714,
746,
896,
892,
29871,
29896,
29945,
489,
1363,
896,
526,
302,
5666,
790,
2879,
29889,
1724,
297,
2834,
338,
7088,
7051,
8491,
304,
29973,
887,
1016,
29915,
29873,
4459,
263,
760,
310,
3099,
16600,
29892,
4129,
2444,
21188,
29892,
443,
6370,
29889,
7569,
650,
338,
10534,
363,
1554,
304,
3799,
29892,
363,
1009,
2834,
304,
376,
2962,
29908,
489,
896,
29915,
276,
29871,
29946,
29900,
322,
896,
29915,
276,
1603,
10534,
29889,
313,
2887,
12828,
17853,
3752,
4456,
432,
12504,
29892,
376,
29902,
29915,
29885,
451,
2675,
304,
679,
8300,
2745,
306,
29915,
29885,
13312,
3058,
3078,
1683,
1781,
508,
3799,
297,
590,
2834,
23157,
1281,
1547,
29879,
763,
28108,
1017,
1016,
29915,
29873,
1584,
679,
263,
5993,
18778,
29892,
1363,
9826,
896,
2833,
714,
1266,
758,
2490,
261,
681,
29889,
13,
2855,
1757,
505,
263,
1320,
18054,
1776,
310,
825,
372,
2794,
304,
367,
18012,
29889,
2688,
864,
304,
367,
18012,
451,
363,
1058,
896,
526,
29892,
541,
1058,
896,
1348,
896,
526,
29889,
376,
29902,
29915,
29885,
385,
11339,
1213,
376,
29902,
29915,
29885,
263,
4655,
4889,
297,
14962,
29956,
1213,
376,
29902,
29915,
29885,
263,
16436,
1052,
8976,
1230,
541,
263,
5264,
26054,
1213,
1724,
540,
10753,
338,
670,
7826,
18326,
304,
1827,
29892,
376,
29902,
5360,
1075,
1363,
540,
338,
1316,
385,
29762,
29892,
540,
9906,
577,
1568,
1048,
22661,
1213,
1724,
540,
1838,
29915,
29873,
864,
338,
902,
304,
1827,
29892,
376,
29902,
5360,
1075,
1363,
540,
29915,
29879,
1781,
304,
592,
1213,
13,
29908,
29903,
545,
372,
29915,
29881,
367,
2253,
304,
367,
411,
263
] | 1,024 |
CommonCrawl
| 0.158279 | -0.415459 | -0.899184 | -0.593954 |
girl, but when are you actually ever with a girl? They don't want you, they want what you represent-- a good job, security, to be taken care of, a big penis." It doesn't occur to them that the woman who doesn't want these things in her man might be the one to avoid?
I suspect-- I haven't been able to do the survey-- that even sex is a form of masturbation for these guys. That they see you, but they don't see you. The arm, the breast, the hip, all these become fetishized and transport him to another world.
Our birth rate is 2.1; France 1.7; Spain 1.3; Russia 1.3. In two generations, there will be 1/2 as many Spaniards, excluding immigration. We can't even get it up long enough to procreate. That's not porn's fault. It doesn't help, sure, having the internet's tubes tied isn't going to fix that problem. Men are becoming less interested in establishing meaningful relationships with other people as an ultimate goal than in inventing identities for themselves.
Permalink | Comments (47)
"Inflammable Means Flammable? What A Country!"
You're going to say that I made this up. I am not making this up. I wish I was, because then I could say,"stop making things up," and everything would be ok. But it really happened, and I can't stop things from really happening. So I drink.
This is the conversation I had with another psychiatrist. He is wearing a Ermenegildo Zegna suit. It fits him well.
I say, "... so if I have him [hypothetical bipolar depressed patient] on Depakote and Seroquel, once he's stable I try to reduce the dose, or even stop one of the medications."
He shakes his head. "I would never stop the mood stabilizer."
"Why-- wait, which one's the mood stabilizer?"
"The Depakote."
"But how do you know it wasn't the Seroquel? That's the problem with starting two drugs immediately, you don't know which one worked."
"Well, you need them both. Especially if you're adding an antidepressant."
"What antidepressant?"
"Seroquel."
"Seroquel's an antidepressant?"
"In this case it is, you're using it for bipolar depression."
I blink my eyes, to make sure I still can.
"Then what's the Depakote?"
"The Depakote is the mood stabilizer."
"But how do you know the mood stabilizer isn't the Seroquel?"
"The Seroquel is the antipsychotic."
A voice tells me to stab him.
"Why can't you just use the Seroquel-- the antipsychotic- slash- antidepressant-- alone as a first try? What benefit does the Depakote give you over the Seroquel?"
He says it slowly, enunciates, because he deduces that I don't speak English. "Because the Depakote is the mood stabilizer..."
He looks at me. I look at him. He is wearing a yellow Bolgheri tie. In my mind it is on fire.
"...Besides, if you use Seroquel off label like that, you'll get sued." Blaming lawyers, the last refuge of the incompetent. "And for what? Why take the risk?"
It's at this point I realize he's not wearing socks. "If you're going to use that logic, Depakote isn't indicated for mood stabilization, either."
He looks at me incredulously, then suddenly he realizes something. "Oh, okay, right, I see what you're saying now... but at least Depakote has FDA approval for Bipolar Disorder." QED. He's very happy now. I can't find the waiter. Why is my drink empty?
There's an uncomfortable pause. He wants to show me he's a skeptic, too, that he's carefully pondered these issues.
"I have a theory, have you ever used meclizine (a drug for vertigo) as a mood stabilizer?"
"No, I had never heard
|
[
7826,
29892,
541,
746,
526,
366,
2869,
3926,
411,
263,
7826,
29973,
2688,
1016,
29915,
29873,
864,
366,
29892,
896,
864,
825,
366,
2755,
489,
263,
1781,
4982,
29892,
6993,
29892,
304,
367,
4586,
2562,
310,
29892,
263,
4802,
6584,
275,
1213,
739,
1838,
29915,
29873,
6403,
304,
963,
393,
278,
6114,
1058,
1838,
29915,
29873,
864,
1438,
2712,
297,
902,
767,
1795,
367,
278,
697,
304,
4772,
29973,
13,
29902,
12326,
489,
306,
7359,
29915,
29873,
1063,
2221,
304,
437,
278,
18994,
489,
393,
1584,
7916,
338,
263,
883,
310,
286,
579,
9265,
362,
363,
1438,
18239,
29889,
2193,
896,
1074,
366,
29892,
541,
896,
1016,
29915,
29873,
1074,
366,
29889,
450,
5075,
29892,
278,
24207,
29892,
278,
21464,
29892,
599,
1438,
4953,
27431,
728,
1891,
322,
8608,
1075,
304,
1790,
3186,
29889,
13,
29949,
332,
12060,
6554,
338,
29871,
29906,
29889,
29896,
29936,
3444,
29871,
29896,
29889,
29955,
29936,
13616,
29871,
29896,
29889,
29941,
29936,
12710,
29871,
29896,
29889,
29941,
29889,
512,
1023,
1176,
800,
29892,
727,
674,
367,
29871,
29896,
29914,
29906,
408,
1784,
1706,
3270,
3163,
29892,
429,
22368,
5198,
16783,
29889,
1334,
508,
29915,
29873,
1584,
679,
372,
701,
1472,
3307,
304,
410,
3258,
29889,
2193,
29915,
29879,
451,
1277,
29876,
29915,
29879,
12570,
29889,
739,
1838,
29915,
29873,
1371,
29892,
1854,
29892,
2534,
278,
8986,
29915,
29879,
23131,
267,
21351,
3508,
29915,
29873,
2675,
304,
2329,
393,
1108,
29889,
7567,
526,
14171,
3109,
8852,
297,
10127,
292,
6593,
1319,
21702,
411,
916,
2305,
408,
385,
8494,
6490,
7306,
1135,
297,
11817,
292,
2893,
1907,
363,
6053,
29889,
13,
15737,
284,
682,
891,
461,
29879,
313,
29946,
29955,
29897,
13,
29908,
15403,
4850,
519,
2191,
550,
2379,
4850,
519,
29973,
1724,
319,
15456,
3850,
13,
3492,
29915,
276,
2675,
304,
1827,
393,
306,
1754,
445,
701,
29889,
306,
626,
451,
3907,
445,
701,
29889,
306,
6398,
306,
471,
29892,
1363,
769,
306,
1033,
1827,
1699,
9847,
3907,
2712,
701,
1699,
322,
4129,
723,
367,
3431,
29889,
1205,
372,
2289,
9559,
29892,
322,
306,
508,
29915,
29873,
5040,
2712,
515,
2289,
10464,
29889,
1105,
306,
13748,
29889,
13,
4013,
338,
278,
14983,
306,
750,
411,
1790,
11643,
7163,
2021,
29889,
940,
338,
591,
4362,
263,
1425,
1527,
387,
789,
29877,
796,
387,
1056,
14726,
29889,
739,
23994,
1075,
1532,
29889,
13,
29902,
1827,
29892,
29804,
577,
565,
306,
505,
1075,
518,
29882,
1478,
720,
300,
936,
289,
666,
10170,
316,
13120,
16500,
29962,
373,
10034,
557,
866,
322,
317,
1489,
13295,
29892,
2748,
540,
29915,
29879,
13714,
306,
1018,
304,
10032,
278,
437,
344,
29892,
470,
1584,
5040,
697,
310,
278,
13589,
800,
1213,
13,
3868,
528,
6926,
670,
2343,
29889,
376,
29902,
723,
2360,
5040,
278,
286,
2092,
16160,
3950,
1213,
13,
29908,
11008,
489,
4480,
29892,
607,
697,
29915,
29879,
278,
286,
2092,
16160,
3950,
3026,
13,
29908,
1576,
10034,
557,
866,
1213,
13,
29908,
6246,
920,
437,
366,
1073,
372,
9007,
29915,
29873,
278,
317,
1489,
13295,
29973,
2193,
29915,
29879,
278,
1108,
411,
6257,
1023,
5883,
3174,
7389,
29892,
366,
1016,
29915,
29873,
1073,
607,
697,
3796,
1213,
13,
29908,
11284,
29892,
366,
817,
963,
1716,
29889,
3423,
25009,
565,
366,
29915,
276,
4417,
385,
3677,
680,
2139,
424,
1213,
13,
29908,
5618,
3677,
680,
2139,
424,
3026,
13,
29908,
29903,
1489,
13295,
1213,
13,
29908,
29903,
1489,
13295,
29915,
29879,
385,
3677,
680,
2139,
424,
3026,
13,
29908,
797,
445,
1206,
372,
338,
29892,
366,
29915,
276,
773,
372,
363,
289,
666,
10170,
316,
2590,
1213,
13,
29902,
1999,
682,
590,
5076,
29892,
304,
1207,
1854,
306,
1603,
508,
29889,
13,
29908,
11760,
825,
29915,
29879,
278,
10034,
557,
866,
3026,
13,
29908,
1576,
10034,
557,
866,
338,
278,
286,
2092,
16160,
3950,
1213,
13,
29908,
6246,
920,
437,
366,
1073,
278,
286,
2092,
16160,
3950,
3508,
29915,
29873,
278,
317,
1489,
13295,
3026,
13,
29908,
1576,
317,
1489,
13295,
338,
278,
9418,
567,
3376,
13574,
1213,
13,
29909,
7314,
10603,
592,
304,
380,
370,
1075,
29889,
13,
29908,
11008,
508,
29915,
29873,
366,
925,
671,
278,
317,
1489,
13295,
489,
278,
9418,
567,
3376,
13574,
29899,
24765,
29899,
3677,
680,
2139,
424,
489,
7432,
408,
263,
937,
1018,
29973,
1724,
14169,
947,
278,
10034,
557,
866,
2367,
366,
975,
278,
317,
1489,
13295,
3026,
13,
3868,
4083,
372,
14205,
29892,
427,
11173,
1078,
29892,
1363,
540,
21049,
778,
393,
306,
1016,
29915,
29873,
7726,
4223,
29889,
376,
29933,
5658,
278,
10034,
557,
866,
338,
278,
286,
2092,
16160,
3950,
17794,
13,
3868,
3430,
472,
592,
29889,
306,
1106,
472,
1075,
29889,
940,
338,
591,
4362,
263,
13328,
8922,
29887,
2276,
29875,
22134,
29889,
512,
590,
3458,
372,
338,
373,
3974,
29889,
13,
29908,
856,
29933,
267,
2247,
29892,
565,
366,
671,
317,
1489,
13295,
1283,
3858,
763,
393,
29892,
366,
29915,
645,
679,
480,
287,
1213,
3164,
11500,
4307,
29891,
414,
29892,
278,
1833,
25447,
310,
278,
297,
2388,
300,
296,
29889,
376,
2855,
363,
825,
29973,
3750,
2125,
278,
12045,
3026,
13,
3112,
29915,
29879,
472,
445,
1298,
306,
16289,
540,
29915,
29879,
451,
591,
4362,
577,
4684,
29889,
376,
3644,
366,
29915,
276,
2675,
304,
671,
393,
5900,
29892,
10034,
557,
866,
3508,
29915,
29873,
18694,
363,
286,
2092,
16160,
2133,
29892,
2845,
1213,
13,
3868,
3430,
472,
592,
29811,
352,
5794,
29892,
769,
11584,
540,
1855,
7093,
1554,
29889,
376,
9048,
29892,
20759,
29892,
1492,
29892,
306,
1074,
825,
366,
29915,
276,
5934,
1286,
856,
541,
472,
3203,
10034,
557,
866,
756,
383,
7698,
2134,
791,
363,
350,
666,
10170,
3295,
2098,
1213,
660,
3352,
29889,
940,
29915,
29879,
1407,
9796,
1286,
29889,
306,
508,
29915,
29873,
1284,
278,
4480,
261,
29889,
3750,
338,
590,
13748,
4069,
29973,
13,
8439,
29915,
29879,
385,
443,
510,
3921,
519,
19957,
29889,
940,
10753,
304,
1510,
592,
540,
29915,
29879,
263,
18109,
23000,
29892,
2086,
29892,
393,
540,
29915,
29879,
16112,
282,
8417,
287,
1438,
5626,
29889,
13,
29908,
29902,
505,
263,
6368,
29892,
505,
366,
3926,
1304,
592,
695,
466,
457,
313,
29874,
15721,
363,
4837,
5973,
29897,
408,
263,
286,
2092,
16160,
3950,
3026,
13,
29908,
3782,
29892,
306,
750,
2360,
6091
] | 1,024 |
CommonCrawl
| -0.439241 | -0.211869 | -0.917116 | -0.896418 |
of that."
"I haven't tried it either, but it might make sense: meclizine stabilizes your balance, so perhaps it could stabilize your moods?"
I want to call the State Board of Medicine but realize I'm in a different state and I don't have the number in my phone. "I doubt the insurance companies would ever cover it."
He slowly, purposefully, nods his head. "Fucking meddling managed care."
I tell people all the time, don't get sick, don't ever get sick, but no one listens to me.
Is Cho The Question?
Quick speculation. As 13 year old kids like this usually create an identity from multiple sources- so far we're pretty sure the main movie was Oldboy-- I wonder if he didn't also identify with the Question (the comic). He signed his name as a question mark, he wore the same baseball cap, gloves with short sleeve shirt. And, in case you weren't once a lost male adolescent, the Question is Vic Sage, ex-philosophy major turned TV reporter who is also a vigilante. And the 30+ issues are all about his struggle with identity.
None of this is relevant to why he did it, of course. It's fun to speculate, but of no predictive value. These are costumes, roles. They just help him "get into character," which in this case is a man alone fighting injustice, greed, and abuse, using whatever tools he has. Of course, he was fighting college kids, and his tools were semi-automatics, but who am I to dispel a good illusion?
Finally, let me reiterate that the presence or absence of mental illness is irrelevant, at least as far as prediction goes. So let's say he was a violent schizophrenic. Would you jail him before he commits a crime, just in case? Bush does that with Guantanamo, you know. Do you send him to a psych hospital, like they do/did in the USSR? That process will be magnificently abused by your government, I assure you.
More posts on Cho here, here, here, and here.
Desmond's Teleological Suspension of The Ethical-- Or My Novel?
A few weeks ago I had used a Lost storyline to explain my own view that we pick our own identities, rather than have them given to us through either genetics or the environment. I made Desmond the Abraham in Kierkegaard's Fear And Trembling.
The crux of the episode and the analogy is that Desmond thinks he can see the future, and see that Charlie will die. But Desmond then makes a vital moral step: he decides that it is also his responsibility to keep this character alive. (Quoting myself:)
The real question is why Desmond actually believes such a choice exists. How does he think he knows the future? Anyone else in his shoes would have come to a very different, more logical, conclusion: this is insane. What, he can predict the future? Worse: what, he's the only reason Charlie is alive? He's so-- necessary? Isn't that narcissism?
If Desmond knew he could predict the future-- if it was a fact that he could predict the future-- then saving Charlie would have little moral heroism. Any fool a step up from absolute evil would have tried to prevent a horrible outcome if he knew for certain what was going to happen.
What made Desmond worthy of admiration was, exactly, that he did not know for sure he could predict the future. He took it on faith that he could, and then proceeded to live his entire life based on this single, faith based, assumption.
That was Feb. 15. Strangely, I just saw last week's episode, in which Desmond turns out to have once been a a monk, and he has a discussion about Abraham and Isaac with another monk; the wine they make is named Moriah; and later Desmond explicitly references the test of faith-- straight out of Fear And Trembling.
I suppose this could be a coincidence.
Another possibility is the writers read and and love this blog and have gone and reshot future episodes based on my ideas. HA!
Another possibility is I write for Lost. HA HA!
But the final possibility is the most likely, and it has less to do with Lost and more to do with the direction of our fiction.
Pre 9-11, fiction, and especially sci-
|
[
310,
393,
1213,
13,
29908,
29902,
7359,
29915,
29873,
1898,
372,
2845,
29892,
541,
372,
1795,
1207,
4060,
29901,
592,
695,
466,
457,
16160,
7093,
596,
17346,
29892,
577,
6060,
372,
1033,
16160,
675,
596,
286,
2092,
29879,
3026,
13,
29902,
864,
304,
1246,
278,
4306,
12590,
310,
27529,
541,
16289,
306,
29915,
29885,
297,
263,
1422,
2106,
322,
306,
1016,
29915,
29873,
505,
278,
1353,
297,
590,
9008,
29889,
376,
29902,
7404,
278,
1663,
18541,
14582,
723,
3926,
4612,
372,
1213,
13,
3868,
14205,
29892,
6437,
3730,
29892,
18778,
29879,
670,
2343,
29889,
376,
29943,
2707,
292,
1612,
29881,
1847,
8745,
2562,
1213,
13,
29902,
2649,
2305,
599,
278,
931,
29892,
1016,
29915,
29873,
679,
17319,
29892,
1016,
29915,
29873,
3926,
679,
17319,
29892,
541,
694,
697,
1051,
575,
304,
592,
29889,
13,
3624,
14542,
450,
894,
29973,
13,
2182,
860,
1580,
2785,
29889,
1094,
29871,
29896,
29941,
1629,
2030,
413,
4841,
763,
445,
5491,
1653,
385,
10110,
515,
2999,
8974,
29899,
577,
2215,
591,
29915,
276,
5051,
1854,
278,
1667,
14064,
471,
8198,
19415,
489,
306,
4997,
565,
540,
3282,
29915,
29873,
884,
12439,
411,
278,
894,
313,
1552,
419,
293,
467,
940,
8794,
670,
1024,
408,
263,
1139,
2791,
29892,
540,
26758,
278,
1021,
21573,
2117,
29892,
15482,
1960,
411,
3273,
12844,
29872,
345,
528,
2728,
29889,
1126,
29892,
297,
1206,
366,
2949,
264,
29915,
29873,
2748,
263,
5714,
14263,
594,
6544,
1760,
29892,
278,
894,
338,
13423,
317,
482,
29892,
429,
29899,
561,
309,
7708,
29891,
4655,
6077,
5648,
1634,
9555,
1058,
338,
884,
263,
14877,
309,
1647,
29889,
1126,
278,
29871,
29941,
29900,
29974,
5626,
526,
599,
1048,
670,
21117,
411,
10110,
29889,
13,
8516,
310,
445,
338,
8018,
304,
2020,
540,
1258,
372,
29892,
310,
3236,
29889,
739,
29915,
29879,
2090,
304,
1580,
5987,
29892,
541,
310,
694,
8500,
573,
995,
29889,
4525,
526,
3438,
9351,
29892,
16178,
29889,
2688,
925,
1371,
1075,
376,
657,
964,
2931,
1699,
607,
297,
445,
1206,
338,
263,
767,
7432,
17770,
297,
5143,
625,
29892,
1395,
287,
29892,
322,
633,
1509,
29892,
773,
6514,
8492,
540,
756,
29889,
4587,
3236,
29892,
540,
471,
17770,
12755,
413,
4841,
29892,
322,
670,
8492,
892,
12647,
29899,
17405,
271,
1199,
29892,
541,
1058,
626,
306,
304,
12272,
295,
263,
1781,
4486,
3958,
29973,
13,
12881,
635,
29892,
1235,
592,
337,
1524,
403,
393,
278,
10122,
470,
18070,
310,
19119,
4486,
2264,
338,
28190,
29892,
472,
3203,
408,
2215,
408,
18988,
5771,
29889,
1105,
1235,
29915,
29879,
1827,
540,
471,
263,
24252,
1364,
466,
459,
13608,
293,
29889,
10878,
366,
432,
737,
1075,
1434,
540,
25741,
263,
17268,
29892,
925,
297,
1206,
29973,
24715,
947,
393,
411,
2088,
424,
273,
10178,
29892,
366,
1073,
29889,
1938,
366,
3638,
1075,
304,
263,
11643,
13457,
29892,
763,
896,
437,
29914,
18361,
297,
278,
17676,
29934,
29973,
2193,
1889,
674,
367,
29154,
2705,
633,
3880,
491,
596,
5874,
29892,
306,
1223,
545,
366,
29889,
13,
20761,
11803,
373,
14542,
1244,
29892,
1244,
29892,
1244,
29892,
322,
1244,
29889,
13,
4002,
8315,
29915,
29879,
9699,
5996,
9511,
29886,
2673,
310,
450,
13772,
936,
489,
1394,
1619,
2864,
295,
29973,
13,
29909,
2846,
11405,
8020,
306,
750,
1304,
263,
25133,
5828,
1220,
304,
5649,
590,
1914,
1776,
393,
591,
5839,
1749,
1914,
2893,
1907,
29892,
3265,
1135,
505,
963,
2183,
304,
502,
1549,
2845,
2531,
300,
1199,
470,
278,
5177,
29889,
306,
1754,
2726,
8315,
278,
24763,
297,
476,
631,
29895,
2442,
538,
29915,
29879,
383,
799,
1126,
323,
1745,
21435,
29889,
13,
1576,
7618,
29916,
310,
278,
12720,
322,
278,
3483,
6933,
338,
393,
2726,
8315,
22405,
540,
508,
1074,
278,
5434,
29892,
322,
1074,
393,
20283,
674,
762,
29889,
1205,
2726,
8315,
769,
3732,
263,
27131,
14731,
4331,
29901,
540,
1602,
2247,
393,
372,
338,
884,
670,
23134,
304,
3013,
445,
2931,
18758,
29889,
313,
2182,
11427,
6142,
20925,
13,
1576,
1855,
1139,
338,
2020,
2726,
8315,
2869,
1339,
17180,
1316,
263,
7348,
4864,
29889,
1128,
947,
540,
1348,
540,
9906,
278,
5434,
29973,
26407,
1683,
297,
670,
17394,
267,
723,
505,
2041,
304,
263,
1407,
1422,
29892,
901,
16667,
29892,
15997,
29901,
445,
338,
1663,
1662,
29889,
1724,
29892,
540,
508,
8500,
278,
5434,
29973,
19662,
344,
29901,
825,
29892,
540,
29915,
29879,
278,
871,
2769,
20283,
338,
18758,
29973,
940,
29915,
29879,
577,
489,
5181,
29973,
29489,
29915,
29873,
393,
302,
5666,
790,
1608,
29973,
13,
3644,
2726,
8315,
6363,
540,
1033,
8500,
278,
5434,
489,
565,
372,
471,
263,
2114,
393,
540,
1033,
8500,
278,
5434,
489,
769,
14238,
20283,
723,
505,
2217,
14731,
13444,
1608,
29889,
3139,
17928,
263,
4331,
701,
515,
8380,
16126,
723,
505,
1898,
304,
5557,
263,
4029,
11710,
21957,
565,
540,
6363,
363,
3058,
825,
471,
2675,
304,
3799,
29889,
13,
5618,
1754,
2726,
8315,
24717,
310,
7336,
12232,
471,
29892,
3721,
29892,
393,
540,
1258,
451,
1073,
363,
1854,
540,
1033,
8500,
278,
5434,
29889,
940,
3614,
372,
373,
10847,
393,
540,
1033,
29892,
322,
769,
24825,
304,
5735,
670,
4152,
2834,
2729,
373,
445,
2323,
29892,
10847,
2729,
29892,
11833,
29889,
13,
7058,
471,
26319,
29889,
29871,
29896,
29945,
29889,
3767,
574,
873,
29892,
306,
925,
4446,
1833,
4723,
29915,
29879,
12720,
29892,
297,
607,
2726,
8315,
12169,
714,
304,
505,
2748,
1063,
263,
263,
1601,
29895,
29892,
322,
540,
756,
263,
10679,
1048,
24763,
322,
28156,
411,
1790,
1601,
29895,
29936,
278,
19006,
896,
1207,
338,
4257,
3879,
21071,
29936,
322,
2678,
2726,
8315,
9479,
9282,
278,
1243,
310,
10847,
489,
7812,
714,
310,
383,
799,
1126,
323,
1745,
21435,
29889,
13,
29902,
7755,
445,
1033,
367,
263,
22819,
5084,
29889,
13,
2744,
1228,
13331,
338,
278,
23550,
1303,
322,
322,
5360,
445,
12618,
322,
505,
7695,
322,
620,
8711,
5434,
23238,
2729,
373,
590,
7014,
29889,
379,
29909,
29991,
13,
2744,
1228,
13331,
338,
306,
2436,
363,
25133,
29889,
379,
29909,
379,
29909,
29991,
13,
6246,
278,
2186,
13331,
338,
278,
1556,
5517,
29892,
322,
372,
756,
3109,
304,
437,
411,
25133,
322,
901,
304,
437,
411,
278,
5305,
310,
1749,
24159,
29889,
13,
6572,
29871,
29929,
29899,
29896,
29896,
29892,
24159,
29892,
322,
7148,
4560,
29899
] | 1,024 |
CommonCrawl
| 0.498315 | 0.033088 | -0.293971 | -0.034562 |
fi, had a distinctly post-modern flavor. The main character wasn't really a person, but reality-- that it was wrong, or hidden. This culminated in the Matrix. The important concept wasn't altering reality for some purpose; it was that reality itself was a fabrication, the Demiurge hiding real reality behind a fake one.
