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Image:SEED HW3-2011.docx
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The Susan & Maile Google Webmaster Hangout Today
Jun 26, 2012 • 8:34 am | (1) by | Filed Under Other Google Topics
Susan Moskwa & Maile Ohye and some of the more well known Googlers who are great out webmaster outreach - they go to many of the US based SEO/SEM events.
Unlike JohnMu, Jonathan Simons, Pierre Far and some other Googlers, when Maile & Susan do Google Webmaster Hangouts, they do so as a team - a duo. So the conversation is a bit more interesting when you have two Googlers as opposed to one. The only issue, since Google Hangouts fit up to 10 people, you can only get 8 webmasters into the Hangout to participate, as opposed to when only one Googler runs it where you can get up to 9 webmasters.
That being said, the hangouts are often fun and if you can get in, it is often worth while.
Today at 1pm PDT or 4pm EDT, Susan and Maile are hosting another hangout. I wanted to give you webmasters and SEOs a heads up so you can join, if you are one of the first.
For more details, see Susan's Google+ post on how to join.
Forum discussion at Google+.
Previous story: YouTube Video Stuck At 301 Views? Here Is Why.
blog comments powered by Disqus
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China: State Security Law Amendment
This post also available in:
Français · Chine : Révision de la loi sur la sécurité de l'état
The latest amendment on State Security Law was submittend to the National People's Congress (NPC) Standing Committee for review yesterday. The drafted law requires telecom operators and Internet service providers to cooperate with public security and State security authorities on investigations of possible State secret leaks.
According to existing practice, Telecom operators and ISPs have been providing the public security and state security authorities users’ information for facilitation investigations of crimes such as criminal libels, rumors, subversive activities and state secrets. For example, Yahoo! provided information on Shi Tao's case in 2005 and Tan Zuoren's case last year 2009. The former was sentenced to 10-year imprisonment under the charge of leaking state secret while the latter was sentence to 5-year imprisonment for defaming the Communist Party of China in email comments about the Tiananmen Square protests of 1989.
International Internet operators, such as Google has criticized the lack of legal bases and transparency in the implementation of law in China, the latest amendment will legitimatize current practice. In the draft law, according to China Daily's report, rather than identifying a clear list / scope of secrets, such as budgetary, finance and military informations, a State secret is defined “as information concerning national security and interests that, if released, would harm the country's security and interests” and the confidentiality of files is defined by the expiration dates set by the government.
China Daily quotes law experts and explains the rationality and context behind the law amendment:
Experts said the changes come as new requirements on solving cases concerning State secrets have emerged in new contexts.
Officials have blamed phone text messages as a tool for criminals to instigate the deadly July 5 riot last year in Northwest China's Xinjiang Uygur autonomous region.
“We now have more media carrying information, rather than just the paper documents of the past. Without cooperation from network carriers and service providers, authorities alone could not collect evidence or sort out the cases,” said Ma Huaide, a law professor of China University of Political Science and Law.
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Friday, 22 February 2013
TUKS Prestige Lecture Program: Louis Harms
The tragedy that is the Blade and The Beautiful is not the only news happening in Pretoria, South Africa at the moment.
We have heard from Dario Tanziani that former Supreme Court of Appeal Judge Louis Harms, an extraordinary professor in the Department of Private Law and the incumbent of the Adams and Adams chair in Intellectual Property at the University of Pretoria (TUKS), is taking part in their Prestige Lecture Program next month.
The TUKS invitation reads:
Prestige lecture Presentation by Judge LTC Harms, Adams and Adams Professor in Intellectual Property Law:
‘Plain packaging and its impact on trademark law’
The Faculty of Law of the University of Pretoria cordially invites you to the abovementioned presentation.
Date Wednesday 13 March 2013 Time 18:30 for 19:00
Venue Senate Chamber, University of Pretoria
RSVP By 1 March 2013, +27 12 362 5184 (f), rsvp.law@up.ac.za
Enquiries Mornay Hassen +27 12 420 4126
Amongst the considerable accolades for the former Judge is his most recent election as an Honorary Master of the Bench of the Middle Temple.
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200 reputation
6
bio website stevefink.net
location Philadelphia, NY
age 31
visits member for 1 year, 9 months
seen Nov 8 '11 at 17:43
stats profile views 19
iPhone/iPad/web dev. Family man. Budding author. Passionate about everything web & mobile & startups. Passionate about building applications and tools that enriches people's lives.
Although I'm platform agnostic and use whatever tool is appropriate for the job, I've taken a huge liking to the Ruby on Rails framework recently. I am working on a couple of web services that utilize Rails in their stack and am enjoying every minute of it.
This user has not answered any questions
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Information for "KFI Docs"
Jump to: navigation, search
Basic information
Display titleKFI Docs
Default sort keyKFI Docs
Page length (in bytes)3,055
Page ID1628
Page content languageEnglish (en)
Search engine statusIndexable
Number of views2,384
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Counted as a content pageYes
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Page creatorRBot (Talk | contribs)
Date of page creation03:34, 6 March 2007
Latest editorAdushistova (Talk | contribs)
Date of latest edit09:57, 27 October 2011
Total number of edits2
Total number of distinct authors2
Recent number of edits (within past 91 days)0
Recent number of distinct authors0
Retrieved from "http://elinux.org/KFI_Docs"
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Information for "RPi CANBus"
Jump to: navigation, search
Basic information
Display titleRPi CANBus
Default sort keyRPi CANBus
Page length (in bytes)10,857
Page ID39740
Page content languageEnglish (en)
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Page creatorZeta (Talk | contribs)
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Latest editorZeta (Talk | contribs)
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Total number of edits3
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Retrieved from "http://elinux.org/RPi_CANBus"
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Paper report
Articles selected by Faculty of 1000: microarrays to study antibiotic resistance; identifying SUMO substrate proteins; parallel SNP genotyping; intron origin and evolution; variation in CpG-island methylation in humans
• Correspondence:
Genome Biology 2004, 5:325 doi:10.1186/gb-2004-5-5-325
The electronic version of this article is the complete one and can be found online at: http://genomebiology.com/2004/5/5/325/
Published:20 April 2004
© 2004 BioMed Central Ltd
Microarrays to study antibiotic resistance
Aminoglycoside microarrays to study antibiotic resistance. Disney MD, Magnet S, Blanchard JS, Seeberger PH. Angew Chem Int Ed Engl 2004, 43:1591-1594.
For the Faculty of 1000 evaluation of this article please see: http://genomebiology.com/reports/F1000/gb-2004-5-5-325.asp#Disney webcite
Identifying SUMO substrate proteins
Broad-spectrum identification of cellular SUMO substrate proteins. Zhao Y, Kwon SW, Anselmo A, Kaur K, White MA. J Biol Chem 2004, Mar 11.
For the Faculty of 1000 evaluation of this article please see: http://genomebiology.com/reports/F1000/gb-2004-5-5-325.asp#Zhao webcite
Parallel SNP genotyping
Parallel genotyping of over 10,000 SNPs using a one-primer assay on a high-density oligonucleotide array. Matsuzaki H, Loi H, Dong S, Tsai YY, Fang J, Law J, Di X, Liu WM, Yang G, Liu G, et al. Genome Res 2004, 14:414-425.
For the Faculty of 1000 evaluation of this article please see: http://genomebiology.com/reports/F1000/gb-2004-5-5-325.asp#Matsuzaki webcite
Intron origin and evolution
Exon junction sequences as cryptic splice sites; implications for intron origin. Sadusky T, Newman AJ, Dibb NJ. Curr Biol 2004, 14:505-509.
For the Faculty of 1000 evaluation of this article please see: http://genomebiology.com/reports/F1000/gb-2004-5-5-325.asp#Sadusky webcite
Variation in CpG-island methylation in humans
A comprehensive analysis of allelic methylation status of CpG islands on human chromosome 21q. Yamada Y, Watanabe H, Miura F, Soejima H, Uchiyama M, Iwasaka T, Mukai T, Sakaki Y, Ito T. Genome Res 2004, 14:247-266.
For the Faculty of 1000 evaluation of this article please see: http://genomebiology.com/reports/F1000/gb-2004-5-5-325.asp#Yamada webcite
Genome Biologypaper reports
Genome Biology aims to highlight significant research articles newly published in other journals. From 2003, individual paper reports have been replaced by summaries of articles of interest to Genome Biology readers, taken from those highlighted by Faculty of 1000, a new online literature awareness tool published by BioMed Central. Faculty of 1000 systematically and comprehensively reviews the most interesting recent articles published in the biological sciences on the basis of recommendations of a faculty of over 1,400 selected leading researchers.
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GlobalVoices in Learn more »
Taiwan: Flying Fish Season on Orchid Island
This post also available in:
Français · Taïwan : La saison des poissons volants sur l’Île de l'orchidée
বাংলা · তাইওয়ানঃ অর্কিড দ্বীপে উড়ন্ত মাছের কাল
русский · Тайвань: сезон летучей рыбы на острове Орхидей
Magyar · Tajvan: Repülőhal szezon az Orchidea-szigeten
Español · Taiwán: Temporada de peces voladores en la isla Orquídea
Every spring, when flying fish are carried to Taiwan by the Kuroshio Current, many fishermen are waiting for them. The most famous traditional flying fish festival is on Orchid Island, where the Tao people have lived for generations.
The influence of the flying fish on Tao culture is immense. Tao people call their island the home of the flying fish (‘ali bang bang’ in the Tao language), and even their calendar is closely related to the flying fish season: Spring is ‘rajun' (the flying fish season), summer and autumn is 'teteka’ (when the flying fish season ends), and winter is ‘aminon’ (when there is no flying fish).
The flying fish season on Orchid Island [zh] starts in March. When Cheilopogon unicolor fish (‘sosowon’ in Tao language) arrive, Tao people catch them at night using light to attract the flying fish. Researchers at the National Museum of Marine Biology and Aquarium in Taiwan described [zh] how only a few other Asian islands share this fishing technique with the Tao people on Orchid Island.
以火把誘引飛魚的捕撈法，除蘭嶼外僅見於菲律賓巴丹諸島及日本；以上島嶼均屬於黑潮海域，因此日本黑潮文化學者國分直一曾提議將這些島嶼列為「火把捕撈飛魚法的文化小國」。
In addition to Orchid Island, catching flying fish by luring them with fire torch is only observed in Batan Island in the Philippines and in Japan. These islands are located along the Kuroshio Current. As a result, the Japanese researcher Kokubu Naoichi, who studies Kuroshio culture, once proposed to group these islands as ‘countries of the catching-flying-fish-by-fire-torch culture.’
Flying fish hung up to dry on Orchid Island. Photo by Grillmagic. CC: BY-NC-SA)
In April and May, the most important flying fish is Cheilopogon spilonotopterus (‘papatawan’ in Tao language). Among all the flying fish swimming to Taiwan, the Tao people consider the Cheilopogon cyanopterus (‘mayaeng’ in Tao language) to be the most respectable because they believe that ‘mayaeng’ is the soul and chief of all flying fish species.
The fishermen preserve the fish by drying them in the sun. Flickr User Grillmagic took some photos of the fish-drying scene on Orchid Island in May 2012.
This year, YouTube user paudull accompanied by several residents of the island recorded the festival on his camera:
Below is a brief description of the video with corresponding time stamps:
1:12 – Throwing out the rope (about 600 meters) for trapping the flying fish (to the net).
1:40 – The fishermen jump into the water.
2:25 – They swim closer to each other to trap the flying fish with the net.
5:11 – Because none of us ate breakfast, someone cut two fish and made sashimi for breakfast.
5:35 – An illustration of the fish-catching strategy.
9:24 – The background music is a little bit boring. Let's listen to a song from Orchid Island (‘A love song of a Tao man’).
12:29 – Wow, do you see that flying fish fly?
15:52 – At last, everyone happily brought some flying fish home.
16:00 – Cleaning and preparing the flying fish that were caught today with family.
As a backpacker to Orchid Island, Chia-Chi Tseng made a short film about the flying fish festival, ‘Libangbang’ (flying fish in Orchid Island), which was entered into the Golden Harvest Awards for films and documentaries. Below is the trailer.
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GWiedeman's bookmarks
"If you do not understand a man you cannot crush him. And if you do understand him, very probably you will not."
Chesterton, Gilbert K. on understanding
3 fans of this quote
"Man seems to be capable of great virtues but not of small virtues; capable of defying his torturer but not of keeping his temper."
Chesterton, Gilbert K. on virtue
"A teacher who is not dogmatic is simply a teacher who is not teaching."
Chesterton, Gilbert K. on teacher
This quotation can be viewed in the context of a book
"Chastity does not mean abstention from sexual wrong; it means something flaming, like Joan of Arc."
Chesterton, Gilbert K. on chastity
This quotation can be viewed in the context of a book
"The honest poor can sometimes forget poverty. The honest rich can never forget it."
Chesterton, Gilbert K. on poverty and the poor
"Those thinkers who cannot believe in any gods often assert that the love of humanity would be in itself sufficient for them; and so, perhaps, it would, if they had it."
Chesterton, Gilbert K. on atheism
"In matters of truth the fact that you don't want to publish something is, nine times out of ten, a proof that you ought to publish it."
Chesterton, Gilbert K. on publishing and publishers
3 fans of this quote
This quotation can be viewed in the context of a book
"The madman is not the man who has lost his reason. He is the man who has lost everything except his reason."
Chesterton, Gilbert K. on reason
6 fans of this quote
"Among the very rich you will never find a really generous man, even by accident. They may give their money away, but they will never give themselves away; they are egoistic, secretive, dry as old bones. To be smart enough to get all that money you must be dull enough to want it."
Chesterton, Gilbert K. on riches
3 fans of this quote
"The ordinary scientific man is strictly a sentimentalist. He is a sentimentalist in this essential sense, that he is soaked and swept away by mere associations."
Chesterton, Gilbert K. on science
"To be clever enough to get all the money, one must be stupid enough to want it."
Chesterton, Gilbert K. on stupidity
3 fans of this quote
"A yawn is a silent shout."
Chesterton, Gilbert K. on bores and boredom
6 fans of this quote
"The chief assertion of religious morality is that white is a color. Virtue is not the absence of vices or the avoidance of moral dangers; virtue is a vivid and separate thing, like pain or a particular smell."
Chesterton, Gilbert K. on virtue
"There are many definite methods, honest and dishonest, which make people rich; the only instinct I know of which does it is that instinct which theological Christianity crudely describes as the sin of avarice."
Chesterton, Gilbert K. on wealth
"There are two kinds of people: those who say to God, Thy will be done, and those to whom God says, All right, then, have it your way."
Lewis, C. S. on dedication
24 fans of this quote
"We were promised sufferings. They were part of the program. We were even told, 'Blessed are they that mourn'"
Lewis, C. S. on difficulties
8 fans of this quote
"Faith... is the art of holding on to things your reason once accepted, despite your changing moods."
Lewis, C. S. on faith
16 fans of this quote
"Friendship is unnecessary, like philosophy, like art. It has no survival value; rather it is one of those things that give value to survival."
Lewis, C. S. on friends and friendship
19 fans of this quote
"Nothing that you have not given away will ever be really yours."
Lewis, C. S. on giving
14 fans of this quote
"I do not believe one can settle how much we ought to give. I am afraid the only safe rule is to give more than we can spare."
Lewis, C. S. on giving
10 fans of this quote
"God cannot give us a happiness and peace apart from Himself, because it is not there. There is no such thing."
Lewis, C. S. on happiness
14 fans of this quote
"If you read history you will find that the Christians who did most for the present world were precisely those who thought most of the next. It is since Christians have largely ceased to think of the other world that they have become so ineffective in this."
Lewis, C. S. on heaven
5 fans of this quote
"Courage is almost a contradiction in terms. It means a strong desire to live taking the form of a readiness to die."
Chesterton, Gilbert K. on courage
4 fans of this quote
"With any recovery from morbidity there must go a certain healthy humiliation."
Chesterton, Gilbert K. on convalescence
"I believe in getting into hot water. I think it keeps you clean."
Chesterton, Gilbert K. on conflict
4 fans of this quote
"The world will never starve for want of wonders, but for want of wonder."
Chesterton, Gilbert K. on wonder
3 fans of this quote
"The vulgar man is always the most distinguished, for the very desire to be distinguished is vulgar."
Chesterton, Gilbert K. on vulgarity
3 fans of this quote
This quotation can be viewed in the context of a book
"One sees great things from the valley; only small things from the peak."
Chesterton, Gilbert K. on adversity
3 fans of this quote
"The Christian ideal has not been tried and found wanting. It has been found difficult; and left untried."
Chesterton, Gilbert K. on christians and christianity
6 fans of this quote
"The doctrine of human equality reposes on this: that there is no man really clever who has not found that he is stupid."
Chesterton, Gilbert K. on cleverness
"I've searched all the parks in all the cities and found no statues of committees."
Chesterton, Gilbert K. on committees and meetings
"The perplexity of life arises from there being too many interesting things in it for us to be interested properly in any of them."
Chesterton, Gilbert K. on complexity
3 fans of this quote
"Compromise used to mean that half a loaf was better than no bread. Among modern statesmen it really seems to mean that half a loaf ;is better than a whole loaf."
Chesterton, Gilbert K. on compromise
"Aim at heaven, and you will get earth thrown in; aim at earth, and you will get neither."
Lewis, C. S. on heaven
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But wait... my book has more: prev 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28 next
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I'm male and made my book on 23rd February 2010.
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It's easy! Just pick the product you like and click-through to buy it from trusted partners of Quotations Book. We hope you like these personalized gifts as much as we do.
Make and then buy your OWN fantastic personalized gift from this quote
Accident is the name of the greatest of all inventors. Twain, Mark
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212 - The Extra Degree
The one extra degree makes the difference. This simple analogy reflects the ultimate definition of excellence. Because it's the one extra degree of effort, in business and life, that can separate the good from the great. This powerful book by S.L. Parker and Mac Anderson gives great examples, great quotes and great stories to illustrate the 212° concept. A warning - once you read it, it will be hard to forget. Your company will have a target for everything you do ... 212°
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It's easy! Just pick the product you like and click-through to buy it from trusted partners of Quotations Book. We hope you like these personalized gifts as much as we do.
Make and then buy your OWN fantastic personalized gift from this quote
He that plants trees loves others besides himself. Proverb, English
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A selection of more great products and gifts!
212 - The Extra Degree
The one extra degree makes the difference. This simple analogy reflects the ultimate definition of excellence. Because it's the one extra degree of effort, in business and life, that can separate the good from the great. This powerful book by S.L. Parker and Mac Anderson gives great examples, great quotes and great stories to illustrate the 212° concept. A warning - once you read it, it will be hard to forget. Your company will have a target for everything you do ... 212°
Click here to buy this »
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Help Wikitravel grow by contributing to an article! Learn how.
Ngorongoro Conservation Area
From Wikitravel
(Redirected from Ngorongoro Conservancy)
Jump to: navigation, search
Ngorongoro Conservation Area is a park in the Northwest Tanzania. It contains an old volcano that has collapsed and formed a crater (caldera). The steep sides of the crater have become a natural enclosure for a wide variety of wildlife.
[edit] Understand
[edit] History
Considered to be "the cradle of civilization".
[edit] Landscape
[edit] Flora and fauna
Fauna: Lions are in the highest density in the world in the crater, so there is a good chance of seeing them. There are herds of wildebeest, zebra, and a lot of buffalo and Grants' gazelles too. This is one of the best places in East Africa to see a Black Rhino. Hippos and flamingos are seen in Lake Magadi. Hyenas are a common predator, and cheetahs are frequently seen. Leopards are in the conservation area but rarely seen, along with wild dogs.
[edit] Climate
[edit] Get in
Most people come to Ngorongoro from Arusha on organized safaris. It is, however, cheaper to organize a safari from Karatu, a town 10km from the gates. Here you can organize a safari with an independent driver. If using an independent driver, the profit will go to an individual Tanzanian instead of a larger safari company, however these vehicles are less reliable and are known to have parts fall off while driving around the crater. That can add or detract from your experience, depending on what you're looking for.
[edit] Fees/Permits
For foreigners: $50 per person/day. $200 per car/day. (not a car full of people, just the car.)
For Tanzanians it's 15,000/= per day. Most of the locals believe that the bulk of the park fees go into pockets.
[edit] Get around
[edit][add listing] See
• Ol Doinyo Lengai - a volcano
[edit][add listing] Do
[edit][add listing] Buy
[edit][add listing] Eat
[edit][add listing] Drink
[edit][add listing] Sleep
[edit] Lodging
[edit] Camping
[edit] Backcountry
[edit] Stay safe
[edit] Get out
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Help Wikitravel grow by contributing to an article! Learn how.
San Luis Obispo County
From Wikitravel
Jump to: navigation, search
San Luis Obispo County [1] is in the Central Coast of California in the United States of America. The county seat is San Luis Obispo (population 46,000), home to California Polytechnic State University (Cal Poly) with 20,000 students. The county is the third largest wine producer in California, behind Sonoma and Napa counties. It has a number of coastal towns. A major attraction is Hearst Castle at San Simeon.
[edit] Regions
[edit] Cities
[edit] Other destinations
[edit] Understand
[edit] Get in
[edit] By plane
The major airport is in San Luis Obispo. Scheduled flights are available to and from San Francisco, Los Angeles, and Phoenix on United [2] (operated by SkyWest [3] as United Express) and USAirways [4] (operated by Mesa [5] as USAirways Express). Prices start around $200 for a ticket purchased separately, but are much less expensive when combined with flights going through San Francisco or Los Angeles. There is an airport in Paso Robles, but it has no commercial service.
An alternate airport lies in Santa Maria, a city just south of the county line. There is weekly service to Honolulu, four times a week service to Las Vegas and frequent service to Los Angeles.
[edit] By train
Amtrak [6] has train stations in San Luis Obispo, Paso Robles and Grover Beach. Coast Starlight goes from Seattle to Los Angeles stopping at Paso Robles and San Luis Obispo within this county. The Pacific Surfliner goes from San Luis Obispo to San Diego and makes all stops. There is connecting bus service to other places statewide including the Bay Area, Los Angeles, and the Central Valley. Make sure you leave plenty of time because it can take up to 7 hours to get to SLO from San Francisco and 8 hours to get to SLO from San Diego, although the spectacular delays frequent in the past have abated significantly following recent agreements with freight rail carriers.
[edit] By bus
Greyhound [7] offers bus routes into SLO County. Once again set aside time because the trips can be long but Greyhound is one of the cheapest ways to go.
[edit] By car
If coming from the north or south, Highway 101 is the fastest route. Travel times from the Bay Area or Los Angeles are about four hours. An alternate route from the Bay Area is to take State Route 1 down the coast. It is a senic drive, but is winding and slow and takes about six hours. Travelers who begin in the Central Valley, like Bakersfield or Fresno, need to take Route 46 or 41 to SLO County.
[edit] Get around
[edit][add listing] See
[edit] Itineraries
[edit][add listing] Do
[edit] Get out
Santa Barbara County is to the south and Monterey County to the north.
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Australian Bureau of Statistics
Celebrating the International Year of Statistics 2013
ABS Home > Statistics > By Release Date
7111.0 - Principal Agricultural Commodities, Australia, Preliminary, 1999-2000
Previous ISSUE Released at 11:30 AM (CANBERRA TIME) 31/10/2000
Page tools: Print Page Print All RSS Search this Product
MEDIA RELEASE
October 31, 2000
Embargoed: 11:30 AM (AEST)
150/2000
Record wheat and canola crops: ABS
Wheat production increased by 17 per cent to a record 25 million tonnes in 1999/2000, according to figures released by the Australian Bureau of Statistics (ABS) today.
Significant increases in wheat production were recorded in New South Wales, Victoria and Western Australia (which reported a state record harvest of 9.2 million tonnes). Wheat plantings were up by 7 per cent to 12.3 million hectares with the main increases recorded in New South Wales and Victoria.
Canola also had another record year in which plantings rose by 54 per cent to 1.9 million hectares and production rose by 44 per cent to 2.4 million tonnes. Significant increases were recorded in all major growing states. Western Australia was the largest canola producing State and recorded the biggest increase, with area sown up by 70 per cent to 909,000 hectares and quantity harvested up by 61 per cent to 989,000 tonnes.
In addition, beef cattle numbers showed signs of recovery from the drop during the previous year, with the herd estimated at 23.6 million head at 30 June 2000. Sheep and lamb numbers remained steady at approximately 116 million at 30 June 2000.
Conversely, preliminary estimates indicated a slight drop in dairy cattle numbers with 3.1 million cattle recorded at 30 June 2000. This was the result of a drop in the estimated number of dairy bulls, heifers and calves as the number of cows remained mainly unchanged.
Pig numbers fell by 7 per cent to 2.4 million head at 30 June 2000 with falls reported in all states.
Details are in Principal Agricultural Commodities Australia (Preliminary) 1999–00 (cat. no. 7111.0) available from ABS bookshops in all capital cities. The summary of the publication can be found on this site. If you wish to purchase a copy of this publication, contact the ABS Bookshop in your capital city.
© Commonwealth of Australia 2013
Unless otherwise noted, content on this website is licensed under a Creative Commons Attribution 2.5 Australia Licence together with any terms, conditions and exclusions as set out in the website Copyright notice. For permission to do anything beyond the scope of this licence and copyright terms contact us.
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This article looks at some classic spreadsheet errors in layman’s terms and suggests some preventative measures.
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Tim Jahn Gives an Intimate Look at Entrepreneurship
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Here is a list of 7 biggest productivity killers that I've come across, and my take on how people can address and tackle each one of them. Read More
Have you failed? Had robust feedback from clients? Did your heart break? This post shares my take on rejection and why it's actually very good for you. Read More
Conflict Management Training: Conflict is inevitable. You Are Bound To Get It. Why? Because people don’t share exactly the same ideas. If we all had the same ideas, then we would always agree, and then the issue of conflict would never come up. But the... Read More
There seems to be a theme over the last few months that the only way leaders are successful or effective is if they get up early. It might be true for small business owners and executives…and it might not.How do you design your schedule and does it really indicate if you are lazy or disengaged? Read More
Why do so many of us feel it's best to avoid sharing our opinions? Learn how opinions can take your next presentation to the next level. Read More
Leadership Training: How Can I Manage My Friend? On a recent training course, in oxford, I was asked the following question, by a delegate called Kim, which I have heard many times before, from... Read More
9 Proven Tips To Control Your Inventory
Posted by sophia2 under Self-Development
From http://www.youngupstarts.com 4 days ago
Made Hot by: zioncampo on May 15, 2013 7:55 am
By Crystal Wells, Product Marketing at Vendio Controlling inventory is the ongoing process of identifying and managing the constant flow of items into and out of an existing inventory. When you don’t know exactly what inventory you have available, where it is located, or where it is for sale, it’s Read More
2014 will mark the 20th anniversary of Amazon’s founding. During the intervening 20-years, Jeff has offered business executives numerous valuable tips, including the following... Read More
7 Ways for Effective Blog Time Management
Posted by TimoK under Self-Development
From http://www.productivesuperdad.com 10 days ago
Made Hot by: maestro68 on May 13, 2013 10:35 pm
Seven effective blog time management tips for building your blog. I’m using these strategies myself and they do work (I’m a dad, an athlete and I have a full-time day job). Read More
You are never paid for hard working or for your efforts, but you are paid for your impact – no matter if you are an entrepreneur or an employee! Read More
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Mike Abasov @Mike_Abasov Serves Business With a Smile
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The Role of Information and Communication Technologies (ICTs) in Delivering Higher Education – A Case of Bangladesh
Shah Md. Safiul Hoque, S. M. Shafiul Alam
Abstract
At present a new era has evolved in the education sector by means of ICTS. Different ICTs are now set to become instrumental to help expand access to education, strengthen the relevance of education to the increasingly digital workplace, and raise educational quality by, among others, helping make teaching and learning into an engaging, active process connected to real life. The application and exposure to and deployment of ICTs fundamentally change the way education is conceived and delivered to students. ICTs are enablers that optimize student-centered pedagogical methods. Due to its easy accessibility this means of education has become very popular all over the world. Distance education has got a thrust after the evolution of ICT-based education system. This paper intends to give an idea about ICT-based higher education all over the globe and its applicability in Bangladesh. Finally, it analyses the responses from different user groups to query about the current status of the ICT-based higher education system Bangladesh.
Full Text: PDF
This work is licensed under a Creative Commons Attribution 3.0 License.
International Education Studies ISSN 1913-9020 (Print), ISSN 1913-9039 (Online)
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advanced search
Category: Educational Resources
Westview on the James Environmental Education Program
A camp with an enviro. education program for any interested group or organization wanting to learn more about the natural world.
Ratings/Review of this resource:
Address:
Westview on the James
Jeff Barry
1231 West View Road
Goochland, Virginia 23063
Phone: (804) 457-4210
Fax: (804) 457-2178
E-Mail: jtbarry@erols.com
Website: http://www.westviewonthejames.org/eeindex.htm
Detailed Information:
Westview on the James facilitates hands-on activities and programs for K-12 grades in order to reinforce in-classroom skills and concepts. In order to capitalize on the experience that the groups receive at Westview, the instructors work with the teachers to create programs and activities around a particular "centerpiece" issue or concern that they are learning about in school or is on the news pertaining to the environment. We are also leading High Adventure trips, canoeing/hiking/etc., where the groups learn about outdoor skills as well as the natural world. Westview also offers workshops with national scopes for interested teachers to learn about various topics such as learning more about setting up an EE program in the school, and learning how EE and the Internet can work together.
Resources that may be related:
Home | Site Map | About EnviroLink | Advanced Search | Suggest a Resource
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Mecklenburg-Strelitz – Herzogtum (duchy)JurisdictionsEdit This Page
From FamilySearch Wiki
Back to Mecklenburg-Strelitz – Großherzogtum (grand duchy) Page
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• This page was last modified on 4 September 2012, at 17:16.
• This page has been accessed 138 times.
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Las Vegas Nevada FSL/Class ScheduleEdit This Page
From FamilySearch Wiki
Revision as of 04:12, 29 November 2012 by Timcdaniel (Talk | contribs)
Contents
Las Vegas FamilySearch Library
509 South 9th Street, Las Vegas, Nevada 702-382-9695 nv_lasvegas@ldsmail.net
Hours
• Mon, Fri, Sat: 9:00 a.m to 5:00 p.m
• Tue-Thu: 9:00 a.m. to 9:00 p.m.
Home
Group Visits
Contact Information
Holiday Schedule
Handouts and Classes
Training
Resources
Services
Local Resources & Newsletters
Links
Volunteer
Please call ahead to reserve your spot 382-9695
Click Here to Sign Up For Our Monthly E-Newsletter
We are just getting started, check back at regularly for updates of new classes that will be added.
Handouts
Classes for 2012
Click on the hyperlink below. It will open in a new window allowing you to download the PDF file to print or save on your computer.
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Montana NewspapersEdit This Page
From FamilySearch Wiki
Revision as of 04:28, 28 August 2012 by CSander (Talk | contribs)
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)
United States U.S. Newspapers Montana Newspapers
Contents
Resources for Montana Newspapers
Libraries and Archives
The Montana Historical Society has the largest collection of Montana newspapers. The society has a card file of obituaries, primarily from pre-1920 newspapers.
The Montana State University Libraryand the Mansfield Library at the University of Montana also have some newspaper collections. The Family History Library has not acquired newspapers from the state.
United States Newspaper Directory
The U.S. Newspaper Directory, 1690-Present is a national directory from the Library of Congress of over 140,000 newspaper titles. The directory provides a list of libraries from around the country that have the newspapers in their collections.
Digital Issues Online
Wikipedia has more about this subject: List of online newspaper archives
Indexes
Websites
Need additional research help? Contact our research help specialists.
Need wiki, indexing, or website help? Contact our product teams.
Did you find this article helpful?
You're invited to explain your rating on the discussion page (you must be signed in).
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Changes related to "Help:User page and preferences"
From FamilySearch Wiki
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"url": "www.fides.org/en/news/32226?idnews=32226&lan=eng",
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http://www.fides.org
Africa
2012-09-14
AFRICA/EGYPT - Between fears of further incidents after the Friday prayers and the attention of the Arab world for the Pope in Lebanon
Cairo (Agenzia Fides) - "We await the end of the Friday prayers, to see if there are new incidents, that everyone a bit expects," says Fr. Rafic Greiche, head of the communications for the Catholic Church in Egypt. "In fact, several groups have announced that, after prayer, they will manifest again in front of the American Embassy in Cairo against the film that offends the Prophet Mohammed. Among other things, one of the most important mosques of Cairo is located in Tahrir Square, which is close to the Embassy," explains Fr. Greiche. "One must also keep in mind that this is an opportunity for the different preachers to show their oratorical skills," adds the priest, suggesting that it is difficult to expect conciliator sermons.
"Messages have been sent to mobile phones inviting Christians to come today to protest peacefully in front of the Coptic Cathedral. The messages were sent by mixed groups of Copts and Muslims," adds the priest.
With regards to the interest in Egypt for the Pope Benedict XVI’s visit in Lebanon, which begins today, Fr. Greiche emphasizes: "Catholic Egyptians are tuned on Telenour (Tele Lumiere), the Lebanon Catholic station, to follow live the Holy Father’s arrival in Beirut. But there is concern throughout the Arab world. For example Al Jazeera broadcast yesterday a long documentary about the papal visit. Our hope is that the Pope will bring a message of reconciliation with the Islamic world," concludes the priest. (L.M.) (Agenzia Fides 14/09/2012)
Share: Facebook Twitter Google Blogger Altri Social Network
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Australian Geodetic Datum
Products
Maps [More]
Data/Applications [More]
Publications [More]
Multimedia [More]
Subscribe
The Australian Geodetic Datum (AGD), both the 1966 and 1984 versions, have been replaced by Geocentric Datum of Australia (GDA94).
Australian Geodetic Datum 1966 (AGD66)
The least squares adjustment of the Australian geodetic network performed in March 1966, used the Australian Geodetic Datum. This adjustment produced a set of coordinates which, in the form of latitudes and longitudes, is known as the Australian Geodetic Datum 1966 coordinate set (AGD66).
Australian Fiducial Network
Johnston Geodetic Station
Stone marker
© Geoscience Australia
The grid coordinates derived from a Universal Transverse Mercator projection of the AGD66 coordinates, using the Australian National Spheroid, is now known as the Australian Map Grid 1966 coordinate set (AMG66).
The Australian Geodetic Datum was proclaimed in the Australian Commonwealth Gazette of 6 October 1966. This proclamation included the parameters of the local ellipsoid, known as the Australian National Spheroid (ANS), which defines the adopted size and shape of the earth, and the position of the origin point - Johnston Geodetic Station. This image displays the Johnston Geodetic Station Stone Marker - S 25° 56' 54.5515", E 133° 12' 30.0771", 571.2 metres (ellipsoid height). (Select image to view larger 85k version)
Australian National Spheroid
Semi-major axis (a): 6 378 160.0 metres
Semi-minor axis (b): 6 356 774.719 metres
Flattening (f): 1/298.25
The adoption of this origin and best fitting local ellipsoid means that the centre of the ANS does not coincide with the centre of mass of the Earth but lies about 200 metres from it.
Extract from: Commonwealth Gazette, No 84, 6th October 1966
Department of National Development
National Mapping Council
At the twenty-fourth meeting of the National Mapping Council held in Melbourne, the Council, on 21 April 1966, adopted the following datum for Australian Geodetic Surveys:
• Designation: The Australian Geodetic Datum
• Reference spheroid: The Australian National Spheroid with a major (equatorial) radius of 6 378 160 metres and a flattening of 1/298.25
• Origin: The Johnston Geodetic Station situated in the Northern Territory at East Longitude 133° 12' 30.0771" and South Latitude 25° 56' 54.5515" and with a ground level elevation of 571.2 metres above the spheroid.
B.P. LAMBERT
Director of National Mapping
Chairman of National Mapping Council
Australian Geodetic Datum 1984 [AGD84]
In 1982, a new national adjustment, referred to as the Geodetic Model of Australia 1982 (GMA82), was performed using all data previously included in the 1966 adjustment as well as additional, modern terrestrial and space-based observations. This new adjustment also used the gazetted Australian Geodetic Datum. The coordinate set resulting from this adjustment was accepted by the National Mapping Council in 1984 and is known as the Australian Geodetic Datum 1984 (AGD84).
Topic contact: geodesy@ga.gov.au Last updated: May 31, 2012
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Noob Seeks Direction
Newbie Member
13Jan2009,13:34 #1
Hi everyone,
Im new to this site, and web programming as a whole. I know Id like to learn php and MySQL. I have some online business ideas that need extensive back end databases built with alot of automated features.
I was wondering what I should learn and what order to learn them.
Someone told me I need to learn HTML language, then CSS, then PHP then MYSQL.
I dont know where to begin and I don't want to waste time. I will need some fundamental knowledge of how to get a webpage up- although I plan to use Dreamweaver and Flash for the design part of it.
Thanks so much,
BobAGraham@gmail.com
Go4Expert Founder
13Jan2009,15:42 #2
First thing first. You should learn PHP and there is nothing in HTML which you can't learn from doing PHP.
Its the programming which you need to learn and how to put images / content in pages is HTML which is very basic stuff if you know somewhat of PHP.
Now after knowing PHP it would have advance where it deals with database and as soon as you have it you would start building your websites and apparently see how HTML and CSS things work.
Definitely starting with HTML/CSS is theoretically correct but I would like to see more practical approach.
Newbie Member
14Jan2009,16:04 #3
Hi there,
I signed up to Lynda.com and got a video on PHP Essentials. Ill start from there. I've used Dreamweaver before and it makes sense so this is the logical next step, correct?
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"warc_url": "http://www.libreoffice.org/download/?type=mac-x86&lang=lv&version=3.6.5"
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The free office suite
Download LibreOffice
LibreOffice Mac OS X (Intel), version 3.6.5, Latvian. Not the version you wanted? Change System, Version or Language
You need to download and install these files in order:
• Source code
LibreOffice is an open source project and you can therefore download the source code to build your own installer.
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{
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"warc_url": "http://www.mariowiki.com/Pipe_Room"
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Pipe Room
From the Super Mario Wiki
Jump to: navigation, search
Pipe Room
Area Area Four
Normal Ghosts Flying Fish; Red and White Grabbing Ghosts
Boo Booffant
Click an area to open the relevant article.
The Pipe Room (Japanese: 配管室 Piping Room) is a room in Area Four of Luigi's Mansion.
Once Luigi enters the Pipe Room, a set of green Flying Fish and Red and White Grabbing Ghosts attack. After Luigi captures them, he uses an Ice Elemental Ghost to get to a valve that stops the flow of water. This causes the Key to the Cold Storage to appear.
[edit] Description
The Pipe Room contains a crate, a bucket, and a frozen barrel. The river of water is untouchable until Luigi freezes it.
[edit] Ghosts in the Pipe Room in the PAL Hidden Mansion
[edit] Trivia
• The Pipe Room and the first floor Washroom are quite possibly the only necessary rooms in the game which are never required to be lit up (unless of course the player is trying to catch all fifty Boos), as Luigi can completely ignore the Grabbing Ghosts, freely freeze the river of water and turn the waterfall off to obtain the key to the next room.
Personal tools
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Low Activity
× You must be logged in to change this data. If you don't have an account, Please join.
Settings : Managers
Analyzed 5 days ago based on code collected 5 days ago.
Stephen Colebourne
1197 commits
I manage this project on Ohloh
I want to edit this project's settings and information on Ohloh. Do I need to sign up as a project manager?
If a project is locked, then project information can only be edited by managers. Otherwise, anyone with an Ohloh account can edit project information.
Who should register as a project manager?
Someone who works on the project. Ideally the owner, founder, lead developer, or release manager.
About Managers
Project managers can:
• Limit edits just to managers.
• Edit project information and settings if community editing has been limited.
• Approve or deny applications to be a manager.
• If a project's edits are limited, then project information can only be edited by managers. Otherwise, anyone with an Ohloh account can edit project information.
Learn More...
Copyright © 2013 Black Duck Software, Inc. and its contributors, Some Rights Reserved. Unless otherwise marked, this work is licensed under a Creative Commons Attribution 3.0 Unported License . Ohloh ® and the Ohloh logo are trademarks of Black Duck Software, Inc. in the United States and/or other jurisdictions. All other trademarks are the property of their respective holders.
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TOP10 chemically competent cells
From OpenWetWare
(Difference between revisions)
Jump to: navigation, search
(Preparing competent cells)
(Preparing seed stocks)
Line 9: Line 9:
===Preparing seed stocks===
===Preparing seed stocks===
-
* streak TOP10 cells on an [[SOB]] plate and grow for single colonies at 23 C
+
* Streak TOP10 cells on an [[SOB]] plate and grow for single colonies at 23°C
** room temperature works well
** room temperature works well
-
* Pick single colonies into 2 ml of SOB medium and shake overnight at 23 C
+
* Pick single colonies into 2 ml of SOB medium and shake overnight at 23°C
** room temperature works well
** room temperature works well
* Add glycerol to 15%
* Add glycerol to 15%
Line 17: Line 17:
* Place tubes into a zip lock bag, immerse bag into a dry ice/ethanol bath for 5 minutes
* Place tubes into a zip lock bag, immerse bag into a dry ice/ethanol bath for 5 minutes
** This step may not be necessary
** This step may not be necessary
-
* Place in -80 freezer indefinitely.
+
* Place in -80°C freezer indefinitely.
===Preparing competent cells===
===Preparing competent cells===
Revision as of 11:52, 4 June 2007
This protocol is a variant of the Hanahan protocol [1] using CCMB80 buffer for DH10B, TOP10 and MachI strains. It builds on Example 2 of the Bloom05 patent as well. This protocol has been tested on TOP10, MachI and BL21(DE3) cells. See Bacterial Transformation for a more general discussion of other techniques. The Jesse '464 patent describes using this buffer for DH5α cells. The Bloom04 patent describes the use of essentially the same protocol for the Invitrogen Mach 1 cells.
Contents
Preparing glassware and media
Detergent is a major inhibitor of competent cell growth and transformation. Glass and plastic must be detergent free for these protocols. The easiest way to do this is to avoid washing glassware, and simply rinse it out. Autoclaving glassware filled 3/4 with DI water is an effective way to remove most detergent residue. Media and buffers should be prepared in detergent free glassware and cultures grown up in detergent free glassware.
Preparing seed stocks
• Streak TOP10 cells on an SOB plate and grow for single colonies at 23°C
• room temperature works well
• Pick single colonies into 2 ml of SOB medium and shake overnight at 23°C
• room temperature works well
• Add glycerol to 15%
• Aliquot 1 ml samples to Nunc cryotubes
• Place tubes into a zip lock bag, immerse bag into a dry ice/ethanol bath for 5 minutes
• This step may not be necessary
• Place in -80°C freezer indefinitely.
Preparing competent cells
• Inoculate 250 ml of SOB medium with 1 ml vial of seed stock and grow at 20°C to an OD600nm of 0.3
• This takes approximately 16 hours.
• Controlling the temperature makes this a more reproducible process, but is not essential.
• Room temperature will work. You can adjust this temperature somewhat to fit your schedule
• Aim for lower, not higher OD if you can't hit this mark
• Centrifuge at 3000g at 4°C for 10 minutes in a flat bottom centrifuge bottle.
• Flat bottom centrifuge tubes make the fragile cells much easier to resuspend
• It is often easier to resuspend pellets by mixing before adding large amounts of buffer
• Gently resuspend in 80 ml of ice cold CCMB80 buffer
• sometimes this is less than completely gentle. It still works.
• Incubate on ice 20 minutes
• Centrifuge again at 4°C and resuspend in 10 ml of ice cold CCMB80 buffer.
• Test OD of a mixture of 200 μl SOC and 50 μl of the resuspended cells.
• Add chilled CCMB80 to yield a final OD of 1.0-1.5 in this test.
• Incubate on ice for 20 minutes
• Aliquot to chilled screw top 2 ml vials or 50 μl into chilled microtiter plates
• Store at -80°C indefinitely.
• Flash freezing does not appear to be necessary
• Test competence (see below)
• Good cells should yield around 100 - 400 colonies
• Thawing and refreezing partially used cell aliquots dramatically reduces transformation efficiency by about 3x the first time, and about 6x total after several freeze/thaw cycles.
CCMB80 buffer
• 10 mM KOAc pH 7.0 (10 ml of a 1M stock/l)
• 80 mM CaCl2.2H2O (11.8 g/l)
• 20 mM MnCl2.4H2O (4.0 g/l)
• 10 mM MgCl2.6H2O (2.0 g/l)
• 10% glycerol (100 ml/l)
• adjust pH DOWN to 6.4 with 0.1N HCl if necessary
• adjusting pH up will precipitate manganese dioxide from Mn containing solutions.
• sterile filter and store at 4C
• slight dark precipitate appears not to affect its function
Measurement of competence
• Transform 50 μl of cells with 1 μl of standard pUC19 plasmid (Invitrogen)
• This is at 10 pg/μl or 10-5 μg
• Hold on ice 0.5 hours
• Heat shock 60 sec at 42C
• Add 250 μl SOC
• Incubate at 37 C for 1 hour in 2 ml centrifuge tubes rotated
• using 2ml centrifuge tubes for transformation and regrowth works well because the small volumes flow well when rotated, increasing aeration.
• For our plasmids (pSB1AC3, pSB1AT3) which are chloramphenicol and tetracycline resistant, we find growing for 2 hours yields many more colonies
• Ampicillin and kanamycin appear to do fine with 1 hour growth
• Plate 20 μl on AMP plates using 3.5 mm glass beads
• Transformation efficiency is 15 x colony count x 105
5x Ligation Adjustment Buffer
• Intended to be mixed with ligation reactions to adjust buffer composition to be near the CCMB80 buffer
• KOAc 40 mM (40 ml/liter of 1 M KOAc solution, pH 7.0)
• CaCl2 400 mM (200 ml/l of a 2 M solution)
• MnCl2 100 mM (100 ml/l of a 1 M solution)
• Glycerol 46.8% (468 ml/liter)
• pH adjustment with 2.3% of a 10% acetic acid solution (12.8ml/liter)
• Previous protocol indicated amount of acetic acid added should be 23 ml/liter but that amount was found to be 2X too much per tests on 1.23.07 --Meaganl 15:50, 25 January 2007 (EST)
• water to 1 liter
• autoclave or sterile filter
• Test pH adjustment by mixing 4 parts ligation buffer + 1 part 5x ligation adjustment buffer and checking pH to be 6.3 - 6.5
References
1. Hanahan D, Jessee J, and Bloom FR. . pmid:1943786. PubMed HubMed [Hanahan91]
2. US Patent 6,709,852 Media:pat6709852.pdf [Bloom04]
3. US Patent 6,855,494 Media:pat6855494.pdf [Bloom05]
4. US Patent 6,960,464 Media:pat6960464.pdf [Jesse05]
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User:Mariana Ruiz Velasco L.
From OpenWetWare
Jump to: navigation, search
Contents
About me!
Mariana Ruiz Velasco Leyva. UNAM-Genomics Mexico.
• Hi! My name is Mariana Ruiz Velasco Leyva, I am 21 years old, and I am currently studying on the Undergraduate Program on Genomic Sciences at the "Universidad Nacional Autonóma de México" (UNAM) campus Morelos. I am a very girly girl who always likes to try new things, tries to keep laughing and smiling, and loves her family and friends very much.
• What I like? Well, I certainly have a big range of activities to get me entertained and busy at all times ;). Right now, my very first love is iGEM <3, but I also make myself time for school, dancing and hanging out with my friends, reading papers for pleasure, exercising, and listening to good music.
Education
UNAM-Center for Genomic Sciences, Mexico.
• My current interest, besides synthetic biology, are epigenetics, and biorremediation.
iGEM 2010
Wet lab journal
Useful links
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[9] “Well,” Xenophon continued, “after I had sent you on ahead, I overtook you again, as I came along with the rearguard, and found you digging a hole to bury the man in, and I stopped and commended you.
This work is licensed under a Creative Commons Attribution-ShareAlike 3.0 United States License.
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CMD sent two reporters to track ALEC in Oklahoma
Click here to help support our future investigations.
Pelican Institute
From SourceWatch
Jump to: navigation, search
Learn more about corporations VOTING to rewrite our laws.
Learn more about how the State Policy Network aids ALEC and spins disinformation in the states.
The Pelican Institute for Public Policy is a "nonpartisan research and educational organization" based in Louisiana.[1] The Institute's mission is to advance "policies based on free enterprise, individual liberty, and constitutionally limited government."[1]It is a member of the State Policy Network (SPN), a network of state think tanks.
Ties to ALEC
The Pelican Institute is a state think tank member of SPN, which is a private sector member of the American Legislative Exchange Council (ALEC). The Pelican Institute also has ties to ALEC through its annual Policy Orientation for the Louisiana Legislature of which ALEC is a sponsor.[2] ALEC members have also sat on policy panels at the event.[3]
Please see SPN Ties to ALEC for more.
About ALEC
ALEC is a corporate bill mill. It is not just a lobby or a front group; it is much more powerful than that. Through ALEC, corporations hand state legislators their wishlists to benefit their bottom line. Corporations fund almost all of ALEC's operations. They pay for a seat on ALEC task forces where corporate lobbyists and special interest reps vote with elected officials to approve “model” bills. Learn more at the Center for Media and Democracy's ALECexposed.org, and check out breaking news on our PRWatch.org site.
Ties to the Franklin Center for Government and Public Integrity
The Pelican Institute is listed as a Franklin Center for Government and Public Integrity "Statehouse News Bureau".[4] The Franklin Center funds reporters in over 40 states.[5] Despite their non-partisan description, many of the websites funded by the Franklin Center have received criticism for their conservative bias.[6][7] On its website, the Franklin Center claims it "provides 10 percent of all daily reporting from state capitals nationwide."[8]
Franklin Center Funding
Franklin Center Director of Communications Michael Moroney told the Center for Public Integrity (CPI) in 2013 that the source of the Franklin Center's funding "is 100 percent anonymous." But 95 percent of its 2011 funding came from DonorsTrust, a spin-off of the Philanthropy Roundtable that functions as a large "donor-advised fund," cloaking the identity of donors to right-wing causes across the country (CPI did a review of Franklin's Internal Revenue Service records).[9] Mother Jones called DonorsTrust "the dark-money ATM of the conservative movement" in a February 2013 article.[10] Franklin received DonorTrust's second-largest donation in 2011.[9]
The Franklin Center also receives funding from the Wisconsin-based Lynde and Harry Bradley Foundation,[11] a conservative grant-making organization.[12]
The Franklin Center was launched by the Chicago-based Sam Adams Alliance (SAM),[13] a 501(c)(3) devoted to pushing free-market ideals. SAM gets funding from the State Policy Network,[14] which is partially funded by the Claude R. Lambe Foundation.[15] Charles Koch, one of the billionaire brothers who co-own Koch Industries, sits on the board of this foundation.[16] SAM also receives funding from the Rodney Fund.
Personnel
Staff
• Kevin Kane, president
• Knud Berthelsen, marketing strategist
• Matt Cole, policy analyst
Core Financials
2010[17]:
• Total Revenue: $435,450
• Total Expenses: $371,682
• Net Assets: $90,200
2009[18]:
• Total Revenue: $377,978
• Total Expenses: $215,938
• Net Assets: $26,432
Contact Details
Pelican Institute for Public Policy
643 Magazine Street, Suite 301
New Orleans, LA 70130
Phone: (504) 267-9404
Email: info@pelicaninstitute.org
Resources
Related SourceWatch Articles
Related PRWatch Articles
External Resources
References
1. 1.0 1.1 Pelican Institute for Public Policy, "About", organizational website, accessed November 2012
2. Pelican Institute, "Sponsors", organizational website, accessed November 2012
3. Pelican Institute for Public Policy, "Pelican Institute For Public Policy Announces Inaugural Policy Orientation for the Louisiana Legislature", organizational website, accessed November 2012
4. Franklin Center for Government and Public Integrity, Franklin Affiliates in Your State, organizational website, accessed March 2013.
5. The Franklin Center for Government and Public Integrity, Think tank Journalism: The Future of Investigative Journalism, organizational website, accessed August 19, 2011.
6. Rebekah Metzler, 'Watchdog' website puts a new spin on politics, The Portland Press Herald, October 2, 2010.
7. Allison Kilkenny, The Koch Spider Web, Truthout, accessed August 19, 2011.
8. Sara Jerving, Franklin Center: Right-Wing Funds State News Source, PRWatch.org, October 27, 2011.
9. 9.0 9.1 Paul Abowd, Center for Public Integrity, Donors use charity to push free-market policies in states, organizational report, February 14, 2013.
10. Andy Kroll, Exposed: The Dark-Money ATM of the Conservative Movement, Mother Jones, February 5, 2013.
11. Daniel Bice, Franklin Center boss wants apology from Democratic staffer, The Milwaukee Journal Sentinel, August 8, 2011.
12. Bradley Foundation, The Bradley Foundation, organizational website, accessed August 19, 2011.
13. Sam Adams Alliance, Sam Adams Alliance Media Kit, organizational PDF, accessed August 19, 2011.
14. Media Matters Action Network, Sam Adams Alliance, Conservative Transparency website, accessed August 19, 2011.
15. Media Matters Action Network. State Policy Network, Conservative Transparency website, accessed August 19, 2011.
16. Media Matters Action Network, Claude R. Lambe Charitable Foundation, Conservative Transparency website, accessed August 19, 2011.
17. Pelican Institute, IRS form 990, 2010. GuideStar.
18. Pelican Institute, IRS form 990, 2009. GuideStar.
This article is a stub. You can help by expanding it.