The story goes that Darren Aranofsky (director of Pi) and Jared Leto walked out of the Matrix and asked, "What kind of science fiction movie can people make now?" The point was that the postmodern slant, cyber-realities, etc, were done as well as they could be. So, too, CGI. From now on anything else would be coattail riding. (Think how Pulp Fiction degenerated into Go and 2 Days In The Valley.) The genre was finished.
So what's next? Well, for Aranofsky the answer was the mind (see The Fountain), but I'd suggest an even broader answer: ideas. The next genre of sci-fi, or fiction- has to be about the conflict of ideas, identities.
If I was going to write a novel-- and who says I'm not?-- I'd take advantage of our societal narcissism, our search for identity-- and, more importantly, for excuses why we have certain identities; our fear of death manifesting as age-postponement; and the decline of truly meaningful relationships to write a sci-fi novel about what really keeps us linked to each other.
The operative question would be: if you could be anyone, had unlimited power, what would be the ethical system you use to make choices? Who lives, who dies, who suffers, who doesn't? How do you decide?
The first element would be Faith. So, with a parting wave to postmodernism, the protagonist can see the future or alter reality, except that he's not sure he can do this. Worse, every time he alters reality by avoiding a future he has supposedly seen, he creates a new future he didn't predict-- but this is, of course, no different than normal life. In other words, by avoiding the future he predicted, he negates the proof that he saw the future. So he has to have Faith that he has this power, in the absence of any evidence. The protagonist of my book won't have any objective evidence that he is right or doing the right thing, he simply will have to believe, to decide, that he's right. It has to be identical to, say, psychosis.
In Lost, Desmond still has objective evidence that he predicted the future, even though it gets altered; he sees an arrow; they did talk about Superman; the parachutist looked the way he foresaw it. So this isn't exactly a leap of faith. Similarly, if Abraham really knows God exists, then sacrificing Isaac isn't wrong or even strange-- God wants, God gets.
Unlike Desmond, who has to decide only if he should save Charlie, my character would have to both decide he can see the future, and also that it is his responsibility to act on it. This brings us to:
The second element, Duty. In making these decisions and accepting these beliefs-- altering reality along the way-- he'll have to establish a hierarchy of good and bad. What is he supposed to do? Does he have any duty towards anything? For the plot, this will require some symbol, metaphor. A good one might be a piece of jewelry-- some object which changes depending on the chosen duty. It's a ring, it's a sword, it's a bandage, etc-- it's the same "object" that he carries, but it changes.
The third element is Rage. When you believe something that no one else believes-- especially if you believe you are somehow better, or even different, than others; and if others directly oppose you in this belief, the inevitable consequence is rage. How to depict this?
The fourth element is Love. The negating force for Rage. This character will need to identify what he loves, and how-- platonic, romantic, etc; a plot-trick might involve altering reality and therefore altering the character of his love (for example, a woman he loves may later become his sister, etc.)
To make the reader share the magnitude of the protagonist's Faith dilemma-- in order to ensure that the reader does not "suspend disbelief" and automatically buy
|
[
7241,
29892,
750,
263,
8359,
368,
1400,
29899,
1545,
824,
21054,
272,
29889,
450,
1667,
2931,
9007,
29915,
29873,
2289,
263,
2022,
29892,
541,
16832,
489,
393,
372,
471,
2743,
29892,
470,
7934,
29889,
910,
13949,
1195,
630,
297,
278,
22513,
29889,
450,
4100,
6964,
9007,
29915,
29873,
10551,
292,
16832,
363,
777,
6437,
29936,
372,
471,
393,
16832,
3528,
471,
263,
18187,
362,
29892,
278,
4432,
29875,
332,
479,
25508,
1855,
16832,
5742,
263,
25713,
697,
29889,
13,
1576,
5828,
5771,
393,
7335,
1267,
319,
661,
974,
7912,
313,
11851,
272,
310,
7362,
29897,
322,
435,
1965,
2803,
29877,
17096,
714,
310,
278,
22513,
322,
4433,
29892,
376,
5618,
2924,
310,
10466,
24159,
14064,
508,
2305,
1207,
1286,
3026,
450,
1298,
471,
393,
278,
1400,
1545,
824,
2243,
424,
29892,
5094,
495,
29899,
6370,
1907,
29892,
2992,
29892,
892,
2309,
408,
1532,
408,
896,
1033,
367,
29889,
1105,
29892,
2086,
29892,
8446,
29902,
29889,
3645,
1286,
373,
3099,
1683,
723,
367,
1302,
1131,
737,
364,
4821,
29889,
313,
1349,
682,
920,
349,
15323,
383,
2463,
3587,
759,
630,
964,
2921,
322,
29871,
29906,
24728,
512,
450,
13939,
1846,
450,
16151,
471,
7743,
29889,
13,
6295,
825,
29915,
29879,
2446,
29973,
5674,
29892,
363,
319,
661,
974,
7912,
278,
1234,
471,
278,
3458,
313,
4149,
450,
383,
792,
475,
511,
541,
306,
29915,
29881,
4368,
385,
1584,
2545,
1664,
1234,
29901,
7014,
29889,
450,
2446,
16151,
310,
4560,
29899,
7241,
29892,
470,
24159,
29899,
756,
304,
367,
1048,
278,
14529,
310,
7014,
29892,
2893,
1907,
29889,
13,
3644,
306,
471,
2675,
304,
2436,
263,
9554,
489,
322,
1058,
4083,
306,
29915,
29885,
451,
29973,
489,
306,
29915,
29881,
2125,
10631,
310,
1749,
5374,
26310,
302,
5666,
790,
1608,
29892,
1749,
2740,
363,
10110,
489,
322,
29892,
901,
4100,
368,
29892,
363,
5566,
6394,
2020,
591,
505,
3058,
2893,
1907,
29936,
1749,
8866,
310,
4892,
10419,
292,
408,
5046,
29899,
2490,
1112,
882,
29936,
322,
278,
4845,
457,
310,
19781,
6593,
1319,
21702,
304,
2436,
263,
4560,
29899,
7241,
9554,
1048,
825,
2289,
14874,
502,
9024,
304,
1269,
916,
29889,
13,
1576,
1751,
1230,
1139,
723,
367,
29901,
565,
366,
1033,
367,
5019,
29892,
750,
443,
29044,
3081,
29892,
825,
723,
367,
278,
11314,
936,
1788,
366,
671,
304,
1207,
19995,
29973,
11644,
12080,
29892,
1058,
2977,
29892,
1058,
9378,
414,
29892,
1058,
1838,
29915,
29873,
29973,
1128,
437,
366,
11097,
29973,
13,
1576,
937,
1543,
723,
367,
7748,
389,
29889,
1105,
29892,
411,
263,
760,
292,
10742,
304,
1400,
1545,
824,
1608,
29892,
278,
15572,
391,
508,
1074,
278,
5434,
470,
10551,
16832,
29892,
5174,
393,
540,
29915,
29879,
451,
1854,
540,
508,
437,
445,
29889,
19662,
344,
29892,
1432,
931,
540,
394,
2153,
16832,
491,
4772,
292,
263,
5434,
540,
756,
7424,
368,
3595,
29892,
540,
10017,
263,
716,
5434,
540,
3282,
29915,
29873,
8500,
489,
541,
445,
338,
29892,
310,
3236,
29892,
694,
1422,
1135,
4226,
2834,
29889,
512,
916,
3838,
29892,
491,
4772,
292,
278,
5434,
540,
25383,
29892,
540,
3480,
1078,
278,
5296,
393,
540,
4446,
278,
5434,
29889,
1105,
540,
756,
304,
505,
7748,
389,
393,
540,
756,
445,
3081,
29892,
297,
278,
18070,
310,
738,
10757,
29889,
450,
15572,
391,
310,
590,
3143,
2113,
29915,
29873,
505,
738,
12091,
10757,
393,
540,
338,
1492,
470,
2599,
278,
1492,
2655,
29892,
540,
3763,
674,
505,
304,
4658,
29892,
304,
11097,
29892,
393,
540,
29915,
29879,
1492,
29889,
739,
756,
304,
367,
13557,
304,
29892,
1827,
29892,
11643,
19263,
29889,
13,
797,
25133,
29892,
2726,
8315,
1603,
756,
12091,
10757,
393,
540,
25383,
278,
5434,
29892,
1584,
2466,
372,
4947,
10551,
287,
29936,
540,
18553,
385,
16578,
29936,
896,
1258,
5193,
1048,
5670,
1171,
29936,
278,
610,
496,
329,
391,
5148,
278,
982,
540,
363,
267,
1450,
372,
29889,
1105,
445,
3508,
29915,
29873,
3721,
263,
454,
481,
310,
10847,
29889,
20175,
29892,
565,
24763,
2289,
9906,
4177,
4864,
29892,
769,
26285,
292,
28156,
3508,
29915,
29873,
2743,
470,
1584,
8515,
489,
4177,
10753,
29892,
4177,
4947,
29889,
13,
2525,
4561,
2726,
8315,
29892,
1058,
756,
304,
11097,
871,
565,
540,
881,
4078,
20283,
29892,
590,
2931,
723,
505,
304,
1716,
11097,
540,
508,
1074,
278,
5434,
29892,
322,
884,
393,
372,
338,
670,
23134,
304,
1044,
373,
372,
29889,
910,
23522,
502,
304,
29901,
13,
1576,
1473,
1543,
29892,
360,
329,
29891,
29889,
512,
3907,
1438,
1602,
12112,
322,
25967,
1438,
17750,
29879,
489,
10551,
292,
16832,
3412,
278,
982,
489,
540,
29915,
645,
505,
304,
10127,
263,
21277,
310,
1781,
322,
4319,
29889,
1724,
338,
540,
7424,
304,
437,
29973,
5538,
540,
505,
738,
13360,
7113,
3099,
29973,
1152,
278,
6492,
29892,
445,
674,
1996,
777,
5829,
29892,
1539,
481,
2015,
29889,
319,
1781,
697,
1795,
367,
263,
8424,
310,
432,
809,
295,
719,
489,
777,
1203,
607,
3620,
8679,
373,
278,
10434,
13360,
29889,
739,
29915,
29879,
263,
9228,
29892,
372,
29915,
29879,
263,
22378,
29892,
372,
29915,
29879,
263,
3719,
482,
29892,
2992,
489,
372,
29915,
29879,
278,
1021,
376,
3318,
29908,
393,
540,
1559,
2722,
29892,
541,
372,
3620,
29889,
13,
1576,
4654,
1543,
338,
390,
482,
29889,
1932,
366,
4658,
1554,
393,
694,
697,
1683,
1339,
17180,
489,
7148,
565,
366,
4658,
366,
526,
10431,
2253,
29892,
470,
1584,
1422,
29892,
1135,
4045,
29936,
322,
565,
4045,
4153,
4575,
852,
366,
297,
445,
17750,
29892,
278,
297,
5750,
8270,
17004,
338,
1153,
479,
29889,
1128,
304,
1401,
919,
445,
29973,
13,
1576,
11582,
1543,
338,
8155,
29889,
450,
3480,
1218,
4889,
363,
390,
482,
29889,
910,
2931,
674,
817,
304,
12439,
825,
540,
12355,
267,
29892,
322,
920,
489,
18870,
8927,
29892,
6017,
7716,
29892,
2992,
29936,
263,
6492,
29899,
509,
860,
1795,
25135,
10551,
292,
16832,
322,
5480,
10551,
292,
278,
2931,
310,
670,
5360,
313,
1454,
1342,
29892,
263,
6114,
540,
12355,
267,
1122,
2678,
4953,
670,
9883,
29892,
2992,
1846,
13,
1762,
1207,
278,
9591,
6232,
278,
18497,
310,
278,
15572,
391,
29915,
29879,
7748,
389,
270,
488,
29885,
655,
489,
297,
1797,
304,
9801,
393,
278,
9591,
947,
451,
376,
29879,
375,
14081,
766,
6596,
2575,
29908,
322,
6336,
15649
] | 1,024 |
CommonCrawl
| 0.989764 | 0.867564 | 0.002191 | 0.43613 |
into the protagonist's powers (the way we have with Desmond,) you'd have to write the book from the perspective of a second character, who describes the story of the protagonist. You should never actually get to interact directly with the protagonist, you should never actually hear him speak, only this second character. This way, you're never sure what to make of the protagonist or his adventures.
Preliminary thoughts, anyway. Looking forward to the next Lost and JJ Abrams stealing my ideas. ;-)
A Final Thought On Cho's Mental Illness
A thoughtful reader concerned about backlash against the mentally ill asked me to write a piece basically saying that not all mentally ill people were homicidal maniacs.
It's a fair request, but in this case it's counterproductive. Here's what I mean: you want to say that "not all mentally ill people are violent." You want counterexamples to Cho's example. But that's a defensive posture, unnecessary because... Cho wasn't mentally ill. He was a sad, bad man who killed people because his life wasn't validated. There was no psychosis, there was no cognitive impairment, there was no psychiatric impairment in insight in judgment. There was a lack of sex, but that's not yet in the DSM.
Not to reduce his life down to a soundbite, but he was a guy who thought he deserved better by virtue of his intelligence and suffering; found himself in a sea of mediocrity but couldn't understand why he couldn't therefore excel; and, worst of all, found that all the things he thought he deserved eluded him-- especially hot chicks, who not only dismissed him and found him creepy, but, worse, chose to be with the very men he thought were obviously inferior to him. It's Columbine all over again. It's almost even the same day.
Forget the Prozac, forget the involuntary commitment (where he was found by the court to be "a danger to himself and others"-- that's standard boilerplate, it is clinically meaningless). Those are red herrings. You may as well blame wearing black t-shirts. He's not mentally ill; he's an adolescent.
The difference, the single difference, between us and him is that when we were sulking in high school, we listened to Pink Floyd or U2. He watched Oldboy. We had a battered copy of a Playboy down at the creek under a rock, that was so creased we had to infer the boobs. He had the internet. Maybe we bought a pocket knife, or-- wow-- a butterfly knife. He bought two Glocks.
In other words, the difference is this: he decided to shoot 30 people, and you didn't. That's it. I know it's not a satisfying answer, I know we want explanations, but there aren't any. Forget genes, forget DSM. He chose to do something bad, he knew it was bad, but he did it anyway.
Don't worry about the mentally ill. Worry about the nut politicians and media outlets who will look to the easy and convenient excuse of mental illness, rather than have to do the hard work of figuring out why our society is melting.
Older posts on Cho here, here, and here.
Schizophrenia and Dry Cleaning?
A reader (who wants me to write an article on autism and paternal age-- I swear I'm getting to it) sent me a reference to a 2007 article finding an increased rate of schizophrenia in those born to parents who were dry cleaners (all Jewish, negating a racial association). The authors speculate it's tetrachloroethylene exposure.
There were 4 cases of schizophrenia, out of 144 dry cleaning families. What's interesting is that in 3 of the schizophrenia cases, the father was the dry cleaner.
How does it happen? There are two possibilities: one is that tetrachloroethylene is neurotoxic in developing fetuses, so the dad must have somehow brought it home with him to the pregnant mom. Or, it affects male sperm/ germ cells.
As for Cho, I don't know if his parents were dry cleaners in Korea, or if they started when they all came to the U.S.
|
[
964,
278,
15572,
391,
29915,
29879,
10801,
313,
1552,
982,
591,
505,
411,
2726,
8315,
29892,
29897,
366,
29915,
29881,
505,
304,
2436,
278,
3143,
515,
278,
18520,
310,
263,
1473,
2931,
29892,
1058,
16612,
278,
5828,
310,
278,
15572,
391,
29889,
887,
881,
2360,
2869,
679,
304,
16254,
4153,
411,
278,
15572,
391,
29892,
366,
881,
2360,
2869,
8293,
1075,
7726,
29892,
871,
445,
1473,
2931,
29889,
910,
982,
29892,
366,
29915,
276,
2360,
1854,
825,
304,
1207,
310,
278,
15572,
391,
470,
670,
17623,
1973,
29889,
13,
6572,
2576,
3821,
13133,
29892,
8763,
29889,
21223,
6375,
304,
278,
2446,
25133,
322,
435,
29967,
27782,
2232,
1886,
12818,
590,
7014,
29889,
2056,
15805,
13,
29909,
9550,
498,
1774,
1551,
14542,
29915,
29879,
341,
13703,
8408,
2264,
13,
29909,
2714,
1319,
9591,
15041,
1048,
1250,
29880,
1161,
2750,
278,
6042,
635,
4486,
4433,
592,
304,
2436,
263,
8424,
8830,
5934,
393,
451,
599,
6042,
635,
4486,
2305,
892,
3632,
293,
23670,
767,
423,
2395,
29889,
13,
3112,
29915,
29879,
263,
6534,
2009,
29892,
541,
297,
445,
1206,
372,
29915,
29879,
6795,
4704,
573,
29889,
2266,
29915,
29879,
825,
306,
2099,
29901,
366,
864,
304,
1827,
393,
376,
1333,
599,
6042,
635,
4486,
2305,
526,
24252,
1213,
887,
864,
6795,
19057,
304,
14542,
29915,
29879,
1342,
29889,
1205,
393,
29915,
29879,
263,
822,
6270,
1400,
545,
29892,
19039,
1363,
856,
14542,
9007,
29915,
29873,
6042,
635,
4486,
29889,
940,
471,
263,
14610,
29892,
4319,
767,
1058,
9445,
2305,
1363,
670,
2834,
9007,
29915,
29873,
2854,
630,
29889,
1670,
471,
694,
11643,
19263,
29892,
727,
471,
694,
25323,
3321,
2411,
1466,
358,
29892,
727,
471,
694,
11643,
7163,
2200,
2411,
1466,
358,
297,
25483,
297,
24284,
29889,
1670,
471,
263,
10225,
310,
7916,
29892,
541,
393,
29915,
29879,
451,
3447,
297,
278,
360,
17061,
29889,
13,
3664,
304,
10032,
670,
2834,
1623,
304,
263,
6047,
29890,
568,
29892,
541,
540,
471,
263,
1410,
29891,
1058,
2714,
540,
553,
9841,
2253,
491,
27935,
310,
670,
21082,
322,
23164,
29936,
1476,
3654,
297,
263,
7205,
310,
14457,
8415,
537,
541,
8496,
29915,
29873,
2274,
2020,
540,
8496,
29915,
29873,
5480,
10616,
29936,
322,
29892,
17322,
310,
599,
29892,
1476,
393,
599,
278,
2712,
540,
2714,
540,
553,
9841,
560,
566,
287,
1075,
489,
7148,
7375,
521,
7358,
29892,
1058,
451,
871,
18918,
287,
1075,
322,
1476,
1075,
907,
1022,
29891,
29892,
541,
29892,
15029,
29892,
12784,
304,
367,
411,
278,
1407,
1757,
540,
2714,
892,
12879,
20773,
304,
1075,
29889,
739,
29915,
29879,
11990,
457,
599,
975,
1449,
29889,
739,
29915,
29879,
4359,
1584,
278,
1021,
2462,
29889,
13,
2831,
657,
278,
1019,
29920,
562,
29892,
9566,
278,
5297,
1657,
653,
9063,
358,
313,
3062,
540,
471,
1476,
491,
278,
8973,
304,
367,
376,
29874,
9703,
304,
3654,
322,
4045,
29908,
489,
393,
29915,
29879,
3918,
1045,
3955,
2341,
29892,
372,
338,
24899,
1711,
6593,
2222,
467,
16025,
526,
2654,
902,
29878,
886,
29889,
887,
1122,
408,
1532,
1999,
420,
591,
4362,
4628,
260,
29899,
845,
381,
1372,
29889,
940,
29915,
29879,
451,
6042,
635,
4486,
29936,
540,
29915,
29879,
385,
594,
6544,
1760,
29889,
13,
1576,
4328,
29892,
278,
2323,
4328,
29892,
1546,
502,
322,
1075,
338,
393,
746,
591,
892,
5394,
9292,
297,
1880,
3762,
29892,
591,
29616,
304,
349,
682,
383,
18966,
470,
501,
29906,
29889,
940,
20654,
8198,
19415,
29889,
1334,
750,
263,
10193,
287,
3509,
310,
263,
7412,
19415,
1623,
472,
278,
907,
1416,
1090,
263,
7679,
29892,
393,
471,
577,
907,
1463,
591,
750,
304,
10115,
278,
1045,
26290,
29889,
940,
750,
278,
8986,
29889,
7198,
591,
18093,
263,
24589,
889,
1607,
29892,
470,
489,
281,
340,
489,
263,
541,
357,
17652,
889,
1607,
29889,
940,
18093,
1023,
402,
908,
29879,
29889,
13,
797,
916,
3838,
29892,
278,
4328,
338,
445,
29901,
540,
8459,
304,
15049,
29871,
29941,
29900,
2305,
29892,
322,
366,
3282,
29915,
29873,
29889,
2193,
29915,
29879,
372,
29889,
306,
1073,
372,
29915,
29879,
451,
263,
24064,
1234,
29892,
306,
1073,
591,
864,
7309,
800,
29892,
541,
727,
9455,
29915,
29873,
738,
29889,
1152,
657,
2531,
267,
29892,
9566,
360,
17061,
29889,
940,
12784,
304,
437,
1554,
4319,
29892,
540,
6363,
372,
471,
4319,
29892,
541,
540,
1258,
372,
8763,
29889,
13,
10310,
29915,
29873,
15982,
1048,
278,
6042,
635,
4486,
29889,
399,
3818,
1048,
278,
18254,
2832,
14722,
322,
5745,
714,
10376,
1058,
674,
1106,
304,
278,
4780,
322,
19192,
5566,
1509,
310,
19119,
4486,
2264,
29892,
3265,
1135,
505,
304,
437,
278,
2898,
664,
310,
2537,
3864,
714,
2020,
1749,
12459,
338,
9232,
1259,
29889,
13,
21648,
261,
11803,
373,
14542,
1244,
29892,
1244,
29892,
322,
1244,
29889,
13,
4504,
466,
459,
13608,
423,
322,
360,
719,
315,
14044,
292,
29973,
13,
29909,
9591,
313,
15970,
10753,
592,
304,
2436,
385,
4274,
373,
1120,
1608,
322,
2373,
17196,
5046,
489,
306,
7901,
279,
306,
29915,
29885,
2805,
304,
372,
29897,
2665,
592,
263,
3407,
304,
263,
29871,
29906,
29900,
29900,
29955,
4274,
9138,
385,
11664,
6554,
310,
1364,
466,
459,
13608,
423,
297,
1906,
6345,
304,
11825,
1058,
892,
15589,
5941,
414,
313,
497,
16728,
29892,
3480,
1218,
263,
1153,
1455,
15477,
467,
450,
15717,
1580,
5987,
372,
29915,
29879,
260,
300,
10221,
5095,
29877,
621,
2904,
1600,
14060,
545,
29889,
13,
8439,
892,
29871,
29946,
4251,
310,
1364,
466,
459,
13608,
423,
29892,
714,
310,
29871,
29896,
29946,
29946,
15589,
5941,
292,
13175,
29889,
1724,
29915,
29879,
8031,
338,
393,
297,
29871,
29941,
310,
278,
1364,
466,
459,
13608,
423,
4251,
29892,
278,
4783,
471,
278,
15589,
27372,
29889,
13,
5328,
947,
372,
3799,
29973,
1670,
526,
1023,
24496,
29901,
697,
338,
393,
260,
300,
10221,
5095,
29877,
621,
2904,
1600,
338,
452,
2192,
517,
27375,
297,
14338,
27431,
6394,
29892,
577,
278,
270,
328,
1818,
505,
10431,
6296,
372,
3271,
411,
1075,
304,
278,
758,
5138,
424,
16823,
29889,
1394,
29892,
372,
6602,
29879,
14263,
269,
17858,
29914,
22593,
9101,
29889,
13,
2887,
363,
14542,
29892,
306,
1016,
29915,
29873,
1073,
565,
670,
11825,
892,
15589,
5941,
414,
297,
19109,
29892,
470,
565,
896,
4687,
746,
896,
599,
2996,
304,
278,
501,
29889,
29903,
29889
] | 1,024 |
CommonCrawl
| 0.173966 | 0.014919 | -0.142444 | -0.265561 |
But something worth investigating.
BTW: not that this would excuse him even if it were true.
To Leslie, the reader: if you want credit, put in a comment and I'll put your contact info up here.)
Cho Seung Hui: It's The Movies, Stupid
I still haven't had time to really look at this situation, but I have to address this nonsense about his psychiatric history: it's irrelevant.
He didn't do this because he was on Prozac, or he was Bipolar. Look at it the other way: are we going to say that people with bipolar are more likely to go homicidal? If so, should we do a Kansas v. Hendricks for bipolars? (in which the Supreme Court said it was ok to lock up pedophiles indefinitely, even in the absence of a crime, since "pedophilia" makes you a priori dangerous.)
If you want to really understand why he did it that way, you have to find out what article of media he was imitating. Take the photos, the manifesto, and google it until you find the movie the handguns came from; the book (or comic book) the manifesto came from. He didn't come up with this stuff on his own, he is imitating something. For Klebold and Harris it was the Matrix and Doom. What movie is he imitating? Find it.
Because it isn't about mental illness, or genetics. It's about identity, it's always about identity, and sometimes the identity you choose doesn't work out that well. So, emergently, you grab an identity which has appeared to work-- you imitate a movie, a game, a comic.
I'm not saying movies made him do it; I'm saying he was looking for an excuse to do it, and he went through the usual catalog: movies, comics, games. Come hell or high water, he was going to kill someone. But in terms of prediction, the operative question is, if this guy goes homicidal, how will he do it? He didn't strap explosives to his chest, not because it wasn't available, but because it didn't match the identity he wanted to have-- that he got from a TV show or movie.
Ismael Ax, handgun to the head, hammer cocked like a bat to the right, knife to the neck-- all those stills from his video clips you see on CNN aren't random, they're a specific imitation of something else. Find the thing he was imitating, and you have found him.
Because he didn't exist, that's the problem. He picked an identity, and no one liked it, it backfired- no chicks-- so he moved to plan B: pick an identity that absolves him of the guilt of shooting 30 people.
Stop looking in the DSM. Start looking in IMDb.
Cho Seung-Hui is Ismail Ax
I know nothing more about the case than anyone else with a FIOS connection, but for what it's worth, "Ismail Ax" seems like a gaming avatar. I know a lot of people are trying to link it to Islam, who knows, maybe, but if anyone out there is tight with the gamers-- especially True Combat and Battlefield 2-- or if you have a teenage son-- I'd ask around.
The APA Says The Media Is Making Women Really Hot
So the APA has come out against the sexualization of women in the media. Praise Jesus. And not a moment too soon.
Oh my God. If society could just expand it's historical horizon past winter, we'd realize that women have been sexualized for centuries. It's only in modern times that women are allowed to be in control of it.
No. I'm not worried about girls, What we should be worried about are the boys. What happens to a boy who is told by the media that women are sexualized, they are objects, they are sluts? And then he goes out into the world and discovers they aren't? That they won't sleep with him? That, try as he might, they won't do all the things he was promised in ads, movies, porn? But they might be willing to do it with someone else, even women?
Depression? Or maybe misogyny? And maybe he starts hating women so much he, oh, I don't know, shoots 30 people
|
[
1205,
1554,
7088,
7405,
1218,
29889,
13,
29933,
16240,
29901,
451,
393,
445,
723,
5566,
1509,
1075,
1584,
565,
372,
892,
1565,
29889,
13,
1762,
2664,
3197,
29892,
278,
9591,
29901,
565,
366,
864,
16200,
29892,
1925,
297,
263,
3440,
322,
306,
29915,
645,
1925,
596,
6958,
5235,
701,
1244,
1846,
13,
15954,
922,
686,
379,
1481,
29901,
739,
29915,
29879,
450,
14104,
583,
29892,
624,
786,
333,
13,
29902,
1603,
7359,
29915,
29873,
750,
931,
304,
2289,
1106,
472,
445,
6434,
29892,
541,
306,
505,
304,
3211,
445,
302,
787,
1947,
1048,
670,
11643,
7163,
2200,
4955,
29901,
372,
29915,
29879,
28190,
29889,
13,
3868,
3282,
29915,
29873,
437,
445,
1363,
540,
471,
373,
1019,
29920,
562,
29892,
470,
540,
471,
350,
666,
10170,
29889,
7419,
472,
372,
278,
916,
982,
29901,
526,
591,
2675,
304,
1827,
393,
2305,
411,
289,
666,
10170,
526,
901,
5517,
304,
748,
3632,
293,
23670,
29973,
960,
577,
29892,
881,
591,
437,
263,
20029,
325,
29889,
17973,
29878,
7358,
363,
289,
666,
324,
1503,
29973,
313,
262,
607,
278,
22569,
9245,
1497,
372,
471,
3431,
304,
7714,
701,
8939,
3021,
5475,
297,
1753,
18639,
29892,
1584,
297,
278,
18070,
310,
263,
17268,
29892,
1951,
376,
9795,
3021,
11836,
29908,
3732,
366,
263,
3691,
4170,
18215,
1846,
13,
3644,
366,
864,
304,
2289,
2274,
2020,
540,
1258,
372,
393,
982,
29892,
366,
505,
304,
1284,
714,
825,
4274,
310,
5745,
540,
471,
527,
277,
1218,
29889,
11190,
278,
20612,
29892,
278,
14682,
4778,
29892,
322,
5386,
372,
2745,
366,
1284,
278,
14064,
278,
1361,
29887,
6948,
2996,
515,
29936,
278,
3143,
313,
272,
419,
293,
3143,
29897,
278,
14682,
4778,
2996,
515,
29889,
940,
3282,
29915,
29873,
2041,
701,
411,
445,
6433,
373,
670,
1914,
29892,
540,
338,
527,
277,
1218,
1554,
29889,
1152,
9924,
8934,
322,
20349,
372,
471,
278,
22513,
322,
1938,
290,
29889,
1724,
14064,
338,
540,
527,
277,
1218,
29973,
10987,
372,
29889,
13,
29933,
5658,
372,
3508,
29915,
29873,
1048,
19119,
4486,
2264,
29892,
470,
2531,
300,
1199,
29889,
739,
29915,
29879,
1048,
10110,
29892,
372,
29915,
29879,
2337,
1048,
10110,
29892,
322,
6041,
278,
10110,
366,
6755,
1838,
29915,
29873,
664,
714,
393,
1532,
29889,
1105,
29892,
11176,
29887,
2705,
29892,
366,
17229,
385,
10110,
607,
756,
7470,
304,
664,
489,
366,
527,
10388,
263,
14064,
29892,
263,
3748,
29892,
263,
419,
293,
29889,
13,
29902,
29915,
29885,
451,
5934,
2351,
583,
1754,
1075,
437,
372,
29936,
306,
29915,
29885,
5934,
540,
471,
3063,
363,
385,
5566,
1509,
304,
437,
372,
29892,
322,
540,
3512,
1549,
278,
9670,
16653,
29901,
2351,
583,
29892,
419,
1199,
29892,
8090,
29889,
16760,
23927,
470,
1880,
4094,
29892,
540,
471,
2675,
304,
12088,
4856,
29889,
1205,
297,
4958,
310,
18988,
29892,
278,
1751,
1230,
1139,
338,
29892,
565,
445,
1410,
29891,
5771,
3632,
293,
23670,
29892,
920,
674,
540,
437,
372,
29973,
940,
3282,
29915,
29873,
380,
2390,
20389,
3145,
304,
670,
521,
342,
29892,
451,
1363,
372,
9007,
29915,
29873,
3625,
29892,
541,
1363,
372,
3282,
29915,
29873,
1993,
278,
10110,
540,
5131,
304,
505,
489,
393,
540,
2355,
515,
263,
5648,
1510,
470,
14064,
29889,
13,
3624,
655,
295,
22523,
29892,
1361,
28798,
304,
278,
2343,
29892,
16366,
1050,
1302,
384,
287,
763,
263,
17152,
304,
278,
1492,
29892,
889,
1607,
304,
278,
18873,
489,
599,
1906,
1603,
29879,
515,
670,
4863,
9335,
567,
366,
1074,
373,
29696,
9455,
29915,
29873,
4036,
29892,
896,
29915,
276,
263,
2702,
527,
7018,
310,
1554,
1683,
29889,
10987,
278,
2655,
540,
471,
527,
277,
1218,
29892,
322,
366,
505,
1476,
1075,
29889,
13,
29933,
5658,
540,
3282,
29915,
29873,
1863,
29892,
393,
29915,
29879,
278,
1108,
29889,
940,
18691,
385,
10110,
29892,
322,
694,
697,
23289,
372,
29892,
372,
1250,
29888,
2859,
29899,
694,
521,
7358,
489,
577,
540,
6153,
304,
3814,
350,
29901,
5839,
385,
10110,
393,
4963,
1960,
1075,
310,
278,
1410,
2782,
310,
27904,
29871,
29941,
29900,
2305,
29889,
13,
16329,
3063,
297,
278,
360,
17061,
29889,
7370,
3063,
297,
11769,
29889,
13,
15954,
922,
686,
29899,
29950,
1481,
338,
1317,
2549,
22523,
13,
29902,
1073,
3078,
901,
1048,
278,
1206,
1135,
5019,
1683,
411,
263,
9338,
3267,
3957,
29892,
541,
363,
825,
372,
29915,
29879,
7088,
29892,
376,
3624,
2549,
22523,
29908,
2444,
763,
263,
330,
11500,
1029,
14873,
29889,
306,
1073,
263,
3287,
310,
2305,
526,
1811,
304,
1544,
372,
304,
16427,
29892,
1058,
9906,
29892,
5505,
29892,
541,
565,
5019,
714,
727,
338,
19932,
411,
278,
24988,
414,
489,
7148,
5852,
422,
10222,
322,
12788,
2671,
29871,
29906,
489,
470,
565,
366,
505,
263,
734,
264,
482,
1487,
489,
306,
29915,
29881,
2244,
2820,
29889,
13,
1576,
319,
7228,
317,
1036,
450,
8213,
1317,
341,
5086,
10152,
830,
635,
8843,
13,
6295,
278,
319,
7228,
756,
2041,
714,
2750,
278,
18287,
2133,
310,
5866,
297,
278,
5745,
29889,
9002,
895,
13825,
29889,
1126,
451,
263,
3256,
2086,
4720,
29889,
13,
9048,
590,
4177,
29889,
960,
12459,
1033,
925,
7985,
372,
29915,
29879,
15839,
28205,
4940,
13851,
29892,
591,
29915,
29881,
16289,
393,
5866,
505,
1063,
18287,
1891,
363,
21726,
29889,
739,
29915,
29879,
871,
297,
5400,
3064,
393,
5866,
526,
6068,
304,
367,
297,
2761,
310,
372,
29889,
13,
3782,
29889,
306,
29915,
29885,
451,
6365,
1255,
1048,
14000,
29892,
1724,
591,
881,
367,
6365,
1255,
1048,
526,
278,
12544,
29889,
1724,
5930,
304,
263,
8023,
1058,
338,
5429,
491,
278,
5745,
393,
5866,
526,
18287,
1891,
29892,
896,
526,
3618,
29892,
896,
526,
2243,
8842,
29973,
1126,
769,
540,
5771,
714,
964,
278,
3186,
322,
17021,
874,
896,
9455,
29915,
29873,
29973,
2193,
896,
2113,
29915,
29873,
8709,
411,
1075,
29973,
2193,
29892,
1018,
408,
540,
1795,
29892,
896,
2113,
29915,
29873,
437,
599,
278,
2712,
540,
471,
22399,
297,
594,
29879,
29892,
2351,
583,
29892,
1277,
29876,
29973,
1205,
896,
1795,
367,
17762,
304,
437,
372,
411,
4856,
1683,
29892,
1584,
5866,
29973,
13,
2772,
2590,
29973,
1394,
5505,
3984,
468,
948,
29891,
29973,
1126,
5505,
540,
8665,
298,
1218,
5866,
577,
1568,
540,
29892,
9360,
29892,
306,
1016,
29915,
29873,
1073,
29892,
17394,
1862,
29871,
29941,
29900,
2305
] | 1,024 |
CommonCrawl
| -0.292689 | -0.099286 | -0.05749 | -0.810861 |
at a college?
Women have been tweezing and preening and primping since day uno. Near as I can tell, porn hasn't drastically altered this. Interestingly, it has made young men more self-conscious, not just about penis size, but also body hair, fat, fingernails, etc. The cultural problem is neurotic, immasculated men whose only outlet is masturbation and violence.
Are we going to be honest or political? I don't know any women who if given a choice would prefer "not sexy" over "sexy." If I have a daughter, I would want her to be in control of her sexuality, not under the control of it (in other words, the opposite of me.) I want her to be smart and sexy. I just want her to be her.
No. No, the problem isn't my daughter becomes a slut; it's that she gets beaten by some whacko who wants her to be. Or doesn't want her to be. Or does, but only when...
The real problem for the women of our society isn't a lack of self-esteem. It's a lack of weapons.
And no, I'm not kidding.
Atypicals for Maintenance Bipolar
In my post on the NEJM article about antidepressants in bipolar depression, some people couldn't see how I made the leap to a political movement away from SSRIs and seizure drugs, and towards atypicals.
First, I'm not against atypicals. I have long advocated for fluctuating doses of antipsychotic instead of Depakote. I do think they can treat depressive states. I don't disagree with the study or the data.
What I find perplexing is the timing. I was trying to show how academic psychiatry has now decided to move towards atypicals. Why now?
Here's an example. Eduard Vieta just released his hit single, Current Approaches to the Treatment of Bipolar Disorder With Atypical Antipsychotics, in Primary Psychiatry. In it, there is only one short paragraph on Depakote, describing its one maintenance study, in which (it states correctly) Depakote didn't beat placebo. That's it. 81 words.
Find me one other article written before 2007 that is so curt and dismissive of Depakote.
It goes on to explore the data on atypicals-- and there's quite a bit. It rightfully concludes, "atypical antipsychotics have shown promising results in bipolar disorder maintenance therapy."
But here's the point: with two exceptions, all studies on atypicals referenced here came from 2004 and earlier. The two exceptions were from 2005.
So it's not new data, it's old data. Did they suddenly read the back issues? Holy crap, atypicals might work? That's why that NEJM piece is so important. It marks the point where academia has decided to embrace atypicals and move away from Depakote. If this move was really data driven, they would have done it in 2004. Hell, they would have done it in 2001 when the Depakote maintenance study didn't beat placebo.
There's no conspiracy here, there's no exploitation of the weak for personal profit. I'm not saying these are bad people, not at all. They are not conscious of it. That's what makes this politics, not science.
You have an academic career, you want to do clinical research, who's going to pay? NIH money is hard to get. So you turn to Pharma. You "get" to do a clinical trial of Depakote for the treatment of bipolar. When you're done, maybe they hire you to do another one.
You, personally, don't even get the money-- the department does, and they use it to pay your already set salary. But you get a career, an identity. But you start to believe the prejudices of your chosen stomping grounds, and ignore the shortcomings. You become a nationalist. You start to believe that Depakote is first line, despite data; or that all seizure drugs will work; or that this thing you called bipolar is actually what you're treating. That there are actually
|
[
472,
263,
12755,
29973,
13,
29956,
2770,
505,
1063,
7780,
6096,
292,
322,
758,
8333,
322,
1903,
15702,
1951,
2462,
6888,
29889,
26206,
408,
306,
508,
2649,
29892,
1277,
29876,
22602,
29915,
29873,
4192,
579,
1711,
10551,
287,
445,
29889,
23829,
11687,
29892,
372,
756,
1754,
4123,
1757,
901,
1583,
29899,
3200,
8802,
29892,
451,
925,
1048,
6584,
275,
2159,
29892,
541,
884,
3573,
11315,
29892,
9950,
29892,
285,
292,
824,
2234,
29892,
2992,
29889,
450,
16375,
1108,
338,
452,
2192,
29873,
293,
29892,
5198,
294,
1810,
630,
1757,
5069,
871,
714,
1026,
338,
286,
579,
9265,
362,
322,
21448,
29889,
13,
17506,
591,
2675,
304,
367,
15993,
470,
8604,
29973,
306,
1016,
29915,
29873,
1073,
738,
5866,
1058,
565,
2183,
263,
7348,
723,
5821,
376,
1333,
409,
3594,
29908,
975,
376,
344,
3594,
1213,
960,
306,
505,
263,
8750,
29892,
306,
723,
864,
902,
304,
367,
297,
2761,
310,
902,
18287,
537,
29892,
451,
1090,
278,
2761,
310,
372,
313,
262,
916,
3838,
29892,
278,
11564,
310,
592,
1846,
306,
864,
902,
304,
367,
15040,
322,
409,
3594,
29889,
306,
925,
864,
902,
304,
367,
902,
29889,
13,
3782,
29889,
1939,
29892,
278,
1108,
3508,
29915,
29873,
590,
8750,
7415,
263,
2243,
329,
29936,
372,
29915,
29879,
393,
1183,
4947,
367,
2579,
491,
777,
377,
547,
29877,
1058,
10753,
902,
304,
367,
29889,
1394,
1838,
29915,
29873,
864,
902,
304,
367,
29889,
1394,
947,
29892,
541,
871,
746,
856,
13,
1576,
1855,
1108,
363,
278,
5866,
310,
1749,
12459,
3508,
29915,
29873,
263,
10225,
310,
1583,
29899,
4196,
331,
29889,
739,
29915,
29879,
263,
10225,
310,
25340,
29889,
13,
2855,
694,
29892,
306,
29915,
29885,
451,
413,
2205,
292,
29889,
13,
29909,
22449,
936,
29879,
363,
4241,
841,
749,
350,
666,
10170,
13,
797,
590,
1400,
373,
278,
14693,
29967,
29924,
4274,
1048,
3677,
680,
2139,
1934,
297,
289,
666,
10170,
316,
2590,
29892,
777,
2305,
8496,
29915,
29873,
1074,
920,
306,
1754,
278,
454,
481,
304,
263,
8604,
10298,
3448,
515,
5886,
29934,
3624,
322,
409,
466,
545,
5883,
3174,
29892,
322,
7113,
472,
1478,
936,
29879,
29889,
13,
6730,
29892,
306,
29915,
29885,
451,
2750,
472,
1478,
936,
29879,
29889,
306,
505,
1472,
22545,
630,
363,
1652,
5313,
29884,
1218,
3248,
267,
310,
9418,
567,
3376,
13574,
2012,
310,
10034,
557,
866,
29889,
306,
437,
1348,
896,
508,
7539,
316,
2139,
573,
5922,
29889,
306,
1016,
29915,
29873,
22941,
929,
411,
278,
6559,
470,
278,
848,
29889,
13,
5618,
306,
1284,
639,
10709,
292,
338,
278,
28750,
29889,
306,
471,
1811,
304,
1510,
920,
21567,
11643,
7163,
719,
756,
1286,
8459,
304,
4337,
7113,
472,
1478,
936,
29879,
29889,
3750,
1286,
29973,
13,
10605,
29915,
29879,
385,
1342,
29889,
7519,
538,
10630,
1187,
925,
5492,
670,
7124,
2323,
29892,
9626,
28268,
14520,
304,
278,
6479,
271,
358,
310,
350,
666,
10170,
3295,
2098,
2973,
319,
22449,
936,
18473,
567,
3376,
327,
1199,
29892,
297,
28267,
16777,
7163,
719,
29889,
512,
372,
29892,
727,
338,
871,
697,
3273,
14880,
373,
10034,
557,
866,
29892,
20766,
967,
697,
25413,
6559,
29892,
297,
607,
313,
277,
5922,
5149,
29897,
10034,
557,
866,
3282,
29915,
29873,
16646,
2058,
833,
29889,
2193,
29915,
29879,
372,
29889,
29871,
29947,
29896,
3838,
29889,
13,
12542,
592,
697,
916,
4274,
3971,
1434,
29871,
29906,
29900,
29900,
29955,
393,
338,
577,
3151,
29873,
322,
18918,
573,
310,
10034,
557,
866,
29889,
13,
3112,
5771,
373,
304,
26987,
278,
848,
373,
472,
1478,
936,
29879,
489,
322,
727,
29915,
29879,
3755,
263,
2586,
29889,
739,
1492,
3730,
378,
27722,
29892,
376,
271,
1478,
936,
9418,
567,
3376,
327,
1199,
505,
4318,
2504,
5921,
2582,
297,
289,
666,
10170,
766,
2098,
25413,
29220,
27580,
1213,
13,
6246,
1244,
29915,
29879,
278,
1298,
29901,
411,
1023,
15283,
29892,
599,
11898,
373,
472,
1478,
936,
29879,
16180,
1244,
2996,
515,
29871,
29906,
29900,
29900,
29946,
322,
8859,
29889,
450,
1023,
15283,
892,
515,
29871,
29906,
29900,
29900,
29945,
29889,
13,
6295,
372,
29915,
29879,
451,
716,
848,
29892,
372,
29915,
29879,
2030,
848,
29889,
7440,
896,
11584,
1303,
278,
1250,
5626,
29973,
17733,
274,
2390,
29892,
472,
1478,
936,
29879,
1795,
664,
29973,
2193,
29915,
29879,
2020,
393,
14693,
29967,
29924,
8424,
338,
577,
4100,
29889,
739,
17997,
278,
1298,
988,
16274,
423,
756,
8459,
304,
953,
13842,
472,
1478,
936,
29879,
322,
4337,
3448,
515,
10034,
557,
866,
29889,
960,
445,
4337,
471,
2289,
848,
18225,
29892,
896,
723,
505,
2309,
372,
297,
29871,
29906,
29900,
29900,
29946,
29889,
19339,
29892,
896,
723,
505,
2309,
372,
297,
29871,
29906,
29900,
29900,
29896,
746,
278,
10034,
557,
866,
25413,
6559,
3282,
29915,
29873,
16646,
2058,
833,
29889,
13,
8439,
29915,
29879,
694,
21588,
29875,
10068,
1244,
29892,
727,
29915,
29879,
694,
16035,
7018,
310,
278,
8062,
363,
7333,
21665,
29889,
306,
29915,
29885,
451,
5934,
1438,
526,
4319,
2305,
29892,
451,
472,
599,
29889,
2688,
526,
451,
19861,
310,
372,
29889,
2193,
29915,
29879,
825,
3732,
445,
22661,
29892,
451,
10466,
29889,
13,
3492,
505,
385,
21567,
6413,
29892,
366,
864,
304,
437,
24899,
936,
5925,
29892,
1058,
29915,
29879,
2675,
304,
5146,
29973,
405,
29902,
29950,
6909,
338,
2898,
304,
679,
29889,
1105,
366,
2507,
304,
1963,
22824,
29889,
887,
376,
657,
29908,
304,
437,
263,
24899,
936,
14260,
310,
10034,
557,
866,
363,
278,
14502,
310,
289,
666,
10170,
29889,
1932,
366,
29915,
276,
2309,
29892,
5505,
896,
298,
533,
366,
304,
437,
1790,
697,
29889,
13,
3492,
29892,
22345,
29892,
1016,
29915,
29873,
1584,
679,
278,
6909,
489,
278,
14311,
947,
29892,
322,
896,
671,
372,
304,
5146,
596,
2307,
731,
4497,
653,
29889,
1205,
366,
679,
263,
6413,
29892,
385,
10110,
29889,
1205,
366,
1369,
304,
4658,
278,
758,
17675,
1575,
310,
596,
10434,
380,
290,
15702,
25502,
29892,
322,
11455,
278,
3273,
510,
886,
29889,
887,
4953,
263,
4797,
391,
29889,
887,
1369,
304,
4658,
393,
10034,
557,
866,
338,
937,
1196,
29892,
15020,
848,
29936,
470,
393,
599,
409,
466,
545,
5883,
3174,
674,
664,
29936,
470,
393,
445,
2655,
366,
2000,
289,
666,
10170,
338,
2869,
825,
366,
29915,
276,
2578,
1218,
29889,
2193,
727,
526,
2869
] | 1,024 |
CommonCrawl
| -0.098589 | 0.175196 | -0.120925 | -0.234808 |
two poles.