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Place:Rendon, Tarrant, Texas, United States
Watchers
NameRendon
TypeCensus-designated place
Coordinates32.577°N 97.24°W
Located inTarrant, Texas, United States
source: Getty Thesaurus of Geographic Names
the text in this section is copied from an article in Wikipedia
Rendon is a census-designated place (CDP) in Tarrant County, Texas, United States. The population was 12,552 at the 2010 census.
Research Tips
This page uses content from the English Wikipedia. The original content was at Rendon, Texas. The list of authors can be seen in the page history. As with WeRelate, the content of Wikipedia is available under the Creative Commons Attribution/Share-Alike License.
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Tell me more ×
Answers OnStartups is a question and answer site for entrepreneurs looking to start or run a new business. It's 100% free, no registration required.
In a founder-funded company, how should the founders treat the cash that they contribute to the company? Should it be convertible debt? Equity (but the founders already have all the equity initially)?
If it makes a difference, I'm interested in the following scenarios:
1. Single founder, cash contributions to the company.
2. Multiple founders, equal cash contributions from each.
3. Multiple founders, different cash contributions from each.
4. Special concerns for non-cash contributions (e.g., computers, furniture, etc.)
Does any of this matter when it comes time to raise money or exit the company? Obviously, it's budget dust if you get a billion dollar valuation like Twitter, but what about in smaller startups?
share|improve this question
4 Answers
up vote 3 down vote accepted
In terms of accounting, any investment by the founders is a part of Owners Equity. If you're doing proper double-entry accounting you'll want to credit an Investments account and debit Cash, or Equipment, or some other asset (assets are increased by debit in accounting... it's confusing, but true).
You could take a look at some accounting basics here to understand this concept better: http://www.moneyinstructor.com/lesson/accountingtransaction.asp
If that doesn't make sense, then definitely consult an accountant. They can get you set up in an hour or two.
share|improve this answer
You should talk to an accountant. In my case, as a S-corp, money that goes into the company is considered funding that is to eventually be paid back with interest. No matter what, keep good records!
share|improve this answer
Structure it as a loan to the company, with interest.
share|improve this answer
What if the founder wants to bring in another person to work part-time and then how does it change for the owners equity. Assuming there is a measure of other person's contribution in terms of time and expertise. Would the founder then issue equity on a time+expertise spent basis?
share|improve this answer
Your Answer
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By posting your answer, you agree to the privacy policy and terms of service.
Not the answer you're looking for? Browse other questions tagged or ask your own question.
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Relationship between Working Capital Management and Profitability in Context of Manufacturing Industries in Bangladesh
Sayeda Tahmina Quayyum
Abstract
This study is an attempt to investigate if there is any relationship between working capital management and
profitability of manufacturing corporations. For this purpose Corporations enlisted with the Dhaka Stock
Exchange has been selected and the analysis covers a time period from year 2005 to 2009. The purpose of this
paper is to figure out if there is statistically significant relationship between the profitability and working capital
management and help explain the necessity of firms optimizing their level of working capital management
efficiency and in that way management taking productive actions to maximize their profitability. The result of
this study clearly shows that except for food industry all other selected industries have a significant level of
relationship between the Profitability Indices and various Working Capital Components. This paper also shows
that the significance level of relationship varies from industry to industry.
Full Text: PDF DOI: 10.5539/ijbm.v7n1p58
This work is licensed under a Creative Commons Attribution 3.0 License.
International Journal of Business and Management ISSN 1833-3850 (Print) ISSN 1833-8119 (Online)
Copyright © Canadian Center of Science and Education
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Effects of Integrated Plant Nutrient Management (IPNM) Practices on the Sustainability of Maize-based Farming Systems in Nepal
Tejendra Chapagain, Gam B. Gurung
Abstract
Maize is a staple summer crop grown in the hilly areas of Nepal, where the soil is fragile and fertility is declining over years due primarily to degradation of natural resource base, high rates of soil erosion, increased cropping intensity and inadequate replenishment of soil nutrients. Forum for Rural Welfare and Agricultural Reform for Development (FORWARD) with the financial support from Hill Maize Research Program (HMRP/CYMMIT) conducted eight Integrated Plant Nutrient Systems (IPNS) trials, 16 Farm Yard Manure (FYM) improvement demonstrations and 16 conservation farming demonstrations each year through two women farmer groups since 2003 in Makawanpur District in order to raise the awareness of farmers on sustainable soil management practices through better utilization of locally available and external resources. The three years' trial results revealed that the maize crop with IPNS (15 t ha-1 FYM + 60:30:30 NPK kg ha-1) was better with respect to crop vigor and grain yields compared to the control treatment (farmers' practice with FYM and urea top dressing). The Improved cultivar with IPNS practices increased the grain yield by 64% (p<0.01) compared to the Local cultivar under farmers' managed condition. The plots with IPNS also exhibited positive effects on the performance of millet, a subsequent crop in the local cropping systems. Farmers were well impressed that FYM along with a balanced dose of Nitrogen (N), Phosphorus (P2O5) and Potash (K2O) is necessary for increased crop yields. There was a good impression of farmers with improved method of FYM preparation in pits over their traditional method of FYM preparation on exposed heaps. Hedgerow system with napier, sunhemp and pigeon pea on terrace edges and riser, was introduced as a new technology to farmers. They have preferred this technology for soil conservation and forage for livestock feeding. These soil fertility management practices could also be extrapolated to the similar recommendation domain so as to promote sustainable soil management practices, and enhanced food security in the region.
Full Text: PDF
This work is licensed under a Creative Commons Attribution 3.0 License.
Journal of Agricultural Science ISSN 1916-9752 (Print) ISSN 1916-9760 (Online)
Copyright © Canadian Center of Science and Education
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Friday, September 30, 2005
Endangered Species Act Gutted
The House of Representatives has passed a bill that weaken the Endangered Species Act substantially. Among other things, it would require payments by the federal government when endangered species stop intended development. It also would place political appointees instead of scientists in charge of determining when a species is endangered. I expect that both of these measures will basically stop further species from being listed as endangered or threatened. Given what happened with the sage grouse earlier this year, I think we know what to expect from political appointees. The bill now goes to the Senate where the committee overseeing the legislation is chaired by Lincoln Chafee (R-RI). Chafee has a reputation as a liberal, but when push comes to shove he usually gets shoved, so I am not too hopeful about what the results will be.
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Wikia
SRD:Destruction Domain
Talk0
9,503pages on
this wiki
Revision as of 06:32, August 11, 2009 by Surgo (Talk | contribs)
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)
This material is published under the OGL
DESTRUCTION DOMAINEdit
Granted Power: You gain the smite power, the supernatural ability to make a single melee attack with a +4 bonus on attack rolls and a bonus on damage rolls equal to your cleric level (if you hit). You must declare the smite before making the attack. This ability is usable once per day.
Destruction Domain Spells
1. Inflict Light Wounds: Touch attack, 1d8 damage +1/level (max +5).
2. Shatter: Sonic vibration damages objects or crystalline creatures.
3. Contagion: Infects subject with chosen disease.
4. Inflict Critical Wounds: Touch attack, 4d8 damage +1/level (max +20).
5. Mass Inflict Light Wounds: Deals 1d8 damage +1/level to any creatures.
6. Harm: Deals 10 points/level damage to target.
7. Disintegrate: Makes one creature or object vanish.
8. Earthquake: Intense tremor shakes 80-ft.-radius.
9. Implosion: Kills one creature/round.
Back to Main PageSystem Reference DocumentSpellsCleric Domains
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Skip to main content Help Control Panel
Lost? Search this Naples Florida website...|Add our search|Login A+ A- 54.226.5.29
Business Directory « Collier County Business Directory «
NORTH COLLIER JUNIOR WOMANS CLUB
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Give your opinion about this listing below:
4680 1ST AVENUE SW, NAPLES, FL 34119
2012-01-21 23:11:03
I am a page for NORTH COLLIER JUNIOR WOMANS CLUB, which is a company located at: 4680 1ST AVENUE SW in the town of NAPLES with the zip code of 34119
I was registered by SVENTEK JAYNE C who is at in NAPLES FL, zipcode 34119
Phone:
Website:
Email:
. Map of NORTH COLLIER JUNIOR WOMANS CLUB - 4680 1ST AVENUE SW, NAPLES, FL 34119
Sventek Jayne C will be the Editor of the 'north-collier-junior-womans-club' page.
hCard:
NORTH COLLIER JUNIOR WOMANS CLUB
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4680 1ST AVENUE SW
NAPLES FL 34119
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Research
Identification of CDK2 substrates in human cell lysates
Yong Chi1,2, Markus Welcker1, Asli A Hizli1, Jeffrey J Posakony3, Ruedi Aebersold2,4 and Bruce E Clurman1*
Author Affiliations
1 Divisions of Clinical Research and Human Biology, Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue N., Seattle, WA 98109, USA
2 Institute for Systems Biology, 1441 N. 34th Street, Seattle, WA 98103, USA
3 Division of Basic Sciences, Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue N., Seattle, WA 98109, USA
4 Institute of Molecular Systems Biology, ETH Zurich and Faculty of Science, University of Zurich, 8093 Zurich, Switzerland
For all author emails, please log on.
Genome Biology 2008, 9:R149 doi:10.1186/gb-2008-9-10-r149
The electronic version of this article is the complete one and can be found online at: http://genomebiology.com/2008/9/10/R149
Received:8 August 2008
Revisions received:29 September 2008
Accepted:13 October 2008
Published:13 October 2008
© 2008 Chi et al.; licensee BioMed Central Ltd.
This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Background
Protein phosphorylation regulates a multitude of biological processes. However, the large number of protein kinases and their substrates generates an enormously complex phosphoproteome. The cyclin-dependent kinases - the CDKs - comprise a class of enzymes that regulate cell cycle progression and play important roles in tumorigenesis. However, despite intense study, only a limited number of mammalian CDK substrates are known. A comprehensive understanding of CDK function requires the identification of their substrate network.
Results
We describe a simple and efficient approach to identify potential cyclin A-CDK2 targets in complex cell lysates. Using a kinase engineering strategy combined with chemical enrichment and mass spectrometry, we identified 180 potential cyclin A-CDK2 substrates and more than 200 phosphorylation sites. About 10% of these candidates function within pathways related to cell division, and the vast majority are involved in other fundamental cellular processes. We have validated several candidates as direct cyclin A-CDK2 substrates that are phosphorylated on the same sites that we identified by mass spectrometry, and we also found that one novel substrate, the ribosomal protein RL12, exhibits site-specific CDK2-dependent phosphorylation in vivo.
Conclusions
We used methods entailing engineered kinases and thiophosphate enrichment to identify a large number of candidate CDK2 substrates in cell lysates. These results are consistent with other recent proteomic studies, and suggest that CDKs regulate cell division via large networks of cellular substrates. These methods are general and can be easily adapted to identify direct substrates of many other protein kinases.
Background
Reversible protein phosphorylation is one of the most common posttranslational modifications and regulates virtually all cellular processes. Protein kinases are among the largest known gene families with more than 500 human kinase genes that comprise nearly 2% of the open reading frames of the human genome [1]. Moreover, approximately 30% of all cellular proteins are phosphorylated [2]. The large number of kinases and their substrates make it very difficult to determine which proteins are phosphorylated by specific kinases in vivo, but this information is critical to understanding kinase functions and the control of biological processes in general. Various strategies have been developed to identify protein kinase substrates, and several have resulted from recent technological advances in substrate detection.
Some approaches have utilized antibodies against phosphomotifs within the substrate proteins as affinity reagents to enrich for phosphorylated peptides. Examples include using antibodies that recognize conserved motifs that are highly specific to a particular kinase [3], as well as the use of phosphomotif antibodies combined with changes in physiological conditions that stimulate kinase function [4]. However, it is difficult to apply these approaches to kinases that phosphorylate broader substrate motifs, since there is less epitope conservation among substrates. Another recent method combined quantitative phosphoproteomics with kinase knock-outs and cellular perturbations to identify kinase targets in yeast [5]. However, with these cell-based approaches, it is often difficult to determine if the putative substrates are direct kinase targets. An in vitro approach employing arrays of proteins phosphorylated by isolated recombinant kinases has been successfully used in a global analysis of yeast kinase substrates, but this strategy may be difficult to apply to organisms with larger proteomes [6].
A 'chemical genetic' approach developed by Kevan Shokat's laboratory addresses many of these potential problems [7]. In this technique, the kinase to be studied is mutated by replacing a conserved bulky residue within the ATP-binding pocket with a smaller residue. This creates an enlarged ATP binding pocket that enables the mutant kinase to utilize bulky ATP analogues that cannot be used by wild-type cellular kinases, thereby isolating the activity of the mutant kinase from all other cellular kinases [8]. This technique is broadly applicable to most protein kinases and has led to important advances in substrate identification, including the description of a large number of potential cyclin-dependent kinase (CDK) substrates in yeast [9-15]. However, several technical hurdles add substantial challenges when applying this approach to mammalian kinases with broad substrate networks.
In this study, we report a method employing the Shokat strategy to identify direct cyclin A-CDK2 substrates in human cell lysates. CDK2 is activated by both the cyclin E and cyclin A subunits, and cyclin A-CDK2 plays critical roles in cell cycle control, primarily in G1 and S-phases [16,17]. We used an engineered cyclin A-CDK2 and ATP-γ-S analogue to label proteins with thiophosphates in cell lysates, and after digestion of the protein mixtures, we employed a single-step chemical enrichment procedure to selectively isolate thiophosphorylated peptides. As these studies were nearing completion, Blethrow et al. [18] independently reported a similar approach employing engineered CDK1 and thiophosphate enrichment methods that they used to identify a group of 68 putative cyclin B-CDK1 substrates within Hela cell lysates.
We identified 180 proteins and over 220 phosphopeptides that were phosphorylated in cell lysates by cyclin A-CDK2, and these proteins represented diverse cellular pathways. To validate these methods, we selected several candidate substrates and confirmed that they were phosphorylated by cyclin A-CDK2 in vitro on the same sites that we identified in the screen. Finally, we selected one novel substrate, the ribosomal protein RL12, for further study: site-directed mutagenesis and phosphopeptide mapping confirmed that CDK2 phosphorylates RL12 in vitro and in vivo on the same site determined by our methods.
Results and discussion
Utilization of ATP analogues by engineered CDKs
We generated mutant 'Shokat' CDKs containing amino acid exchanges at a conserved bulky residue in their ATP binding pockets. In the case of CDK2 and CDK3 this was a phenylalanine to alanine exchange at position 80, designated CDK2 (F80A). We also synthesized 12 ATP analogues to determine if the engineered CDKs can use these analogues to phosphorylate recombinant Retinoblastoma (Rb) protein in vitro, and found that they utilized N6-(2-phenylethyl)-ATP (PE-ATP) most efficiently (Figure 1a, and data not shown). Although both wild-type and cyclin E-CDK2 (F80A) used normal ATP to phosphorylate glutathione-S-transferase (GST)-Rb protein, only the F80A kinase could use PE-ATP. Similar results were obtained with cyclin E-CDK3, but in this case the F80A mutant could no longer use normal ATP, although the structural basis for this observation is unclear (Figure 1a). These studies confirmed that the wild-type and engineered kinases exhibit the desired ATP specificities.
Figure 1. Characterization of the engineered CDKs. (a) Cyclin E-CDK2/3 complexes, or their F80A engineered counterparts (indicated by asterisks), were immunoprecipitated from transfected U2OS cell lysates via the HA-tag on the CDK subunit and subjected to in vitro kinases assays with 10 μM of either normal ATP or PE-ATP analogue. Phosphorylation of GST-Rb was monitored by immunoblotting with a phosphospecific anti-pS780-Rb antibody (New England Biolabs). The wild-type kinases cannot use PE-ATP. (b) Thin-layer chromatography - analysis reveals hydrolysis of ATP in cell lysate and transfer to acceptor nucleotides (left). The positions of the free phosphate and ATP are indicated. In contrast, ATP-γ-S is not hydrolyzed in cell lysates (right). (c) Kinase assays were performed using wild-type and cyclin A-CDK2 (F80A) complexes purified from E. coli and GST-Rb in the presence of 200 μM of ATP, ATP-γ-S and PE-ATP-γ-S at room temperature for 2 h. Kinase reactions were analyzed by SDS gel electrophoresis and visualized by Coomassie staining. Extent of the GST-Rb phosphorylation was monitored by the electromobility shift of GST-Rb.
Whereas the mutant CDKs efficiently used PE-ATP to phosphorylate Rb in vitro, similar experiments utilizing radiolabeled PE-ATP in cell lysates failed because the labeled phosphate was cleaved from PE-ATP by an ATPase activity in the lysates (data not shown). We therefore switched to the thiophosphate form of the ATP analogue (PE-ATP-γ-S), which was not hydrolyzed by lysates (Figure 1b). Although kinases often use ATP-γ-S less efficiently than normal ATP, thiophosphorylation has several advantages in this context. First, thiophosphates are more stable and resistant to phosphatases [19]. Second, since there are no pre-existing thiophosphorylation events in the cells, thiophosphate labeling provides unique markers for proteins phosphorylated by the mutant kinase. Finally, the thiophosphate group has similar chemical properties as the sulfhydryl group and is amenable to chemical modifications. We expressed and purified soluble wild-type and cyclin A-CDK2 (F80A) complexes from bacteria and carried out a similar Rb kinase assay to test their ability to use PE-ATP-γ-S. As shown in Figure 1c, although both kinases can use ATP or ATP-γ-S to phosphorylate GST-Rb protein (as indicated by its electromobility shift) only the F80A mutant can use PE-ATP-γ-S. We thus used PE-ATP-γ-S for all of our subsequent studies.
Single-step purification of thiophosphorylated peptides
The use of engineered CDKs and ATP analogues facilitates highly specific substrate phosphorylation: the next challenge is how to identify them within a complex lysate. We sought to utilize the thiophosphate tags to covalently capture and enrich thiophosphorylated peptides after phosphorylation and digestion of the lysates. However, a key problem was to chemically distinguish thiophosphopeptides and cysteine-containing peptides. Although a chemoselective method for enriching thiophosphopeptides has been described, the overwhelming abundance of cysteine residues in a complex protein mixture makes this approach difficult [20]. We used a simple capture-and-release method to selectively isolate thiophosphorylated peptides within trypsinized cell lysates (Figure 2a). Proteins within the lysate were phosphorylated in vitro with cyclin A-CDK2 (F80A) and PE-ATP-γ-S. The protein mixture was subsequently digested and the resulting peptides were mixed with Thiopropyl Sepharose 6B [21], an activated disulfide resin that captures both cysteine-containing peptides and thiophosphopeptides through a disulfide exchange reaction. Because traditional dithiothreitol (DTT) elution releases both types of bound peptides, we utilized the qualitative differences between resin-bound thiophosphate peptides and cysteine-containing peptides at the phosphorothiolatesulfide linkage and alkyldisulfide linkage, respectively. At high pH values, the phosphorothiolatesulfide linkage is hydrolyzed and the alkyldisulfide remains intact [22]. Treatment of the resin with a strong base (such as sodium hydroxide) specifically releases thiophosphopeptides (and also converts them to normal phosphopeptides), but not the cysteine-containing peptides, by hydrolyzing the phosphorothiolatesulfide linkage (Figure 2b). Because peptides bound via cysteine are not eluted in the final step, we cannot recover cysteine-containing thiophosphopeptides. Moreover, the elution results in loss of the thiophosphate signature by converting the thiophosphopeptide to a phosphopeptide. Our thiophosphopeptide isolation method differs modestly from that of Blethrow et al. [18] in that we capture the thiophosphopeptides using disulfide exchange chemistry (disulfide resin) instead of alkylation (iodoacetamide resin), and we selectively elute them with base hydrolysis rather than oxidation.
Figure 2. Single-step purification of thiophosphopeptides. (a) General scheme for thiophosphospeptide isolation. Proteins were labeled with cyclin A-CDK2 (F80A) and PE-ATP-γ-S and subjected to tryptic digest. The resulting peptides were mixed with disulfide beads, which capture both thiophosphopeptides and cysteine-containing peptides. The beads were then treated with basic solution to selectively release only the phosphopeptides. (b) The chemistry underlying the thiophosphopeptide selectivity. Both thiophosphate and cysteine moieties contain reactive thiol groups that can be covalently captured by disulfide beads. At high pH values, the phosphorothiolatesulfide linkages (near the upper arrow) are hydrolyzed to allow the release of the bead-bound peptides while the alkyldisulfide linkages (near the lower arrow) are stable and thus peptides are retained on the beads. Note that during the hydrolysis of the phosphorothiolatesulfide linkage, thiophosphate is converted to normal phosphate.
To test the feasibility of this approach, we phosphorylated GST-Rb with cyclin A-CDK2 (F80A) and PE-ATP-γ-S, and applied our purification procedure to a trypsin digest of the reaction mixture. The isolated peptides were analyzed by electrospray tandem mass spectrometry (ESI-MS/MS) using an ion trap mass spectrometer. Peptides were identified by matching the tandem mass spectra to a human protein sequence database (with GST-Rb sequence added) using SEQUEST software [23]. The GST-Rb substrate contains five cysteines as well as seven SP/TP sites, which are sites favored for CDK phosphorylation [24]. We recovered multiple phosphopeptides containing only and all the expected phosphorylation sites (Table 1). Furthermore, we recovered no cysteine-containing peptides and very few non-specific peptides. This provided a proof-of-principle for our large-scale assays.
Table 1. Phosphopeptides identified from in vitro phosphorylated GST-Rb
Identification of human cyclin A-CDK2 substrates in cell lysates
Our goal was to identify potential cyclin A-CDK2 substrates on a proteome-wide scale. To reduce the sample complexity, we fractionated the whole cell lysate of HEK293 cells into 11 fractions using ion-exchange chromatography and ammonium sulfate precipitation (Additional data file 1). We then carried out in vitro kinase assays on each fraction. As a positive control, we also added a small amount of GST-Rb to each reaction. After digesting the reaction mixture with trypsin, we applied our purification protocol to isolate the thiophosphopeptides from the peptide mixtures. The recovered peptides were subjected to liquid chromatography-MS/MS analysis and database searching. We recovered varying numbers of peptides and at least one Rb phosphopeptide from each of the lysate fractions (Additional data file 2).
CDKs phosphorylate proteins in a proline-directed manner on either serine or threonine, and numerous studies support the idea that the motif S/T-P-X-R/K represents the CDK consensus motif. From the phosphopeptides we identified a total of 203 proteins: 180 candidates were phosphorylated within SP or TP motifs (proline-directed; Additional data file 3). These candidate substrates represent a wide range of biological processes, including cell cycle control, DNA and RNA metabolism, translation and cellular structures (Figure 3). A total of 96 out of 222 (43%) of the proline-directed sites conformed to the known CDK consensus (with a positively charged residue in the +3 position). Interestingly, about 24% of the proline-directed sites (53/222) contained a positively charged residue in either the +4 or +5 positions, suggesting that this motif may also be favored by CDK2. Indeed, Blethrow et al. [18] also noted that a substantial number of cyclin B-CDK1 substrates contain non-consensus sites. For the peptides that contained non-consensus sites, we found that about 50% of the corresponding proteins carried at least one K/RXLφ or K/RXLXφ motif (where φ is a large hydrophobic residue and X is any amino acid) distal to the phosphorylation sites, and almost all of them carried at least one minimal RXL motif (Additional data file 3). This is consistent with the well-established idea that these motifs promote cyclin A-CDK2 binding to substrates [16]. In addition to selecting for phosphorylations with CDK consensus motifs, we identified 28 proteins that have been previously implicated as CDK substrates (marked in bold in Additional data file 3) [25-52]. Thus, nearly 15% of our candidates have been previously found as CDK targets, further supporting the idea that our methods captured and enriched for CDK2 substrates. Finally, 43% of the phosphorylations we found have been previously identified in large-scale, in vivo phosphoproteome analyses, indicating that these phosphorylations are not limited to our in lysate conditions [53-58].
Figure 3. Classification of proteins by functional category. Numbers indicate identified proteins in each category.
Both our studies and those reported by Blethrow et al. [18] used similar phosphopeptide isolation schemes and related cyclin-CDKs, and we anticipated that there might be substantial overlap in the substrates revealed by both studies. Indeed, we found nearly 50% (30/68) of the cyclin B-CDK1 substrates in our list of cyclin A-CDK2 candidates; thus, these methods are robust and reproducible (Additional data file 4). However, there are also substantial differences between the two lists, and these likely resulted from many factors, including procedural differences, different cell types, incomplete peptide identification by MS, and substrate specificity conferred by the cyclin and/or kinase subunits. Some of these differences may also reflect the different biological functions of cyclin A-CDK2 and cyclin B-CDK1. For example, we found nine proteins involved in protein translation and/or ribosome function, but none of these proteins were found with cyclin B-CDK1, despite their relatively high abundance.
Although we identified a number of known CDK2 substrates, we did not identify some previously described CDK2 substrates. Some of the factors listed above may also account for the failure to find known CDK2 substrates in our analyses. In addition, substrates already phosphorylated by endogenous CDKs would not have been thiophosphorylated in vitro. It is also possible that some proteins were not solubilized during lysate preparation and/or the sonication step and excluded from our analyses. Finally, it is possible that large protein complexes may have been disrupted by the fractionation procedures prior to the kinase reaction, and that proteins that are phosphorylated by CDK2 only in the context of these complexes may not be discovered by our methods.
We also recovered four phosphopeptides corresponding to cyclin A and CDK2 and 27 additional phosphopeptides with non-proline directed sites (Additional data file 5). We suspected these phosphopeptides resulted from auto-phosphorylation of cyclin A-CDK2 and background phosphorylation by other kinases, and others have reported similar background phosphorylation [15]. To test these possibilities, we carried out a control kinase reaction using cyclin A-CDK2 (F80A) and PE-ATP-γ-S with no cell lysate added and recovered three of the four cyclin A-CDK2 peptides (Additional data file 5). Furthermore, when we performed a similar 'kinase-only' reaction in the presence of γ-32P-ATP, we observed 32P incorporation into both of these proteins in a dose-dependent manner (Additional data file 6). These experiments confirmed that there was background auto-phosphorylation of cyclin A-CDK2 in the original assays.
We also performed control kinase reactions using the lysate fractions, GST-Rb 'spike-in', and PE-ATP-γ-S without the addition of cyclin A-CDK2. These 'no-kinase' control reactions phosphorylated 7 of the 27 non-proline directed phosphopeptides on our list (Additional data file 5), suggesting that most, if not all, of these phosphopeptides resulted from background phosphorylation by kinases that are able to use the ATP analogue to a limited extent. For example, most of these peptides contain acidic residue-directed phosphorylation sites that are casein kinase 2 motifs. Casein kinase 2 is unique in that it can utilize GTP as well as ATP; thus, the active site may accommodate the bulky ATP analogues, such as PE-ATP [59]. Importantly, we did not recover any Rb phosphopeptides from these control experiments, indicating that there was no non-specific CDK activity in our assays. We also recovered 44 unmodified peptides, 12 of which contained cysteine residues. The majority of these peptides originated from several lysate fractions (Additional data file 2). We suspect these resulted from low-level non-specific binding of peptides to the resin despite stringent wash conditions, and in the case of the cysteine-containing peptides, from a small amount of hydrolysis of the alkyldisulfide linkage during the elution step. In summary, our methods were highly selective, and our studies identified a surprisingly large group of candidate cyclin A-CDK2 substrates, most of which have not been previously identified as CDK targets.
Validation of candidate substrates as cyclin A-CDK2 targets
We employed several strategies to validate some of the novel candidates in our list as cyclin A-CDK2 substrates. Because our protein identifications were based on peptide sequences, we began by confirming that cyclin A-CDK2 phosphorylated three full-length and native candidates that were immunoprecipitated via epitope tags from transfected 293 cells (EF2, TRF2, and RAP1). Because these proteins were not present on the cyclin B-CDK1 list [18], we determined if they are also phosphorylated by cyclin B-CDK1. Each CDK2 candidate was phosphorylated by both cyclin A-CDK2 and cyclin B-CDK1, although EF2 was phosphorylated to a lesser extent than either TRF2 or RAP1 (Figure 4a). We also expressed TRF2, RAP1, and the ribosomal protein RL12 as GST-fusions and purified them from Escherichia coli. When we used cyclin A-CDK2 to phosphorylate TRF2 and RAP1 in vitro, the proteins were highly phosphorylated, and MS analyses revealed that these phosphorylations occurred on the same sites we initially identified (Figure 4b). We also used cyclin A-CDK2 and cyclin B-CDK1 to phosphorylate GST-RL12, and found that both CDKs also phosphorylated RL12 in vitro (Figure 4c). These studies thus confirm that the peptides identified in our screen represent proteins that can be phosphorylated by cyclin A-CDK2, at least in vitro. Although we did find qualitative differences in the ability of cyclin A-CDK2 and cyclin B-CDK1 to phosphorylate specific proteins, in each case the candidates were phosphorylated by both CDKs. Because the enzyme preparation we used in these studies contained an excess of free CDK2 (F80A), we considered the possibility that some substrate phosphorylations might result from the association of endogenous cyclins with CDK2 (F80A) that was either monomeric, or that may have dissociated from cyclin A during the assay conditions. We found that the amount of cyclin B-CDK2 (F80A) activity in these extracts was negligible compared with cyclin A-CDK2 (F80A), and we could not detect any cyclin E-CDK2 (F80A) activity (Additional data file 7). Nonetheless, we cannot exclude the possibility that some peptides may have been phosphorylated by CDK2 (F80A) in complex with an endogenous cyclin, and the specificity of any candidate substrate for cyclin A versus other cyclins that activate CDK2 needs to be validated as described below.
Figure 4. In vitro validation of selective candidate CDK2 substrates. (a) HEK293 cells were transiently transfected with vectors expressing FLAG-tagged EF2, TRF2, and RAP1. Anti-FLAG antibody immunoprecipitates were in vitro phosphorylated with cyclin A-CDK2 or cyclin B-CDK1 in the presence of γ-32P-ATP (upper left panels). In parallel reactions, histone H1 was phosphorylated as a control to normalize the activities of cyclin A-CDK2 and cyclin B-CDK1 (right panel). 'C' denotes 'kinase only' reactions without transfected substrates (left panel) and 'no kinase' reaction (right panel). Protein samples were separated by SDS PAGE and the gels were transferred onto PVDF membranes. Phospho-signals were visualized by autoradiography. The membrane was subsequently probed with anti-FLAG antibody (Sigma-Aldrich) to confirm the identity of the phospho-signal bearing band (lower left panels). The asterisk represents a non-specific band from the commercial cyclin B-CDK2 preparation. (b) Kinase reaction was carried out using γ-32P-ATP, and GST-TRF2, GST-RAP1, GST-Rb (positive control) and GST (negative control) as substrates in the presence or absence of wild-type cyclin A-CDK2 kinase. Reactions were visualized by SDS PAGE followed by Coomassie staining and autoradiography (left panel). A similar kinase assay was carried out using wild-type cyclin A/CDK2 and ATP-γ-S, and subsequently subjected to a phosphopeptide isolation scheme. MS analysis confirmed that TRF2 and RAP1 were each phosphorylated on the exact sites we identified from the screen with one additional site for RAP1 (right panel). (c) Kinase assay was carried out using γ-32P-ATP, cyclin A-CDK2 or cyclin B-CDK1 with increasing amounts of purified GST-RL12. Samples were separated by SDS PAGE and the gel was stained with Coomassie (lower panel) followed by autoradiography (upper panel). 'C' denotes 'kinase only' reactions without transfected substrates.
Validation of RL12 as an in vivo CDK2 substrate
The above studies validated several novel candidates identified in our screen as CDK2 substrates in vitro. However, to determine if a novel substrate is also phosphorylated by CDK2 in vivo, we performed a more comprehensive analysis of the ribosomal protein RL12. We first mixed immunoprecipitates of epitope-tagged cyclin A-CDK2 and RL12 expressed in human cells in the presence of γ-32P-ATP and found that cyclin A-CDK2 phosphorylated RL12 in vitro (Figure 5a). The phosphorylation was largely abolished in a mutant RL12 where the identified phosphoserine S38 was replaced with an alanine, and it was restored when S38 was replaced with a threonine (Figure 5b). We then used phosphopeptide mapping to identify the peptide containing S38, and confirmed that it was directly phosphorylated by CDK2 in vitro (Figure 5c). To test if RL12 is also phosphorylated in vivo in a CDK2-dependent manner at the same site, we metabolically labeled cells with 32P-orthophosphate, and immunoprecipitated wild-type or RL12-S38A from cells overexpressing either cyclin E-CDK2, catalytically inactive cyclin E-CDK2, or the CDK inhibitor p21 (to inhibit endogenous CDKs) [60]. Because we cannot study endogenous RL12 phosphorylation due to the lack of a suitable anti-RL12 antibody, these studies examined the phosphorylation of ectopic RL12 in vivo (these transfection conditions led to an approximately five- to ten-fold overexpression of RL12 mRNA (Additional data file 8). We found that wild-type RL12, but not RL12-S38A, was phosphorylated in vivo (Figure 5d). This phosphorylation was enhanced in cells overexpressing cyclin E-CDK2, but not inactive cyclin E-CDK2, and was diminished in cells overexpressing the p21 CDK inhibitor (which inhibits endogenous cyclin-CDKs; Figure 5d). Finally, we used phosphopeptide mapping and phosphoamino acid analyses of RL12 protein immunoprecipitated from the labeled cells and confirmed that RL12 phosphorylation by cyclin E-CDK2 in vivo also occurred on S38 (Figure 5e). RL12 S38 phosphorylation in vivo has also been reported in phosphoproteome studies [54,55].
Figure 5. Phosphorylation of RL12 in vitro and in vivo. (a) HA-tagged RL12 or vector control ('vec') were transiently transfected into U2OS cells and immunoprecipitated using 12CA5 antibody. Cyclin A-CDK2 complexes were also transiently expressed separately in U2OS cells and immunoprecipitated using an antibody against cyclin A. Kinase assays were carried out using the RL12 (or control) immunoprecipitate with or without the cyclin A-CDK2 immunoprecipitate in the presence of γ-32P-ATP. (b) Similar assays were conducted using cyclin A-CDK2 and RL12 immunoprecipitates containing wild-type (wt) and the indicated RL12 phosphosite mutants. The asterisk denotes the light chain of the antibody. (c) Phosphopeptide mapping and phosphoamino acid analysis of the radiolabeled wild-type (left panel) and S38T mutant (right panel) of RL12. (d) Wild-type RL12, RL12-S38A, or vector control was transiently co-transfected with cyclin E-CDK2, catalytically inactive (dn) cyclin E-CDK2, or p21. All cells were subjected to 32P orthophosphate labeling. RL12 was immunoprecipitated from cell lysates and visualized by SDS PAGE followed by autoradiography. (e) Phosphopeptide mapping analysis was also carried out on the radiolabeled RL12 shown in (d). The bottom arrow shows a second and minor phosphorylation site detected in vivo.
Although we have validated each of the novel candidates that we have tested thus far by showing that the full-length proteins are phosphorylated by CDK2, some candidates on our list will likely prove not to be physiologically relevant cyclin A-CDK2 substrates. For example, the kinase reactions were performed in lysates, and in vivo subcellular compartmentalization may restrict the access of CDK2 to some candidates. Moreover, it is possible that other cellular kinases are either redundant with, or more important than, CDK2 with respect to individual substrates in vivo. It is thus critical that candidates be rigorously evaluated in as physiological a context as possible. Towards this end, in ongoing studies we are using a gene targeting approach to mutate a subset of these phosphorylation sites in the endogenous genes to study their physiological significance.
Conclusions
In summary, we describe a rapid and efficient method to identify candidate CDK substrates in cell lysates. We identified 180 candidate cyclin A-CDK2 substrates and found that our method is robust, sensitive, and capable of identifying novel CDK2 targets. Since most protein kinases have conserved ATP binding domains and the kinetics of thiophosphorylation can be optimized [61], these methods should be broadly applicable to the study of many kinases and their substrate networks. Moreover, thiophosphorylation-based phosphopeptide isolation should also facilitate the mapping of phosphorylation sites within individual proteins or protein complexes in vitro.
Materials and methods
Reagents, cell culture, recombinant protein expression and purification
All standard chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA). Triphosphate synthesis was carried out according the method of Ludwig [62]. PE-ATP was synthesized similar to as previously described [63] with N6-(2-phenylethyl)-adenosine as a precursor. PE-ATP-γ-S was custom synthesized by TriLink Biotechnologies (San Diego, CA, USA). All CDK (CDK2 (F80A) and CDK3 (F80A)) and RL12 phosphosite mutants were generated by site-directed mutagenesis with the Quick Change method (Stratagene, La Jolla, CA, USA). All cDNAs used in this study except TRF2 and RAP1 were generated from human mRNA via RT-PCR, and clones were sequenced. RAP1 cDNA was purchased from Open Biosystems (Huntsville, AL, USA). A TRF2 clone was obtained from Addgene (Addgene plasmid 16066, Cambridge, MA, USA). U2OS and HEK293 cell lines were maintained in Dulbecco's modified Eagle's medium with 10% fetal calf serum and expression of wild-type and mutant CDKs and other proteins in U2OS cells were performed by transient transfections or co-transfections via calcium phosphate precipitation using standard procedures. CDKs expressed in human cells were purified by immunoprecipitation using 12CA5 antibody for HA-tagged Cdk subunits or using antibodies against the cyclin subunit. GST-Rb (GST fused to the carboxy-terminal 156 amino acids of Rb), GST-RL12, GST-TRF2, and GST-RAP1 were expressed in bacteria and purified using standard glutathione resin. Purified cyclin A-CDK2 and cyclin B-CDK1 kinases used in Figure 4 were purchased from New England Biolabs (Beverly, MA, USA).
Preparation of cyclin A-CDK2 complexes expressed in bacteria
Active and soluble wild-type and F80A cyclin A-CDK2 complexes were produced by co-expressing Saccharomyces cerevisiae GST-CAK1, full-length untagged cyclin A and His6-tagged CDK2 in bacteria similar to the commercial version (New England Biolabs). Cyclin A and His6-CDK2 were cloned into the pRSFDuet-1 vector (Novagen, San Diego, CA, USA) and GST-CAK1 was cloned into pGEX-2T vector (GE Healthcare, Piscataway, NJ, USA). All three proteins were co-expressed by transforming an E. coli BL21 strain with both plasmids. For large scale preparation, cells expressing F80A cyclin A-CDK2 were grown in 500 ml of LB medium to OD600 of 0.8 and isopropyl β-D-thiogalactoside (IPTG) was added to 0.5 mM. Cells were then cultured overnight at 37°C before being harvested by centrifugation. The cell pellet was lysed by incubating with 20 ml lysis buffer (50 mM Tris, pH 7.5, 1 mM DTT, 1 mM MgCl2, 25 U/ml Benzonase (Novagen), 2 mg/ml lysozyme (Sigma-Aldrich) and protease inhibitors cocktail (Sigma-Aldrich)) at room temperature for 1 h. After NaCl was added to 250 mM and Triton X-100 to 0.025%, the cell slurry was sonicated followed by centrifugation. The supernatant was dialyzed against phosphate-buffered saline (PBS) before incubation with 3 ml of Ni-NTA resin (Qiagen, Valencia, CA, USA), which was washed sequentially with 15 ml PBS and low imidazole buffer (30 mM Tris, pH 7.5, 150 mM NaCl, 50 mM imidazole), and eluted with 6 ml high imidazole buffer (30 mM Tris, pH 7.5, 150 mM NaCl, 300 mM imidazole).
Immunoblotting, immunoprecipitation, orthophosphate labeling, and phosphoamino acid analysis
These procedures were described previously [64].
Preparation and fractionation of 293 native cell lysates
HEK293 cells were grown on 15 cm plates to near confluency and harvested in PBS buffer. Cell pellets were resuspended in hypotonic lysis buffer (50 mM Tris, pH 7.5, 1 mM DTT, 1 mM MgCl2, 0.1% Triton X-100, 25 U/ml Benzonase (Novagen), and protease inhibitors cocktail (Sigma-Aldrich)) and incubated at 4°C, followed by sonication in 150 mM NaCl. Cell debris was pelleted by centrifugation and the supernatant was diluted such that the salt concentration was below 25 mM. The whole cell lysate was then loaded onto a SP Sepharose (GE Healthcare) column manually by atmospheric pressure and the flowthrough was collected. After washing the column with loading buffer (30 mM Tris, pH 7.5, 25 mM NaCl, 1 mM DTT), bound proteins were eluted sequentially with load buffer containing 100 mM, 200 mM, 300 mM, 400 mM, and 600 mM NaCl. The flowthrough was loaded onto a Q Sepharose (GE Healthcare) column and similar procedures were carried out to collect flowthrough and elute the column. Proteins from Q Sepharose flowthrough were pelleted by ammonium sulfate precipitation at 60% and resuspended in load buffer. All fractions were concentrated with salt concentration adjusted between 100 and 200 mM by serial dilution and concentration. These fractions were used in the first set of experiments and subsequently dialyzed extensively against a Tris buffer (30 mM Tris, pH 7.5, 150 mM NaCl) to be used in a second set of experiments.
In vitro kinase assays and purification of thiophosphorylated peptides
Kinase assays using kinases purified from human cells have been described previously [64]. For kinase assays using cell lysate fractions (or GST-Rb) and PE-ATP-γ-S, recombinant wild-type and F80A cyclin A-CDK2 complexes purified from E. coli (as described above) were used. In each of the lysate reactions, about 100-200 μg of lysate fraction (and 100-200 ng of GST-Rb as positive control) was mixed with approximately 1-2 μg of cyclin A-CDK2 (F80A) complex, 250 mM PE-ATP-γ-S in kinase reaction buffer (40 mM Tris, pH 7.5, 10 mM MgCl2, 50 mM NaCl). The reaction was incubated at 30°C for 5 h and the protein mixture was denatured by adding acetonitrile to 15% and digested with trypsin (1/20 mass ratio) at 37°C for at least 6 h. Peptides were then incubated with 20 μl of disulfide beads Thiopropyl Sepharose 6B (GE Healthcare) with rotation at room temperature overnight. The beads were loaded onto a Micro Bio-Spin column (Bio-Rad Laboratories, Hercules, CA, USA), and washed sequentially with 3 ml water, 5 ml of 30% acetonitrile in 0.1% formic acid, 5 ml of 2 M NaCl, and 3 ml water. Beads were collected and incubated with 20 μl of 20 mM NaOH at room temperature for 2 h. The eluate was neutralized and acidified with 1% formic acid to pH 3 for direct analysis. Similar phosphopeptide capturing protocol was carried out for the GST-Rb kinase assay using 5 μg of GST-Rb and 0.5 μg of cyclin A-CDK2 (F80A) complex.
Mass spectrometry analysis and database search
Phosphopeptides samples were analyzed by microcapillary high performance liquid chromatography-electrospray ionization-MS/MS using an ion-trap mass spectrometer (LCQ, ThermoFinnigan, San Jose, CA, USA). Peptides were pressure-loaded onto a 75 μM × 12 cm self-packed C18 column and resolved by a non-linear gradient of 5-28% acetonitrile containing 0.1% formic acid at the flow rate of 200 nl/minute over the course of 2 h. Tandem spectra acquired were searched against the human NCI database (07.20.2006) using SEQUEST. Search parameters included one tryptic end and differential mass modification to serine and threonine due to phosphorylation. For listing purposes, all entries were manually updated using current Swiss-Prot nomenclature. Search results from two independent experiments on each lysate fraction were pooled and filtered using the statistical tool PeptideProphet [65]. Peptides with probabilities higher than 0.9 (error rate <1.8%) were manually validated to further exclude ones with poor MS/MS spectra before inclusion in the final list.
Abbreviations
CDK, cyclin-dependent kinase; DTT, dithiothreitol; GST, glutathione-S-transferase; MS, mass spectrometry; MS/MS, tandem MS; PBS, phosphate-buffered saline; PE-ATP, N6-(2-phenylethyl)-ATP; Rb, Retinoblastoma.
Authors' contributions
YC designed the method, performed the method validation, in-lysate kinase assays, and substrate validation experiments, carried out MS and data analyses. MW performed the kinase mutagenesis and characterization, ATP analogue synthesis and substrate validation experiments. AAH contributed to substrate validation experiments. JJP contributed to the method design. RA provided MS and software resources. YC, RA, and BEC designed the research project. YC and BEC wrote the manuscript.
Additional data files
The following additional data are available. Additional data file 1 is a figure showing the scheme of HEK293 cell lysate fractionation. Additional data file 2 is a table listing the numbers of peptides identified in the lysate fractions. Additional data file 3 is a table listing the identified proline-directed phosphopeptide sequences, the corresponding protein names, and their functional categorizations. Additional data file 4 is a table comparing the proteins and phosphopeptides identified for cyclin A-CDK2 and cyclin B-CDK1. Additional data file 5 is a table listing the phosphopeptides with non-proline-directed sites and from cyclin A-CDK2 autophosphorylation. Additional data file 6 is a figure showing the autophosphorylation of cyclin A-CDK2 in vitro. Additional data file 7 is a figure showing that cyclin B-CDK2 (F80A) and cyclin E-CDK2 (F80A) complexes that may have formed in the lysate represent a negligible fraction of the total amount of CDK2 (F80A) activity. Additional data file 8 is a figure showing the Northern analysis of RL12 expression.
Additional data file 1. Scheme of HEK293 cell lysate fractionation.
Format: PDF Size: 919KB Download file
This file can be viewed with: Adobe Acrobat Reader
Additional data file 2. Numbers of peptides identified in the lysate fractions.
Format: XLS Size: 18KB Download file
This file can be viewed with: Microsoft Excel Viewer
Additional data file 3. Identified proline-directed phosphopeptide sequences, the corresponding protein names, and their functional categorizations.
Format: XLS Size: 66KB Download file
This file can be viewed with: Microsoft Excel Viewer
Additional data file 4. Proteins and phosphopeptides identified for cyclin A-CDK2 and cyclin B-CDK1.
Format: XLS Size: 25KB Download file
This file can be viewed with: Microsoft Excel Viewer
Additional data file 5. Phosphopeptides with non-proline-directed sites and from cyclin A-CDK2 autophosphorylation.
Format: XLS Size: 22KB Download file
This file can be viewed with: Microsoft Excel Viewer
Additional data file 6. Autophosphorylation of cyclin A-CDK2 in vitro.
Format: PDF Size: 933KB Download file
This file can be viewed with: Adobe Acrobat Reader
Additional data file 7. Cyclin B-CDK2 (F80A) and cyclin E-CDK2 (F80A) complexes that may have formed in the lysate represent a negligible fraction of the total amount of CDK2 (F80A) activity.
Format: PDF Size: 4.5MB Download file
This file can be viewed with: Adobe Acrobat Reader
Additional data file 8. Northern analysis of RL12 expression.
Format: PDF Size: 2MB Download file
This file can be viewed with: Adobe Acrobat Reader
Acknowledgements
We thank David Morgan, Kevan Shokat, Justin Blethrow, and Jonathan Cooper for helpful discussions and Titia de Lange for the TRF2 clone. This work was supported by grants from the NIH (to BEC), the Burroughs Welcome Foundation (to BEC), the Hearst Foundation (to YC), the Leukemia and Lymphoma Society (to MW); the National Heart, Lung, and Blood Institute, National Institutes of Health, under contract No: N01-HV-28179 (to RA) and by SystemsX.ch (to RA).
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Syria: “Lens of a Young Homsi”, Photographs of a City under Siege
This post also available in:
Español · Siria: "Lentes de un joven de Homs", fotografías de una ciudad sitiada
Malagasy · Syria: "Lens of a Young Homsi", Sarin'ny Tanàna Iray Atao Fahirano
বাংলা · সিরিয়াঃ- লেন্স অফ এ ইয়াং হোমিস- অবোরধে থাকা একটি শহরের আলোকচিত্র
Dansk · Syrien: "kameralinsen bag en ung borger fra Homs", Fotografier af en storby under belejring
Català · Síria: "L'objectiu d'un jove d'Homs", fotografies d'una ciutat sota setge
Deutsch · Syrien: „Lens of a Young Homsi“, Fotos einer Stadt im Belagerungszustand
简体中文 · 叙利亚：年轻荷姆斯人镜头下的围城
繁體中文 · 敘利亞：年輕荷姆斯人鏡頭下的圍城
Français · Syrie : "Derrière l'objectif d'un jeune de Homs", des photos d'une ville assiégée
عربي · عدسة شاب حمصي، صور من مدينة سورية تحت الحصار
When demonstrations started in Syria in March 2011, a group of friends from Homs, all under 25, took their cameras to photograph these unprecedented protests in their city. Soon after, they were photographing the destruction of the city´s streets and neighborhoods as a result of the regime crackdown. Through their lenses, they aim to show the world what Homs was like, and what it has become, documenting memorial revolutionary moments from “the city of the revolution”. They receive daily requests from people who had to flee the city and want to know if their houses have been destroyed or not.
They insist that they are just a group of volunteers, with no professional training, but their work is outstanding. They are the “Lens of a Young Homsi”:
Photo by the “Lens of a Young Homsi”, taken from their Facebook page
This video shows some of their work and why Lens of a Young Homsi makes a difference to those who have been forced to flee their homes in Homs:
“Sandbags, barricades, soldiers armed to their teeth, bullets, bombs, checkpoints cutting out one district from the other…” This is the scenario these young Homsis show through their lenses:
You can follow their work on Facebook and Twitter.
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Studier Lysate Prep
From OpenWetWare
Revision as of 15:13, 3 June 2005 by Skosuri (Talk | contribs)
(diff) ←Older revision | Current revision (diff) | Newer revision→ (diff)
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Summary
How to make a lysate from a plaque preparation.
Protocol
1. Add 5mL fresh overnight (BL21 if wild-type T7) into 15 mL of T broth (125 mL flask)
2. Add a single plaque to above
3. Shake at 30°C until lysis is visually apparent
4. As soon as lysis is observed and appears to be complete, add 1g of NaCL to the flask and dissolve by shaking.
• Studier has observed that leaving released phage in the cell debris causes a reduction in the kinetics of phage binding (personal communication). It becomes difficult to get good time courses in this case. It is recommended when making a large stock that will be used in timing experiments, that you check that most of your phage adsorb to the cell in the first 30 seconds.
References
Studier, FW Virology 39:562 (1969) Link: Pubmed
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The indispensable first step to getting the things you want out of life is this: Decide what you want. Stein, Ben
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Coal is a portable climate. It carries the heat of the tropics to Labrador and the polar circle; and it is the means of transporting itself whithersoever it is wanted. Watt and Stephenson whispered in the ear of mankind their secret, that a half-ounce of coal will draw two tons a mile, and coal carries coal, by rail and by boat, to make Canada as warm as Calcutta, and with its comfort brings its industrial power. Emerson, Ralph Waldo
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RationalWiki:Requests to change username
From RationalWiki
Jump to: navigation, search
Please place below you request, and the username which you would like to possess, along with your reasons for the change.
Contents
[edit] Requests
[edit] July 2007
[edit] September 2010
• Usurp the never used User:Admin, I think that was mine anyway! Admin2 (talk) 01:35, 7 September 2010 (UTC)
That's the default username created upon MW installation. -- Nx / talk 06:44, 7 September 2010 (UTC)
No its not, that account was User:RWadmin, controlled by ColinR[1] Admin2 (talk) 15:28, 7 September 2010 (UTC)
My bad. It's done. -- Nx / talk 15:38, 7 September 2010 (UTC)
[edit] May 2011
Done. -- Nx / talk 14:44, 29 May 2011 (UTC)
[edit] July
Hi me again. Yeah, just another tweak... let's go with "Rationalizer bunny" as a pun on the Energizer bunny. What do you think? Good/bad? RatMaster háblame 21:56, 20 July 2011 (UTC)
Are you asking for my opinion or are you asking me to actually do it? -- Nx / talk 07:43, 21 July 2011 (UTC)
[edit] August
From User:Ullhateme to User:UHM, just because I want it shorter. --ʤɱ structuralist 08:46, 6 August 2011 (UTC)
Done. -- Nx / talk 10:26, 6 August 2011 (UTC)
Hey, Nx. Could you please move me from BbMaj7 to B♭maj7#11, if you would be so kind. (I couldn't get the non-alphanumerics in when I signed up). Thanks, sir. BbMaj7 Doin' to you in your ear hole. 15:33, 15 August 2011 (UTC)
[edit] September '11
I have - metaphorically speaking - sobered up and decided to ditch the name 'eyeonicr' on this site (if nothing else I'd rather this not be seen to categorise me) and change it to User:PeterL. You know, just in case a RFNALI policy is ever implemented ;)
If you're not too busy/deposed, of course...