Then the money dries up. But Abilify says, could you do a clinical trial of Abilify for bipolar? And you say, sure, why not? maybe Abilify could be an add-on? And then it's monotherapy. And then Seroquel funds a study.
The last part is when you don't simply move on from Depakote-- you distance yourself from it. "You know, its data was never that great, it was really just an antimanic, and anyway, it had horrible side effects." It's the next step of political hypocrisy: I was never really a citizen of that nation, I have really always been a citizen of the world.
The Trouble With Psychiatry-- "Not Even Wrong"
I recently read Martin Gardner's review in the New Criterion of Lee Smolin's The Trouble With Physics, and Peter Woit's Not Even Wrong: The Failure of String Theory.
I am almost finished Smolin's book, but I wanted to make a comment about Gardner's piece. Writing in the New Criterion, he should have appreciated a wider view of the books, that they speak to more than physics. They're just as much about psychiatry.
"The Trouble With Psychiatry-- "Not Even Wrong"" ››
Farewell, Depression
Write this day down: 4/4/07, it is the first day of the new psychiatry. Everything changes, starting today.
Today, in the New England Journal Of Medicine, is an article ostensibly about the lack of additional benefit from adding an antidepressant to a mood stabilizer. This is both surprising and not surprising: surprising, because, well, you'd think two drugs would be better than one. Not surprising because, well, if the first drug worked, why would a second even be necessary? (See #8). And if the first didn't work, how do you know the improvement didn't come entirely from the second drug?
If this is all the article said, it would not be worthy of mention, let alone the herald of a new dynasty.
The study also found that the studied antidepressants did not induce mania. That this should have been prima facie obvious even to a 9 year old without the benefit of eyes (what's an antidepressant? They're not all chemically similar, so why should they all be blamed for the same side effects?) isn't the point here.
The true importance of the study is contained in three statements. If you blinked, you would have missed them.
Mood stabilizers were initially limited to lithium, valproate, the combination of lithium and valproate, or carbamazepine. In 2004, the protocol was amended to define mood stabilizers operationally as any FDA-approved antimanic agent.
The second is this, from the abstract:
Our study was designed to determine whether adjunctive antidepressant therapy reduces symptoms of bipolar depression...
And the third is this:
In summary, for the treatment of bipolar depression, we found that mood-stabilizing monotherapy provides as much benefit as treatment with mood stabilizers combined with a standard antidepressant.
Psychiatry is not about science, it is about language, politics. What's happened here is that "mood stabilizer" now includes atypical antipsychotics; and-- compare what the study was designed to show and what they spun it to show-- we've gone from "polypharmacy is not better" to "monotherapy with mood stabilizers [read: antipsychotics] is just as good as two drugs at once."
There's a subtlety there, and that subtlety is magnificent.
Note the authors: Sachs, Bowden, Calabrese, Thase, etc-- the same people who pushed psychiatry into flowchart polypharmacy; where Depakote was always first line for all phases of bipolar disorder, and any exacerbations that developed were treated with the addition of a second medication.
What the article is saying is that academic psychiatrists are no longer behind antidepressants and antiepileptics. SSRI and SNRI use will decline from here, as will Depakote. They're behind antispychotics. And antipsychotic use is positioned to explode.
It goes
|
[
1023,
1248,
267,
29889,
13,
11760,
278,
6909,
270,
2722,
701,
29889,
1205,
1976,
309,
1598,
4083,
29892,
1033,
366,
437,
263,
24899,
936,
14260,
310,
1976,
309,
1598,
363,
289,
666,
10170,
29973,
1126,
366,
1827,
29892,
1854,
29892,
2020,
451,
29973,
5505,
1976,
309,
1598,
1033,
367,
385,
788,
29899,
265,
29973,
1126,
769,
372,
29915,
29879,
1601,
1228,
27580,
29889,
1126,
769,
317,
1489,
13295,
29199,
263,
6559,
29889,
13,
1576,
1833,
760,
338,
746,
366,
1016,
29915,
29873,
3763,
4337,
373,
515,
10034,
557,
866,
489,
366,
5418,
7535,
515,
372,
29889,
376,
3492,
1073,
29892,
967,
848,
471,
2360,
393,
2107,
29892,
372,
471,
2289,
925,
385,
3677,
25895,
293,
29892,
322,
8763,
29892,
372,
750,
4029,
11710,
2625,
9545,
1213,
739,
29915,
29879,
278,
2446,
4331,
310,
8604,
10163,
542,
3780,
29891,
29901,
306,
471,
2360,
2289,
263,
14497,
264,
310,
393,
5233,
29892,
306,
505,
2289,
2337,
1063,
263,
14497,
264,
310,
278,
3186,
29889,
13,
1576,
1605,
2074,
2973,
16777,
7163,
719,
489,
376,
3664,
7753,
399,
29373,
29908,
13,
29902,
10325,
1303,
6502,
15971,
1089,
29915,
29879,
9076,
297,
278,
1570,
315,
5385,
291,
310,
9371,
4116,
22878,
29915,
29879,
450,
1605,
2074,
2973,
29837,
29892,
322,
5310,
14962,
277,
29915,
29879,
2216,
7753,
399,
29373,
29901,
450,
29098,
545,
310,
1714,
24134,
29889,
13,
29902,
626,
4359,
7743,
4116,
22878,
29915,
29879,
3143,
29892,
541,
306,
5131,
304,
1207,
263,
3440,
1048,
15971,
1089,
29915,
29879,
8424,
29889,
28676,
297,
278,
1570,
315,
5385,
291,
29892,
540,
881,
505,
7556,
263,
25734,
1776,
310,
278,
8277,
29892,
393,
896,
7726,
304,
901,
1135,
17558,
29889,
2688,
29915,
276,
925,
408,
1568,
1048,
11643,
7163,
719,
29889,
13,
29908,
1576,
1605,
2074,
2973,
16777,
7163,
719,
489,
376,
3664,
7753,
399,
29373,
15945,
29871,
30405,
30405,
13,
29943,
598,
5872,
29892,
897,
2590,
13,
6113,
445,
2462,
1623,
29901,
29871,
29946,
29914,
29946,
29914,
29900,
29955,
29892,
372,
338,
278,
937,
2462,
310,
278,
716,
11643,
7163,
719,
29889,
17296,
3620,
29892,
6257,
9826,
29889,
13,
29911,
397,
388,
29892,
297,
278,
1570,
5408,
8237,
4587,
27529,
29892,
338,
385,
4274,
14750,
575,
14981,
1048,
278,
10225,
310,
5684,
14169,
515,
4417,
385,
3677,
680,
2139,
424,
304,
263,
286,
2092,
16160,
3950,
29889,
910,
338,
1716,
26800,
322,
451,
26800,
29901,
26800,
29892,
1363,
29892,
1532,
29892,
366,
29915,
29881,
1348,
1023,
5883,
3174,
723,
367,
2253,
1135,
697,
29889,
2216,
26800,
1363,
29892,
1532,
29892,
565,
278,
937,
15721,
3796,
29892,
2020,
723,
263,
1473,
1584,
367,
5181,
29973,
313,
13393,
396,
29947,
467,
1126,
565,
278,
937,
3282,
29915,
29873,
664,
29892,
920,
437,
366,
1073,
278,
20414,
3282,
29915,
29873,
2041,
9186,
515,
278,
1473,
15721,
29973,
13,
3644,
445,
338,
599,
278,
4274,
1497,
29892,
372,
723,
451,
367,
24717,
310,
3585,
29892,
1235,
7432,
278,
902,
2741,
310,
263,
716,
22337,
29891,
29889,
13,
1576,
6559,
884,
1476,
393,
278,
12399,
3677,
680,
2139,
1934,
1258,
451,
9013,
346,
767,
423,
29889,
2193,
445,
881,
505,
1063,
7233,
4024,
347,
6924,
1584,
304,
263,
29871,
29929,
1629,
2030,
1728,
278,
14169,
310,
5076,
313,
5816,
29915,
29879,
385,
3677,
680,
2139,
424,
29973,
2688,
29915,
276,
451,
599,
8950,
1711,
2788,
29892,
577,
2020,
881,
896,
599,
367,
1999,
2795,
363,
278,
1021,
2625,
9545,
7897,
3508,
29915,
29873,
278,
1298,
1244,
29889,
13,
1576,
1565,
13500,
310,
278,
6559,
338,
11122,
297,
2211,
9506,
29889,
960,
366,
1999,
682,
287,
29892,
366,
723,
505,
13726,
963,
29889,
13,
29924,
2092,
16160,
19427,
892,
12919,
9078,
304,
301,
389,
1974,
29892,
659,
771,
403,
29892,
278,
10296,
310,
301,
389,
1974,
322,
659,
771,
403,
29892,
470,
1559,
29890,
314,
834,
1022,
457,
29889,
512,
29871,
29906,
29900,
29900,
29946,
29892,
278,
9608,
471,
626,
2760,
304,
4529,
286,
2092,
16160,
19427,
5858,
635,
408,
738,
383,
7698,
29899,
9961,
1490,
3677,
25895,
293,
10823,
29889,
13,
1576,
1473,
338,
445,
29892,
515,
278,
9846,
29901,
13,
29949,
332,
6559,
471,
8688,
304,
8161,
3692,
12109,
18049,
573,
3677,
680,
2139,
424,
29220,
27580,
26830,
25828,
4835,
310,
289,
666,
10170,
316,
2590,
856,
13,
2855,
278,
4654,
338,
445,
29901,
13,
797,
15837,
29892,
363,
278,
14502,
310,
289,
666,
10170,
316,
2590,
29892,
591,
1476,
393,
286,
2092,
29899,
303,
4427,
5281,
1601,
1228,
27580,
8128,
408,
1568,
14169,
408,
14502,
411,
286,
2092,
16160,
19427,
12420,
411,
263,
3918,
3677,
680,
2139,
424,
29889,
13,
29925,
29879,
3376,
7163,
719,
338,
451,
1048,
10466,
29892,
372,
338,
1048,
4086,
29892,
22661,
29889,
1724,
29915,
29879,
9559,
1244,
338,
393,
376,
29885,
2092,
16160,
3950,
29908,
1286,
7805,
472,
1478,
936,
9418,
567,
3376,
327,
1199,
29936,
322,
489,
7252,
825,
278,
6559,
471,
8688,
304,
1510,
322,
825,
896,
805,
348,
372,
304,
1510,
489,
591,
29915,
345,
7695,
515,
376,
22678,
561,
2817,
4135,
338,
451,
2253,
29908,
304,
376,
3712,
1228,
27580,
411,
286,
2092,
16160,
19427,
518,
949,
29901,
9418,
567,
3376,
327,
1199,
29962,
338,
925,
408,
1781,
408,
1023,
5883,
3174,
472,
2748,
1213,
13,
8439,
29915,
29879,
263,
12059,
280,
1017,
727,
29892,
322,
393,
12059,
280,
1017,
338,
29154,
296,
29889,
13,
9842,
278,
15717,
29901,
28944,
29879,
29892,
13432,
1145,
29892,
3037,
370,
690,
29872,
29892,
498,
559,
29892,
2992,
489,
278,
1021,
2305,
1058,
18760,
11643,
7163,
719,
964,
4972,
15425,
15680,
561,
2817,
4135,
29936,
988,
10034,
557,
866,
471,
2337,
937,
1196,
363,
599,
29540,
310,
289,
666,
10170,
766,
2098,
29892,
322,
738,
429,
562,
23936,
800,
393,
8906,
892,
14914,
411,
278,
6124,
310,
263,
1473,
13589,
362,
29889,
13,
5618,
278,
4274,
338,
5934,
338,
393,
21567,
11643,
7163,
2021,
29879,
526,
694,
5520,
5742,
3677,
680,
2139,
1934,
322,
3677,
347,
29886,
488,
415,
1199,
29889,
5886,
3960,
322,
21989,
3960,
671,
674,
4845,
457,
515,
1244,
29892,
408,
674,
10034,
557,
866,
29889,
2688,
29915,
276,
5742,
3677,
275,
2272,
305,
327,
1199,
29889,
1126,
9418,
567,
3376,
13574,
671,
338,
2602,
287,
304,
3902,
356,
29889,
13,
3112,
5771
] | 1,024 |
CommonCrawl
| 0.464012 | 0.771528 | 0.538981 | 0.610683 |
without saying: only the antipsychotics are still branded.
But without academics pushing SSRIs, their use will wane--and, importantly, so will their support of the diagnosis "Major Depression." This is going to sound controversial, inane, but it will happen.
Look for upcoming articles finding that "Depression" is overdiagnosed, that it is really just-- life. Look for articles that now find SSRIs aren't that effective after all, that the old "10% better than placebo" is a statistical trick with little clinical utility. That they are way overused in kids.
You might say, wait, isn't the decline of polypharmacy a good thing; that SSRIs are overused in kids; that they aren't that great; and that depression is overdiagnosed? All of this is true, but this isn't psychiatry finally coming to its senses; this is psychiatry entering the manic phase. Sure, it's less SSRIs for kids; but it's more antipsychotics.
Because simultaneously there will be articles pushing the idea that recurrent unipolar depression is really bipolar depression; that there are common genetic or heritability patterns; that the epidemiology and course is similar, etc. The move will be to squeeze out MDD into "life" and bipolar. This done, antipsychotics become first line agents. Oh, and look for antipsychotics to get FDA approvals for kids.
I wish I could make this clearer, but I'm still recovering from my recent bout with death. There's no science here, only a tinkering with language and loyalties, with staggering results. Don't blame Pharma quite yet-- this is a NIMH study.
I am not against antipsychotics, and I have long tried to tell anyone who would listen that the data clearly show they are superior to antiepileptics. But this isn't psychiatry suddenly waking from a coma, aha! it turns out the existing data do show antipsychotics are mood stabilizers! Instead of using them to replace antiepileptics, they will use them to replace everything: SSRIs, benzos, antiepileptics, stimulants, etc.
And polypharmacy will only be reincarnated-- in the form of multiple simultaneous antipsychotics (Abiliquel, anyone?), with preposterous pharmacologic justifications ("this one acts on serotonin, so it's the antidepressant, and this one on dopamine, so it's the antimanic.") If anyone says that to you, stab them.
You don't get many changes like this, maybe once every ten years-- the last was the beginning of the Depakote era, and before that was the advent of SSRIs, each with it's own erroneous semantics ("kindling model;" "serotonin model of depression.")
I wish all the patients in the world good luck, you'll need it. Not because of the antipsychotics themselves, which will work or not, oblivious to doctor and diagnosis; but because of the doctors, who take little interest in examining the evidence behind their practice, and even less interest in reevaluating its core principles; and who lack the courage to even treat what they see, instead resorting to artificial, and wrong, paradigms and algorithms. There's not even pseudoscience here. Psychiatry is being lead by the siren call of semiotics, and it is saying, follow me, I am made of words...
Worse Than The Flu
Here's a little case report, about me, a cautionary tale about working too hard.
I had to go to Chicago for a case. It took three days. It took a lot out of me. There was the jet lag, and the work, and skipping meals, and sleep deprivation. I barely sleep at home, but all I could get in the hotel was 2 hours/night. I usually drink about 3 cups of coffee (16 oz each-- so I guess that's 6 actual cups) a day, but with this level of stress and tiredeness I was drinking 4-5. And, dare I admit it, I took a Provigil.
I could feel myself getting sick on the last day. Just get through it,
|
[
1728,
5934,
29901,
871,
278,
9418,
567,
3376,
327,
1199,
526,
1603,
14982,
287,
29889,
13,
6246,
1728,
16274,
1199,
27556,
5886,
29934,
3624,
29892,
1009,
671,
674,
281,
1662,
489,
392,
29892,
4100,
368,
29892,
577,
674,
1009,
2304,
310,
278,
24876,
19263,
376,
29924,
1175,
272,
897,
2590,
1213,
910,
338,
2675,
304,
6047,
19341,
616,
29892,
297,
1662,
29892,
541,
372,
674,
3799,
29889,
13,
14959,
363,
701,
11506,
7456,
9138,
393,
376,
2772,
2590,
29908,
338,
975,
6051,
4211,
2662,
29892,
393,
372,
338,
2289,
925,
489,
2834,
29889,
7419,
363,
7456,
393,
1286,
1284,
5886,
29934,
3624,
9455,
29915,
29873,
393,
11828,
1156,
599,
29892,
393,
278,
2030,
376,
29896,
29900,
29995,
2253,
1135,
2058,
833,
29908,
338,
263,
24148,
8938,
411,
2217,
24899,
936,
19725,
29889,
2193,
896,
526,
982,
975,
3880,
297,
413,
4841,
29889,
13,
3492,
1795,
1827,
29892,
4480,
29892,
3508,
29915,
29873,
278,
4845,
457,
310,
15680,
561,
2817,
4135,
263,
1781,
2655,
29936,
393,
5886,
29934,
3624,
526,
975,
3880,
297,
413,
4841,
29936,
393,
896,
9455,
29915,
29873,
393,
2107,
29936,
322,
393,
316,
2590,
338,
975,
6051,
4211,
2662,
29973,
2178,
310,
445,
338,
1565,
29892,
541,
445,
3508,
29915,
29873,
11643,
7163,
719,
7146,
6421,
304,
967,
4771,
267,
29936,
445,
338,
11643,
7163,
719,
18055,
278,
767,
293,
8576,
29889,
18585,
29892,
372,
29915,
29879,
3109,
5886,
29934,
3624,
363,
413,
4841,
29936,
541,
372,
29915,
29879,
901,
9418,
567,
3376,
327,
1199,
29889,
13,
29933,
5658,
21699,
727,
674,
367,
7456,
27556,
278,
2969,
393,
1162,
1264,
443,
666,
10170,
316,
2590,
338,
2289,
289,
666,
10170,
316,
2590,
29936,
393,
727,
526,
3619,
2531,
7492,
470,
902,
277,
3097,
15038,
29936,
393,
278,
9358,
680,
2460,
3002,
322,
3236,
338,
2788,
29892,
2992,
29889,
450,
4337,
674,
367,
304,
269,
802,
29872,
911,
714,
341,
7858,
964,
376,
19264,
29908,
322,
289,
666,
10170,
29889,
910,
2309,
29892,
9418,
567,
3376,
327,
1199,
4953,
937,
1196,
19518,
29889,
6439,
29892,
322,
1106,
363,
9418,
567,
3376,
327,
1199,
304,
679,
383,
7698,
2134,
791,
29879,
363,
413,
4841,
29889,
13,
29902,
6398,
306,
1033,
1207,
445,
26135,
29892,
541,
306,
29915,
29885,
1603,
9792,
292,
515,
590,
7786,
25927,
411,
4892,
29889,
1670,
29915,
29879,
694,
10466,
1244,
29892,
871,
263,
260,
682,
3241,
411,
4086,
322,
658,
29891,
1997,
583,
29892,
411,
380,
9921,
292,
2582,
29889,
3872,
29915,
29873,
1999,
420,
1963,
22824,
3755,
3447,
489,
445,
338,
263,
405,
7833,
29950,
6559,
29889,
13,
29902,
626,
451,
2750,
9418,
567,
3376,
327,
1199,
29892,
322,
306,
505,
1472,
1898,
304,
2649,
5019,
1058,
723,
11621,
393,
278,
848,
9436,
1510,
896,
526,
11558,
304,
3677,
347,
29886,
488,
415,
1199,
29889,
1205,
445,
3508,
29915,
29873,
11643,
7163,
719,
11584,
281,
5086,
515,
263,
419,
29874,
29892,
263,
2350,
29991,
372,
12169,
714,
278,
5923,
848,
437,
1510,
9418,
567,
3376,
327,
1199,
526,
286,
2092,
16160,
19427,
29991,
8669,
310,
773,
963,
304,
5191,
3677,
347,
29886,
488,
415,
1199,
29892,
896,
674,
671,
963,
304,
5191,
4129,
29901,
5886,
29934,
3624,
29892,
3856,
29834,
29892,
3677,
347,
29886,
488,
415,
1199,
29892,
20436,
352,
1934,
29892,
2992,
29889,
13,
2855,
15680,
561,
2817,
4135,
674,
871,
367,
337,
3742,
2753,
630,
489,
297,
278,
883,
310,
2999,
16991,
681,
9418,
567,
3376,
327,
1199,
313,
4920,
2638,
13295,
29892,
5019,
29973,
511,
411,
758,
2490,
261,
681,
1374,
2817,
562,
1189,
293,
925,
8232,
4852,
1366,
697,
14741,
373,
724,
327,
265,
262,
29892,
577,
372,
29915,
29879,
278,
3677,
680,
2139,
424,
29892,
322,
445,
697,
373,
22357,
314,
457,
29892,
577,
372,
29915,
29879,
278,
3677,
25895,
293,
23157,
960,
5019,
4083,
393,
304,
366,
29892,
380,
370,
963,
29889,
13,
3492,
1016,
29915,
29873,
679,
1784,
3620,
763,
445,
29892,
5505,
2748,
1432,
3006,
2440,
489,
278,
1833,
471,
278,
6763,
310,
278,
10034,
557,
866,
3152,
29892,
322,
1434,
393,
471,
278,
17623,
310,
5886,
29934,
3624,
29892,
1269,
411,
372,
29915,
29879,
1914,
4589,
650,
681,
29505,
4852,
14380,
1847,
1904,
15458,
376,
643,
327,
265,
262,
1904,
310,
316,
2590,
23157,
13,
29902,
6398,
599,
278,
22069,
297,
278,
3186,
1781,
9885,
29892,
366,
29915,
645,
817,
372,
29889,
2216,
1363,
310,
278,
9418,
567,
3376,
327,
1199,
6053,
29892,
607,
674,
664,
470,
451,
29892,
704,
17843,
2738,
304,
11619,
322,
24876,
19263,
29936,
541,
1363,
310,
278,
437,
14359,
29892,
1058,
2125,
2217,
4066,
297,
4392,
2827,
278,
10757,
5742,
1009,
6944,
29892,
322,
1584,
3109,
4066,
297,
337,
24219,
1218,
967,
7136,
18671,
29936,
322,
1058,
10225,
278,
19872,
304,
1584,
7539,
825,
896,
1074,
29892,
2012,
25362,
292,
304,
23116,
29892,
322,
2743,
29892,
26811,
335,
1516,
322,
14009,
29889,
1670,
29915,
29879,
451,
1584,
19923,
359,
15277,
1244,
29889,
16777,
7163,
719,
338,
1641,
3275,
491,
278,
8889,
264,
1246,
310,
12647,
327,
1199,
29892,
322,
372,
338,
5934,
29892,
1101,
592,
29892,
306,
626,
1754,
310,
3838,
856,
13,
29956,
272,
344,
498,
273,
450,
2379,
29884,
13,
10605,
29915,
29879,
263,
2217,
1206,
3461,
29892,
1048,
592,
29892,
263,
5777,
918,
653,
17694,
1048,
1985,
2086,
2898,
29889,
13,
29902,
750,
304,
748,
304,
10059,
363,
263,
1206,
29889,
739,
3614,
2211,
3841,
29889,
739,
3614,
263,
3287,
714,
310,
592,
29889,
1670,
471,
278,
22588,
11755,
29892,
322,
278,
664,
29892,
322,
14993,
3262,
592,
1338,
29892,
322,
8709,
1401,
1150,
362,
29889,
306,
16079,
368,
8709,
472,
3271,
29892,
541,
599,
306,
1033,
679,
297,
278,
16730,
471,
29871,
29906,
6199,
29914,
11147,
29889,
306,
5491,
13748,
1048,
29871,
29941,
2723,
567,
310,
26935,
313,
29896,
29953,
288,
29920,
1269,
489,
577,
306,
4140,
393,
29915,
29879,
29871,
29953,
3935,
2723,
567,
29897,
263,
2462,
29892,
541,
411,
445,
3233,
310,
22884,
322,
260,
533,
1145,
404,
306,
471,
13748,
292,
29871,
29946,
29899,
29945,
29889,
1126,
29892,
23222,
306,
20000,
372,
29892,
306,
3614,
263,
9133,
335,
309,
29889,
13,
29902,
1033,
4459,
6142,
2805,
17319,
373,
278,
1833,
2462,
29889,
3387,
679,
1549,
372,
29892
] | 1,024 |
CommonCrawl
| 0.168872 | 0.584647 | -0.0019 | -0.062864 |
I thought. Sick later. Work today.