Eyeonicr talk, or type, or whatever... 04:39, 18 September 2011 (UTC)
Done. -- Nx / talk 04:59, 18 September 2011 (UTC)
Thank you. Peter talk, or type, or whatever... 05:14, 18 September 2011 (UTC)
[edit] December 2011
Can I please change my username to "Restorationist"? Thank you and Goddess Bless! Restoration Christian Church (talk) 07:00, 26 December 2011 (UTC)
Done. -- Nx / talk 08:59, 26 December 2011 (UTC)
"Socal212" is incredibly tacky and a testament to my lack of creativity when it comes to usernames. May I have changed to just "Sam"? Sam Tally-ho! 00:27, 27 December 2011 (UTC)
Hecho. Acer Blue 00:34, 27 December 2011 (UTC)
¡Muchas gracias, Azul! Sam Tally-ho! 00:39, 27 December 2011 (UTC)
[edit] April 2012
Could I please change my name to "StacyB"? I've decided that I (a) just don't like my current name and (b) do not want to be confused with another person using the same name on another site. --Danfly (talk) 16:02, 8 April 2012 (UTC)
Done. Liquid Blue 06:10, 16 April 2012 (UTC)
Could anybody change my username to User:Maximaman? I think my username sounds stupid. Additionally, the new one is an alias I use to log into most sites which I'm registered in. Qocheedy daiin (talk) 05:34, 16 April 2012 (UTC)
Done. Acer Blue 06:08, 16 April 2012 (UTC)
Thanks. Maximaman (talk) 06:34, 16 April 2012 (UTC)
[edit] June 2012
Please rename User:Maratrean to User:ZackMartin. I don't want people to confuse my own personal blibber-blatherings about nothing in particular with the official positions of the religion which I lead. Zack Martin 23:03, 14 June 2012 (UTC)
Done. AceModerator 23:08, 14 June 2012 (UTC)
Thanks Ace! Zack Martin 23:12, 14 June 2012 (UTC)
[edit] July 2012
I'd like to be renamed to User:Ochotonaprinceps because I seem to have typoed it at the time of my account creation and didn't notice it until now. I'm a freaking IDIOT . Incidentally, it's the Latin name for these cute little guys. Ochotonaprincepsnot a pokémon 02:37, 15 July 2012 (UTC)
Done. Тytalk 03:30, 15 July 2012 (UTC)
Many thanks, good sir! Ochotonaprincepsnot a pokémon 03:54, 15 July 2012 (UTC)
I'd like to change my name to Andy Franlinson...I misspelled my name too... --Andy Frankinson (talk) 13:00, 23 July 2012 (UTC)
Oh wow I misspelled it again! Stupid autocorrect. I meant Franklinson, with a k.... *Sigh*...I'm an idiot --Andy Franlinson (talk) 16:59, 23 July 2012 (UTC)
Done.--talk 21:21, 23 July 2012 (UTC)
Thank you very much! --Andy Franklinson (talk) 23:47, 23 July 2012 (UTC)
[edit] February 2013
It would be very nice if I could usurp the unused account of Special:Contributions/Alice because Alice is what people usually call me. Thanks in advance if that would be possible... --singaporeAlice 04:53, 2 February 2013 (UTC)
It's done. Peter Droid whisperer 05:13, 2 February 2013 (UTC)
Wow! That was fast.
Thank you very much indeed, Peter. --Alice 05:20, 2 February 2013 (UTC)
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Australian Bureau of Statistics
Celebrating the International Year of Statistics 2013
ABS Home > Statistics > By Catalogue Number
4430.1.40.001 - Disability, Ageing and Carers, Summary Tables, New South Wales, 1998
Latest ISSUE Released at 11:30 AM (CANBERRA TIME) 05/05/1999
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• About this Release
ABOUT THIS RELEASE
Contains tables for New South Wales from the 1998 Survey of Disability, Ageing and Carers with data on people with disabilities, their carers and people aged 65 years and over. Tables include information on the age, sex, living arrangements and educational and labour force experience of people with disabilities and people aged 65 and over. Information is provided on their need for and receipt of assistance with a range of everyday activities. Carer information indicates how much of this assistance is provided by family and friends.
© Commonwealth of Australia 2013
Unless otherwise noted, content on this website is licensed under a Creative Commons Attribution 2.5 Australia Licence together with any terms, conditions and exclusions as set out in the website Copyright notice. For permission to do anything beyond the scope of this licence and copyright terms contact us.
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Software
Random allocation software for parallel group randomized trials
Mahmood Saghaei
Author Affiliations
Department of Anaesthesia, Isfahan University of Medical Sciences, Isfahan, Iran
BMC Medical Research Methodology 2004, 4:26 doi:10.1186/1471-2288-4-26
The electronic version of this article is the complete one and can be found online at: http://www.biomedcentral.com/1471-2288/4/26
Received:17 August 2004
Accepted:9 November 2004
Published:9 November 2004
© 2004 Saghaei; licensee BioMed Central Ltd.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Background
Typically, randomization software should allow users to exert control over the different aspects of randomization including block design, provision of unique identifiers and control over the format and type of program output. While some of these characteristics have been addressed by available software, none of them have all of these capabilities integrated into one package. The main objective of the Random Allocation Software project was to enhance the user's control over different aspects of randomization in parallel group trials, including output type and format, structure and ordering of generated unique identifiers and enabling users to specify group names for more than two groups.
Results
The program has different settings for: simple and blocked randomizations; length, format and ordering of generated unique identifiers; type and format of program output; and saving sessions for future use. A formatted random list generated by this program can be used directly (without further formatting) by the coordinator of the research team to prepare and encode different drugs or instruments necessary for the parallel group trial.
Conclusions
Random Allocation Software enables users to control different attributes of the random allocation sequence and produce qualified lists for parallel group trials.
Background
An important aspect of any trial that should be clearly stated in the final report is the method used to assign treatments (or other interventions) to participants [1]. In the final report of the trial, authors should specify the method of sequence generation, i.e. whether they have used mechanical means, a computer generated random list or random number table. After preparing a random sequence, subjects will be allocated to the trial groups using an implementation method such as numbered containers, central telephone line, or allocation by a person who is not involved in the main research and patient care (the encoder). During the process of allocation each subject will be given a unique identification code (Unique Identifier, UI). This UI will be used as a label to uniquely identify the patient's group after completion of the study. During the study period this UI will be given to the main researchers together with the necessary treatment (e.g. drug or placebo). Usually these treatments are prepared by the encoder with the same physical characteristics (shape, color, size, etc), differing only by the UI labels to blind the main researchers about the actual patient group. Following enrollment of all subjects into the study, these UIs are decoded to determine the patient group. Depending on the preference of the researchers or facilities of the research environment, subjects are randomly allocated to intervention groups using either a random list prepared before the study (In Advance method) or a randomized allocation at the moment of intended intervention (Just In Time method; JIT). Both the JIT and In Advance methods produce acceptable allocations, and the actual choice depends on the availability of certain facilities for each method. Usually the randomization components of these two methods are produced by running a randomization software on either an Internet service provider or a local computer. The local encoder will obtain the next allocation (JIT method) or the entire random list (In Advance method) from the service provider or from the software on local computer and prepare the necessary blinded equipments.
Without the use of computer software or Internet services the maintenance of the whole process of randomization and allocation is difficult. In addition, in the case of any necessary restrictions on the process of randomization (i.e. block randomization [2]) the complexity of the process will be increased even further and be prone to errors. Randomization software may run on a local computer or may be hosted by an Internet server. A complete list of these software and services can be found on Martin Bland's web site [3]. Most of randomization software are hosted by websites for both JIT and In Advance methods, which require access to the Internet [4-8]. However, most of these Internet services have restricted capabilities with respect to the block design specification, control over the output format and flexibility of UIs. Among these available services, the tool in Randomization.com [4] seems to be more advanced than the others. It allows users to specify the number of subjects per block, the number of blocks and up to 20 treatment labels. Therefore, this service produces simple and block randomization using fixed and equal block sizes. Unfortunately, this service does not allow further restriction on block design (e.g., multiple block lengths or random variation in block number or size). The generated random list is in the form of UI and group name pairs, formatted in a single column, which in cases of large sample sizes may require further work to fit it in multiple columns for fine printing. Moreover, the block borders are not visible to allow for easy visual inspection of block sizes and equality of cases. Although a minor problem, Randomization.com only produces sequential numeric UIs with variable lengths (e.g. 1, 10, 100). The variability in lengths of UIs may disturb the visual impact of the generated list compared to the fixed length UIs (e.g. 001, 010, 100). Some researchers prefer to use random UIs in mixed alphanumeric format to decrease the likelihood of memorization and to improve the blindness of the study and concealment of allocations. Available randomization software have more restrictions in their capabilities than the Internet randomization services. They have limitations in their output format [9] and users can not specify the number and naming of the treatment groups [10,11]. In addition, these software are not designed with the capability to produce flexible UIs for participants. Therefore, the main objective of Random Allocation Software was to construct a randomization software for parallel group trials with the following characteristics:
1. Independent running on a local computer without any need to access the Internet
2. Different types of program output: to file (html or text files), window and system clipboard
3. Provisions for different block design
4. Capability to deal with a larger number of groups
5. Specifying a name for each group
6. Control over the format, length and ordering of the generated UIs
7. Control over the format of generated sequence
8. Saving or loading the randomization settings
9. Viewing previously generated randomized sequences
Implementation
Random Allocation Software is a program created in Microsoft Visual Basic 6, and it installs in the same way as ordinary Windows software (i.e. running setup.exe and following on screen instructions). Once installed and run, there are some controls in the main window for specifying the number of groups (2 to 16), sample size and the name of each group. It also contains menu items to determine the program output and randomization settings. The default program output is saved into either html or text files, and it may also have output to a window or to the system clipboard. A variety of randomization options can be set in the options window. The length of generated UIs (named as Code in the program) can be between 3 to 10 characters and there are options for different alphanumeric structures. In addition, these UIs can appear in sequential or random order in the generated random list. The program can generate simple or block randomization in different types, including equal size blocks, multiple block lengths with random variation among the specified block sizes and complete randomized blocks (random number and size of blocks). The generated sequence will appear in a multiple columns format and the number of columns (1 to 10) can be changed in the options window. Output to html file will be formatted in the form of one block per table. Borders of the tables may be shown or hidden. By clicking the 'Generate' button in the main window the random sequence will be generated and opened by the default viewer for the output file (e.g. Internet Explorer for html files). Previously created output files can also be viewed from inside the program. Additional options include saving the current randomization settings, loading a previous setting and enabling the program to save the last setting upon program exit.
During execution, the program produces a random sequence of allocation using the Rnd function that generates a floating point random number. The Rnd function uses the linear-congruent method for pseudo-random number generation as depicted by the following formula:
x1 = (x0 * a + c) MOD (2^24) [12]
where:
x1 = new value
x0 = previous value (an initial value of 327680 is used by Visual Basic unless the Randomize X function is used to specify a different seed as X)
a = 1140671485
c = 12820163
The seed of the Rnd will be the Timer function, which will return the number of seconds elapsed since midnight. Although this version of the program does not produce repeatable lists, it is possible to revise the program in subsequent versions to save the value of the seed to reproduce the same random list. The output consisted of shuffled allocations each of which is a UI, group name pair. The program checks for the uniqueness of the UIs and generates an error message if the specified UI length is insufficient to hold the entire sample size.
Runs test was used to check randomness of the output list with sample sizes from 10 to 190 (10, 30, 50, ..., 190) and from 200 to 3000 (200, 600, 1000, ..., 3000). Each runs test was carried out for the group number of 2, 3, 5 and 6. SPSS 10 software was used to perform the runs test.
Results
The program starts running with the default settings. Users may run the program with the default settings or set the number of groups, the name of each group and the sample size. Clicking the 'Generate' button (figure 1) produces the random sequence. Before generating the random sequence, the option window will be displayed and different randomization settings can be entered (figure 2 and 3). Consider, for example, that we want to produce a simple randomized list for a sample size of 30 subjects into three groups of Case, Control and Placebo with numeric sequential UIs. After setting different options and clicking the 'Generate' button, the generated list will appear in columns (Table 1). Each entry in the list consists of a UI, and a group name pair. Alternatively numeric UIs may appear in random order (Table 2). Table 3 shows the output of the program for a block randomization with blocks of equal sizes.
Figure 1. Main window. The main window showing different options for number of groups, sample size, and group names.
Figure 2. Options window: Blocks. Options window, settings for block design.
Figure 3. Options window: Code. Options window, setting the format of unique identifier (UI) specified in the program as Code.
Table 1. A simple randomized list produced by the software for a sample size of 30 subjects into three groups of Case, Control and Placebo with numeric sequential unique identifiers
Table 2. The same setting as in table 1, but with the numeric UIs in random order
Table 3. A block randomization list with four blocks of equal sizes
Figure 4 is the printed output of the program for a block randomization with random block sizes.
Figure 4. Sample output. Sample output for a block randomization with random block sizes.
In block randomization the final sample size is usually larger than the specified one.
A total of 18 runs test were performed to check the randomness of the program output, which resulted in P values of 0.22 to 0.81.
Discussion
The main use of Random Allocation Software is to produce simple or block randomized sequences for parallel group trials. Its use is restricted to parallel group randomized trials. Compared with similar software, it enables the user to control the length, order and format of the UIs; and the type and format of the output. It allows specifying up to 16 groups for parallel trials.
Conclusions
Random Allocation Software has been designed to produce random sequences consisting of UI, group name pairs with additional control over the output format and type. Available randomization software generally has limitations in the number of groups, naming each group, generating UIs and control over the output. Many of these problems have been addressed in the present software. As has been stated in previous sections, the main use of this software is for randomization in parallel group trials. The software can be revised to support crossover and other types of randomized trials. The experienced user may test the randomness of the program output by selecting numeric labels for group names and then exporting the generated list into a statistical software such as SPSS to execute a runs test on the exported data.
Availability and requirements
Project name: Random Allocation Software
Public use access:
http://www.msaghaei.com/Softwares/dnld/RA.zip webcite (Latest version)
http://ftp.mui.ac.ir/RA.zip webcite
Operating systems: Windows 98, Me, 2000, XP. It should be noted that on some Windows operating systems (especially Windows 2000) during installation of the program an error message like "Setup Cannot Continue... System Files Are Out of Date" may be displayed. If this happens, click OK and restart the system. Then run the setup.exe again. This is due to a known bug in the installation programs of Microsoft Visual Basic [13]. This problem has been removed from the newer versions of the program. Users are recommended to download the latest version from the first address.
Programming Language: Visual Basic 6
Other requirements: Internet Browser (Internet Explorer 5 or higher is recommended)
License: Free for academic use.
Abbreviations
JIT = Just in Time
UI = Unique Identifier
Competing interests
The author declares that he has no competing interests.
Acknowledgements
The author thanks Azra Kianinejad, Department of Human Development, Kobe University, Kobe, Japan for her kind review of the manuscript.
References
1. Schulz KF: Randomized controlled trials.
Clin Obstet Gynecol 1998, 41:245-56. PubMed Abstract | Publisher Full Text
2. Altman DG, Schulz KF, Moher D, Egger M, Davidoff F, Elbourne D, Gøtzsche PC, Lang T, CONSORT GROUP (Consolidated Standards of Reporting Trials): The Revised CONSORT Statement for Reporting Randomized Trials: Explanation and Elaboration.
Ann of Intern Med 2001, 134:663-694.
3. Directory of randomization software and services [http://www-users.york.ac.uk/~mb55/guide/randsery.htm] webcite
4. Randomization.com [http://www.randomization.com/] webcite
5. GraphPad QuickCalcs [http://www.graphpad.com/quickcalcs/randomize1.cfm] webcite
6. EDGAR [http://www.jic.bbsrc.ac.uk/services/statistics/edgar.htm] webcite
7. PARADIGM Registration-Randomisation Software [http://telescan.nki.nl/paradigm.html] webcite
8. Randomizer [https://medinfo.uni-graz.at/randemo/web/about.php] webcite
9. Simple Statistical Software by Martin Bland [http://www-users.york.ac.uk/~mb55/soft/soft.htm] webcite
10. Randomization Generator download [http://www.som.soton.ac.uk/staff/jrg/files/Randomisation Generator download/] webcite
11. KEYFINDER [http://lib.stat.cmu.edu/designs/] webcite
12. Microsoft Knowledge Base Article – 231847 [http://support.microsoft.com/default.aspx?scid=kb;en-us;231847] webcite
13. Microsoft Knowledge Base Article – 174135 [http://support.microsoft.com/default.aspx?scid=kb;en-us;174135#appliesto] webcite
Pre-publication history
The pre-publication history for this paper can be accessed here:
http://www.biomedcentral.com/1471-2288/4/26/prepub
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An inspirational, true-life story of online marketing, revisionism, idiocy and redemption. And saving money.
Maybe you have some old marketing assets that can be quickly and cheaply reused!
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<D <M <Y
Y> M> D>
(2) Leonard's Household Tips: Don't make caramel sauce without a really well thought-out plan for storing it, or you will burn and melt things and the caramel will become useless.
(1) NewsBruiser: Aggregated Assault: I have some 5x7 notecards and I write down my big projects on them so that if I ever feel lackadaisical like I don't have any big projects, I can look at the notecards and remember that I'm just lazy.
This is one of the projects and to be honest there's not much on its notecard except the name--that's how lazy I am. But phase one is now complete: you can now take arbitrary RSS feeds and aggregate them into a NewsBruiser weblog, a la my arch-rival Scott's Planet.
Viola! I give you the long-promised Richardson/Chadwick/Matkin/Whitney/Walch recipe weblog! Right now it only has me and Susanna since we're the only ones with a category where our recipes go. If you've got a weblog on this site and you want in, send me mail and I'll show you how to create a category. You don't have to do anything special to get recipes to show up in this weblog; just post to your category and they'll show up within an hour.
Butter Pecan Ice Cream: As a test of the automatic aggregation, I'll post the most recent recipe I've made. I'm having a little dinner party tomorrow and tonight I made butter pecan ice cream. This is a good time to formally state the pound-cake-like Generic Ice Cream Mneumonic I've come up with:
I'm experimenting with heating the sugar along with the cream and milk. It worked out well this time, so heat all that up in a pan.
Now, the rest of my mneumonic (patent pending) is '1 cup flavor stuff'. However, the flavor of butter pecan ice cream depends in large part on the brown sugar we're using, so that kind of counts as 'flavor stuff' and we actually want less than 1 cup. What I used was:
If you're offended by the idea of deviation from the mneumonic I made up, then 1) that's kind of weird, and 2) you can probably do 1 cup of nuts and 3 tablespoons of butter without ruining the ice cream.
Melt the butter in a saucepan. Add the pecans and toss to coat. Toast the pecans. I dunno when you stop exactly; generally you stop toasting nuts as soon as you smell the oils being released, but something is wrong with my sense of smell today so I just stopped when they looked right. Dump the nuts and butter into the cream/milk/sugar mixture. Stir in The Old Standbys:
When you've got nuts and sugar and cold, you know there's got to be salt.
There's your mix. Let it cool and then machine it. Very tasty.
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Kleiber, Max
Kleiber, Max
This article has been reviewed by the following Topic Editor: Peter Saundry
Max Kleiber. (Source: UC Davis Magazine)
Max Kleiber (1893-1976), a Swiss-American animal scientist who was one of the first to make accurate measurements of the rate of energy use relative to body size among different species of mammals (1932). He claimed that the 3/4 power of body weight was the most reliable basis for predicting the basal metabolic rate of mammals and for comparing nutrient requirements among mammals of different sizes. Klieber’s Law: MR =aM0.75, where MR is metabolic rate (watts), M is body mass (kg), and a is a constant. However, although Kleiber's law has been widely used and thought to apply to all of life, significant variation in metabolic scaling has been observed among various taxonomic groups of animals and plants, and among various physiological states. Kleiber also provided the basis for the conclusion that total efficiency of energy utilization is independent of body size and pioneered the use of isotopes to study metabolic processes associated with lactation in cattle (1947).
Further Reading
Citation
Douglas Glazier (Lead Author);Cutler J. Cleveland (Contributing Author);Peter Saundry (Topic Editor) "Kleiber, Max". In: Encyclopedia of Earth. Eds. Cutler J. Cleveland (Washington, D.C.: Environmental Information Coalition, National Council for Science and the Environment). [First published in the Encyclopedia of Earth August 21, 2008; Last revised Date August 21, 2008; Retrieved May 18, 2013 <http://www.eoearth.org/article/Kleiber,_Max>
The Author
Douglas Glazier is a professor of biology at Juniata College. He is presently the leader of Juniata College’s Environmental Task Force and is very dedicated to the research pursuits of his students. Dr. Glazier was chair of Juniata College’s Environmental Science Program from 1995-1998. His research interests include functional biology and life-history evolution of mammals (especially murid rodents) and crustaceans (especially cladocerans and pericaridans), ecology of freshwater springs, eco ... (Full Bio)
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Reviewing the Results of Calorie Restriction Primate Studies
Permalink | View Comments (0) | Post Comment | | Posted by Reason
In the past few years two ongoing studies of long term calorie restriction (CR) in primates have started to publish their results on longevity. Both research programs have been underway for more than 20 years, one run by the National Institute on Aging and the other by the University of Wisconsin-Madison. Researchers have followed small groups of rhesus monkeys to see how the benefits to health and life expectancy resulting from a restricted calorie intake compare with those obtained in mice and other short-lived species. At this point the results are ambiguous, unfortunately: one study shows a modest gain in life expectancy that has been debated, while the other shows no gain in life expectancy, and that result has also been debated.
Calorie restriction does produce considerable benefits in short term measures of health in rhesus monkeys and humans, that much is definitive, but the present consensus in the research community is that it doesn't greatly extend life in longer-lived primates - perhaps a few years at most in humans. Differences and issues in the two primate studies mean that effects of this size on longevity may never be clear from the data generated. Other factors will wash it out, such as differences in the diet fed to the control groups, or the different age at which calorie restriction started. Certainly the results so far support the conjecture that calorie restriction is exceedingly good for health but doesn't have the same impressive effects on longevity as it does in short-lived animals. Why that is the case is a puzzle to be solved - but not one that has a great deal of relevance to the future of human longevity. One would hope that we'll be a long way down the road to rejuvenation therapies by the time another set of better constructed primate studies are nearing completion.
You'll find a long article over at the SENS Research Foundation that examines the NIA and Wisconsin primate studies, their differences, and their results in great detail - but I'm just going to skip ahead and quote some of the conclusions:
CR in Nonhuman Primates: A Muddle for Monkeys, Men, and Mimetics
In this post, I have sketched out in detail two major possible interpretations of the disparate mortality outcomes in the NIA and WNPRC nonhuman primate CR studies. The "Diminishing Returns" hypothesis posits that the health and longevity benefits of "CR" reported in the WNPRC study were merely the unsurprising results of one group of animals being fed a high-sucrose, low-nutrient chow on a literally ad libitum basis, and another group being kept to portions of that diet low enough to avoid the deranged metabolisms flowing from obesity and (possibly) fructose toxicity. In this interpretation, the more severe restrictions of energy intake imposed at the NIA - particularly when the chow to which access was restricted may have been healthier to begin with - led to no further health benefit, because there are none to be gained: the dramatic age-retarding effects of CR observed in laboratory rodents and other species do not translate into longevous species such as primates, and the sole benefit of controlling energy intake is avoidance of overweight and obesity.
The "Dose-Response" hypothesis begins from the same interpretation of the WNPRC study, but posits that far from being excessive (or, at best, superfluous) to that required for good health, the additional energy restriction imposed at NIA were too little, and imposed during too narrow a window, to elicit a clear signal in health and lifespan benefits; this is supported by the evidence that the NIA primates were not especially hungry, and only weakly and inconsistently exhibited improvements in risk factors and endocrine signatures of CR that are seen both in life-extending CR in rodents, and in humans under rigorous CR.
Unfortunately, it seems very unlikely that this question will be resolved. Even the narrow question of whether the age-retarding effects of CR in laboratory rodents translate into nonhuman primates could only be established with confidence after yet another trial in nonhuman primates. [Such] a study is extremely unlikely in light of the enormous expense of the first two trials, disappointment (and possibly embarrassment) with the results, [and] the ill winds for nonhuman primate research. [Even] if such a well-designed and well-executed study were initiated: what then? Supposing that support were maintained for the duration of the experiment [it] would be a further three decades before the earliest point at which survival data could be reported.
The timescales involved in resolving these questions cannot be reconciled with the immediate imperatives that drive us to pose them. With the scale of the humanitarian, economic, and social crisis that looms in the coming decades due to global demographic aging and associated ill-health, the near-term need for effective interventions against the aging process could not be greater. Whether CR can retard aging in nonhuman primates or not; whether it can retard aging in humans or not; whether even effective CR mimetics can somehow be shepherded through clinical trials - even the most optimistic projection for retarding aging through such approaches is inadequate to the needs and suffering of aging world.
The point made in the article is the same one that should be made for all means of slowing the pace of aging by altering metabolism, whether by the use of drugs to replicate some of the changes caused by calorie restriction or via other mechanisms. These are very difficult and challenging projects, certainly very expensive in time and funds, and which will produce poor and uncertain end results even if successful. Ways to modestly slow aging do nothing for people who are already old, and we will grow old waiting for success in the development of drugs that can safely tinker our metabolisms into a state of slower aging.
The better approach is that outlined by the SENS Research Foundation: targeted therapies to repair the known forms of cellular and molecular damage that cause aging. This path is cheaper, more certain, and the resulting therapies will be capable of rejuvenation - of reversing degenerative aging, not just slowing it down a little. They will be greatly beneficial for the old, and extend the length of life lived in health and vigor. This is why I say that calorie restriction studies are irrelevant to the future of our health and longevity: the only thing that really matters is whether or not the SENS vision or similar repair therapies are prioritized, funded, and developed.
A Possible Biomarker for Senescent Cells
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There are any number of techniques under development that allow individual cells to be destroyed provided that you can distinguish them from their neighbors: the challenge is in finding characteristic differences in the cells you want destroyed, such as cancer cells or senescent cells. Most of the efforts aimed at producing targeted cell destruction therapies are taking place in the cancer research community, but senescent cells accumulate with age and contribute to degenerative aging - they must also be destroyed. Unfortunately good ways to target senescent cells are somewhat lacking. Candidate mechanisms are emerging, however, and here is another of them:
Due to its role in aging and antitumor defense, cellular senescence has recently attracted increasing interest. However, [the] detection of senescent cells remains difficult due to the lack of specific biomarkers. ndeed, most determinants of cellular senescence, such as the upregulation of p53, p16Ink4a, p21WAF/CIP1 or SASP-associated cytokines, are not exclusively observed in senescence, but can also occur in other types of stress responses. In addition, alterations like SAHF or DNA-SCARS formation are frequently observed, but not necessarily a mandatory feature or exclusive to senescent cells.
The current gold standard for the detection of senescence is the so-called senescence-associated β-galactosidase (SA-β-Gal) activity. Although SA-β-Gal has been first suggested as a distinct enzyme, its activity is derived from lysosomal β-Gal encoded by the GLB1 gene. β-Gal is an accepted marker of senescence, but its reliability and specificity have been questioned, as a positive β-Gal reaction has also been detected in human cancer cells that were chemically induced to differentiate, or upon contact inhibition. Moreover, several cell types, such as epithelial cells and murine fibroblasts generally show a weak β-Gal staining.
In the present study, we investigated several lysosomal hydrolases for their suitability as senescence markers and identified α-fucosidase, a lysosomal glycosidase involved in the breakdown of glycoproteins, oligosaccharides and glycolipids, as a novel biomarker for senescence. We demonstrate that α-fucosidase is upregulated [in] all canonical types of cellular senescence, including replicative, DNA damage- and oncogene-induced senescence. Our results suggest that detection of α-fucosidase might be a highly valuable biomarker for senescence in general and in particular in those cases where SA-β-Gal activity fails to properly discriminate between senescent- and non-senescent cells.