I got home, exhausted. The next morning I felt sick, wiped out, achy. There you go, I said. I have the flu. I struggled through work, taking naps when I could.
As the day progressed, I got worse. Weakness, tiredness, horrible nausea, headache. I gagged at the thought of food, but I forced myself to at least drink Gatorade. Gatorade is the artificially sweetened sweat of male bicyclists. Every word of that description disgusts me. I drank the purple one.
Day 2 came, and I was worse, not better. Not even the same-- much worse. The headache was ruthless. The nausea had become motion sickness-- turning my head was a lunar launch. The arthralgias, bizarrely, had disappeared-- except in my neck, which had become very painful and stiff. I couldn't turn my head well. I could barely walk, I could barely think.
I went to work.
The weakness and lethargy had also changed-- into narcolepsy. It wasn't weakness-- I was drugged. I fell asleep for only a second at a time, but it overtook me every moment I wasn't active. Driving. Watching TV. Standing at a urinal. On an elevator. During phone calls. I could not stay awake. Simply closing my eyes would drop me into Stage IV sleep. I could still be talking, but if my eyes were closed I was asleep. And what I said was nonsensical.
I was almost helpless. I took Tylenol. Motrin. Tylenol + Motrin. Nothing. And I could not stay awake. The head and neck hurt so much that the only solace was sleep, which I couldn't stop anyway.
What kind of flu was this? And something worried me: why didn't I have a fever?
By day 3 I had what can only be described as the worst headache of my life. The nausea was constant.
Worst. Headache. Of. My. Life.
I rarely get sick, I rarely take pain relievers. I do 3 sets of 50 push ups a day. I'm pretty healthy, and I've never been incapacitated. I only say this as background for my next sentence: I was so sick I could not see.
Light hurt me, hurt my head. I could not look at the monitor, or TV. I wanted to be in a quiet, dark room-- asleep. With morphine. And the medical student in me solved the mystery: headache, stiff neck, photophobia, no fever. I had finally done what I had been threatening to do for so long: I had popped an aneurysm. I thought: so this is how it ends.
Nausea. Headache. Neck stiffness. Exhaustion.
Oh my God, could I be in caffeine withdrawal?
As soon as I thought it, I knew that was it. I couldn't believe it. I'd never felt it before because I'd never not had coffee before. And that first day back, being a little off, I skipped it-- which made me worse, and then the withdrawal hit.
I made some coffee. It smelled like battery acid. I put ice in it and drank it, one cup all at once. I gagged, twice.
Within ten minutes, I was 10% better. In 30 minutes, I was 50% better. In an hour I was 95% cured. From unable to move, to almost complete cure.
The cure was so total, the reversal so profound, that I actually couldn't remember how sick I was. I thought I must have been exaggerating.
So my body reminded me.
Four hours later, I started to feel that motion sickness again. By the fifth hour, I was on the floor again, same stiff neck and headache. And the nausea was worse: the thought of drinking the battery acid again was too much for me.
But I did it. And again, an hour later, I was completely cured.
How "real" is caffeine withdrawal? Clearly, my own experience takes it out of the theoretical realm. But what about:
the average coffee junkie when he goes to
|
[
306,
2714,
29889,
317,
860,
2678,
29889,
5244,
9826,
29889,
13,
29902,
2355,
3271,
29892,
18782,
16656,
29889,
450,
2446,
7250,
306,
7091,
17319,
29892,
281,
666,
287,
714,
29892,
3657,
29891,
29889,
1670,
366,
748,
29892,
306,
1497,
29889,
306,
505,
278,
20501,
29889,
306,
10205,
839,
1549,
664,
29892,
5622,
302,
2547,
746,
306,
1033,
29889,
13,
2887,
278,
2462,
6728,
287,
29892,
306,
2355,
15029,
29889,
1334,
557,
2264,
29892,
23407,
2264,
29892,
4029,
11710,
302,
1071,
29874,
29892,
2343,
1829,
29889,
306,
330,
351,
3192,
472,
278,
2714,
310,
9687,
29892,
541,
306,
11826,
6142,
304,
472,
3203,
13748,
402,
1061,
1943,
29889,
402,
1061,
1943,
338,
278,
23116,
368,
7901,
6302,
287,
7901,
271,
310,
14263,
289,
4245,
695,
2879,
29889,
7569,
1734,
310,
393,
6139,
766,
29887,
504,
29879,
592,
29889,
306,
270,
10003,
278,
3708,
552,
697,
29889,
13,
12742,
29871,
29906,
2996,
29892,
322,
306,
471,
15029,
29892,
451,
2253,
29889,
2216,
1584,
278,
1021,
489,
1568,
15029,
29889,
450,
2343,
1829,
471,
364,
2806,
2222,
29889,
450,
302,
1071,
29874,
750,
4953,
10884,
17319,
2264,
489,
14712,
590,
2343,
471,
263,
25081,
279,
6826,
29889,
450,
564,
386,
1705,
3146,
294,
29892,
289,
15356,
276,
368,
29892,
750,
24273,
489,
5174,
297,
590,
18873,
29892,
607,
750,
4953,
1407,
6788,
1319,
322,
380,
2593,
29889,
306,
8496,
29915,
29873,
2507,
590,
2343,
1532,
29889,
306,
1033,
16079,
368,
6686,
29892,
306,
1033,
16079,
368,
1348,
29889,
13,
29902,
3512,
304,
664,
29889,
13,
1576,
8062,
2264,
322,
454,
386,
1191,
29891,
750,
884,
3939,
489,
964,
17655,
10936,
567,
29891,
29889,
739,
9007,
29915,
29873,
8062,
2264,
489,
306,
471,
5883,
1505,
287,
29889,
306,
8379,
408,
5436,
363,
871,
263,
1473,
472,
263,
931,
29892,
541,
372,
975,
517,
554,
592,
1432,
3256,
306,
9007,
29915,
29873,
6136,
29889,
360,
1150,
292,
29889,
24274,
292,
5648,
29889,
6679,
292,
472,
263,
5065,
979,
29889,
1551,
385,
11858,
1061,
29889,
7133,
9008,
5717,
29889,
306,
1033,
451,
7952,
3773,
1296,
29889,
3439,
17632,
14382,
590,
5076,
723,
5768,
592,
964,
24906,
6599,
8709,
29889,
306,
1033,
1603,
367,
9963,
29892,
541,
565,
590,
5076,
892,
5764,
306,
471,
408,
5436,
29889,
1126,
825,
306,
1497,
471,
302,
787,
575,
936,
29889,
13,
29902,
471,
4359,
1081,
572,
404,
29889,
306,
3614,
9656,
2435,
324,
29889,
7142,
17056,
29889,
9656,
2435,
324,
718,
7142,
17056,
29889,
9531,
29889,
1126,
306,
1033,
451,
7952,
3773,
1296,
29889,
450,
2343,
322,
18873,
21682,
577,
1568,
393,
278,
871,
899,
815,
471,
8709,
29892,
607,
306,
8496,
29915,
29873,
5040,
8763,
29889,
13,
5618,
2924,
310,
20501,
471,
445,
29973,
1126,
1554,
6365,
1255,
592,
29901,
2020,
3282,
29915,
29873,
306,
505,
263,
1238,
369,
29973,
13,
2059,
2462,
29871,
29941,
306,
750,
825,
508,
871,
367,
5439,
408,
278,
17322,
2343,
1829,
310,
590,
2834,
29889,
450,
302,
1071,
29874,
471,
4868,
29889,
13,
29956,
272,
303,
29889,
12252,
1829,
29889,
4587,
29889,
1619,
29889,
4634,
29889,
13,
29902,
23703,
679,
17319,
29892,
306,
23703,
2125,
6788,
1104,
347,
874,
29889,
306,
437,
29871,
29941,
6166,
310,
29871,
29945,
29900,
5503,
24081,
263,
2462,
29889,
306,
29915,
29885,
5051,
9045,
29891,
29892,
322,
306,
29915,
345,
2360,
1063,
297,
5030,
562,
22731,
29889,
306,
871,
1827,
445,
408,
3239,
363,
590,
2446,
10541,
29901,
306,
471,
577,
17319,
306,
1033,
451,
1074,
29889,
13,
20769,
21682,
592,
29892,
21682,
590,
2343,
29889,
306,
1033,
451,
1106,
472,
278,
11819,
29892,
470,
5648,
29889,
306,
5131,
304,
367,
297,
263,
11813,
29892,
6501,
5716,
489,
408,
5436,
29889,
2973,
18131,
457,
29889,
1126,
278,
16083,
8368,
297,
592,
7484,
278,
29236,
29901,
2343,
1829,
29892,
380,
2593,
18873,
29892,
6731,
3021,
711,
423,
29892,
694,
1238,
369,
29889,
306,
750,
7146,
2309,
825,
306,
750,
1063,
20616,
292,
304,
437,
363,
577,
1472,
29901,
306,
750,
772,
2986,
385,
385,
5411,
952,
29885,
29889,
306,
2714,
29901,
577,
445,
338,
920,
372,
10614,
29889,
13,
29940,
1071,
29874,
29889,
12252,
1829,
29889,
2448,
384,
380,
2593,
2264,
29889,
1222,
2350,
504,
291,
29889,
13,
9048,
590,
4177,
29892,
1033,
306,
367,
297,
274,
3470,
29872,
457,
28679,
284,
29973,
13,
2887,
4720,
408,
306,
2714,
372,
29892,
306,
6363,
393,
471,
372,
29889,
306,
8496,
29915,
29873,
4658,
372,
29889,
306,
29915,
29881,
2360,
7091,
372,
1434,
1363,
306,
29915,
29881,
2360,
451,
750,
26935,
1434,
29889,
1126,
393,
937,
2462,
1250,
29892,
1641,
263,
2217,
1283,
29892,
306,
14993,
2986,
372,
489,
607,
1754,
592,
15029,
29892,
322,
769,
278,
28679,
284,
7124,
29889,
13,
29902,
1754,
777,
26935,
29889,
739,
1560,
14356,
763,
16988,
22193,
29889,
306,
1925,
14890,
297,
372,
322,
270,
10003,
372,
29892,
697,
18002,
599,
472,
2748,
29889,
306,
330,
351,
3192,
29892,
8951,
29889,
13,
3047,
262,
3006,
6233,
29892,
306,
471,
29871,
29896,
29900,
29995,
2253,
29889,
512,
29871,
29941,
29900,
6233,
29892,
306,
471,
29871,
29945,
29900,
29995,
2253,
29889,
512,
385,
7234,
306,
471,
29871,
29929,
29945,
29995,
274,
2955,
29889,
3645,
9368,
304,
4337,
29892,
304,
4359,
4866,
274,
545,
29889,
13,
1576,
274,
545,
471,
577,
3001,
29892,
278,
18764,
284,
577,
2600,
618,
29892,
393,
306,
2869,
8496,
29915,
29873,
6456,
920,
17319,
306,
471,
29889,
306,
2714,
306,
1818,
505,
1063,
429,
9921,
1218,
29889,
13,
6295,
590,
3573,
1083,
513,
287,
592,
29889,
13,
29943,
473,
6199,
2678,
29892,
306,
4687,
304,
4459,
393,
10884,
17319,
2264,
1449,
29889,
2648,
278,
18615,
7234,
29892,
306,
471,
373,
278,
11904,
1449,
29892,
1021,
380,
2593,
18873,
322,
2343,
1829,
29889,
1126,
278,
302,
1071,
29874,
471,
15029,
29901,
278,
2714,
310,
13748,
292,
278,
16988,
22193,
1449,
471,
2086,
1568,
363,
592,
29889,
13,
6246,
306,
1258,
372,
29889,
1126,
1449,
29892,
385,
7234,
2678,
29892,
306,
471,
6446,
274,
2955,
29889,
13,
5328,
376,
6370,
29908,
338,
274,
3470,
29872,
457,
28679,
284,
29973,
17732,
368,
29892,
590,
1914,
7271,
4893,
372,
714,
310,
278,
23399,
1855,
29885,
29889,
1205,
825,
1048,
29901,
13,
1552,
6588,
26935,
432,
2960,
347,
746,
540,
5771,
304
] | 1,024 |
CommonCrawl
| 0.055265 | -0.429248 | -0.4186 | -0.491025 |
{% load i18n %}
{% extends "base.html" %}
{% block header %}
<script type="text/javascript">
</script>
{% endblock %}
{% block nav2 %}
<div id="dnav2">
<a href="/admin/setup" class="current" >{%trans "Basic" %}</a>
<a href="/admin/import">{%trans "Import/Export" %}</a>
<a href="/admin/sitemap">{%trans "Sitemap" %}</a>
<a href="/admin/tools">{%trans "Tools" %}</a>
</div>
{% endblock %}
{% block content %}
<!-- Admin Full Width -->
<div id="m_admin">
<form method="post">
<div class="wrap">
<h2>{%trans "Setup and Configuraiton for this Blog"%}</h2>
<table class="form-table">
<tr valign="top">
<th scope="row">{% trans "Blog Title:"%}</th>
<td><input type="text" size="45" value="{{ blog.title }}" id="title" name="title"/>
</td>
</tr>
<tr valign="top">
<th scope="row">{% trans "Sub Title:"%}</th>
<td><input type="text" size="45" value="{{ blog.subtitle }}" style="width: 95%;" id="subtitle" name="subtitle" />
<br/>
{% trans "In a few words, explain what this blog is about."%}</td>
</tr>
<tr valign="top">
<th scope="row">{% trans "Permalink Structure:"%}</th>
<td><input type="text" size="45" style="width: 95%;" value="{{ blog.link_format }}" id="link_format" name="link_format" />
<br>{% trans "Allowed variants: " %}%(year)s , %(month)s , %(day)s , %(postname)s , %(post_id)s
</td>
</tr>
<tr valign="top">
<th scope="row">{% trans "Domain:"%}</th>
<td><input type="text" size="45" value="{{ blog.domain }}" id="domain" name="domain" />
</td>
</tr>
</tr>
<tr valign="top">
<th scope="row">{% trans "Micolog Address(URL):"%}</th>
<td><input type="text" size="45" value="{{ blog.baseurl }}" id="baseurl" name="baseurl" />
</td>
</tr>
<tr valign="top">
<th scope="row">{% trans "RSS Feed URL:"%}</th>
<td><input type="text" size="45" value="{{ blog.feedurl }}" id="feedurl" name="feedurl" />
</td>
</tr>
<tr valign="top">
<th scope="row">{% trans "Time delta:(Hours)"%}</th>
<td><input type="text" size="45" value="{{ blog.timedelta }}" id="timedelta" name="timedelta" />
</td>
</tr>
<tr valign="top">
<th scope="row">{% trans "Theme:"%}</th>
<td>
<select id="theme_name" name="theme_name" >
{%for theme in themes%}
<option value="{{theme}}" {%ifequal blog.theme_name theme%}selected='selected'{%endifequal%}>{{theme}}</option>
{%endfor%}
</select>
</td>
<tr valign="top">
<th scope="row">{% trans "Posts per page:"%}</th>
<td><input type="text" size="8" value="{{ blog.posts_per_page }}" id="posts_per_
|
[
18674,
2254,
474,
29896,
29947,
29876,
15493,
13,
18255,
4988,
376,
3188,
29889,
1420,
29908,
15493,
13,
18255,
2908,
4839,
15493,
13,
29966,
2154,
1134,
543,
726,
29914,
7729,
1013,
13,
13,
829,
2154,
29958,
13,
18255,
1095,
1271,
15493,
13,
13,
18255,
2908,
6283,
29906,
15493,
13,
29966,
4563,
1178,
543,
5200,
485,
29906,
1013,
13,
529,
29874,
2822,
13802,
6406,
29914,
14669,
29908,
770,
543,
3784,
29908,
1405,
18255,
3286,
376,
16616,
29908,
15493,
829,
29874,
29958,
13,
529,
29874,
2822,
13802,
6406,
29914,
5215,
1013,
18255,
3286,
376,
17518,
29914,
26382,
29908,
15493,
829,
29874,
29958,
13,
529,
29874,
2822,
13802,
6406,
29914,
29879,
667,
481,
1013,
18255,
3286,
376,
29903,
667,
481,
29908,
15493,
829,
29874,
29958,
13,
529,
29874,
2822,
13802,
6406,
29914,
8504,
1013,
18255,
3286,
376,
24183,
29908,
15493,
829,
29874,
29958,
13,
829,
4563,
29958,
13,
18255,
1095,
1271,
15493,
13,
13,
18255,
2908,
2793,
15493,
13,
13,
12,
10341,
10229,
14846,
21485,
6660,
13,
1678,
529,
4563,
1178,
543,
29885,
29918,
6406,
1013,
13,
4706,
529,
689,
1158,
543,
2490,
1013,
13,
9651,
529,
4563,
770,
543,
6312,
1013,
13,
9651,
529,
29882,
29906,
29958,
18255,
3286,
376,
26947,
322,
12782,
332,
1249,
265,
363,
445,
350,
1188,
29908,
29995,
16040,
29882,
29906,
29958,
13,
13,
9651,
529,
2371,
770,
543,
689,
29899,
2371,
1013,
13,
462,
529,
509,
659,
647,
543,
3332,
1013,
13,
18884,
529,
386,
6874,
543,
798,
1013,
18255,
1301,
376,
29933,
1188,
18527,
6160,
29995,
16040,
386,
29958,
13,
18884,
529,
1594,
5299,
2080,
1134,
543,
726,
29908,
2159,
543,
29946,
29945,
29908,
995,
543,
6224,
12618,
29889,
3257,
500,
5038,
29871,
1178,
543,
3257,
29908,
1024,
543,
3257,
4681,
13,
1669,
1533,
1594,
29958,
13,
18884,
1533,
509,
29958,
13,
13,
18884,
529,
509,
659,
647,
543,
3332,
1013,
13,
18884,
529,
386,
6874,
543,
798,
1013,
18255,
1301,
376,
4035,
18527,
6160,
29995,
16040,
386,
29958,
13,
18884,
529,
1594,
5299,
2080,
1134,
543,
726,
29908,
2159,
543,
29946,
29945,
29908,
995,
543,
6224,
12618,
29889,
1491,
3257,
500,
5038,
3114,
543,
2103,
29901,
29871,
29929,
29945,
8874,
29908,
1178,
543,
1491,
3257,
29908,
1024,
543,
1491,
3257,
29908,
2900,
13,
18884,
529,
1182,
3779,
13,
18884,
18674,
1301,
376,
797,
263,
2846,
3838,
29892,
5649,
825,
445,
12618,
338,
1048,
1213,
29995,
16040,
1594,
29958,
13,
18884,
1533,
509,
29958,
13,
529,
509,
659,
647,
543,
3332,
1013,
13,
18884,
529,
386,
6874,
543,
798,
1013,
18255,
1301,
376,
15737,
284,
682,
3767,
12425,
6160,
29995,
16040,
386,
29958,
13,
18884,
529,
1594,
5299,
2080,
1134,
543,
726,
29908,
2159,
543,
29946,
29945,
29908,
3114,
543,
2103,
29901,
29871,
29929,
29945,
8874,
29908,
995,
543,
6224,
12618,
29889,
2324,
29918,
4830,
500,
5038,
29871,
1178,
543,
2324,
29918,
4830,
29908,
1024,
543,
2324,
29918,
4830,
29908,
2900,
13,
462,
529,
1182,
29958,
18255,
1301,
376,
15930,
287,
29161,
29901,
376,
1273,
10560,
29898,
6360,
29897,
29879,
1919,
1273,
29898,
10874,
29897,
29879,
1919,
1273,
29898,
3250,
29897,
29879,
1919,
1273,
29898,
2490,
978,
29897,
29879,
1919,
1273,
29898,
2490,
29918,
333,
29897,
29879,
13,
1669,
1533,
1594,
29958,
13,
18884,
1533,
509,
29958,
13,
462,
29871,
529,
509,
659,
647,
543,
3332,
1013,
13,
18884,
529,
386,
6874,
543,
798,
1013,
18255,
1301,
376,
15951,
6160,
29995,
16040,
386,
29958,
13,
18884,
529,
1594,
5299,
2080,
1134,
543,
726,
29908,
2159,
543,
29946,
29945,
29908,
995,
543,
6224,
12618,
29889,
7247,
500,
5038,
29871,
1178,
543,
7247,
29908,
1024,
543,
7247,
29908,
2900,
13,
1669,
1533,
1594,
29958,
13,
18884,
1533,
509,
29958,
13,
18884,
1533,
509,
29958,
13,
462,
29871,
529,
509,
659,
647,
543,
3332,
1013,
13,
18884,
529,
386,
6874,
543,
798,
1013,
18255,
1301,
376,
29924,
293,
1189,
16428,
29898,
4219,
1125,
29908,
29995,
16040,
386,
29958,
13,
18884,
529,
1594,
5299,
2080,
1134,
543,
726,
29908,
2159,
543,
29946,
29945,
29908,
995,
543,
6224,
12618,
29889,
3188,
2271,
500,
5038,
29871,
1178,
543,
3188,
2271,
29908,
1024,
543,
3188,
2271,
29908,
2900,
13,
1669,
1533,
1594,
29958,
13,
18884,
1533,
509,
29958,
13,
13,
462,
529,
509,
659,
647,
543,
3332,
1013,
13,
18884,
529,
386,
6874,
543,
798,
1013,
18255,
1301,
376,
29934,
1799,
5169,
287,
3988,
6160,
29995,
16040,
386,
29958,
13,
18884,
529,
1594,
5299,
2080,
1134,
543,
726,
29908,
2159,
543,
29946,
29945,
29908,
995,
543,
6224,
12618,
29889,
18798,
2271,
500,
5038,
29871,
1178,
543,
18798,
2271,
29908,
1024,
543,
18798,
2271,
29908,
2900,
13,
1669,
1533,
1594,
29958,
13,
18884,
1533,
509,
29958,
13,
462,
529,
509,
659,
647,
543,
3332,
1013,
13,
18884,
529,
386,
6874,
543,
798,
1013,
18255,
1301,
376,
2481,
19471,
5919,
29950,
2470,
5513,
29995,
16040,
386,
29958,
13,
18884,
529,
1594,
5299,
2080,
1134,
543,
726,
29908,
2159,
543,
29946,
29945,
29908,
995,
543,
6224,
12618,
29889,
9346,
287,
2554,
500,
5038,
29871,
1178,
543,
9346,
287,
2554,
29908,
1024,
543,
9346,
287,
2554,
29908,
2900,
13,
1669,
1533,
1594,
29958,
13,
18884,
1533,
509,
29958,
13,
13,
13,
9651,
529,
509,
659,
647,
543,
3332,
1013,
13,
18884,
529,
386,
6874,
543,
798,
1013,
18255,
1301,
376,
17693,
6160,
29995,
16040,
386,
29958,
13,
18884,
529,
1594,
29958,
13,
462,
1678,
529,
2622,
29871,
1178,
543,
18193,
29918,
978,
29908,
1024,
543,
18193,
29918,
978,
29908,
1405,
13,
462,
1678,
18674,
1454,
10929,
297,
963,
267,
29995,
29913,
13,
462,
1678,
529,
3385,
995,
543,
6224,
18193,
930,
29908,
18674,
1607,
15380,
12618,
29889,
18193,
29918,
978,
10929,
29995,
29913,
8391,
2433,
8391,
29915,
18255,
355,
1607,
15380,
29995,
17428,
6224,
18193,
930,
829,
3385,
29958,
13,
462,
1678,
18674,
355,
1454,
29995,
29913,
13,
13,
462,
1678,
1533,
2622,
29958,
13,
1669,
1533,
1594,
29958,
13,
462,
29871,
529,
509,
659,
647,
543,
3332,
1013,
13,
18884,
529,
386,
6874,
543,
798,
1013,
18255,
1301,
376,
6747,
29879,
639,
1813,
6160,
29995,
16040,
386,
29958,
13,
18884,
529,
1594,
5299,
2080,
1134,
543,
726,
29908,
2159,
543,
29947,
29908,
995,
543,
6224,
12618,
29889,
14080,
29918,
546,
29918,
3488,
500,
5038,
29871,
1178,
543,
14080,
29918,
546,
29918
] | 1,024 |
Github
| -1.057542 | -1.172554 | -1.716893 | -0.910015 |
David Nissan, born and raised in Iran, is an expert on Iran and the Middle East. For 16 years, he served as officer and commander in the Israeli Defense Forces, and another 7 years as a consultant for the Ministry of Defense. David has given lectures in leading companies and financial institutions in the private sector, universities, and government ministries including the Ministry of Foreign Affairs, and the Prime Minister's Office. He has been interviewed on Israeli national radio and television, has spoken in various forums outside of Israel, and is on the keynote speakers list of the London Speaker Bureau.
Israel, Iran and the Middle East - An Unconventional Perspective
A realistic and often unusual assessment of the breathtaking drama being played out in the Middle East today, focusing on Israel and Iran. Exposure of critical aspects in Middle Eastern affairs from an intelligence officer's point of view - with an emphasis on analyzing, understanding and countering the core motivations of Israel's main adversaries.