Link: http://www.landesbioscience.com/journals/cc/article/24944/?show_full_text=true
Inhibiting ICMT as a Progeria Therapy
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Progress towards a therapy for the rare accelerated aging condition progeria continues. It remains unclear as to whether the mechanisms responsible for progeria exist in normal aging to a level that is in any way significant. Progeria is caused by malformed prelamin A, and tiny amounts of broken prelamin A can be found in old tissues - but it would really require a therapy for progeria that addressed the issues with prelamin A to easily find out whether this has any meaningful contribution to normal aging.
The classical form of progeria, called Hutchinson-Gilford Progeria Syndrome (HGPS), is caused by a spontaneous mutation, which means that it is not inherited from the parents. Children with HGPS usually die in their teenage years from myocardial infarction and stroke.
The progeria mutation occurs in the protein prelamin A and causes it to accumulate in an inappropriate form in the membrane surrounding the nucleus. The target enzyme, called ICMT, attaches a small chemical group to one end of prelamin A. Blocking ICMT, therefore, prevents the attachment of the chemical group to prelamin A and significantly reduced the ability of the mutant protein to induce progeria. "We are collaborating with a group in Singapore that has developed candidate ICMT inhibitor drugs and we will now test them on mice with progeria. Because the drugs have not yet been tested in humans, it will be a few years before we know whether these drugs will be appropriate for the treatment of progeria."
"The resemblance between progeria patients and normally-aged individuals is striking and it is tempting to speculate that progeria is a window into our normal aging process. The children develop osteoporosis, myocardial infarction, stroke, and muscle weakness. They display poor growth and lose their hair, but interestingly, they do not develop dementia or cancer." [The researchers are] also studying the impact of inhibiting ICMT on the normal aging process in mice.
Link: http://www.eurekalert.org/pub_releases/2013-05/uog-ptf051413.php
Are the Most Influential Futurists Those Who Put in the Work to Make Their Visions Real?
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We'll take a short excursion into ranking futurists for today, prompted by a recent article that offers a (transhumanism-slanted) opinion on the identity of the most important futurists of the past few decades.
The Most Significant Futurists of the Past 50 Years
Our visions of the future tend to be forged in the pages of science fiction. But for the past half-century, a number of prominent thinkers, activists, and scientists have made significant contributions to our understanding of what the future could look like. Here are 10 recent futurists you absolutely need to know about. Needless to say, there were dozens upon dozens of amazing futurists who could have been included in this article, so it wasn't easy to pare down this list. But given the width and breadth of futurist discourse, we decided to select thinkers whose contributions should be considered seminal and highly influential to their field of study.
Those selected include Robert Ettinger, one of the founders of modern cryonics, and Aubrey de Grey, who presently works to make his SENS roadmap to human rejuvenation a reality. Ray Kurzweil is notably absent from the list.
It isn't mentioned as a selection criteria in the article, but I think that ranking the importance of futurists by how effectively they help to create the future that they envisage isn't all that bad of an idea. Advocates and popularists play a needed role in moving from vision to reality, but progress also needs people to perform and orchestrate the actual work of research and development. Kurzweil, for example, is a popularist and an advocate with respect to his futurism: beyond the books and films and persuasion his day job as an inventor and entrepreneur is so far largely irrelevant to the future he envisages. I don't think anyone can argue that he isn't important in the arena of ideas regarding machine intelligence, accelerating change, and how this will all play out in the decades ahead. But how much more important would Kurzweil be if, for example, he had decided a decade or two back to create a company like Zyvex as a long term play to advance molecular manufacturing, or something equivalent in AI work?
In contrast Ettinger and de Grey both founded successful organizations devoted to realizing their particular visions: the Cryonics Institute and the SENS Research Foundation. Both were instrumental in creating the groundwork and the early community of supporters to enable a new industry and branch of research in applied medicine. That seems like the best approach to futurism to me: not just persuasion, but also working to create the change you want to see in the world.
Excess Body Fat Hardens Arteries
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There are all sorts of good reasons to avoid becoming fat. Excess fat tissue is linked to an increased risk of all the common diseases of aging, and correlates well with a shorter life expectancy and higher lifetime medical expenditures. Fat tissue creates higher levels of chronic inflammation and alters the signaling environment in the body, causing a wide range of changes. Here is another of them:
Having too much body fat makes arteries become stiff after middle age, a new study has revealed. In young people, blood vessels appear to be able to compensate for the effects of obesity. But after middle age, this adaptability is lost, and arteries become progressively stiffer as body fat rises - potentially increasing the risk of dying from cardiovascular disease. The researchers suggest that the harmful effects of body fat may be related to the total number of years that a person is overweight in adulthood. Further research is needed to find out when the effects of obesity lead to irreversible damage to the heart and arteries, they said.
Researchers [scanned] 200 volunteers to measure the speed of blood flow in the aorta, the biggest artery in the body. Blood travels more quickly in stiff vessels than in healthy elastic vessels, so this allowed them to work out how stiff the walls of the aorta were using an MRI scanner. In young adults, those with more body fat had less stiff arteries. However, after the age of 50 increasing body fat was associated with stiffer arteries in both men and women. Body fat percentage, which can be estimated by passing a small electric current through the body, was more closely linked with artery stiffness than body mass index, which is based just on weight and height.
"We don't know for sure how body fat makes arteries stiffer, but we do know that certain metabolic products in the blood may progressively damage the elastic fibres in our blood vessels. Understanding these processes might help us to prevent the harmful effects of obesity."
Link: http://www.sciencedaily.com/releases/2013/05/130515085333.htm
Therapeutic Cloning Attained
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Therapeutic cloning or somatic cell nuclear transfer are names given to a method of producing embryonic stem cells from a patient's own cells. These embryonic stem cells could then be used to generate cells of any type as a basis for regenerative therapies. Making the process work has proven to be challenging, however, both from a technical point of view and thanks to misguided attempts to make it illegal. In recent years the focus shifted towards work on induced pluripotent stem cells instead, but a research group now claims success in the original goal:
Scientists [have] successfully reprogrammed human skin cells to become embryonic stem cells capable of transforming into any other cell type in the body. It is believed that stem cell therapies hold the promise of replacing cells damaged through injury or illness. The technique used [is] a variation of a commonly used method called somatic cell nuclear transfer, or SCNT. It involves transplanting the nucleus of one cell, containing an individual's DNA, into an egg cell that has had its genetic material removed. The unfertilized egg cell then develops and eventually produces stem cells.
Previous unsuccessful attempts by several labs showed that human egg cells appear to be more fragile than eggs from other species. Therefore, known reprogramming methods stalled before stem cells were produced. To solve this problem, the [researchers] studied various alternative approaches first developed in monkey cells and then applied to human cells. Through moving findings between monkey cells and human cells, the researchers were able to develop a successful method. The key to this success was finding a way to prompt egg cells to stay in a state called "metaphase" during the nuclear transfer process. Metaphase is a stage in the cell's natural division process (meiosis) when genetic material aligns in the middle of the cell before the cell divides. The research team found that chemically maintaining metaphase throughout the transfer process prevented the process from stalling and allowed the cells to develop and produce stem cells.
Link: http://www.eurekalert.org/pub_releases/2013-05/ohs-ort051313.php
SENS Research Foundation is the Watering Hole, Not the Herd
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If you visit Fight Aging! on a regular basis you'll know that I strongly favor the SENS Research Foundation and the approach taken by its founders, advisors, and staff to speed the development of human rejuvenation. I think we could do with another ten or twenty similar organizations, and certainly a hundredfold increase in the funding for rejuvenation research, but right now we have just the one. So send the Foundation a donation if you're feeling flush today, because there's no-one else out there at the moment who can do as much for your future longevity with that money.
Or rather I should say that there are dozens and possibly hundreds of people out there who can do as much for your future longevity with those funds - it's just that you don't know who they are. Would you know enough to chase down William Bains in the UK and ask him to work on AGE-breaker drugs for glucosepane, for example? Or pick the group at the Buck Institute best placed work on ways to selectively destroy senescent cells by interfering in their characteristic biology? Or have Janko Nikolich-Žugich in Arizona work on restoring the aged immune system by removing unwanted T cells? Of course not. But there is a whole world of researchers out there with useful specialist knowledge and who are these days quite willing to work on the foundation technologies needed for human rejuvenation - provided that the funding can be found.
Organizations like the SENS Research Foundation are the interface between you and the research community: the Foundation staff provide domain knowledge and relationships needed in order to direct funds effectively. Without their work it would be impossible for folk like you or I to help make this field of science move faster - we wouldn't know where to start or who to talk to, never mind where to send funds, and finding out would be so costly in comparison to what we could donate as to make the whole exercise pointless.
The SENS Research Foundation is the watering hole, not the herd. It is the gateway, not the city. It is the door to a network of researchers who are interested in human rejuvenation, but that network is a greater and broader thing than the Foundation. I bring up this point because many people look no further than the gateway: they see the SENS Research Foundation and think of an enclosed group, off to one side of the scientific community, doing its own thing in isolation, and therefore easy to dismiss. For all that this point of view is absolutely incorrect, it is not uncommon. You'll see it liberally applied to biotechnology companies, noted laboratories, and other organizations that are also gateways to broader scientific networks. People look at an organization, see its staff performing some research work in its own domain, but fail to see beyond that to take in the great tree of relationships and connections behind the name plate.
The greatest achievement of the folk behind the SENS Research Foundation (and the Methuselah Foundation before it) is their construction of a lasting and growing network of supporters of rejuvenation research within the life sciences. This was quite the task over the past decade and involved a lot of persuasion, changing the culture of the research community to become more receptive towards longevity science, building relationships, holding conferences, and tireless advocacy. It is that web of relationships, and not the existence of the Foundation per se, that enables growth in funding and progress towards the goal of ending aging. As for all areas of human endeavor, it is relationships and networking that make the world turn: the Foundation is a mailbox, a guidebook, and a banner for a larger community, an outgrowth of that community even, and it is the community that gets things done.
This is worth bearing in mind, because it's all to easy to focus on organizations rather than people and thus miss the whole point of the exercise.
The Immune System Ages More Slowly in Women
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Women tend to live longer than men, and there are any number of competing explanations as to why this is the case. They range from risk of mortality relating to lifestyle choices to evolutionary selection operating on the male role in reproduction to various differences in biochemistry that exist between the genders. That the female immune system ages more slowly shouldn't be terribly surprising - but it might be cause or consequence.
Women's immune systems age more slowly than men's, [and] the slower decline in a woman's immune system may contribute to women living longer than men. Researchers looked at the blood of healthy volunteers in Japan, ranging in age between 20 and 90 years old; in both sexes the total number of white blood cells per person decreased with age. The number of neutrophils decreased for both sexes and lymphocytes decreased in men and increased in women. Younger men generally have higher levels of lymphocytes than similarly aged women, so as aging happens, the number of lymphocytes becomes comparable.
Looking in more detail it became apparent that the rate in decline in T cells and B cells was slower for women than men. Both CD4+ T cells and NK cells increased with age, and the rate of increase was higher in women than men. Similarly an age-related decline in IL-6 and IL-10 was worse in men. There was also a age-dependent decrease in red blood cells for men but not women.
"The process of aging is different for men and women for many reasons. Women have more oestrogen than men which seems to protect them from cardiovascular disease until menopause. Sex hormones also affect the immune system, especially certain types of lymphocytes. Because people age at different rates a person's immunological parameters could be used to provide an indication of their true biological age."
Link: http://www.alphagalileo.org/ViewItem.aspx?ItemId=131061&CultureCode=en
Considering Anti-Amyloid Immunotherapy
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Amyloids are solid masses that form in tissues as a result of misfolded proteins. The amount of amyloid increases with age, perhaps due to a failure of mechanisms that keep the levels of damaged or misfolded proteins under control, and this is thought to cause harm and contribute to degenerative aging. In most cases researchers are still lacking a full understanding of the mechanisms involved, however. At the very least having solid clumps and fibrils present where they shouldn't exist can disrupt tissue integrity or even cause larger scale issues such as clogging blood vessels.
One approach to removing amyloid involves the use of the immune system. Immune therapies direct immune cells to attack and break down a specific target, and much of the innovation in their use as a therapy to remove amyloid is happening in the Alzheimer's research community. That condition is associated with amyloid beta, but we can hope that any successful therapies will prove adaptable to other forms of amyloid and thus applicable to human rejuvenation.
Alzheimer's disease (AD) is the most common dementia in the industrialized world, with prevalence rates well over 30% in the over 80-years-old population. AD is strongly associated with Amyloid-beta (Abeta) protein aggregation, which results in extracellular plaques in the brain, and according to the amyloid cascade hypothesis appeared to be a promising target for the development of AD therapeutics.
Within the past decade convincing data has arisen positioning the soluble prefibrillar Abeta-aggregates as the prime toxic agents in AD. However, different Abeta aggregate species are described but their remarkable metastability hampers the identification of a target species for immunization. Passive immunotherapy with monoclonal antibodies (mAbs) against Abeta is in late clinical development but recently the two most advanced mAbs, Bapineuzumab and Solanezumab, targeting an N-terminal or central epitope, respectively, failed to meet their target of improving or stabilizing cognition and function.
Preliminary data from off-label treatment of a small cohort for 3 years with intravenous polyclonal immunoglobulins (IVIG) that appear to target different conformational epitopes indicate a cognitive stabilization. Thus, it might be the more promising strategy reducing the whole spectrum of Abeta-aggregates than to focus on a single aggregate species for immunization.
Link: http://www.immunityageing.com/content/10/1/18/abstract
Telomere Length: Cause of Aging or Marker of Aging?
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Telomeres are repeating sequences of nucleic acids that cap the ends of chromosomes in the cell nucleus and stop actual gene-coding DNA from being chopped off when a cell divides. The mechanisms of DNA replication require extra leg room at the ends of the strand, a trailing sequence that is not copied over to the new strand under assembly - and the primary role of telomeres is to be the part that is dropped on the floor. A little of their length is thus lost with every cell division. This shortening acts as a clock to count cell divisions, and cells with very short telomeres stop replicating - they either enter cellular senescence (which ideally then causes the immune system to destroy them) or destroy themselves directly via programmed cell death mechanisms.
Telomere length is more dynamic than this simple picture, however. In some cell populations, such as the various types of stem cell that maintain tissues and produce new cells to replace those lost or damaged, an enzyme called telomerase continually lengthens telomeres so as to allow a cell lineage to continue dividing indefinitely.
Ordinary, non-stem cell populations exhibit a range of telomere lengths, some short, some long. You might imagine that a population of cells replenished more frequently or recently by stem cells will have longer telomeres on average. A population that is receiving less support might have shorter telomeres. Researchers have shown that a higher proportion of short telomeres in white blood cells correlates well with ill health or stress, and somewhat correlates with age. Some more complex measures of telomere length, a step above just taking the average, have been shown to correlate well with age, however, and other techniques do a fair job of predicting future life expectancy in laboratory animals.
A few years back a brace of startup biotech companies were aiming to address aspects of aging by lengthening telomeres through the use of telomerase. None of that went anywhere, unfortunately, but it's possible that they were just too early - it is frequently the case that all of the first batch of companies in a new area of biotechnology fail. It's a tough business to be in. I was a skeptic at the time regarding their potential for success based on my expectation that telomere length will prove not to be a root cause of aging.
Nonetheless, researchers are demonstrating extension of life in mice through telomerase these days, but it is as yet unknown as to exactly why this works. Perhaps it makes stem cells work harder to maintain tissues, perhaps there is just one critically limiting type of stem cell or tissue that benefits from more telomerase, or perhaps it involves other effects causes by increased levels of telomerase that have nothing to do with telomere length. It is worth bearing in mind that there are considerable differences in natural levels of telomerase and the resulting telomere dynamics between mice and people, however. Telomerase therapy is probably not something you'd want to just up and try without the research community first obtaining a much greater understanding of why it works to extend life in mice.
Why? Well, the risk of telomere lengthening in humans is cancer. Any mechanism that globally, or possibly even narrowly, extends telomere length in people will raise the risk of suffering cancer. The whole system of telomere dynamics and cellular senescence is intimately tied to the processes of cancer suppression, while all cancers evolve ways of lengthening their telomeres to allow unlimited cell division. Boosting your telomerase levels looks a lot more risky to me than, say, undergoing first generation stem cell transplants.
There continues to be a lot of activity in telomere research and development. The present brace of telomere-related biotech startups are commercializing ways to measure telomere length rather than extend it. The products are tests that will at first add another measure to inform patients on the state of their health, then possibly act as an effective biomarker of biological age, and perhaps later prove useful in further research if it turns out that telomerase-based therapies can be beneficial in humans.
How Long Will You Live?
A growing number of researchers say telomere length is a critically important indicator of how old we really are, and of how many healthy years we may have in front of us. A new industry is sprouting up around the science of longevity, offering telomere testing to the public - and Nobel laureate Elizabeth Blackburn is a notable part of it. Her company, Telome Health, is set to launch a telomere test later this year, joining a handful of others that already do. Like a cholesterol or blood-pressure test, telomere testing could one day become standard in doctors' offices.
And maybe in the future, we'll be able to slow or reverse the effects of aging -the vision of researchers searching for ways to boost telomerase, a goal already achieved in lab mice. Some are already marketing so-called "telomerase activators" to a public hungry for ways to stop the clock, although no such drugs have been approved. With so many companies rushing to come on board, "there's a lot of weird stuff going on out there," cautions Jerry W. Shay of the University of Texas Southwestern Medical Center, an expert on cell biology and telomere length.
Certainly you should be looking askance at any group that's selling herbal "telomerase activators" - it's the standard garbage from the supplement marketplace, and sadly that's the place that formerly funded companies doing original research often end up. It's hard to make money doing something useful in medical research, but depressingly easy to make money doing something useless in the supplement business. The traditional model here is to grab a little research that's somewhat relevant, scare up a bunch of Chinese herb extracts, and then hope that if you market the thing hard enough it'll overcome the obvious ineffectiveness and pointlessness. If you can buy out the shell of a company formerly doing research to try to profit from its one-time reputation, then all the better. Caveat emptor is the watchword, as ever.
So where do telomeres fit in the taxonomy of cause versus secondary effect in aging? Because of the dynamic nature of telomere length I'm given to think that it's a secondary effect: get sick and average telomere length in white blood cells shortens; get well and it lengthens again. This sounds very much like a system responding to circumstances, and those circumstances most likely include the general level of cellular damage, inflammation, and metabolic waste products - all of which grow with age. As for so many other similar questions about aging, the fastest and cheapest way to answer this question about telomere length is to implement the Strategies for Engineered Negligible Senescence (SENS): build the biotechnologies to repair these forms of damage and then see what happens to telomere length once its done. That is a good deal easier at this point than obtaining a full understanding of the aging of human biology.
None of the above precludes short telomeres from causing further damage or changes of their own, of course. Aging proceeds as a cascade of harmful effects as damage causes further damage and flailing biological systems cope badly with the new circumstances they find themselves in. Here is a recent article on how telomere length can impact gene expression and thus the operation of metabolism in a previously unsuspected way, for example:
Telomeres Affect Gene Expression
DUX4, a gene responsible for the genetic disease facioscapulohumeral muscular dystrophy (FSHD), is normally silenced because it sits next to a telomere - a protective DNA sequence that caps the ends of chromosomes, according to [a recent study]. But as telomeres shorten, as they do with age, DUX4 expression climbs, which may explain the late onset of FSHD. Another gene, called FRG2, which sits 100 kilobases away from the telomere, is also affected by telomere length.
"This was completely unexpected. We think that DUX4 and FRG2 are the tip of an iceberg." Due to shrinking telomeres, many genes might gradually become more active as we get older, which may be important for several diseases of old age. "This represents a very significant general advance in our understanding of how telomere shortening may affect human biology."
Membrane Pacemaker Hypothesis and Ames Dwarf Mice
Permalink | View Comments (0) | Post Comment | | Posted by Reason
Ames dwarf mice lack growth hormone and as a consequence live much longer than their peers. Here the biochemistry of this lineage is considered in light of the membrane pacemaker hypothesis of aging, which suggests that the degree of resistance to oxidative damage in cell membranes is a driving factor in determining longevity. Thus similar species with different proportions of more resistant and less resistant molecules making up their cell membranes have different life spans. Is it possible that this can happen within a species thanks to genetic engineering of the sort that produced the Ames dwarf mouse lineage?
Membrane fatty acid (FA) composition is correlated with longevity in mammals. The "membrane pacemaker hypothesis of ageing" proposes that animals which cellular membranes contain high amounts of polyunsaturated FAs (PUFAs) have shorter life spans because their membranes are more susceptible to peroxidation and further oxidative damage. It remains to be shown, however, that long-lived phenotypes such as the Ames dwarf mouse have membranes containing fewer PUFAs and thus being less prone to peroxidation, as would be predicted from the membrane pacemaker hypothesis of ageing.
Here, we show that across four different tissues, i.e., muscle, heart, liver and brain as well as in liver mitochondria, Ames dwarf mice possess membrane phospholipids containing between 30 and 60 % PUFAs (depending on the tissue), which is similar to PUFA contents of their normal-sized, short-lived siblings. However, we found that that Ames dwarf mice membrane phospholipids were significantly poorer in n-3 PUFAs. While lack of a difference in PUFA contents is contradicting the membrane pacemaker hypothesis, the lower n-3 PUFAs content in the long-lived mice provides some support for the membrane pacemaker hypothesis of ageing, as n-3 PUFAs comprise those FAs being blamed most for causing oxidative damage. By comparing tissue composition between 1-, 2- and 6-month-old mice in both phenotypes, we found that membranes differed both in quantity of PUFAs and in the prevalence of certain PUFAs. In sum, membrane composition in the Ames dwarf mouse supports the concept that tissue FA composition is related to longevity.
At some point a research group will find a way to alter only membrane constituent molecules and no other factors in laboratory mice, which should go some way towards quantifying the effect on aging and longevity. The challenge with using any of the well known long-lived lineages of mice is that many aspects of their metabolism are different - it is difficult to point to any one of those and talk about how important it may or may not be to extended longevity given the presence of the others.
Link: http://www.ncbi.nlm.nih.gov/pubmed/23640425
On Methionine Restriction
Permalink | View Comments (0) | Post Comment | | Posted by Reason
Levels of the essential amino acid methionine in the diet appear to be involved in generating the beneficial effects of calorie restriction on health and longevity. Some portion of the resulting changes in the operation of metabolism is based on sensing low levels of methionine. It is thus possible that humans might obtain benefits comparable to those generated by calorie restriction from a sensibly constructed low-methionine diet with a normal calorie intake. The research in support of this supposition is still sparse in comparison to that for calorie restriction, however.
It was first reported in 1993 that rats subjected to a diet restricted in methionine (MR) enjoyed comparable life spans to rats that were on caloric restriction (CR). In the first experiments, methionine was reduced to ⅕ its normal level in the diet, and growth of the rats was severely stunted. We can't live entirely without methionine - the body would not be able to make any proteins at all. Restricting methionine is likely to have impacts on growth, health, and wellbeing that are as yet unstudied in humans. Rats fed a diet without methionine developed steatohepatitis (fatty liver), anemia and lost two thirds of their body weight over 5 weeks. In one experiment where methionine was severely restricted but not eliminated entirely, ⅕ of the mice died, and the other ⅘ went on to live longer than control mice.
Here's a clue about why methionine is special. The instructions for making proteins is coded into DNA, via the genetic code, which specifies words of 3 DNA letters, each corresponding to one of the 20 amino acids. The genetic code also contains "punctuation", instructions to start and stop. The "start codon" is also the word for methionine. Every chain of amino acids that the body constructs begins with methionine. No methionine - no protein synthesis. A shortage of methionine means that the body is inhibited in making every kind of protein. More genes are expressed (more proteins synthesized) as the body grows older. Perhaps methionine restriction is putting a brake on this production of extra proteins that are not produced when we're young, and that contribute to aging.
Methionine restriction in practice involves eating foods that are low in methionine. Though all protein has methionine, some protein sources are much lower in methionine than others. All animal sources (including milk and especially eggs) are high in methionine. So a methionine-restricted diet is a vegan diet, not just any vegan diet, but a subset of vegan protein sources. There appear to be no general rules. For example, almonds are a good source of low-methionine protein, but Brazil nuts are terrible. Even a strict vegan diet would only reduce methionine intake by about 1/2. Extrapolating from the rodent experiments, we may need to reduce by ~ 3/4 before crossing a threshold where benefits kick in.
Link: http://joshmitteldorf.scienceblog.com/2013/05/13/could-cutting-this-one-nutrient-make-you-live-longer/
Be Dubious About Longevity Hotspots
Permalink | View Comments (0) | Post Comment | | Posted by Reason
"Cui bono?", "to whose benefit?", is a question that should never be far from mind. It is rarely the case that the loudest threads in our grand, connected cultural conversation represent the best, the most useful, or the most virtuous of what is possible. That is just as true in any subculture as it is in the mainstream: follow the money and much becomes clear.
Longevity hotspots might not be a term familiar to you, but Blue Zones might be thanks to a fair degree of publicity for that latter term. They mean the same thing, but the latter is a brand rather than a description. A small industry associated with this brand is devoted to promoting the idea that some parts of the world exhibit pockets of exceptional human longevity. It is convenient for various businesspeople to act as though this is proven beyond a doubt and that the root causes involve aspects of local culture, diet, and lifestyle that can be packaged up and sold. So the world goes on: this sort of thing is a textbook example of how small science projects on minor aspects of human longevity can spawn commercial monstrosities set on muddying the waters, promoting myths, and profiting from the credulous.
It is by no means certain that longevity hotspots exist in actuality, or at least not in the sense that Blue Zone business ventures would like you to think, but those most interested in carrying on a dialog on this topic - i.e. marketing folk involved in tourism, diet, lifestyle coaching, and so forth - don't really care to hear that message. Nonetheless:
Designating longevity hotspots: cautions concerning the instability of per capita centenarian estimates
Estimates of per capita centenarians in a Utah population varied between one per 12,864 and one per 4,675, depending on the data that were used, the population assumptions that were made, and the boundary limits that were employed. In general, caution is warranted in claims about the existence of longevity hotspots.
Performing any sort of statistical study on human populations in a given geographical area, even on something as apparently simple as age, is enormously complex. People move and data is ever incomplete or outright false. Some locations attract the wealthy in large numbers, a demographic already well correlated with greater life expectancy. When a region in the US with good demographic data can produce a threefold range of results for a simple population question, one has to wonder about the accuracy of other studies - and the smaller the group the less helpful that statistical procedures become.
This is not to say that there is nothing to be learned by comparing different populations with different lifestyles, but I would be extremely surprised to see the end results be anything other than additional support for the value of exercise and calorie restriction (and derived measures such as body mass index). These line items strongly correlate with health in large statistical studies.
Neither exercise nor calorie restriction will let you reliably live to see 100, however. The only thing that can achieve that goal is significant progress in new medical science. Longevity hotspots are, like so much of what is discussed in relation to aging these days, nothing but a sideshow - something that occupies time and energy and attention, and all to no good end. That the data is most likely flawed and what little science there was is now largely buried beneath an industry that strives to make money by promoting magical thinking and ignorance just makes the joke a little more black.
Amphibian Species with a Chemical Defence Live Longer
Permalink | View Comments (0) | Post Comment | | Posted by Reason
When it comes to evolutionary influences on longevity, the evidence supports the idea that species with a high mortality rate due to external causes (e.g. being eaten) will tend to be short-lived. There is no evolutionary pressure to develop the biological mechanisms that will lead to longer reproductive lives if near all individuals are killed comparatively early in life. This study is a novel way to add further supporting evidence to this point of view:
Evolutionary hypotheses for ageing generally predict that delayed senescence should evolve in organisms that experience lower extrinsic mortality. Thus, one might expect species that are highly toxic or venomous (i.e. chemically protected) will have longer lifespans than related species that are not likewise protected. This remarkable relationship has been suggested to occur in amphibians and snakes.
First, we show that chemical protection is highly conserved in several lineages of amphibians and snakes. Therefore, accounting for phylogenetic autocorrelation is critical when conservatively testing evolutionary hypotheses because species may possess similar longevities and defensive attributes simply through shared ancestry. Herein, we compare maximum longevity of chemically protected and nonprotected species, controlling for potential nonindependence of traits among species using recently available phylogenies.
Our analyses confirm that longevity is positively correlated with body size in both groups which is consistent with life-history theory. We also show that maximum lifespan was positively associated with chemical protection in amphibian species but not in snakes. Chemical protection is defensive in amphibians, but primarily offensive (involved in prey capture) in snakes. Thus, we find that although chemical defence in amphibians favours long life, there is no evidence that chemical offence in snakes does the same.
Link: http://www.ncbi.nlm.nih.gov/pubmed/23638626
Children of Long-Lived Parents Resistant to Dementia
Permalink | View Comments (0) | Post Comment | | Posted by Reason
Some degree of human longevity is genetic rather than the result of environment and lifestyle choice; researchers have guessed that perhaps 25% of variations are genetic, but this is hardly a firm number. It appears to be the case that survival at extreme old age is more influenced by genetic variations than it is in early old age, for example. Given that some predisposition to longevity is thus inherited, it isn't surprising to find that risk levels for specific conditions of aging also correlate with familial longevity:
Based on comparisons of people in their 90s, their spouses, siblings, children and their children's spouses, researchers found that the offspring of people with exceptional longevity were about 40 percent less likely than peers to be cognitively impaired between ages 65 and 79. "It's not necessarily that these individuals never become cognitively impaired, but what it seems like is that there is a delayed onset of cognitive impairment."
For the new study, the researchers used data on cognitive impairment from 1,870 people who are part of the Long Life Family Study, which includes volunteer participants in New York, Massachusetts, Pennsylvania and Denmark. The study included 1,510 people with a family history of longevity and 360 of their spouses, but for this study, researchers used information on just the volunteers who were 89 years old or older when they were recruited.
Overall, the researchers found that about 6 percent of the volunteers' children were cognitively impaired between ages 65 and 79 years old, compared to 13 percent of their spouses and about 11 percent of their cousins. Among the study's long-lived older generation, participants were just as likely to be cognitively impaired by about age 90 as their siblings or spouses. "These families seem relatively protected, but once they reach extreme old age - say after 90 (years old) - their rates of cognitive impairment become comparable."
Link: http://www.reuters.com/article/2013/05/06/us-family-dementia-idUSBRE9450VL20130506
First Steps
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• All of Fight Aging!, with the exception of the introductory articles, is published under the Creative Commons Attribution 3.0 license. In short, this means that you are encouraged to republish and rewrite Creative Commons licensed Fight Aging! content in any way you see fit, the only requirements being that you (a) link to the original, (b) attribute the author, and (c) attribute Fight Aging!.
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A Reminder Of Kurzweil's Projections
Permalink | View Comments (0) | Post Comment | | Posted by Reason
InformationWeek provides a reminder of Ray Kurzweil's trend-based predictions for the next few decades: "Fifteen years from now, it'll be a very different world. We'll have cured cancer and heart disease, or at least rendered them to manageable chronic conditions that aren't life threatening. We'll get to the point where we can stop the aging process and stave off death ... By the 2020s, we'll be adding a year of longevity or more for every year that passes ... scientists will be able to regrow our own cells, tissues, and even whole organs, and then introduce them into our bodies, all without surgery. As part of what he calls the 'emerging field of rejuvenation medicine,' new tissue and organs will be built out of cells that have been made younger." These end results of present trends are not pulled from thin air; they will come to pass if we work to ensure these trends continue - although I suspect the incompressible length of a business cycle run by humans means that the timeline is optimistic by a decade or more.
Link: http://www.informationweek.com/shared/printableArticle.jhtml?articleID=195200003
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Upcoming events
From Forensics Wiki
Revision as of 15:40, 30 March 2009 by Frnzxguy (Talk | contribs)
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PLEASE READ BEFORE YOU EDIT THE LISTS BELOW
Events should be posted in the correct section, and in date order. An event should NEVER be listed in more than one section (i.e. Ongoing/Continuous events should not be listed in Scheduled Training). When events begin the same day, events of a longer length should be listed first. New postings of events with the same date(s) as other events should be added after events already in the list. If a provider offers the same event at several locations simultaneously, the listing should have a single (ONE) entry in the list with the date(s) and ALL locations for the event. Please use three-letter month abbreviations (i.e. Sep, NOT Sept. or September), use two digit dates (i.e. Jan 01 NOT Jan 1), and use date ranges rather than listing every date during an event(i.e. Jan 02-05, NOT Jan 02, 03, 04, 05).
Some events may be limited to Law Enforcement Only or to a specific audience. Such restrictions should be noted when known.
This is a BY DATE listing of upcoming events relevant to digital forensics. It is not an all inclusive list, but includes most well-known activities. Some events may duplicate events on the generic conferences page, but entries in this list have specific dates and locations for the upcoming event.
This listing is divided into four sections (described as follows):
1. Calls For Papers - Calls for papers for either Journals or for Conferences, relevant to Digital Forensics (Name, Closing Date, URL)
2. Conferences - Conferences relevant for Digital Forensics (Name, Date, Location, URL)
3. On-Going / Continuous Training - Training opportunities that are either always available online/distance learning format (start anytime) or that are offered the same time every month (Name, date-if applicable, URL)
4. Scheduled Training Courses - Training Classes/Courses that are scheduled for specific dates/locations. This would include online (or distance learning format) courses which begin on specific dates, instead of the "start anytime" courses listed in the previous section. (Provider, URL) (note: this has been moved to its own page.)
The Conference and Training List is provided by the American Academy of Forensic Sciences (AAFS) Digital and Multimedia Sciences Section Listserv. (Subscribe by sending an email to listserv@lists.mitre.org with message body containing SUBSCRIBE AAFS-DIGITAL-MULTIMEDIA-LIST) Requests for additions, deletions or corrections to this list may be sent by email to David Baker (bakerd AT mitre.org).
Contents
Calls For Papers
Please help us keep this up-to-date with deadlines for upcoming conferences that would be appropriate for forensic research.
Title Due Date Notification Date Website
1st International Workshop on Managing Insider Security Threats Mar 22, 2009 Apr 20, 2009 http://isyou.hosting.paran.com/mist09/data/MIST09-CFP.pdf
Systematic Approaches to Digital Forensic Engineering International Workshop Mar 25, 2009 Apr 15, 2009 http://conf.ncku.edu.tw/sadfe/sadfe09/cfp.html
Layer One - 2009 Apr 01, 2009 Apr 15, 2009 http://layerone.info/
Recent Advances in Intrusion Detection (RAID) International Symposium Apr 05, 2009 Jun 08, 2009 http://www.rennes.supelec.fr/RAID2009/cfp.html
ACM CCS 2009 Apr 2009 http://www.sigsac.org/ccs
3rd International Workshop on Computational Forensics Apr 24, 2009 May 22, 2009 http://iwcf09.arsforensica.org/
ICDF2C 2009 Apr 30, 2009 Jun 30, 2009 http://www.d-forensics.org/
Usenix Lisa 2009 Apr 30, 2009 Jul 07, 2009 http://www.usenix.org/events/lisa09/cfp/
New Security Paradigms Conference 2009 Apr 2009 http://www.nspw.org/current/
International Workshop on Information Forensics and Security (WIFS) May 22, 2009 Aug 21, 2009 http://www.wifs09.org/index.php?option=com_content&view=article&id=15&Itemid=42
ACSAC 25 Jun 01, 2009 Aug 17, 2009 http://www.acsac.org
IMF 2009 Jun 01, 2009 Jul 22, 2009 http://www.imf-conference.org/
American Academy of Forensic Sciences 2010 Annual Meeting Aug 01, 2009 Nov, 2009 http://www.aafs.org/default.asp?section_id=meetings&page_id=aafs_annual_meeting
IEEE Symposium on Security and Privacy 2010 Nov 2009
ShmooCon 2010 Dec 2009 Jan 2010 http://www.shmoocon.org/cfp.html
AusCERT Conference 2010 Dec 2009 Jan 2010 http://conference.auscert.org.au/conf2010/cfp2010.html
Conferences
Title Date/Location Website
5th Cyber Security and Information Intelligence Research Workshop Apr 13-15
Oak Ridge National Laboratory, TN
http://www.csiir.ornl.gov/csiirw/
Blackhat Europe Apr 14-17
Amsterdam, The Netherlands
https://www.blackhat.com/html/bh-europe-09/bh-eu-09-main.html
IEEE Canadian Conference on Electrical and Computer Engineering(CCECE 2009) May 03-06
St. John's, Newfoundland & Labrador, Canada
http://www.ieee.ca/ccece09/index_en.php
Ohio HTCIA Spring Training Conference May 12-14
Kirtland, OH
http://ohiohtcia.org/conference.html
AusCERT2009 May 17-22
Gold Coast, Australia
http://conference.auscert.org.au/conf2009/
Computer Security Institute: Security Exchange May 17-22
Las Vegas, NV
http://www.csisx.com/
ADFSL 2009 Conference on Digital Forensics, Security and Law May 20-22
Burlington, VT
http://www.digitalforensics-conference.org
Fourth International Workshop on Systematic Approaches to Digital Forensic Engineering (co-located with IEEE Security & Privacy) May 21
Oakland, CA
http://conf.ncku.edu.tw/sadfe/sadfe09/
LayerOne 2009 Security Conference May 23-24
Anaheim, CA
http://layerone.info/
Mobile Forensics World 2009 May 26-30
Chicago, IL
http://www.mobileforensicsworld.com
2009 Techno Security Conference May 31-Jun 03
Myrtle Beach, SC
http://www.techsec.com/index.html
USENIX 2009 Jun 14-19
San Diego, CA
http://www.usenix.org/events/usenix09/
IEEE ICC Communication and Information Systems Security (CISS) Symposium Jun 14-18
Dresden, Germany
http://www.ieee-icc.org/2009/
1st International Workshop on Managing Insider Security Threats Jun 15-19
Purdue University, IN
http://isyou.hosting.paran.com/mist09/
SANS SANSFIRE 2009 Jun 13-22
Baltimore, MD
http://www.sans.org/sansfire09/
1st Workshop on Internet Multimedia Search and Mining (IMSM'09) Jul 03
Cancun, Mexico
http://research.microsoft.com/en-us/um/people/xshua/imsm/index.html
Blackhat USA 2009 Jul 25-30
Las Vegas, NV
https://www.blackhat.com/
DefCon 17 Jul 31-Aug 02
Las Vegas, NV
http://www.defcon.org/
Usenix Security Sypmosium Aug 10-14
Montreal, Quebec, Canada
http://www.usenix.org/events/sec09/
3rd International Workshop on Computational Forensics Aug 13-14
The Hague, The Netherlands
http://iwcf09.arsforensica.org/
Digital Forensic Research Workshop Aug 17-19
Montreal, Quebec, Canada
http://www.dfrws.org
Triennial Meeting of the European Academy of Forensic Science Sep 08-11
Glasgow, Scotland, UK
http://www.eafs2009.com/
IMF 2009 Sep 15-17
Stuttgart, Germany
http://imf-conference.org/
Hacker Halted USA 2009 Sep 20-24
Miami, FL
http://www.hackerhalted.com
Recent Advances in Intrusion Detection (RAID) International Symposium Sep 23-25
Saint-Malo, Brittany, France
http://www.rennes.supelec.fr/RAID2009/index.html
The 1st International ICST Conference on Digital Forensics & Cyber Crime (ICDF2C) Sep 30 - October 02
Albany, NY
http://www.d-forensics.org/
First IEEE Workshop on Information Forensics and Security Dec 06-09
London, England
http://www.wifs09.org/
AAFS Annual Meeting Feb. 22-27
Seattle, WA
http://www.aafs.org/default.asp?section_id=meetings&page_id=aafs_annual_meeting
On-going / Continuous Training
Title Date/Location Website
DISTANCE LEARNING
Basic Computer Examiner Course - Computer Forensic Training Online Distance Learning Format http://www.cftco.com
Linux Data Forensics Training Distance Learning Format http://www.crazytrain.com/training.html
SANS On-Demand Training Distance Learning Format http://www.sans.org/ondemand/?portal=69456f95660ade45be29c00b0c14aea1
Champlain College - CCE Course Online / Distance Learning Format http://extra.champlain.edu/cps/wdc/alliances/cce/landing/
Las Positas College Online Computer Forensics Courses http://www.laspositascollege.edu
RECURRING TRAINING
MaresWare Suite Training First full week every month
Atlanta, GA
http://www.maresware.com/maresware/training/maresware.htm
Evidence Recovery for Windows Vista™ First full week every month
Brunswick, GA
http://www.internetcrimes.net
Evidence Recovery for Windows Server® 2003 R2 Second full week every month
Brunswick, GA
http://www.internetcrimes.net
Evidence Recovery for the Windows XP™ operating system Third full week every month
Brunswick, GA
http://www.internetcrimes.net
Computer Forensics Training and CCE™ Testing for Litigation Support Professionals Third weekend of every month(Fri-Mon)
Dallas, TX
http://www.md5group.com
See Also
References
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Deallocation of static STL i.e. map
Newbie Member
24Oct2007,17:55 #1
I am getting coredump on linux (Red Hat 3.4.6-8).
I am using gcc (GCC) 3.4.6 20060404 compiler.
The problem is I have used a static map inside a function (making the map local to the function). This function is in libStreet.so Now at run time another host application is loading this XYZ.so, using it and then unloading it.Everything works fine,But when host application closes[i.e. exits] It is giving coredump.If I change static map to static vector then it works fine[no coredump while closing host application.].
I know that the static variables get deallocated after exiting from main() of host application, please let me know if my understanding is correct.
In my case host application crashes after exiting main, however replacing the map with a vector solves the problem. Can anyone please help on this issue, Thanks.
Please note that the same code works fine on Sun Solaris, gcc <> version
Go4Expert Founder
24Oct2007,21:13 #2
Moved to C-C++ forum for better responses.
Team Leader
24Oct2007,22:03 #3
Try using Valgrind, see if that gives you any info.
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About this Journal Submit a Manuscript Table of Contents
Stroke Research and Treatment
Volume 2012 (2012), Article ID 391538, 15 pages
doi:10.1155/2012/391538
Review Article
Stroke Prevention: Managing Modifiable Risk Factors
1Stroke Unit, Department of Neuroscience, University of Tor Vergata, Viale Oxford 81, 00133 Rome, Italy
2Santa Lucia Foundation, IRCCS, Viale Ardeatina 306, 00134 Rome, Italy
Received 30 July 2012; Accepted 8 October 2012
Academic Editor: Cristina Sierra-Benito
Copyright © 2012 Silvia Di Legge et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Prevention plays a crucial role in counteracting morbidity and mortality related to ischemic stroke. It has been estimated that 50% of stroke are preventable through control of modifiable risk factors and lifestyle changes. Antihypertensive treatment is recommended for both prevention of recurrent stroke and other vascular events. The use of antiplatelets and statins has been shown to reduce the risk of recurrent stroke and other vascular events. Angiotensin-converting enzyme inhibitors (ACEIs) and angiotensin II receptor blockers (ARBs) are indicated in stroke prevention because they also promote vascular health. Effective secondary-prevention strategies for selected patients include carotid revascularization for high-grade carotid stenosis and vitamin K antagonist treatment for atrial fibrillation. The results of recent clinical trials investigating new anticoagulants (factor Xa inhibitors and direct thrombin inhibitors) clearly indicate alternative strategies in stroke prevention for patients with atrial fibrillation. This paper describes the current landscape and developments in stroke prevention with special reference to medical treatment in secondary prevention of ischemic stroke.
1. Introduction
It is estimated that 530,000 people experience each year a new ischemic stroke (IS) in the USA and on average every 40 seconds someone in the same country has a stroke [1]. In terms of mortality, stroke ranks number 4 among all causes of death after heart disease, cancer, and chronic lower respiratory disease [2]. However, it remains the first cause of adult neurological disability in developed countries [3]. About 80% of patients come back home, but about half of them needs permanent or temporary help in the home setting [4]. Data from the Framingham Heart Study showed that stroke incidence is declining over time: in particular, the age-adjusted incidence of first stroke per 1000 person-years has decreased from 7.6 for men and 6.2 for women in the period 1950–1977 to 6.2 for men and 5.1 for women in the period 1990–2004 [5]. However, a recent systematic review has shown a 42% decrease in stroke incidence in the past four decades in high-income countries and a greater than 100% increase in stroke incidence in low-to-middle income countries [6]. On the contrary, stroke severity did not vary across these periods [5].
Prevention plays a crucial role in counteracting morbidity and mortality related to IS. It has been estimated that 50% of stroke are preventable through control of modifiable risk factors and lifestyle changes. Recently, stroke prevention has been set as one of the priorities by an international community of leaders involved in this field [7], and the American Heart Association (AHA) and the American Stroke Association (ASA) have published updated guidelines for secondary prevention of stroke [8]. Among stroke risk factors, transient ischemic attacks (TIAs) confer an important short-term risk of stroke (10% within 90 days and 5% within 2 days) [9]; hypertension plays a crucial role in the risk of both ischemic stroke and intracranial hemorrhage [10]. Diabetes mellitus nearly triples while current cigarette smoking doubles this risk [11]. Atrial fibrillation, although often asymptomatic and undetected, is an important risk factor for stroke, increasing stroke risk about 5-fold throughout all ages so that its relevance could be underestimated [12, 13]. Patients with low concentrations of HDL cholesterol have been found to be at higher risk of stroke [14]. Further, depressive symptoms have been increasingly recognized as a risk factor (4-fold higher) for stroke/TIA [15]. Primary prevention strategies that work in primary prevention of IS are treating hypertension (HTN), using statins and angiotensin-converting enzyme inhibitors (ACEIs), and anticoagulation in nonvalvular atrial fibrillation. Attention to lifestyle factors is routinely warranted in both primary and secondary IS prevention: aerobic exercise to counteract inactivity, weight loss in obesity, glucose control in diabetics, smoking cessation, and diet. Antihypertensive treatment is recommended for both prevention of recurrent stroke and other vascular events. Cholesterol lowering with statins and antiplatelets have been shown to reduce the risk of recurrent stroke and other vascular events; ACEIs or angiotensin II receptor blockers (ARBs) are indicated in stroke prevention because they promote vascular health; effective secondary-prevention strategies for selected patients include carotid revascularization for high-grade carotid stenosis and vitamin K antagonist (i.e., warfarin) treatment for atrial fibrillation. Among potentially modifiable risk factors, consensus does not exist on the role of treating, among others, hyperhomocysteinemia, coagulation disorders, and patent foramen ovale. The results of recent clinical trials investigating new anticoagulants (factor Xa inhibitors and direct thrombin inhibitors) clearly indicate alternative strategies in stroke prevention for patients with atrial fibrillation. Recently, the American College of Chest Physicians [16] and the AHA/ASA [17] have published evidence-based clinical practice guidelines for prevention of stroke in nonvalvular atrial fibrillation and antithrombotic therapy for valvular disease based on the optimal balance of thrombotic and hemorrhagic risk. The results of RCTs testing safety and efficacy of antiplatelet treatments alternative to aspirin as cilostazol, sarpogrelate, and triflusal are discussed, as well as clinical indications for combined antithrombotic medications.
This paper describes the current landscape and developments in stroke prevention with special reference to medical treatment in secondary IS prevention.
2. Hypertension Control
Arterial hypertension (HTN) is the single most important modifiable risk factor for stroke. HTN contributes to 60% of all strokes (through the following mechanisms: atheroma in carotids, vertebral arteries and aortic arch; friability of small cerebral arteries; left ventricular dysfunction and atrial fibrillation). There is a close, continuous, and approximately linear relationship between blood pressure (BP) levels and primary incidence of stroke in both hypertensive and normotensive populations. A 5-year reduction of 5-6 mm Hg diastolic blood pressure (DBP) (with mainly diuretics and beta blockers) has been associated with a 42% relative risk reduction (RRR) of first stroke [18]. Randomized trials on primary stroke prevention in middle-aged and elderly populations confirmed that treating HTN reduces the incidence of stroke. Two placebo-controlled trials (the first using chlorthalidone and atenolol, the second using nitrendipine, with the possible addition of enalapril and hydrochlorothiazide) have demonstrated 36% [19] and 42% [20] RRR for first ischemic stroke in treating systolic blood pressure (SBP) as compared to placebo. More recently, the LIFE study [21] compared losartan-based regimen versus atenolol-based regimen in 9,193 hypertensive patients with a mean followup of 4.8 years. While comparable reductions in BP were observed, losartan treatment was associated with a significant 25% risk reduction versus atenolol of fatal and nonfatal stroke. Losartan was better tolerated and seemed to confer benefits beyond reduction in BP. A recent analysis of RCTs compared first-line calcium channel blockers (CCBs) with other antihypertensive classes, in order to determine whether CCBs reduced the incidence of major adverse cardiovascular events compared to the other antihypertensive classes [22]. Eighteen RCTs, with at least 100 randomized hypertensive participants and with a followup of at least two years, with a total of 141,807 participants, were included. Although some results were not enough robust to change practice, CCBs reduced stroke as compared to beta blockers, ACEIs, and ARBs. However, diuretics were the preferred first-line treatment over CCBs to optimize the reduction of cardiovascular events and congestive heart failure. Few trials have focused on antihypertensive therapy for prevention of recurrent stroke. Two studies using ACEIs have demonstrated that, for patients with hypertension, effective control of BP reduces the risk of recurrent stroke. The Perindopril Protection against Recurrent Stroke Study (PROGRESS) [23] was started in 1996 to answer this question. In this study, about 6,000 hypertensive patients with a prior TIA or stroke in the last 5 years were randomized to receive aggressive HTN treatment (perindopril 4 mg ± indapamide 2.5 mg) versus usual care (1 drug) or placebo (1 or 2 placebo). Patients were followed up for 4 years, and the primary outcome was total strokes. Aggressive treatment was associated with a 43% RRR in stroke risk versus usual care (mean BP reduction was 12/5 and 5/3 mm Hg). The HOPE trial [24] was a RCT, with 267 participating hospitals in 19 countries. There were 9,297 patients with vascular disease or diabetes plus an additional risk factor, followed up for 4.5 years (11% of them had prior TIA or IS). Patients were randomized to receive ramipril 10 mg versus placebo. The rate of stroke and TIA were assessed. Reduction in BP was modest (3.8 SBP and 2.8 DBP). The RRR for any stroke was 32%, and the RRR of fatal stroke was 61% (17 versus 44 events) favouring ramipril. Benefits were consistent across patients’ subgroups.
More recently, other trials have tested safety and efficacy of ARBs, when used alone or in combination with an ACEI, in preventing stroke recurrence in high-risk populations. In the Prevention Regimen for Effectively Avoiding Second Strokes (PRoFESS) trial [25], 20,332 ischemic stroke patients were randomized to telmisartan 80 mg or placebo. After a mean treatment of 2 years, telmisartan showed a nonsignificant lower rate of recurrent stroke versus placebo (880 versus 934; hazard ratio (HR) 0.95; 95% confidence interval (CI), 0.86–1.04). However, in a post hoc analysis, a significantly reduced number of strokes was observed in the telmisartan group compared to placebo (533 versus 608; HR 0.88; 95% CI 0.78–0.99; ) [26]. The Ongoing Telmisartan Alone and in Combination with Ramipril Global Endpoint Trial (ONTARGET) and The Telmisartan Randomized Assessment Study in aCE-iNtolerant subjects with Cardiovascular Disease (TRANSCEND), compared telmisartan 80 mg versus ramipril 10 mg, and telmisartan 80 mg versus placebo (in patients intolerant to ACEIs), respectively. In the stroke subgroup, telmisartan 80 mg showed a trend toward reducing recurrent stroke versus ramipril 10 mg (HR 0.91; 95% CI, 0.79–1.05). In a combined analysis of PRoFESS and TRANSCEND, the incidence of the composite of stroke, myocardial infarction, or vascular death was 12.8% for telmisartan versus 13.8% for placebo (HR 0.91; 95% CI, 0.85–0.98; ) [27]. The MOSES study showed for the first time superiority of an ARB (eprosartan) compared with a calcium channel antagonist (nitrendipine) in antihypertensive treatment for secondary stroke prevention [28]. In this study, 1,405 high-risk hypertensive stroke patients were randomized. BP was reduced to a comparable extent without any significant differences between the 2 groups (from 150.7/84 mm Hg and 152.0/87.2 mm Hg with eprosartan and nitrendipine therapy to 137.5/80.8 mm Hg and 136.0/80.2 mm Hg, resp.). For the same level of BP control, eprosartan was significantly more effective than nitrendipine in reducing cerebrovascular morbidity and mortality (102 strokes in the eprosartan and134 in the nitrendipine group; ). Based on all these studies, the renin-angiotensin system (RAS) blockers (ACEIs and ARBs) are guideline-recognized, highly effective antihypertensive agents which offer benefits that extend beyond BP reduction alone. Experimental and clinical data suggest that reducing the activity of the RAS may have cerebroprotective effects. The Angiotensin II has hemodynamic properties (potent peripheral vasoconstrictor, stimulates aldosteron), action on endothelium (mediates endothelium dysfunction, vascular smooth cells hypertrophy), stimulates oxidative stress, including oxidation of low-density lipoprotein cholesterol (LDL-C), and inflammation (associated with expression of cellular adhesion molecules, chemotactic and proinflammatory cytokines), thus contributing to endothelial damage and arterial wall injury [29]. In a recent meta-analysis, after controlling for effects on BP control, ARBs appeared to be more effective than either ACEIs or β-blockers in stroke prevention; however, CCBs were superior to RAS blockers in stroke prevention [30]. In conclusion, epidemiological studies and clinical trials confirmed the hypothesis of managing HTN to counteract the risk of stroke. The recommendation [8] to prevent recurrent stroke is to treat HTN aggressively (Class I, Level of Evidence A); although an absolute target of BP level has not been clearly defined, benefit has been associated with an average reduction of 10/5 mm Hg, and normal BP levels have been defined as <120/80 mm 5 Class IIa, Level of Evidence B). These recommendations extend to all patients with prior IS or TIA, irrespective of history or HTN, if BP reduction is considered appropriate (Class IIa, Level of Evidence B). Studies aimed at comparing different antihypertensive drugs are insufficient and have not reached clear results, but what is clear is that the effects of antihypertensive go beyond the simple control of BP. Diuretics alone or in combination with an ACEI are indicated (Class I, Level of Evidence A). The choice of a specific drug should be individualized based on drug and patient characteristics (extracranial occlusive disease, renal impairment, cardiac disease, and diabetes) (Class IIa, Level of Evidence B).