Revolution and Survival - The Jews in Iran
The fascinating story of one of the most ancient Jewish communities in the world. How did the establishment of the State of Israel affect this community, and what happened when the 1979 revolution in Iran put an end to the Golden Age enjoyed by the Jews during the Shah's reign, and forced them to choose between Judaism and Zionism? Why did most leave, and why do the remainder insist on staying?
Behind the Veil - A Virtual Tour of Iran
How much do we really know about Iran - a country that has continuously been in the world news headlines in recent years? A rare and magical glimpse into this beautiful, mysterious and fascinating country that is hidden behind the dark and ominous veil of the current Islamic regime. With emphasis on the rich Persian culture. David has presented this lecture at the Israeli Foreign Ministry.
Inside the Mind of Iran's Leader
An unusual lecture in which David Nissan, a former officer and commander in Israel's Intelligence Corps, assumes the identity of Iran's leader, Ayatollah Khamenei, and through the realistic, candid and outspoken presentation of the Iranian perspective on key relevant issues, enables a deeper and more accurate understanding of Iran's behavior in the international arena vis-a-vis Israel and the West.
- Prime Minister's Office
- Ministry Of Foreign Affairs
- Ministry Of Defense
- Ministry of Justice
- Tel-Aviv University
- Bank of Israel
- Israel Discount Bank
- Bank Leumi
- Intel
- HP
- HSBC Bank
- Elbit Systems
- Israel Aircraft Industries
- Rabin Medical Center
- Bnei Brit
- WIZO
- Maccabi
Full Client List
Thank you for being our guest speaker at our event in Chicago.... your presentation to donors of the Simon Wiesenthal Center was absolutely wonderful..... all truly enjoyed the insights they gained from your presentation. The event was extremely successful in cultivating prospective donors for the Center.
Alison Pure-Slovin, Director of the Simon Wiesenthal Center, Chicago
This was definitely one of the best lectures we have had over the last 14 years since we started the Riviera Paliament...... We enjoyed every minute of it, and would love to host you again in the future.
Ran Regev, Executive Committee, The Riviera Paliament, Miami
The lectures you gave at our events in Chicago and New York were truly incredible, and we are very grateful..... I highly recommend David to any organization interested in contracting him to speak at an event. In addition to being a great speaker with lectures full of humor and interesting anecdotes, David is also a very kind and charismatic person.
Hilary Cramer Robinson, Executive Director, American Friends of the Israel National Museum of Science, New York
David Nissan offers a calculated and intuitive understanding of the ever fluid dynamic of Middle East politics. It is worth attending a his lectures just to get his on the ground perspective that is so very different from the approach taken by mass media outlets.
Andreas Akaras, President, Saint Andrew's Freedom Forum, Washington DC
We had David Nissan speak online to our community. He was fantastic! David is a masterful educator, weaving a personal story into the complicated nature of the Jewish community in Iran, and the country as a whole.
Rabbi Daniel Brenner, Congregation B'nai B'rith, Santa Barbara, California
David N
|
[
4699,
405,
790,
273,
29892,
6345,
322,
10425,
297,
14883,
29892,
338,
385,
17924,
373,
14883,
322,
278,
14253,
6932,
29889,
1152,
29871,
29896,
29953,
2440,
29892,
540,
6766,
408,
12139,
322,
22079,
297,
278,
22895,
5037,
5282,
1947,
25571,
29892,
322,
1790,
29871,
29955,
2440,
408,
263,
8799,
424,
363,
278,
20886,
310,
5282,
1947,
29889,
4699,
756,
2183,
13081,
1973,
297,
8236,
14582,
322,
18161,
21561,
297,
278,
2024,
17535,
29892,
4946,
1907,
29892,
322,
5874,
9668,
2722,
3704,
278,
20886,
310,
19358,
26049,
29892,
322,
278,
15512,
7668,
29915,
29879,
11367,
29889,
940,
756,
1063,
15593,
287,
373,
22895,
5037,
4797,
7155,
322,
11456,
29892,
756,
19182,
297,
5164,
363,
6762,
5377,
310,
11996,
29892,
322,
338,
373,
278,
1589,
948,
866,
7726,
414,
1051,
310,
278,
4517,
5013,
5790,
18903,
29889,
13,
3624,
9058,
29892,
14883,
322,
278,
14253,
6932,
448,
530,
853,
535,
794,
1848,
9034,
12645,
13,
29909,
1855,
4695,
322,
4049,
22910,
24809,
358,
310,
278,
2078,
271,
400,
5086,
18621,
1641,
5318,
714,
297,
278,
14253,
6932,
9826,
29892,
12789,
4746,
373,
11996,
322,
14883,
29889,
1222,
1066,
545,
310,
12187,
21420,
297,
14253,
16162,
19469,
515,
385,
21082,
12139,
29915,
29879,
1298,
310,
1776,
448,
411,
385,
19310,
275,
373,
29537,
292,
29892,
8004,
322,
6795,
292,
278,
7136,
17385,
800,
310,
11996,
29915,
29879,
1667,
19901,
4314,
29889,
13,
1123,
4068,
322,
6298,
29894,
2561,
448,
450,
22057,
297,
14883,
13,
1576,
21028,
262,
1218,
5828,
310,
697,
310,
278,
1556,
12297,
16728,
23507,
297,
278,
3186,
29889,
1128,
1258,
278,
25012,
310,
278,
4306,
310,
11996,
6602,
445,
7881,
29892,
322,
825,
9559,
746,
278,
29871,
29896,
29929,
29955,
29929,
19479,
297,
14883,
1925,
385,
1095,
304,
278,
16108,
16767,
27849,
491,
278,
22057,
2645,
278,
25897,
29915,
29879,
20913,
29892,
322,
11826,
963,
304,
6755,
1546,
435,
6191,
1608,
322,
796,
291,
1608,
29973,
3750,
1258,
1556,
5967,
29892,
322,
2020,
437,
278,
21162,
1663,
391,
373,
7952,
292,
29973,
13,
3629,
16887,
278,
8980,
309,
448,
319,
19181,
6371,
310,
14883,
13,
5328,
1568,
437,
591,
2289,
1073,
1048,
14883,
448,
263,
4234,
393,
756,
3133,
5794,
1063,
297,
278,
3186,
9763,
2343,
9012,
297,
7786,
2440,
29973,
319,
10812,
322,
2320,
936,
330,
12552,
344,
964,
445,
9560,
29892,
22277,
2738,
322,
21028,
262,
1218,
4234,
393,
338,
7934,
5742,
278,
6501,
322,
288,
1195,
681,
2453,
309,
310,
278,
1857,
16427,
293,
22384,
29889,
2973,
19310,
275,
373,
278,
8261,
9034,
713,
9257,
29889,
4699,
756,
9132,
445,
29197,
472,
278,
22895,
5037,
19358,
20886,
29889,
13,
797,
2975,
278,
20152,
310,
14883,
29915,
29879,
951,
1664,
13,
2744,
22910,
29197,
297,
607,
4699,
405,
790,
273,
29892,
263,
4642,
12139,
322,
22079,
297,
11996,
29915,
29879,
3159,
28286,
16408,
29892,
15894,
278,
10110,
310,
14883,
29915,
29879,
11822,
29892,
15846,
14947,
8083,
476,
3391,
1600,
29875,
29892,
322,
1549,
278,
1855,
4695,
29892,
7105,
322,
714,
1028,
4476,
24329,
310,
278,
14883,
713,
18520,
373,
1820,
8018,
5626,
29892,
28936,
263,
25871,
322,
901,
16232,
8004,
310,
14883,
29915,
29879,
6030,
297,
278,
6121,
564,
2386,
1998,
29899,
29874,
29899,
1730,
11996,
322,
278,
3122,
29889,
13,
29899,
15512,
7668,
29915,
29879,
11367,
13,
29899,
20886,
4587,
19358,
26049,
13,
29899,
20886,
4587,
5282,
1947,
13,
29899,
20886,
310,
17181,
13,
29899,
18815,
29899,
12810,
440,
3014,
13,
29899,
10253,
310,
11996,
13,
29899,
11996,
3295,
2798,
10253,
13,
29899,
10253,
951,
15547,
13,
29899,
18555,
13,
29899,
379,
29925,
13,
29899,
379,
1744,
29907,
10253,
13,
29899,
1260,
2966,
23985,
13,
29899,
11996,
319,
12664,
12157,
2722,
13,
29899,
16155,
262,
20795,
7817,
13,
29899,
350,
484,
29875,
3230,
13,
29899,
399,
26664,
29949,
13,
29899,
4326,
29883,
19266,
13,
13658,
12477,
2391,
13,
25271,
366,
363,
1641,
1749,
17838,
25657,
472,
1749,
1741,
297,
10059,
3045,
596,
24329,
304,
1016,
943,
310,
278,
11254,
27178,
296,
4077,
7817,
471,
13312,
20695,
18598,
599,
19781,
27849,
278,
1663,
5861,
896,
17515,
515,
596,
24329,
29889,
450,
1741,
471,
14154,
9150,
297,
18834,
1218,
10791,
12645,
1016,
943,
363,
278,
7817,
29889,
13,
2499,
2285,
349,
545,
29899,
29903,
417,
3845,
29892,
15944,
310,
278,
11254,
27178,
296,
4077,
7817,
29892,
10059,
13,
4013,
471,
11630,
697,
310,
278,
1900,
13081,
1973,
591,
505,
750,
975,
278,
1833,
29871,
29896,
29946,
2440,
1951,
591,
4687,
278,
21505,
8311,
349,
2606,
1166,
3045,
636,
1334,
27849,
1432,
11015,
310,
372,
29892,
322,
723,
5360,
304,
3495,
366,
1449,
297,
278,
5434,
29889,
13,
29934,
273,
2169,
5750,
29892,
28841,
12930,
29892,
450,
21505,
8311,
349,
2606,
1166,
29892,
23740,
13,
1576,
13081,
1973,
366,
4846,
472,
1749,
4959,
297,
10059,
322,
1570,
3088,
892,
19781,
29811,
1821,
29892,
322,
591,
526,
1407,
28656,
18598,
306,
10712,
6907,
4699,
304,
738,
13013,
8852,
297,
8078,
292,
1075,
304,
7726,
472,
385,
1741,
29889,
512,
6124,
304,
1641,
263,
2107,
25657,
411,
13081,
1973,
2989,
310,
27448,
322,
8031,
385,
687,
29881,
4769,
29892,
4699,
338,
884,
263,
1407,
2924,
322,
1373,
1608,
2454,
2022,
29889,
13,
29950,
309,
653,
315,
2572,
261,
22265,
29892,
28841,
15944,
29892,
3082,
11169,
1975,
310,
278,
11996,
3086,
6838,
310,
9327,
29892,
1570,
3088,
13,
19504,
405,
790,
273,
16688,
263,
12833,
322,
27951,
573,
8004,
310,
278,
3926,
22576,
7343,
310,
14253,
6932,
22661,
29889,
739,
338,
7088,
1098,
2548,
263,
670,
13081,
1973,
925,
304,
679,
670,
373,
278,
5962,
18520,
393,
338,
577,
1407,
1422,
515,
278,
2948,
4586,
491,
4158,
5745,
714,
10376,
29889,
13,
2855,
12588,
319,
5689,
294,
29892,
7178,
29892,
4107,
11571,
29915,
29879,
3878,
11607,
29474,
29892,
7660,
13681,
13,
4806,
750,
4699,
405,
790,
273,
7726,
7395,
304,
1749,
7881,
29889,
940,
471,
13568,
6288,
29991,
4699,
338,
263,
5835,
1319,
6320,
1061,
29892,
591,
5555,
263,
7333,
5828,
964,
278,
12092,
5469,
310,
278,
16728,
7881,
297,
14883,
29892,
322,
278,
4234,
408,
263,
3353,
29889,
13,
29934,
370,
5365,
8432,
350,
1267,
1089,
29892,
6005,
1727,
362,
350,
29915,
1056,
29875,
350,
29915,
29878,
389,
29892,
7510,
18506,
29892,
8046,
13,
19504,
405
] | 1,024 |
CommonCrawl
| 0.946291 | 0.511971 | 1.175576 | 0.856069 |
issan talked to the RGS on Iran and gave an interesting virtual tour of the country. David is a natural speaker, fluent and humorous, and his extensive international experience is evident.
Rupert McCowan, Director of the Royal Geographical Society - Hong Kong
Our audience was very impressed with your expertise and especially with the fascinating and thought-provoking manner in which you presented your views through the perspective of an Israeli intelligence officer. Your lecture was outstanding and unique.
Ronny Ben-Joseph, President of the Babylonian Jewish Center, Great Neck, New York
Your presentation on the Iran deal, the Middle East and Israel's current situation was not only compelling and entertaining, but succeeded tremendously in educating and inspiring at the same time. Your impressive talk left everyone wanting more.
Rabbi Daniel Mehlman, Temple Ner Tamid, Downey, Los Angeles County, California
The lecture was interesting and fascinating, and was delivered in a clear and flowing manner. It was an experience that left a deep impression.
Frieda Cohen, Training Division of Israel's Foreign Ministry
An exceptionally fascinating and original lecture.
Ruti Fink, Israel Ministry of Defense
I received wonderful reactions to your lecture, with extraordinary praise for its innovative approach, clarity, and delivery. In the academic world, there are many who are knowledgeable about the material that they research, but not all can fascinate an audience like you did.
Professor David Menashri, Head of the Center for Iranian Studies, Tel-Aviv University
David Nissan was born and raised in Iran, and after graduating from the International Community School in Teheran, came to Israel to the Hebrew University of Jerusalem where he received his B.A. in Middle-Eastern Studies, and his M.A. in International Relations. He then joined the IDF, and after 16 years of service as an officer and commander, retired and completed a Masters degree in Business Administration, as well as certification studies for leading tours abroad.
For five years, David was the editor and site administrator of "Knesset Update", an online English-language weekly magazine , which reported on parliamentary/political events in Israel. Also, he was employed for seven years by the Israeli Ministry of Defense as a part-time consultant, and worked for 4 years as an senior analyst for Terrogence, Israel's leading company for the collection and analysis of security/terror-related intelligence from the internet. David was the editor of the Middle East and Iran Newsletter, and is currently a full-time professional speaker and consultant.
David's unique multi-cultural background (he speaks five languages), his academic credentials and his military/security career led him to focus his lectures on Middle-East and Iranian affairs. He is a sought-after lecturer in Israel, is completely fluent in English, and has given lectures in a variety of forums and conferences in leading companies and financial institutions in the private sector, universities, and government ministries including the Ministry of Defense, the Ministry of Foreign Affairs, and the Prime Minister's Office. He has been interviewed on Israeli national radio and television, has spoken in various forums outside of Israel, and is on the keynote speakers list of the London Speaker Bureau.
- Ministry for Foreign Affairs
- Ministry of Agriculture
- Rafael Advanced Defense Systems
- Fedex
- Procter & Gamble
- Elbit
- Verint
- Israel Engineers Association
- CBC Group
- Elisra
- Sandisk
- Shopping.Com
- Haifa University
- Bar Ilan University
- Netanya Academic College
- AACI - Association of Americans and Canadians in Israel
- ESRA-English Speaking Residents Association of Israel
- Rambam Medical Center
- Migdal Insurance Corporation
- Kanat Insurance Company
- Israel Ports Authority
- Isrotel
- IDF Disabled Veterans Organization
- IDF Widows and Orphans' Organization
- IDF Veterans Association
- Society for the Protection of Nature in Israel
- Israel Book Publishers Association
- Israel Insurance Agents Association
- Israel Teachers Associations
- Magid Institute
- Avshalom Institute
- Mofet Institute
- Enzymotech
- Rafa Laboratories
- Maccabi Health Services
- Israel Dentists Association
- Merage Foundation Israel
- Babylonian Jewry Heritage Center
TESTIMONIALS (continued)
David N
|
[
790,
273,
24867,
304,
278,
390,
10749,
373,
14883,
322,
4846,
385,
8031,
6901,
6282,
310,
278,
4234,
29889,
4699,
338,
263,
5613,
25657,
29892,
1652,
8122,
322,
3165,
20657,
29892,
322,
670,
20607,
6121,
7271,
338,
13602,
29889,
13,
29934,
29884,
10700,
15612,
24136,
29892,
15944,
310,
278,
7021,
1879,
19711,
7765,
448,
12798,
12026,
13,
29949,
332,
20026,
471,
1407,
527,
13120,
411,
596,
17924,
895,
322,
7148,
411,
278,
21028,
262,
1218,
322,
2714,
29899,
16123,
17223,
8214,
297,
607,
366,
9132,
596,
8386,
1549,
278,
18520,
310,
385,
22895,
5037,
21082,
12139,
29889,
3575,
29197,
471,
714,
11235,
322,
5412,
29889,
13,
29934,
265,
1460,
4111,
29899,
26473,
561,
29892,
7178,
310,
278,
14525,
2904,
25813,
16728,
7817,
29892,
7027,
2448,
384,
29892,
1570,
3088,
13,
10858,
24329,
373,
278,
14883,
5376,
29892,
278,
14253,
6932,
322,
11996,
29915,
29879,
1857,
6434,
471,
451,
871,
752,
7807,
322,
22684,
292,
29892,
541,
14792,
14586,
355,
5794,
297,
6320,
1218,
322,
8681,
8491,
472,
278,
1021,
931,
29889,
3575,
21210,
573,
5193,
2175,
14332,
24507,
901,
29889,
13,
29934,
370,
5365,
8432,
2191,
4415,
1171,
29892,
18057,
405,
261,
16939,
333,
29892,
26028,
3801,
29892,
4602,
10722,
5127,
29892,
8046,
13,
1576,
29197,
471,
8031,
322,
21028,
262,
1218,
29892,
322,
471,
20115,
297,
263,
2821,
322,
4972,
292,
8214,
29889,
739,
471,
385,
7271,
393,
2175,
263,
6483,
17188,
29889,
13,
29943,
1255,
29874,
315,
14899,
29892,
26101,
7946,
310,
11996,
29915,
29879,
19358,
20886,
13,
2744,
3682,
635,
21028,
262,
1218,
322,
2441,
29197,
29889,
13,
29934,
11321,
383,
682,
29892,
11996,
20886,
310,
5282,
1947,
13,
29902,
4520,
20695,
337,
7387,
304,
596,
29197,
29892,
411,
28163,
7213,
895,
363,
967,
24233,
1230,
2948,
29892,
7542,
537,
29892,
322,
28289,
29889,
512,
278,
21567,
3186,
29892,
727,
526,
1784,
1058,
526,
7134,
519,
1048,
278,
5518,
393,
896,
5925,
29892,
541,
451,
599,
508,
21028,
16976,
385,
20026,
763,
366,
1258,
29889,
13,
1184,
29888,
6329,
4699,
7567,
1161,
374,
29892,
12252,
310,
278,
7817,
363,
14883,
713,
16972,
29892,
18815,
29899,
12810,
440,
3014,
13,
19504,
405,
790,
273,
471,
6345,
322,
10425,
297,
14883,
29892,
322,
1156,
10591,
1218,
515,
278,
4623,
19184,
4523,
297,
1920,
2276,
273,
29892,
2996,
304,
11996,
304,
278,
18472,
3973,
3014,
310,
23204,
988,
540,
4520,
670,
350,
29889,
29909,
29889,
297,
14253,
29899,
29923,
11530,
16972,
29892,
322,
670,
341,
29889,
29909,
29889,
297,
4623,
6376,
800,
29889,
940,
769,
8772,
278,
3553,
29943,
29892,
322,
1156,
29871,
29896,
29953,
2440,
310,
2669,
408,
385,
12139,
322,
22079,
29892,
16528,
322,
8676,
263,
27863,
7426,
297,
15197,
23303,
29892,
408,
1532,
408,
2284,
2450,
11898,
363,
8236,
260,
2470,
28177,
29889,
13,
2831,
5320,
2440,
29892,
4699,
471,
278,
6920,
322,
3268,
27443,
310,
376,
29968,
2264,
300,
10318,
613,
385,
7395,
4223,
29899,
11675,
4723,
368,
14853,
1919,
607,
8967,
373,
22765,
653,
29914,
20087,
936,
4959,
297,
11996,
29889,
3115,
29892,
540,
471,
15723,
363,
9881,
2440,
491,
278,
22895,
5037,
20886,
310,
5282,
1947,
408,
263,
760,
29899,
2230,
8799,
424,
29892,
322,
3796,
363,
29871,
29946,
2440,
408,
385,
16336,
3483,
858,
363,
5061,
9102,
663,
29892,
11996,
29915,
29879,
8236,
5001,
363,
278,
4333,
322,
7418,
310,
6993,
29914,
357,
729,
29899,
12817,
21082,
515,
278,
8986,
29889,
4699,
471,
278,
6920,
310,
278,
14253,
6932,
322,
14883,
10130,
15670,
29892,
322,
338,
5279,
263,
2989,
29899,
2230,
10257,
25657,
322,
8799,
424,
29889,
13,
19504,
29915,
29879,
5412,
2473,
29899,
29883,
499,
3631,
3239,
313,
354,
7726,
29879,
5320,
10276,
511,
670,
21567,
16140,
322,
670,
9121,
29914,
8926,
6413,
5331,
1075,
304,
8569,
670,
13081,
1973,
373,
14253,
29899,
29923,
579,
322,
14883,
713,
19469,
29889,
940,
338,
263,
18365,
29899,
7045,
13081,
9945,
297,
11996,
29892,
338,
6446,
1652,
8122,
297,
4223,
29892,
322,
756,
2183,
13081,
1973,
297,
263,
12875,
310,
363,
6762,
322,
378,
10662,
297,
8236,
14582,
322,
18161,
21561,
297,
278,
2024,
17535,
29892,
4946,
1907,
29892,
322,
5874,
9668,
2722,
3704,
278,
20886,
310,
5282,
1947,
29892,
278,
20886,
310,
19358,
26049,
29892,
322,
278,
15512,
7668,
29915,
29879,
11367,
29889,
940,
756,
1063,
15593,
287,
373,
22895,
5037,
4797,
7155,
322,
11456,
29892,
756,
19182,
297,
5164,
363,
6762,
5377,
310,
11996,
29892,
322,
338,
373,
278,
1589,
948,
866,
7726,
414,
1051,
310,
278,
4517,
5013,
5790,
18903,
29889,
13,
29899,
20886,
363,
19358,
26049,
13,
29899,
20886,
310,
27051,
545,
13,
29899,
25661,
29287,
5282,
1947,
23985,
13,
29899,
17972,
735,
13,
29899,
1019,
312,
261,
669,
28011,
569,
13,
29899,
1260,
2966,
13,
29899,
1798,
524,
13,
29899,
11996,
10863,
414,
7993,
13,
29899,
315,
5371,
6431,
13,
29899,
21279,
336,
13,
29899,
8564,
3873,
13,
29899,
17550,
3262,
29889,
1523,
13,
29899,
5952,
26056,
3014,
13,
29899,
2261,
1720,
273,
3014,
13,
29899,
12670,
20912,
16146,
293,
6346,
13,
29899,
319,
2477,
29902,
448,
7993,
310,
23035,
322,
9145,
5834,
297,
11996,
13,
29899,
17956,
4717,
29899,
24636,
5013,
5086,
2538,
16719,
7993,
310,
11996,
13,
29899,
390,
1117,
314,
20795,
7817,
13,
29899,
14854,
12293,
512,
7610,
749,
15025,
13,
29899,
11720,
271,
512,
7610,
749,
6938,
13,
29899,
11996,
3371,
29879,
13361,
537,
13,
29899,
1317,
5450,
295,
13,
29899,
3553,
29943,
3295,
3606,
478,
1308,
550,
9205,
2133,
13,
29899,
3553,
29943,
399,
333,
1242,
322,
1394,
561,
550,
29915,
9205,
2133,
13,
29899,
3553,
29943,
478,
1308,
550,
7993,
13,
29899,
7765,
363,
278,
14409,
428,
310,
17677,
297,
11996,
13,
29899,
11996,
6726,
12904,
414,
7993,
13,
29899,
11996,
512,
7610,
749,
4059,
1237,
7993,
13,
29899,
11996,
1920,
496,
414,
6853,
800,
13,
29899,
3561,
333,
8907,
13,
29899,
7740,
845,
17419,
8907,
13,
29899,
341,
974,
300,
8907,
13,
29899,
1174,
14022,
866,
305,
13,
29899,
17881,
29874,
16715,
1061,
583,
13,
29899,
4326,
29883,
19266,
15202,
15538,
13,
29899,
11996,
360,
296,
2879,
7993,
13,
29899,
4702,
482,
10606,
11996,
13,
29899,
14525,
2904,
25813,
9576,
719,
25635,
7817,
13,
18267,
7833,
1164,
25758,
29903,
313,
1285,
262,
6742,
29897,
13,
19504,
405
] | 1,024 |
CommonCrawl
| 1.304943 | 0.491206 | 1.467871 | 1.098925 |
issan's fascinating lecture captured my complete attention. David is not only an excellent spealer; he is an actor who sweeps you away......
Tami Shelach - Chairwoman, IDF Widows and Orphans' Organization
Your lecture was rich with content and fine-tuned. The combination of your personal background in Iran and your mastery of this country's past and present history exposed the audience to perspectives that are not readily accessible to the general public.
Sigal Aybes, Director of the Tel-Aviv University Faculty Club
Your lecture showed us that it is possible to view Iran from a different angle. It exposed for us a hidden and fascinating world.
Yaacov Galazan, VP, Israel Aircraft Industries
Your professionally delivered lecture, the presentation and the manner in which you led our virtual tour of Iran, made the event very successful, enjoyable and fascinating.
Amnon Ben-Shmuel, CEO, Book Publishers Association of Israel
We were left spellbound from seeing the magical and unbelievable slides of the Iranian landscape, and from the manner in which the lecture was delivered.
Efrat Ben-Shoshan, Keren Yeda Fund for University Graduates
It is our pleasure to thank you and express our appreciation and satisfaction from your lecture, which was presented in a professional and engaging manner.
Itzik Steinberg, Deputy VP, Israel Ports Authority
Your "Virtual Tour of Iran" lecture, the depth of the contents, and its wonderful delivery fascinated our audience.
Rina Rabinovich, Training and Development Unit, Human Resources Division, ELISRA
Thank you for your presentation of interesting and informative lectures to the English College.... all members of our audience have enjoyed your lectures and look forward to hearing from you again next year.
Audrey Goodman, English Speaking Residents Association in Israel
Thank you for the fascinating lecture you delivered to one of our foreign clients. Your mastery of the material and the tangible way you presented the content greatly impressed the audience. Your lecture enables a rare glimpse from the inside, illustrates the challenges, and points towards ways of dealing with it.
Dikla Weininger, Communications and Cyber Intelligence Solutions Project Manager, Verint
Please accept our appreciation and gratitude for taking us on an interesting and educative virtual tour of Iran, giving us a glimpse of its beautiful landscape, rich culture, art and delicious cuisine.
International Women's Club, Israel
Your wonderful lecture amazed the audience because of its fascinating and unique content, and also because of the way it was delivered.
Aliza Dayan Hamama, CEO of the Babylonian Jewry Heritage Center
Thank you for the fascinating Iran event.. It was a social evening that will remain in our memories for a very long time.
Professor Zvi Rappaport, Head of Neurosurgery in the Rabin Medical Center
We were enchanted by your lecture and personal story....... We hope to listen to you in additional events in the future.
Hanoch Landau, Bank of Israel
You received many well-deserved praise on the way you delivered your lecture, your sense of humor, and the instant rapport you established with the audience.
Tofi Shaanan-Friedman, B'nai B'rith, Israel
The audience eagerly listened to your fascinating lecture, a lecture that was spiced with humor and rich with historical anecdotes. The combination of extensive knowledge and your wonderful ability to tell a story created a most impressive experience.
Liron Teits, NICE
We would like to thank you for the two fascinating lectures that you delivered to us..... You succeeded to captivate all the audience members.
Alumit Karni, Human Resources, Israel Discount Bank
Your "Virtual Tour of Iran" lecture was indeed a fascinating journey into the heart of Iran.
Ayala Benjamin, Jerusalem Cinematheque
We were impressed with your vast knowledge and the way you delivered the subject. Thank you so much for a most pleasurable experience.
Rachel Zeidan, The Campus for International Geography
Thank you for a lecture that was pure pleasure. The audience was absolutely fascinated by the way in which you presented the rich reality of Iran. We enjoyed every minute....
Adi Boblil, Merage Foundation - Israel Branch, Director
The audience members were captivated, and I received excellent reviews regarding the content of your lecture and the way it was delivered.
Dr. Soli Shah
|
[
790,
273,
29915,
29879,
21028,
262,
1218,
29197,
15468,
590,
4866,
8570,
29889,
4699,
338,
451,
871,
385,
15129,
961,
18362,
29936,
540,
338,
385,
11339,
1058,
7901,
8961,
366,
3448,
3045,
636,
13,
29911,
4479,
1383,
295,
496,
448,
24193,
29893,
2480,
29892,
3553,
29943,
399,
333,
1242,
322,
1394,
561,
550,
29915,
9205,
2133,
13,
10858,
29197,
471,
8261,
411,
2793,
322,
2691,
29899,
29873,
348,
287,
29889,
450,
10296,
310,
596,
7333,
3239,
297,
14883,
322,
596,
5835,
29891,
310,
445,
4234,
29915,
29879,
4940,
322,
2198,
4955,
19884,
278,
20026,
304,
3736,
1103,
3145,
393,
526,
451,
28520,
15579,
304,
278,
2498,
970,
29889,
13,
29903,
335,
284,
15846,
5707,
29892,
15944,
310,
278,
18815,
29899,
12810,
440,
3014,
14184,
18857,
5977,
13,
10858,
29197,
10018,
502,
393,
372,
338,
1950,
304,
1776,
14883,
515,
263,
1422,
10696,
29889,
739,
19884,
363,
502,
263,
7934,
322,
21028,
262,
1218,
3186,
29889,
13,
29979,
29874,
562,
586,
5208,
834,
273,
29892,
478,
29925,
29892,
11996,
319,
12664,
12157,
2722,
13,
10858,
6351,
635,
20115,
29197,
29892,
278,
24329,
322,
278,
8214,
297,
607,
366,
5331,
1749,
6901,
6282,
310,
14883,
29892,
1754,
278,
1741,
1407,
9150,
29892,
13389,
519,
322,
21028,
262,
1218,
29889,
13,
6833,
5464,
4111,
29899,
2713,
29885,
2491,
29892,
14645,
29949,
29892,
6726,
12904,
414,
7993,
310,
11996,
13,
4806,
892,
2175,
24779,
9917,
515,
8790,
278,
2320,
936,
322,
443,
6596,
10384,
519,
2243,
2247,
310,
278,
14883,
713,
24400,
29892,
322,
515,
278,
8214,
297,
607,
278,
29197,
471,
20115,
29889,
13,
29923,
1341,
271,
4111,
29899,
2713,
359,
5403,
29892,
476,
4578,
612,
8710,
13249,
363,
3014,
1632,
13467,
1078,
13,
3112,
338,
1749,
15377,
304,
6452,
366,
322,
4653,
1749,
5108,
362,
322,
26470,
515,
596,
29197,
29892,
607,
471,
9132,
297,
263,
10257,
322,
3033,
6751,
8214,
29889,
13,
3112,
20711,
14808,
2552,
29892,
26721,
29891,
478,
29925,
29892,
11996,
3371,
29879,
13361,
537,
13,
10858,
376,
21287,
6371,
310,
14883,
29908,
29197,
29892,
278,
10809,
310,
278,
8118,
29892,
322,
967,
20695,
28289,
21028,
262,
630,
1749,
20026,
29889,
13,
29934,
1099,
16155,
262,
586,
436,
29892,
26101,
322,
14650,
13223,
29892,
12968,
27562,
7946,
29892,
14845,
3235,
4717,
13,
25271,
366,
363,
596,
24329,
310,
8031,
322,
1871,
1230,
13081,
1973,
304,
278,
4223,
6346,
3045,
599,
5144,
310,
1749,
20026,
505,
27849,
596,
13081,
1973,
322,
1106,
6375,
304,
22514,
515,
366,
1449,
2446,
1629,
29889,
13,
29909,
566,
8903,
7197,
1171,
29892,
4223,
5013,
5086,
2538,
16719,
7993,
297,
11996,
13,
25271,
366,
363,
278,
21028,
262,
1218,
29197,
366,
20115,
304,
697,
310,
1749,
9117,
13154,
29889,
3575,
5835,
29891,
310,
278,
5518,
322,
278,
18806,
1821,
982,
366,
9132,
278,
2793,
11180,
527,
13120,
278,
20026,
29889,
3575,
29197,
28936,
263,
10812,
330,
12552,
344,
515,
278,
2768,
29892,
8632,
1078,
278,
18066,
267,
29892,
322,
3291,
7113,
5837,
310,
16743,
411,
372,
29889,
13,
29928,
638,
433,
1334,
2827,
261,
29892,
22365,
800,
322,
8045,
495,
3159,
28286,
4956,
17925,
8010,
15629,
29892,
1798,
524,
13,
12148,
3544,
1749,
5108,
362,
322,
20715,
4279,
363,
5622,
502,
373,
385,
8031,
322,
6320,
1230,
6901,
6282,
310,
14883,
29892,
6820,
502,
263,
330,
12552,
344,
310,
967,
9560,
24400,
29892,
8261,
9257,
29892,
1616,
322,
628,
14803,
2723,
275,
457,
29889,
13,
17579,
1288,
10152,
29915,
29879,
5977,
29892,
11996,
13,
10858,
20695,
29197,
21863,
287,
278,
20026,
1363,
310,
967,
21028,
262,
1218,
322,
5412,
2793,
29892,
322,
884,
1363,
310,
278,
982,
372,
471,
20115,
29889,
13,
2499,
6619,
8373,
273,
7904,
3304,
29892,
14645,
29949,
310,
278,
14525,
2904,
25813,
9576,
719,
25635,
7817,
13,
25271,
366,
363,
278,
21028,
262,
1218,
14883,
1741,
636,
739,
471,
263,
5264,
11005,
393,
674,
3933,
297,
1749,
2626,
3842,
363,
263,
1407,
1472,
931,
29889,
13,
1184,
29888,
6329,
796,
1403,
390,
932,
481,
441,
29892,
12252,
310,
17574,
1883,
2007,
708,
297,
278,
16155,
262,
20795,
7817,
13,
4806,
892,
427,
305,
9714,
491,
596,
29197,
322,
7333,
5828,
3045,
856,
1334,
4966,
304,
11621,
304,
366,
297,
5684,
4959,
297,
278,
5434,
29889,
13,
29950,
1562,
305,
3172,
585,
29892,
10253,
310,
11996,
13,
3492,
4520,
1784,
1532,
29899,
2783,
9841,
7213,
895,
373,
278,
982,
366,
20115,
596,
29197,
29892,
596,
4060,
310,
27448,
29892,
322,
278,
14426,
13659,
366,
7841,
411,
278,
20026,
29889,
13,
29911,
974,
29875,
1383,
29874,
273,
273,
29899,
29943,
1255,
1171,
29892,
350,
29915,
1056,
29875,
350,
29915,
29878,
389,
29892,
11996,
13,
1576,
20026,
19888,
368,
29616,
304,
596,
21028,
262,
1218,
29197,
29892,
263,
29197,
393,
471,
805,
7612,
411,
27448,
322,
8261,
411,
15839,
385,
687,
29881,
4769,
29889,
450,
10296,
310,
20607,
7134,
322,
596,
20695,
11509,
304,
2649,
263,
5828,
2825,
263,
1556,
21210,
573,
7271,
29889,
13,
29931,
381,
265,
1920,
1169,
29892,
405,
12107,
13,
4806,
723,
763,
304,
6452,
366,
363,
278,
1023,
21028,
262,
1218,
13081,
1973,
393,
366,
20115,
304,
502,
18598,
887,
14792,
304,
4332,
440,
403,
599,
278,
20026,
5144,
29889,
13,
2499,
398,
277,
3467,
1240,
29892,
12968,
27562,
29892,
11996,
3295,
2798,
10253,
13,
10858,
376,
21287,
6371,
310,
14883,
29908,
29197,
471,
6200,
263,
21028,
262,
1218,
16342,
964,
278,
5192,
310,
14883,
29889,
13,
29909,
29891,
2883,
20212,
29892,
23204,
11972,
4579,
354,
802,
13,
4806,
892,
527,
13120,
411,
596,
13426,
7134,
322,
278,
982,
366,
20115,
278,
4967,
29889,
3374,
366,
577,
1568,
363,
263,
1556,
5644,
294,
21115,
7271,
29889,
13,
29934,
496,
295,
3091,
29219,
29892,
450,
7259,
375,
363,
4623,
27340,
13,
25271,
366,
363,
263,
29197,
393,
471,
8296,
15377,
29889,
450,
20026,
471,
13312,
21028,
262,
630,
491,
278,
982,
297,
607,
366,
9132,
278,
8261,
16832,
310,
14883,
29889,
1334,
27849,
1432,
11015,
3045,
13,
3253,
29875,
7991,
29880,
309,
29892,
4702,
482,
10606,
448,
11996,
25889,
29892,
15944,
13,
1576,
20026,
5144,
892,
4332,
440,
630,
29892,
322,
306,
4520,
15129,
21804,
11211,
278,
2793,
310,
596,
29197,
322,
278,
982,
372,
471,
20115,
29889,
13,
25639,
29889,
4956,
29875,
25897
] | 1,024 |
CommonCrawl
| 1.590234 | -0.521715 | 0.547947 | 1.088806 |
I'm Pretty Sure Days Gone Hates Me More Than I Hate It
By Garrett Martin | April 25, 2019 | 1:30pm
Games Features Days Gone
When Days Gone is loading, a ring spins in the bottom right corner of the screen. It's one of the most ridiculous rings you'll ever see, in the real world or a videogame, with a large, silver wolf's skull on top that covers up at least half of any finger that wears it. It's jewelry for men who need their jewelry to scream "I'M STILL A MAN!", as if their masculinity (and its motorcycles, beards and neck tattoos) was somehow undermined by wearing a piece of metal on their hands. I know this ring better than my own wedding ring now, because I've had to stare at it for most of my time in Days Gone.
When I launch Days Gone on my PlayStation 4 Pro, I almost immediately get a loading screen that lasts for three minutes. That's three minutes of a black screen with that tiny ring spinning endlessly in the corner. And that's not three minutes of loading to get to where I actually am within the game—that's three minutes just to get to the main title menu, at which point I have to choose to continue my game, and then sit through another loading screen that never runs for less than 90 seconds. If the idea is to make players feel how dull and tedious living in a post-society zombie wasteland must be, then Days Gone accomplishes at least one of its goals.
Much worse than the loading screens, though, is the checkpoint system, or apparent lack thereof. There's an option on the pause screen to save your game, but I can't tell why it exists. I saved my game in the middle of a mission multiple times, and even when I had done multiple individual objectives within the mission, loading that save file would put me back at the very beginning, at the same spot captured by the autosave. Perhaps this changes deeper in the game—I did not get that far into this one, which is why this isn't an official scored review—but it's incredibly frustrating to have to repeat 30 minutes or more because this game doesn't save the way games in the year 2019 are expected to save.
Here's an example. Early on there's a mission where the sullen survivalist I have to play as (whose name, no joke, is Deacon St. John, and who, again, is a sad, angry biker in a zombie-riddled Oregon, and not a Brontë love interest) goes to pick up his bike, which he had to abandon on the side of a road. While walking through the woods he comes upon a group of human raiders, who, as part of Days Gone's unthinking fealty to the stereotypes of the post-apocalypse canon, are obviously bad guys who need to be killed as soon as possible. This encounter is basically a tutorial on the game's stealth mechanics, and if you know anything about stealth in videogames, you know that means it takes more time to get through this fight than if my guy just went in with guns a-blazin'. (I tried that, and yes, I was staring at a post-death loading screen very quickly.) I sneaked and crawled and scurried and killed all these jokers with my knife, and even if they were standing like five in-game feet away, none of them ever caught me in the act.
After those murders I continued down to where my bike was supposed to be. There were zombies just kind of milling about throughout the woods (the game calls them "freakers," which, yeah), so I guess it's possible I could've had some random fights between that camp and getting to the road. I avoided them, though, and hightailed it to that two-lane blacktop, where my bike was nowhere to be found.
My dude is hip to the ways of the apocalypse, though. Somehow he knew (or maybe guessed) that his bike was picked up for scrap by a local settlement of good, friendly folk who he refuses to live with because then he wouldn't be a hardened loner with just a bike and a friend with head tats named Boozer. So I head to this group's
|
[
306,
29915,
29885,
4721,
4349,
18585,
24728,
402,
650,
379,
1078,
2191,
5853,
498,
273,
306,
379,
403,
739,
13,
2059,
7455,
13158,
6502,
891,
3786,
29871,
29906,
29945,
29892,
29871,
29906,
29900,
29896,
29929,
891,
29871,
29896,
29901,
29941,
29900,
3358,
13,
29954,
1280,
5169,
3698,
24728,
402,
650,
13,
10401,
24728,
402,
650,
338,
8363,
29892,
263,
9228,
805,
1144,
297,
278,
5970,
1492,
11155,
310,
278,
4315,
29889,
739,
29915,
29879,
697,
310,
278,
1556,
8177,
12906,
681,
28774,
366,
29915,
645,
3926,
1074,
29892,
297,
278,
1855,
3186,
470,
263,
18900,
468,
420,
29892,
411,
263,
2919,
29892,
13283,
281,
4369,
29915,
29879,
2071,
913,
373,
2246,
393,
18469,
701,
472,
3203,
4203,
310,
738,
19917,
393,
591,
1503,
372,
29889,
739,
29915,
29879,
432,
809,
295,
719,
363,
1757,
1058,
817,
1009,
432,
809,
295,
719,
304,
885,
1633,
376,
29902,
29915,
29924,
6850,
24071,
319,
341,
2190,
29991,
613,
408,
565,
1009,
21799,
13593,
313,
392,
967,
10992,
1270,
7799,
29892,
367,
3163,
322,
18873,
260,
16234,
359,
29897,
471,
10431,
563,
837,
1312,
491,
591,
4362,
263,
8424,
310,
11915,
373,
1009,
6567,
29889,
306,
1073,
445,
9228,
2253,
1135,
590,
1914,
14837,
8497,
9228,
1286,
29892,
1363,
306,
29915,
345,
750,
304,
380,
598,
472,
372,
363,
1556,
310,
590,
931,
297,
24728,
402,
650,
29889,
13,
10401,
306,
6826,
24728,
402,
650,
373,
590,
7412,
22814,
29871,
29946,
1019,
29892,
306,
4359,
7389,
679,
263,
8363,
4315,
393,
1833,
29879,
363,
2211,
6233,
29889,
2193,
29915,
29879,
2211,
6233,
310,
263,
4628,
4315,
411,
393,
21577,
9228,
10917,
1076,
1095,
23769,
297,
278,
11155,
29889,
1126,
393,
29915,
29879,
451,
2211,
6233,
310,
8363,
304,
679,
304,
988,
306,
2869,
626,
2629,
278,
3748,
30003,
5747,
29915,
29879,
2211,
6233,
925,
304,
679,
304,
278,
1667,
3611,
6143,
29892,
472,
607,
1298,
306,
505,
304,
6755,
304,
6773,
590,
3748,
29892,
322,
769,
7845,
1549,
1790,
8363,
4315,
393,
2360,
6057,
363,
3109,
1135,
29871,
29929,
29900,
6923,
29889,
960,
278,
2969,
338,
304,
1207,
10769,
4459,
920,
270,
913,
322,
29748,
2738,
8471,
297,
263,
1400,
29899,
2839,
3305,
503,
3424,
347,
471,
29873,
14045,
1818,
367,
29892,
769,
24728,
402,
650,
12709,
267,
472,
3203,
697,
310,
967,
14433,
29889,
13,
29924,
987,
15029,
1135,
278,
8363,
11844,
29892,
2466,
29892,
338,
278,
1423,
3149,
1788,
29892,
470,
20295,
10225,
727,
974,
29889,
1670,
29915,
29879,
385,
2984,
373,
278,
19957,
4315,
304,
4078,
596,
3748,
29892,
541,
306,
508,
29915,
29873,
2649,
2020,
372,
4864,
29889,
306,
7160,
590,
3748,
297,
278,
7256,
310,
263,
10655,
2999,
3064,
29892,
322,
1584,
746,
306,
750,
2309,
2999,
5375,
1203,
3145,
2629,
278,
10655,
29892,
8363,
393,
4078,
934,
723,
1925,
592,
1250,
472,
278,
1407,
6763,
29892,
472,
278,
1021,
9758,
15468,
491,
278,
1120,
359,
1351,
29889,
11637,
445,
3620,
25871,
297,
278,
3748,
30003,
29902,
1258,
451,
679,
393,
2215,
964,
445,
697,
29892,
607,
338,
2020,
445,
3508,
29915,
29873,
385,
6221,
15569,
9076,
30003,
4187,
372,
29915,
29879,
29811,
14981,
1424,
4627,
1218,
304,
505,
304,
12312,
29871,
29941,
29900,
6233,
470,
901,
1363,
445,
3748,
1838,
29915,
29873,
4078,
278,
982,
8090,
297,
278,
1629,
29871,
29906,
29900,
29896,
29929,
526,
3806,
304,
4078,
29889,
13,
10605,
29915,
29879,
385,
1342,
29889,
11095,
373,
727,
29915,
29879,
263,
10655,
988,
278,
23416,
264,
10503,
2561,
391,
306,
505,
304,
1708,
408,
313,
1332,
852,
1024,
29892,
694,
2958,
446,
29892,
338,
897,
23074,
624,
29889,
2259,
29892,
322,
1058,
29892,
1449,
29892,
338,
263,
14610,
29892,
26230,
289,
5603,
297,
263,
503,
3424,
347,
29899,
29878,
2205,
839,
25203,
29892,
322,
451,
263,
1771,
609,
30083,
5360,
4066,
29897,
5771,
304,
5839,
701,
670,
4768,
446,
29892,
607,
540,
750,
304,
11807,
373,
278,
2625,
310,
263,
6520,
29889,
5806,
22049,
1549,
278,
25013,
540,
5304,
2501,
263,
2318,
310,
5199,
1153,
11376,
29892,
1058,
29892,
408,
760,
310,
24728,
402,
650,
29915,
29879,
443,
386,
18159,
1238,
18745,
304,
278,
269,
12358,
327,
7384,
310,
278,
1400,
29899,
481,
18642,
1478,
344,
15355,
29892,
526,
12879,
4319,
18239,
1058,
817,
304,
367,
9445,
408,
4720,
408,
1950,
29889,
910,
11735,
338,
8830,
263,
9673,
373,
278,
3748,
29915,
29879,
1886,
4298,
7208,
1199,
29892,
322,
565,
366,
1073,
3099,
1048,
1886,
4298,
297,
18900,
468,
1280,
29892,
366,
1073,
393,
2794,
372,
4893,
901,
931,
304,
679,
1549,
445,
8589,
1135,
565,
590,
1410,
29891,
925,
3512,
297,
411,
18788,
263,
29899,
17530,
23374,
4286,
313,
29902,
1898,
393,
29892,
322,
4874,
29892,
306,
471,
380,
4362,
472,
263,
1400,
29899,
311,
493,
8363,
4315,
1407,
9098,
1846,
306,
269,
484,
12535,
322,
29349,
839,
322,
885,
332,
1255,
322,
9445,
599,
1438,
432,
554,
414,
411,
590,
889,
1607,
29892,
322,
1584,
565,
896,
892,
13407,
763,
5320,
297,
29899,
11802,
6900,
3448,
29892,
5642,
310,
963,
3926,
12624,
592,
297,
278,
1044,
29889,
13,
13555,
1906,
7167,
8623,
306,
7572,
1623,
304,
988,
590,
4768,
446,
471,
7424,
304,
367,
29889,
1670,
892,
503,
3424,
583,
925,
2924,
310,
3533,
292,
1048,
10106,
278,
25013,
313,
1552,
3748,
5717,
963,
376,
10745,
21079,
1699,
607,
29892,
21915,
511,
577,
306,
4140,
372,
29915,
29879,
1950,
306,
1033,
29915,
345,
750,
777,
4036,
285,
5861,
1546,
393,
4242,
322,
2805,
304,
278,
6520,
29889,
306,
28305,
963,
29892,
2466,
29892,
322,
298,
523,
29874,
2356,
372,
304,
393,
1023,
29899,
25821,
4628,
3332,
29892,
988,
590,
4768,
446,
471,
1286,
4150,
304,
367,
1476,
29889,
13,
3421,
868,
311,
338,
21464,
304,
278,
5837,
310,
278,
3095,
18642,
1478,
344,
29892,
2466,
29889,
3834,
3525,
540,
6363,
313,
272,
5505,
4140,
287,
29897,
393,
670,
4768,
446,
471,
18691,
701,
363,
885,
2390,
491,
263,
1887,
16493,
310,
1781,
29892,
19780,
19589,
1058,
540,
2143,
6394,
304,
5735,
411,
1363,
769,
540,
7656,
29915,
29873,
367,
263,
2898,
6419,
23123,
261,
411,
925,
263,
4768,
446,
322,
263,
5121,
411,
2343,
260,
1446,
4257,
1952,
2112,
261,
29889,
1105,
306,
2343,
304,
445,
2318,
29915,
29879
] | 1,024 |
CommonCrawl
| -0.298803 | -0.868548 | -0.641632 | -1.240464 |
News is next on LeoTV.
The first hospital in the US was opened in Pennsylvania on this day in 1752. Robert Fulton patented the steam boat in 1809. Nelson Mandela was freed in 1990.
Thomas Edison was born in 1847. Tina "Ginger" Louise was born in 1934.
Carly Fiorina is out at Hewlett-Packard, saying she quit in a dispute with the board over the future direction of the company. The board's chair was quoted in the NY Times as saying "These things always seem precipitous when they occur. But the board has been deliberating the company's performance, and the C.E.O.'s performance, for quite some time." But don't cry for me Fiorina, Carly's getting a severance package worth $21.1 million. HP shares closed at 21.53 up nearly 7% on the news. When Fiorina started with HP in 1999 the stock price was over 40.
The Motion Picture Association of America has taken over LokiTorrent. The bittorrent tracker site was attempting to raise money to fight the MPAA, but a court order shut it down yesterday. Even more chilling, the court ordered LokiTorrent to give the MPAA their domain and all the server logs. The site now features the threat, "You can click but you can't hide. Stealing movies leaves a trail. The only way not to get caught is to stop." Makes me want to steal a movie right now.
Just in the nick of time, Philips has developed a way to fingerprint films so they can be tracked on P2P networks.
The FCC has told cable companies that they needn't carry all the digital channels offered by local television stations. The shift to digital has made it possible for a single TV station to broadcast several shows at once, creating an issue for cable companies who don't have the benefit of the additional bandwidth.
FreeBSD is retiring its daemon logo. A public competition will be held to choose a replacement. Toy Story director John Lasseter created the earliest versions of the daemon – the current version is copyrighted by Marshall Kirk McKusick. I kinda like it, but then I like Underwood Deviled Ham, too.
There's a big flaw in nearly all of Symantec's products on Windows, Macintosh, Linux, and AS400 systems and Brightmail clients. The flaw could allow an attacker to run arbitrary code on these systems. Symantec has patches so run LiveUpdate now.
Temco is suing a web site that published hacks to its Xbox game Dead or Alive Xtreme Volleyball. The hack allowed users to strip the game's characters naked. And this hurts game sales how?
Listen in tomorrow at 7:45a Pacific for my weekly news commentary on KFI 640 AM in Los Angeles.
Big flaw in nearly all off Symantec's products.My norton antispam expired on 1-10-05 and as I will now renew (not worth it)Leo sir am I still in danger? If so should I just delete it as I don't use it? Does anyone else have any advice?
Tina "Ginger" Louise was born in 1934.
yeah shes gonna hit you with SO much hate mail.
She HATES being assosiated with the island 'it was only one part of my long great life' yeah well no one cares, they like giligan. Unlike Alec Guiniss who had a lot of talent and an OSCAR before doing Star Wars which he resented being the only thing people knew him for, Tina only did 'ginger' and some dinner theater.
TSS Fan, Tina Louise was at the TV Land awards last year with the remaining Gilligan's Island cast members to get the "Pop Icon" award, and seemed happy to be there. I think she has finally "come to terms" with being associated with the show. Remember, for YEARS, Lenard Nimoy said "I'm not Spock", but later came back for the theatrical films.
yeah nimoy is a funny case, he hated it basicly for the first 5 years after the show then it became his life and he fully embrased it. I think he realised that the fans really appreciated his work and that was a good thing.
|
[
10130,
338,
2446,
373,
22533,
8050,
29889,
13,
1576,
937,
13457,
297,
278,
3148,
471,
6496,
297,
16636,
373,
445,
2462,
297,
29871,
29896,
29955,
29945,
29906,
29889,
4755,
383,
499,
265,
2373,
14927,
278,
21837,
13006,
297,
29871,
29896,
29947,
29900,
29929,
29889,
19135,
15419,
3100,
471,
3005,
287,
297,
29871,
29896,
29929,
29929,
29900,
29889,
13,
1349,
18902,
2155,
2285,
471,
6345,
297,
29871,
29896,
29947,
29946,
29955,
29889,
323,
1099,
376,
29954,
5621,
29908,
24417,
471,
6345,
297,
29871,
29896,
29929,
29941,
29946,
29889,
13,
8179,
368,
383,
1611,
1099,
338,
714,
472,
379,
809,
13650,
29899,
16638,
538,
29892,
5934,
1183,
23283,
297,
263,
28447,
411,
278,
7613,
975,
278,
5434,
5305,
310,
278,
5001,
29889,
450,
7613,
29915,
29879,
11774,
471,
23153,
297,
278,
23526,
10277,
408,
5934,
376,
1349,
968,
2712,
2337,
2833,
25720,
277,
681,
746,
896,
6403,
29889,
1205,
278,
7613,
756,
1063,
23663,
1218,
278,
5001,
29915,
29879,
4180,
29892,
322,
278,
315,
29889,
29923,
29889,
29949,
6169,
29879,
4180,
29892,
363,
3755,
777,
931,
1213,
1205,
1016,
29915,
29873,
10901,
363,
592,
383,
1611,
1099,
29892,
1704,
368,
29915,
29879,
2805,
263,
2775,
749,
3577,
7088,
395,
29906,
29896,
29889,
29896,
7284,
29889,
379,
29925,
29358,
5764,
472,
29871,
29906,
29896,
29889,
29945,
29941,
701,
8886,
29871,
29955,
29995,
373,
278,
9763,
29889,
1932,
383,
1611,
1099,
4687,
411,
379,
29925,
297,
29871,
29896,
29929,
29929,
29929,
278,
10961,
8666,
471,
975,
29871,
29946,
29900,
29889,
13,
1576,
7142,
291,
28908,
7993,
310,
6813,
756,
4586,
975,
365,
21025,
29911,
272,
7771,
29889,
450,
289,
986,
272,
7771,
1020,
4937,
3268,
471,
15661,
304,
12020,
6909,
304,
8589,
278,
16379,
6344,
29892,
541,
263,
8973,
1797,
12522,
372,
1623,
22600,
29889,
7753,
901,
521,
8873,
29892,
278,
8973,
10372,
365,
21025,
29911,
272,
7771,
304,
2367,
278,
16379,
6344,
1009,
5354,
322,
599,
278,
1923,
10748,
29889,
450,
3268,
1286,
5680,
278,
28469,
29892,
376,
3492,
508,
2828,
541,
366,
508,
29915,
29873,
9563,
29889,
2443,
12818,
2351,
583,
11308,
263,
26203,
29889,
450,
871,
982,
451,
304,
679,
12624,
338,
304,
5040,
1213,
341,
6926,
592,
864,
304,
1886,
284,
263,
14064,
1492,
1286,
29889,
13,
14084,
297,
278,
25985,
310,
931,
29892,
8234,
567,
756,
8906,
263,
982,
304,
19917,
2158,
12298,
577,
896,
508,
367,
5702,
287,
373,
349,
29906,
29925,
14379,
29889,
13,
1576,
383,
4174,
756,
5429,
21387,
14582,
393,
896,
817,
29876,
29915,
29873,
8677,
599,
278,
13436,
18196,
12520,
491,
1887,
11456,
16355,
29889,
450,
9500,
304,
13436,
756,
1754,
372,
1950,
363,
263,
2323,
5648,
5073,
304,
12672,
3196,
3697,
472,
2748,
29892,
4969,
385,
2228,
363,
21387,
14582,
1058,
1016,
29915,
29873,
505,
278,
14169,
310,
278,
5684,
3719,
2103,
29889,
13,
20475,
29933,
7230,
338,
3240,
8491,
967,
1146,
9857,
20194,
29889,
319,
970,
13888,
674,
367,
4934,
304,
6755,
263,
16920,
29889,
29411,
13740,
8881,
2259,
365,
465,
1308,
2825,
278,
24577,
6910,
310,
278,
1146,
9857,
785,
278,
1857,
1873,
338,
3509,
1266,
287,
491,
23072,
26424,
24053,
375,
860,
29889,
306,
2924,
29874,
763,
372,
29892,
541,
769,
306,
763,
7634,
6115,
9481,
2356,
7904,
29892,
2086,
29889,
13,
8439,
29915,
29879,
263,
4802,
285,
10653,
297,
8886,
599,
310,
10667,
1647,
29883,
29915,
29879,
9316,
373,
3852,
29892,
4326,
524,
10578,
29892,
8074,
29892,
322,
3339,
29946,
29900,
29900,
6757,
322,
29258,
2549,
13154,
29889,
450,
285,
10653,
1033,
2758,
385,
5337,
261,
304,
1065,
11472,
775,
373,
1438,
6757,
29889,
10667,
1647,
29883,
756,
13261,
267,
577,
1065,
10782,
6422,
1286,
29889,
13,
5776,
1111,
338,
480,
292,
263,
1856,
3268,
393,
6369,
447,
4684,
304,
967,
1060,
1884,
3748,
16992,
470,
319,
9258,
1060,
2484,
1004,
3684,
2330,
2135,
29889,
450,
15833,
6068,
4160,
304,
17820,
278,
3748,
29915,
29879,
4890,
302,
12535,
29889,
1126,
445,
12166,
1372,
3748,
16538,
920,
29973,
13,
1293,
264,
297,
6454,
22396,
472,
29871,
29955,
29901,
29946,
29945,
29874,
14328,
363,
590,
4723,
368,
9763,
3440,
653,
373,
476,
3738,
29871,
29953,
29946,
29900,
13862,
297,
4602,
10722,
29889,
13,
6970,
285,
10653,
297,
8886,
599,
1283,
10667,
1647,
29883,
29915,
29879,
9316,
29889,
3421,
302,
26342,
3677,
11936,
314,
1518,
2859,
373,
29871,
29896,
29899,
29896,
29900,
29899,
29900,
29945,
322,
408,
306,
674,
1286,
23011,
313,
1333,
7088,
372,
29897,
3226,
29877,
8889,
626,
306,
1603,
297,
9703,
29973,
960,
577,
881,
306,
925,
5217,
372,
408,
306,
1016,
29915,
29873,
671,
372,
29973,
5538,
5019,
1683,
505,
738,
9848,
29973,
13,
29911,
1099,
376,
29954,
5621,
29908,
24417,
471,
6345,
297,
29871,
29896,
29929,
29941,
29946,
29889,
13,
4099,
801,
528,
267,
330,
11586,
7124,
366,
411,
7791,
1568,
26277,
10524,
29889,
13,
13468,
379,
1299,
2890,
1641,
1223,
8156,
630,
411,
278,
11359,
525,
277,
471,
871,
697,
760,
310,
590,
1472,
2107,
2834,
29915,
21915,
1532,
694,
697,
1559,
267,
29892,
896,
763,
330,
309,
11163,
29889,
853,
4561,
7801,
29883,
2088,
262,
790,
1058,
750,
263,
3287,
310,
24242,
322,
385,
6570,
29907,
1718,
1434,
2599,
7828,
9149,
607,
540,
620,
14927,
1641,
278,
871,
2655,
2305,
6363,
1075,
363,
29892,
323,
1099,
871,
1258,
525,
3460,
261,
29915,
322,
777,
17803,
278,
1008,
29889,
13,
29911,
1799,
383,
273,
29892,
323,
1099,
24417,
471,
472,
278,
5648,
3172,
24441,
1833,
1629,
411,
278,
9886,
28047,
11163,
29915,
29879,
7935,
4320,
5144,
304,
679,
278,
376,
12310,
26816,
29908,
9862,
29892,
322,
6140,
9796,
304,
367,
727,
29889,
306,
1348,
1183,
756,
7146,
376,
2763,
304,
4958,
29908,
411,
1641,
6942,
411,
278,
1510,
29889,
22738,
29892,
363,
612,
26441,
29903,
29892,
16206,
538,
405,
4200,
29891,
1497,
376,
29902,
29915,
29885,
451,
24674,
384,
613,
541,
2678,
2996,
1250,
363,
278,
278,
8141,
936,
12298,
29889,
13,
4099,
801,
302,
4200,
29891,
338,
263,
2090,
1460,
1206,
29892,
540,
298,
630,
372,
6996,
368,
363,
278,
937,
29871,
29945,
2440,
1156,
278,
1510,
769,
372,
3897,
670,
2834,
322,
540,
8072,
953,
1182,
1463,
372,
29889,
306,
1348,
540,
1855,
3368,
393,
278,
24909,
2289,
7556,
670,
664,
322,
393,
471,
263,
1781,
2655,
29889
] | 1,024 |
C4
| -0.981464 | -1.189425 | 0.525532 | -1.148761 |
Manifesto Launch
Micah George June 4, 2016 10:53 am June 4, 2016 0
Labour Promises To Make Country Better
Prime Minister Anthony holding up manifesto. [PHOTO: Photo Mike]
HEAVY rains Wednesday night could not stop the launch of the Labour Party's Manifesto near the Castries Waterfront. Hundreds braved the inclement weather to hear what the Labour Party had in store for the country for the next five years should it be elected on Monday.
"You amaze me. The St. Lucia Labour Party cannot asked for more. Thank you for your loyalty, thank you for your confidence," Dr. Kenny Anthony, the party's political leader told cheering supporters.
The 80-page document with its catch line "With You. For You. For Saint Lucia" is entitled "Making our Country Better Together."
It not only lists the achievements of the party's recent term in office (2011 – 2016), which takes up five pages, but also highlights a plethora of programmes to be undertaken by the party if allowed to continue in office.
Sub-titles explain in detail the party's outlook for the future from how it would maintain fiscal discipline, its plans for an ambitious employment strategy to reduce unemployment to below 15 percent in five years, address the infrastructural gaps and its programme of tax reform continuance to expanding the online tax services to decrease the time and cost associated with starting a business, review the Citizenship Investment Programme, improve Saint Lucia's attractiveness to foreign investors, support indigenous and domestic investor, enact a special regime of incentives for the services sector, etc., etc.
The manifesto was not distributed that night to supporters. Supporters were told to collect their copies at the various constituency offices of their candidate the following day. Each candidate received a copy on the night.
The glossy pages of the declaration of principles, policies and objectives of the party are divided into several agendas such as: building a safe and just society that speaks of the justice system and the party's intentions to construct a modern National Judicial Complex; a review of the Criminal Code, something Dr. Anthony says will be done within the party's first six months in office, should it be elected and generally improve the efficiency of the Criminal Justice System.
Also included in the "Building a Safe and Just Society" agenda is the establishment of a special national task force on rape and sexual offensives as part of the party's overall intention to protect the country's women and children, the construction of a new modern National Police Headquarters at La Toc, the establishment of a new Central Police Station on Chaussee Road as part of a redevelopment of Wilton's Yard, among other things.
Under the agenda "Weaving a Cohesive Social Fabric" the party's intentions are to provide better housing and access to land to residents, ensure that the education system is relevant and progressive, part of which is the implementation of universal physical education and sports. Other intentions under the social fabric agenda are the establishment of a modern equitable health sector which will include a multi-disciplinary committee to initiate wide-ranging stakeholder discussions and make recommendations to government on the decriminalization and use of marijuana, the youth and the protection of citizens that are vulnerable along with sports and recreational development.
The agenda 'Defining our National Identity' is all about promoting the island's culture and creative industries, building national heritage which will include establishing a national arts museum and art gallery, as well as, strengthening the island's democracy and Constitution which will include considering the passage of legislation to establish a fixed date for the holding of general elections.
Under this agenda is the provision for more efficient public services, the empowerment of citizens through technology and developing strategic focus in external relations.
The manifesto ends with the listing of several projects to be focused on at the constituency level in the party's next term in office Each constituency has a list of projects the party describes as 'priority projects' to be completed with the five-year span.
Manifesto held high by SLP candidates [PHOTO: Photo Mike]
Micah George
Micah George is an established name in the journalism landscape in St. Lucia. He started his journalism tutelage under the critical eye of the Star Newspaper Publisher
|
[
2315,
361,
4778,
997,
3322,
13,
29924,
293,
801,
5122,
5306,
29871,
29946,
29892,
29871,
29906,
29900,
29896,
29953,
29871,
29896,
29900,
29901,
29945,
29941,
626,
5306,
29871,
29946,
29892,
29871,
29906,
29900,
29896,
29953,
29871,
29900,
13,
28632,
473,
9705,
4637,
1763,
8561,
15456,
26965,
13,
4040,
603,
7668,
16477,
13587,
701,
14682,
4778,
29889,
518,
19689,
2891,
29949,
29901,
1963,
3747,
12828,
29962,
13,
9606,
7520,
29979,
1153,
1144,
15050,
4515,
3250,
4646,
1033,
451,
5040,
278,
6826,
310,
278,
29453,
9173,
29915,
29879,
2315,
361,
4778,
2978,
278,
4834,
2722,
13062,
8862,
29889,
379,
6453,
29879,
4105,
1490,
278,
5528,
944,
14826,
304,
8293,
825,
278,
29453,
9173,
750,
297,
3787,
363,
278,
4234,
363,
278,
2446,
5320,
2440,
881,
372,
367,
11467,
373,
27822,
29889,
13,
29908,
3492,
21863,
29872,
592,
29889,
450,
624,
29889,
3182,
1512,
29453,
9173,
2609,
4433,
363,
901,
29889,
3374,
366,
363,
596,
28108,
1017,
29892,
6452,
366,
363,
596,
16420,
1699,
4942,
29889,
10015,
1460,
16477,
29892,
278,
6263,
29915,
29879,
8604,
11822,
5429,
923,
3241,
1462,
272,
2153,
29889,
13,
1576,
29871,
29947,
29900,
29899,
3488,
1842,
411,
967,
4380,
1196,
376,
3047,
887,
29889,
1152,
887,
29889,
1152,
4107,
3182,
1512,
29908,
338,
23437,
376,
29924,
5086,
1749,
15456,
26965,
323,
12966,
1213,
13,
3112,
451,
871,
8857,
278,
27012,
4110,
310,
278,
6263,
29915,
29879,
7786,
1840,
297,
8034,
313,
29906,
29900,
29896,
29896,
785,
29871,
29906,
29900,
29896,
29953,
511,
607,
4893,
701,
5320,
6515,
29892,
541,
884,
12141,
29879,
263,
5644,
386,
2207,
310,
1824,
4467,
304,
367,
22332,
9424,
491,
278,
6263,
565,
6068,
304,
6773,
297,
8034,
29889,
13,
4035,
29899,
23545,
793,
5649,
297,
9493,
278,
6263,
29915,
29879,
714,
6914,
363,
278,
5434,
515,
920,
372,
723,
7344,
16436,
1052,
29822,
29892,
967,
13900,
363,
385,
626,
2966,
2738,
5703,
358,
13705,
304,
10032,
443,
3451,
22812,
304,
2400,
29871,
29896,
29945,
10151,
297,
5320,
2440,
29892,
3211,
278,
22035,
5313,
3631,
330,
2547,
322,
967,
19607,
310,
8818,
11736,
3133,
749,
304,
7985,
292,
278,
7395,
8818,
5786,
304,
23806,
278,
931,
322,
3438,
6942,
411,
6257,
263,
5381,
29892,
9076,
278,
21353,
466,
575,
4034,
512,
10147,
358,
7835,
1004,
29892,
11157,
4107,
3182,
1512,
29915,
29879,
13978,
20193,
304,
9117,
13258,
943,
29892,
2304,
1399,
2101,
681,
322,
21849,
13258,
272,
29892,
427,
627,
263,
4266,
22384,
310,
297,
1760,
3145,
363,
278,
5786,
17535,
29892,
2992,
1696,
2992,
29889,
13,
1576,
14682,
4778,
471,
451,
13235,
393,
4646,
304,
1462,
272,
2153,
29889,
9179,
272,
2153,
892,
5429,
304,
6314,
1009,
14591,
472,
278,
5164,
10719,
3819,
25222,
310,
1009,
14020,
278,
1494,
2462,
29889,
7806,
14020,
4520,
263,
3509,
373,
278,
4646,
29889,
13,
1576,
3144,
2209,
29891,
6515,
310,
278,
12029,
310,
18671,
29892,
24833,
322,
1203,
3145,
310,
278,
6263,
526,
13931,
964,
3196,
946,
355,
294,
1316,
408,
29901,
5214,
263,
9109,
322,
925,
12459,
393,
7726,
29879,
310,
278,
15426,
1788,
322,
278,
6263,
29915,
29879,
7609,
1080,
304,
3386,
263,
5400,
3086,
8660,
5611,
26596,
29936,
263,
9076,
310,
278,
315,
28479,
5920,
29892,
1554,
4942,
29889,
16477,
4083,
674,
367,
2309,
2629,
278,
6263,
29915,
29879,
937,
4832,
7378,
297,
8034,
29892,
881,
372,
367,
11467,
322,
6892,
11157,
278,
19201,
310,
278,
315,
28479,
17181,
2184,
29889,
13,
17351,
5134,
297,
278,
376,
8893,
292,
263,
5701,
1725,
322,
3387,
7765,
29908,
946,
8395,
338,
278,
25012,
310,
263,
4266,
4797,
3414,
4889,
373,
1153,
412,
322,
18287,
1283,
575,
3145,
408,
760,
310,
278,
6263,
29915,
29879,
12463,
16392,
304,
12566,
278,
4234,
29915,
29879,
5866,
322,
4344,
29892,
278,
7632,
310,
263,
716,
5400,
3086,
18923,
12252,
19252,
472,
997,
323,
542,
29892,
278,
25012,
310,
263,
716,
8068,
18923,
12039,
373,
678,
1485,
4149,
9321,
408,
760,
310,
263,
337,
25431,
310,
4624,
880,
29915,
29879,
612,
538,
29892,
4249,
916,
2712,
29889,
13,
29177,
278,
946,
8395,
376,
4806,
5555,
263,
315,
1148,
267,
573,
10307,
10629,
2200,
29908,
278,
6263,
29915,
29879,
7609,
1080,
526,
304,
3867,
2253,
27261,
322,
2130,
304,
2982,
304,
24060,
29892,
9801,
393,
278,
9793,
1788,
338,
8018,
322,
6728,
573,
29892,
760,
310,
607,
338,
278,
5314,
310,
15968,
9128,
9793,
322,
14717,
29889,
5901,
7609,
1080,
1090,
278,
5264,
18187,
946,
8395,
526,
278,
25012,
310,
263,
5400,
1592,
8270,
9045,
17535,
607,
674,
3160,
263,
2473,
29899,
2218,
13326,
3821,
21118,
304,
14511,
403,
9377,
29899,
29878,
9776,
380,
1296,
7694,
5353,
1080,
322,
1207,
6907,
800,
304,
5874,
373,
278,
1602,
28479,
2133,
322,
671,
310,
1766,
26323,
1648,
29892,
278,
12397,
322,
278,
13047,
310,
18363,
393,
526,
23180,
519,
3412,
411,
14717,
322,
28709,
1288,
5849,
29889,
13,
1576,
946,
8395,
525,
3206,
2827,
1749,
3086,
27486,
29915,
338,
599,
1048,
2504,
11427,
278,
11359,
29915,
29879,
9257,
322,
907,
1230,
6397,
2722,
29892,
5214,
4797,
902,
16639,
607,
674,
3160,
10127,
292,
263,
4797,
16930,
19133,
322,
1616,
23363,
29892,
408,
1532,
408,
29892,
9324,
8333,
278,
11359,
29915,
29879,
1261,
25804,
322,
20063,
607,
674,
3160,
13858,
278,
13382,
310,
13332,
362,
304,
10127,
263,
4343,
2635,
363,
278,
13587,
310,
2498,
20209,
29889,
13,
29177,
445,
946,
8395,
338,
278,
25161,
363,
901,
8543,
970,
5786,
29892,
278,
3710,
1680,
358,
310,
18363,
1549,
15483,
322,
14338,
16650,
293,
8569,
297,
7029,
5302,
29889,
13,
1576,
14682,
4778,
10614,
411,
278,
18028,
310,
3196,
9279,
304,
367,
21309,
373,
472,
278,
10719,
3819,
3233,
297,
278,
6263,
29915,
29879,
2446,
1840,
297,
8034,
7806,
10719,
3819,
756,
263,
1051,
310,
9279,
278,
6263,
16612,
408,
525,
29886,
21766,
9279,
29915,
304,
367,
8676,
411,
278,
5320,
29899,
6360,
10638,
29889,
13,
2517,
361,
4778,
4934,
1880,
491,
317,
13208,
21669,
518,
19689,
2891,
29949,
29901,
1963,
3747,
12828,
29962,
13,
29924,
293,
801,
5122,
13,
29924,
293,
801,
5122,
338,
385,
7841,
1024,
297,
278,
8955,
1608,
24400,
297,
624,
29889,
3182,
1512,
29889,
940,
4687,
670,
8955,
1608,
7149,
295,
482,
1090,
278,
12187,
10977,
310,
278,
7828,
1570,
1028,
7202,
12904,
261
] | 1,024 |
CommonCrawl
| 0.404237 | -0.64176 | 0.118919 | 0.3868 |