3. Anticoagulants in Atrial Fibrillation
Patients with nonvalvular atrial fibrillation (AF) are at increased risk of stroke [12, 13]. The efficacy of warfarin over placebo has been consistently demonstrated across studies yielding an overall RRR of 68% (95% CI, 50% to 79%) and an absolute risk reduction (ARR) in annual stroke rate from 4.5% for controls to 1.4% in patients with adjusted-dose warfarin: 31 ischemic strokes will be prevented each year for 1000 patients treated [8]. Warfarin use has been also associated with a modest 1.3% annual rate of major bleeding as compared with 1% for patients on placebo or aspirin. As compared to warfarin, a weaker efficacy of aspirin has been demonstrated in a pooled analysis of three clinical trials [31]. Analysis of an additional arm of the Atrial Fibrillation Clopidogrel Trial with Irbesartan for Prevention of Vascular Events (ACTIVE A) in patients unsuitable for vitamin K antagonists therapy [32] showed superiority of combination therapy with aspirin and clopidogrel over aspirin alone in reducing the rate of stroke (2.4% versus 3.3% per year; ). However, based on the observation that the rate of major vascular events combined with major hemorrhages did not significantly differe between the two groups, aspirin remains the treatment of choice in patients with AF and a clear contraindication to vitamin K antagonists. The European Atrial Fibrillation Trial (EAFT) [33] demonstrated superiority of anticoagulation over aspirin in preventing stroke recurrence in patients with history of IS or TIA: anticoagulation was significantly more effective than aspirin (HR 0.60; 95% CI, 0.41–0.87). The incidence of major bleeding events was higher in the anticoagulation group, but low in both groups (2.8% and 0.9% per year); in absolute terms, 90 vascular events (mainly strokes) could be prevented if 1000 patients were treated with anticoagulation for one year. Aspirin demonstrated to be a valid, although less effective, alternative when anticoagulation was contraindicated, preventing 40 vascular events each year for every 1000 treated patients. More recently, a systematic review of primary prevention studies in patients with nonvalvular AF demonstrated that adjusted-dose warfarin and related oral anticoagulants reduced stroke, disabling stroke and other major vascular events by about one-third when compared with antiplatelet therapy [34]. For patients with AF who suffer an IS or TIA despite therapeutic anticoagulation, increasing the intensity of anticoagulation or adding an antiplatelet does not provide additional protection in preventing stroke while increases the risk of bleeding [31, 34]. Several new antithrombotic agents have been developed for stroke prevention in patients with nonvalvular AF. New drugs for oral anticoagulation that do not exhibit the limitations of vitamin K antagonists include direct factor Xa inhibitors and direct thrombin inhibitors. In the Stroke Prevention Using Oral Thrombin Inhibitor in Atrial Fibrillation (SPORTIF) III (open label, ) [35] and V (double blind, ) [36], safety and efficacy of the oral direct thrombin inhibitor ximelagatran (fixed dose, 36 mg twice daily) were compared to warfarin (adjusted dose, target international normalized ratio (INR) 2.0-3.0) in patients with nonvalvular AF and at least 1 risk factor for stroke. Pooled analysis showed that the efficacy of ximelagatran was comparable (noninferior) with extremely well-controlled warfarin therapy in preventing stroke and systemic embolic events; the primary event rates were 1.65% per year and 1.62% per year in the warfarin and ximelagatran groups, respectively (). In patients with a history of stroke or TIA (about 20% of the SPORTIF population), the event rates were 3.27% per year and 2.83% per year in the warfarin and ximelagatran groups, respectively (). Intracranial hemorrhage occurred at a rate of 0.20% per year with warfarin and 0.11% per year with ximelagatran. Combined rates of minor and major bleeding were significantly lower with ximelagatran than with warfarin (32% per year versus 39% per year; ). The authors concluded that ximelagatran administered without coagulation monitoring or dose adjustment was as effective as well-controlled, adjusted-dose warfarin for prevention of stroke and systemic embolic events and was associated with significantly less total bleedings [36]. More recent trials have compared warfarin to dabigatran (RE-LY) [37], rivaroxaban (ROCKET-AF) [38], and apixaban (ARISTOTLE) [39], while in the AVERROES [40], safety and efficacy of apixaban (at a dose of 5 mg twice daily) were investigated as alternative treatment to aspirin (81 to 324 mg per day) in patients who were not suitable candidates for or were unwilling to receive vitamin K antagonist therapy. Taken together, these trials have demonstrated similar efficacy in patients with AF of these novel anticoagulants in both primary and secondary stroke prevention, with significantly lower incidences of intracranial bleeding, compared with warfarin. Of note, the AVERROES was stopped prematurely because a clear benefit in favor of apixaban was observed: apixaban reduced the risk of stroke or systemic embolism without significantly increasing the risk of major bleeding or intracranial hemorrhage. Based on the results of the RE-LY study [37], and with a modest absolute reduction of stroke or systemic embolism (1.7% versus 1.1%, ), the most successful alternative anticoagulant is dabigatran, given at a dose of 150 mg twice daily. A systematic review and meta-analysis of RCT has been recently performed to compare the efficacy and safety of new oral anticoagulants to those of warfarin in patients with AF [41]. The authors identified 3 studies, including 44,563 patients. Patients randomized to new oral anticoagulants had a decreased risk for all-cause stroke and systemic embolism (relative risk (RR) 0.78, 95% CI, 0.67 to 0.92), ischemic and unidentified stroke (RR 0.87, 95% CI, 0.77 to 0.99), hemorrhagic stroke (RR 0.45, 95% CI, 0.31 to 0.68), all-cause mortality (RR 0.88, 95% CI 0.82 to 0.95), and vascular mortality (RR 0.87, 95% CI, 0.77 to 0.98). Randomization to a new oral anticoagulant was associated with a lower risk for intracranial bleeding (RR 0.49, 95% CI, 0.36 to 0.66). Based on these findings, new oral anticoagulants seem to be superior to warfarin in preventing stroke and systemic embolism in patients with AF. Further, they appear to have a favorable safety profile, making them promising alternatives to warfarin. However, a recent meta-analysis of RCTs has assessed safety and efficacy outcomes in patients with AF treated with warfarin for stroke prevention compared with an alternative thromboprophylaxis strategy [42]. Eight high-quality RCTs published in the last 10 years were selected, with a total of 32,053 patients included. The pooled analysis yielded 55,789 patient-years of followup. Overall, the time spent in the therapeutic range was 55% to 68%. The annual incidence of stroke or systemic embolism in patients with AF taking warfarin was estimated to be 1.66% (95% CI, 1.41%–1.91%). Major bleeding rates varied from 1.40% to 3.40% per year across the studies. The risk of stroke per year was significantly higher in elderly patients (2.27%), female patients (2.12%), patients with a history of stroke (2.64%), and patients reporting no previous exposure to vitamin K antagonists (1.96%). The authors concluded that warfarin used as a stroke prevention agent in patients with AF was associated with a significantly lower rate of recurrent stroke or systemic embolism estimated compared to other antithrombotic treatments.
Conclusions: patients with IS or TIA with paroxysmal (intermittent) or permanent nonvalvular AF should receive anticoagulation with a vitamin K antagonist (target INR 2.5, range 2-3) (Class I, Level of Evidence A); for patients unable to take oral anticoagulants, aspirin alone is recommended (Class I, Level of Evidence A); combination therapy with aspirin plus clopidogrel carries a risk of bleeding similar to that of warfarin and therefore is not recommended for patients with a hemorrhagic contraindication to warfarin (Class III, Level of Evidence B) [8, 17].
Dabigatran is a useful alternative to warfarin for stroke prevention in patients with AF who do not have a prosthetic hearth valve or hemodynamically significant valve disease, severe renal failure (CrCl <15 mL/min), or advanced liver disease (impaired baseline clotting function) (Class I, Level of Evidence B) [8]. New recommendations for the clinical use of dabigatran, rivaroxaban, and apixaban as alternative treatment to warfarin have been recently released [17], also taking into account unresolved issues as lack of data directly comparing dabigatran, rivoroxaban, and apixaban to one another, the duration of followup in clinical trials, and lack of information on the increased risk of thromboembolism in noncompliant patients due to the short half-lives of these new treatments as well as to the inability to presently test their drug activity.
4. Statins in Treating Hyperlipidemia
Cholesterol levels represent an important and modifiable risk factor for coronary artery disease (CAD). However, the epidemiological association between cholesterol and stroke is controversial: direct and moderately strong, direct but fairly weak, J shaped unclear, and absent. Observational studies may be limited because cholesterol may have different effects on different stroke types; further, the incidence of stroke is lower and occurs later as compared to CAD. An association between serum cholesterol levels and both incident and recurrent stroke rate has not been clearly demonstrated. The Prospective Studies Collaboration [43] evaluated this association in pooled data of 45 prospective observational cohorts, with a total of 450,000 individuals, a mean followup of 16 years, and a total of 13,397 strokes recorded. The analysis detected a “flat effect” of increasing cholesterol levels on stroke risk. Despite only week or no association of cholesterol levels with stroke, treatment with statins has consistently shown positive effects. The Scandinavian Simvastatin Survival Study (4S) [44] was a placebo-controlled trial which detected a 30% RRR in incidence of any stroke in patients treated with simvastatin as compared to placebo. In the Heart Protection Study (HPS) [45], 20,536 high-risk patients (CAD, occlusive arterial disease, diabetes mellitus) were enrolled in 69 UK hospitals: 13,379 (65%) with CAD, 3,280 (16%) with stroke+CAD, and 1,822 (9%) with stroke only. Patients were randomized to receive 40 mg simvastatin or placebo, and they had a followup of 5.5 years. Statin treatment was associated with a 27% RRR for all strokes, and a 25% RRR for ischemic stroke. However, no clear RRR in stroke recurrence was observed in patients with a prior stroke and no known CAD. A meta-analysis that included 16 statin trials with 34,000 patients and 860 strokes evidenced an overall 25% RRR for stroke (14% to 35%), with a nonsignificant 15% RRR (−28% to 43%) for primary, and a significant 35% RRR (18% to 49%) for secondary stroke prevention [46]. The Stroke Prevention by Aggressive Reduction in Cholesterol Levels (SPARCL) [47] evaluated secondary stroke prevention. The study demonstrated a 16% risk reduction of recurrent fatal or nonfatal stroke in patients randomized to 80 mg atorvastatin versus placebo.
In secondary prevention of stroke, the use of statins has shown a positive effect in both decreasing progression and/or inducing regression of carotid artery plaque and stroke recurrence. By analyzing data from all available studies on statin treatment and stroke risk, Amarenco et al. [48] demonstrated a linear relationship between low-density lipoprotein-cholesterol (LDL-C) values and RRR of stroke. Each 10% reduction in LDL-C was estimated to reduce the risk of all strokes by 15.6% (95% CI, 6.7 to 23.6). This analysis also demonstrated a strong correlation between progression of carotid intima-media thickness (IMT) and LDL-C reduction. The question is why should statins prevent ischemic stroke? The possible explanations are two: (i) lipid effects (LDL-C lowering) and (ii) nonlipid effects. Among these the following have been demonstrated: (i) stabilization of atherosclerotic plaque, (ii) improvement of the endothelial function, (iii) decrease in the inflammation, (iv) decrease of platelet aggregation, (v) a direct lowering effect of blood pressure, (vi) a decrease in cardiac emboli, and (vii) miscellaneous (reduced left ventricular hypertrophy, HTN, effects on endothelial progenitor cells). Experimental data suggests that LDL-C may damage the vascular endothelium by oxidation of lipids, glycation, and oxidation of proteins. Its accumulation under the endothelium leads to activation of inflammation which triggers movements of macrophages and T cells into the intima, plaque development, and progression, as well as development of a fibrous cap over the lipid core, which ultimately leads to plaque rupture and thrombus formation. Direct evidence of the molecular mechanisms modulating the plaque composition and its instability, and of a direct effect of statins on the inflammatory processes that are supposed to be involved in the plaque rupture have been demonstrated [49].
The association of total and high-density lipoprotein cholesterol (HDL-C) with stroke risk is unclear. This association has been recently investigated in among 58,000 Finnish people aged from 25 to 74 years. During a mean follow-up period of 20.1 years, 3,914 participants developed incident stroke (3,085 were ischemic). Low levels of HDL-C and high total/HDL cholesterol ratio were associated with increased risks of ischemic stroke in both sexes. These associations attenuated after adjustment for body mass index, blood pressure, and history of diabetes [50]. Two recent meta-analyses have shown that ezetimibe coadministration with a statin provides significant additional lipid-lowering effect, allowing more patients to achieve low density lipoprotein cholesterol (LDL-C) target values [51, 52]. Although reduction of LDL-C remains the primary goal for lipid-lowering interventions, other targets (e.g., HDL-C and triglycerides) may also be important. However, there is no definitive evidence showing that raising HDL-C levels in patients on statins will result in a significant reduction in vascular events [53, 54].
Conclusions: statins are recommended to prevent the first stroke in high-risk patients to lower LDL-C level <100 mg/dL; an LDL-C < 70 mg/dL is recommended for highest risk (high risk: any of LDL > 4.1, age >45 M/55 F, positive family history, smoking, HTN, left ventricular hypertrophy; highest risk: DM or established atherosclerosis) (Class I, Level of Evidence A). A new recommendation for secondary stroke prevention: on the basis of the SPARCL trial [8], statin treatment with intensive lipid-lowering effects is recommended to reduce the risk of stroke and cardiovascular events for patients with IS or TIA, evidence of atherosclerosis, an LDL-C level ≥100 mg/dL, and without known coronary artery disease (Class I, Level of Evidence B). In these patients, a target reduction of at least 50% in LDL-C or a target LDL-C level <70 mg/dL is recommended (Class IIa, Level of Evidence B). Patients with IS or TIA with low HDL-C may be considered for treatment with niacin or gemfibrozil (Class IIb, level B).
5. Diabetes Mellitus
Epidemiological studies show that diabetes is a risk factor for first ischemic stroke, while data on stroke recurrence are more sparse [8, 11]. Further, the role of tight glycemic control in reducing the risk of stroke is still uncertain [55]. Patients with diabetes have higher mortality, more severe disability, and slower recovery after a stroke, as well as higher rates of stroke recurrence at 1 month (4.9% versus 2.6%) and at 2.6 years (15.2% versus 11.4%) compared to nondiabetic stroke patients [56, 57]. In addition, the ten-year risk of ipsilateral ischemic stroke after carotid endarterectomy is higher in the presence of diabetes (HR 2.24; 95% CI 1.35–3.74; ) [58]. The United Kingdom Prospective Diabetes Study (UKPDS) was a landmark study in the treatment of type 2 diabetes from the time of diagnosis [59]. This study has demonstrated that intensive treatment of type 2 diabetes versus standard treatment determined a 12% RRR the event rate (including stroke). In the Heart Outcomes Prevention Evaluation (HOPE) [60], 3,577 people with diabetes, who had a previous cardiovascular event or at least one other cardiovascular risk factor, were randomly assigned to ramipril (10 mg/day) or placebo. The combined primary outcome was myocardial infarction, stroke, or cardiovascular death. The study was stopped 6 months early (after 4.5 years) because of a consistent benefit of ramipril compared with placebo: ramipril lowered the risk of the combined primary outcome by 25% (95% CI, 12–36, ), stroke by 33% (10–50), and, among other outcomes, total mortality by 24% (8–37). After adjustment for the changes in SBP and DBP, ramipril still lowered the risk of the combined primary outcome by 25% (12–36, ). The study demonstrated that the vasculoprotective effect of ramipril in diabetic patients was greater than that attributable to the decrease in blood pressure. A secondary analysis of the Stroke Prevention by Aggressive Reduction in Cholesterol Levels (SPARCL) trial, which tested the effect of treatment with atorvastatin in reducing stroke in subjects with a recent stroke or TIA, investigated the effects of treatment in subjects with type 2 DM or metabolic syndrome (MetS) [61]. In this subanalysis, subjects with type 2 DM () had increased risks of stroke (HR 1.62; 95% CI, 1.33–1.98; ) and major cardiovascular events (HR 1.66; 95% CI, 1.39–1.97; ) compared with patients with neither diabetes nor MetS (,295). This exploratory analysis found no difference in the effect of statins in reducing these events in subjects with or without type 2 DM. Intensive glucose therapy did not prove effective treatment in reducing the rate of cardiovascular events or death in patients with type 2 DM and prior history of cardiovascular disease, stroke, or vascular risk factors. In the Action to Control Cardiovascular Risk in Diabetes (ACCORD) trial [62], 10,251 diabetic patients were randomly assigned to intensive glucose control (HbA1c < 6%) versus standard treatment (HbA1c 7–7.9%). The study was interrupted after 3.5 years of followup because of higher mortality in the intensive treatment group. No difference in rate of nonfatal stroke was observed between the two groups (HR 1.06; 95% CI, 0.75–1.50; ). In the Action in Diabetes and Vascular Disease (ADVANCE) trial [63], 11,140 diabetic patients were randomized to intensive treatment (HbA1c ≤ 6.5%) versus standard treatment (HbA1c ≤7%). The two groups did not differ in occurrence of nonfatal stroke (HR 0.94; 95% CI, 0.84–1.06; ). Also in the Veterans Affairs Diabetes Trial (VADT) trial [64], intensive glucose control did not reduce combined vascular outcomes compared to standard care (HR 1.07; 95% CI, 0.81–1.42; ).
The Prospective PioglitAzone Clinical Trial in Macrovascular Events (PROactive) was designed to evaluate the efficacy of pioglitazone efficacy in preventing vascular events in patients with type 2 DM. In the subset of patients with history of stroke enrolled in the study ( in the pioglitazone group and in the placebo group), pioglitazone was associated with a 47% RRR in recurrent fatal and nonfatal stroke (HR, 0.53; 95% CI, 0.34 to 0.85; ), and 25% RRR in stroke, MI, or vascular death (HR, 0.72; 95% CI, 0.53 to 1.00; ) [65]. Intensive hypertensive treatment (SBP < 120 mm Hg) in type 2 DM has not been supported by a recent analysis of the ACCORD patients. In this study, the annual rate of the primary outcome (i.e., composite outcome was nonfatal myocardial infarction, nonfatal stroke, or death from cardiovascular causes) was 1.87% in the intensive BP treatment and 2.09% in the standard-therapy group (HR, 0.88; 95% CI, 0.73 to 1.06; ). The annual rates of stroke were 0.32% and 0.53%, respectively (HR, 0.59; 95% CI, 0.39 to 0.89; ). Serious adverse events attributed to antihypertensive treatment were significantly more frequent in the intensive-therapy group (3.3% versus 1.3%; ) [62]. Conclusions: in patients with type 2 DM with history of TIA and stroke, glucose control is recommended (Class I, Level of Evidence B) [66, 67]. Levels of HbA1c < 6.5% should not be achieved in diabetic patients with history of cardiovascular disease or vascular risk factors [6264]. Based on the current knowledge, a target BP < 130/80 mm Hg for patients with type 2 DM is recommended [66, 67]. An ongoing trial will provide further insight on pioglitazone usefulness in preventing recurrent stroke in diabetic patients.
6. Antiplatelet Therapy
Within the established efficacy in stroke prevention through pharmacological and lifestyle control of modifiable vascular risk factors, a central role is played by antiplatelets. Aspirin represents the prototype of all antiplatelets. It acts by inhibiting the cyclooxygenase pathway with subsequent reduction in platelet thromboxane A2 synthesis and partial block of the final step of platelet aggregation. In a recent meta-analysis of 16 secondary prevention trials (17,000 individuals at high average risk, 43,000 person-years, 3,306 serious vascular events) comparing long-term aspirin versus control, treatment with aspirin reduced of about a fifth the rate of total stroke (2.08% versus 2.54% per year, ), with a nonsignificant increase in haemorrhagic stroke [68]. Second generation platelet inhibitors such as thienopyridines (e.g., ticlopidine or clopidogrel) work by blocking the platelet adenosine diphosphate (ADP) receptor; they may offer greater preventive efficacy, especially in specific populations of atherothrombotic disease patients. In the Clopidogrel versus Aspirin in Patients at Risk of Ischemic Events (CAPRIE) study [69], over 19,000 patients at high risk of recurrent stroke were randomized to either treatment with clopidogrel 75 mg/day or aspirin 325 mg/day for a mean follow-up period of 1.9 years. Clopidogrel reduced the risk of major cardiovascular events (IS, MI, or vascular death) in a small but statistically significant manner compared with aspirin (RRR 8.7%; 95% CI, 0.3 to 16.5; ). However, subgroup analysis revealed that the RRR was statistically significant only in PAD but not in stroke nor in myocardial infarction (MI) patients.
In the Management of Atherothrombosis with Clopidogrel in High-Risk Patients with Recent Transient Ischaemic Attack or Ischaemic Stroke (MATCH) study [70], 7,599 patients were randomized to receive either clopidogrel 75 mg plus aspirin 75 mg or clopidogrel 75 mg alone. Combination therapy was not superior to clopidogrel alone in preventing primary composite outcomes (IS, MI, vascular death, or rehospitalization for any ischemic event), with significant increase in major bleeding complications. The Clopidogrel for High Atherothrombotic Risk and Ischemic Stabilization, Management, and Avoidance (CHARISMA) trial [71] evaluated whether the addition of clopidogrel to aspirin better prevented recurrent stroke. In this trial, 15,603 patients with cardiovascular disease or multiple vascular risk factors for cardiovascular disease (35% had history of cerebrovascular diseases in the previous 5 years) were randomized to clopidogrel 75 mg plus low-dose aspirin (75–162 mg) or placebo plus aspirin (75–162 mg). The two groups did not differ in the rates of nonfatal IS (1.7% versus 2.1%; ) and had similar rates of intracerebral hemorrhage (0.3%). Patients in the combination arm treatment had a higher rate of moderate bleeding (not of severe or fatal). The study did not demonstrate superiority of combination treatment over aspirin in the subgroup of patients with prior history of IS or TIA. Dipyridamole is a phosphodiesterase inhibitor which inhibits platelet aggregation. The European/Australasian Stroke Prevention in Reversible Ischemia Trial (ESPRIT) [72] was a randomized, nonblinded study, comparing aspirin (30–325 mg) and dipyridamole (200 mg twice daily; 83% extended-release dipyridamole) versus aspirin (30–325 mg) alone in 2,763 patients who had TIA, monocular blindness, and minor stroke (modified Rankin score ≤3) in the 6 months prior to enrollment. The mean follow-up was 3.5 years. Combined treatment conferred an absolute risk reduction of 1% for primary outcome (death from all vascular causes, nonfatal stroke, nonfatal myocardial infarction, or fatal bleeding complications). Despite few study weaknesses have been detected (nonblinded, nonstandard aspirin doses, divergence of significance between the on-treatment and intention-to-treat analyses, high rate of discontinuation therapy in the combination treatment arm), this study provided additional evidence of superiority of combined therapy with aspirin plus extended-release dipyridamole compared to aspirin alone in stroke prevention in patients with noncardioembolic IS.
Further, similar rates of recurrent stroke were observed with combination dipyridamole plus aspirin compared with clopidogrel, with no evidence that either of the two treatments was superior to the other in the prevention of recurrent stroke [73]. In the PERFORM study [74], the selective thromboxane-prostaglandin receptor antagonist terutroban (30 mg per day) was compared with aspirin (100 mg per day) in prevention of IS and cardiovascular events in patients with a recent noncardioembolic IS. The primary efficacy endpoint was a composite of any IS, any MI, or other vascular death. The study was stopped prematurely for futility, because it showed similar rates of the primary endpoint with terutroban and aspirin, without safety advantages for terutroban. Recommendations for patients with history of noncardioembolic stroke or TIA are aspirin (50–325 mg/die) monotherapy (Class I, Level of Evidence A), the combination of aspirin 25 mg and extended-release dipyridamole 200 mg twice daily (Class I, Level of Evidence B), and clopidogrel 75 mg monotheraphy (Class IIa, Level of Evidence B), which are all acceptable options as initial therapy for prevention of recurrent stroke or other cardiovascular events. Clopidogrel is a reasonable alternative option in patients allergic to aspirin (Class IIa, Level of Evidence C). The addition of aspirin to clopidogrel increases the risk of hemorrhage, and the combination therapy is not recommended unless specific indications exist (i.e., coronary and other vascular stent or acute coronary syndrome) (Class III, Level of Evidence A). For patients who have an IS while on aspirin, alternative antiplatelets may be considered although this has not been assessed yet by RCTs (Class IIb, level of Evidence C) [8].
6.1. Double Antiplatelet Therapy or Treatment with both an Anticoagulant and Aspirin
Dual antiplatelet treatment (aspirin plus clopidogrel or, less frequently, ticlopidine) has assumed a central role in the setting of cardiovascular disease and its use is increasing in the field of cerebrovascular disease as well. Safety and efficacy of dual- (aspirin+dipyridamole and aspirin+clopidogrel) versus mono-antiplatelet therapy have been compared in a recent systematic review and meta-analysis of RCTs [75]. This analysis included 12 RCTs involving 3,766 patients with noncardioembolic acute (≤3 days) IS or TIA. In comparison with mono-antiplatelet therapy, dual antiplatelets were associated with reduced early stroke recurrence, composite vascular events (stroke, MI, and vascular death), and combined stroke, TIA, acute coronary syndrome, and all death. Dual therapy was also associated with a trend to increase major bleeding. However, a recent RCT involving 3,020 patients with recent symptomatic lacunar infarcts has shown that the addition of clopidogrel to aspirin 325 mg daily did not significantly reduce the risk of recurrent stroke as compared with aspirin alone (2.5% per year versus 2.7% per year; hazard ratio, 0.92; 95% CI, 0.72 to 1.16), while significantly increased the risk of bleeding and death (2.1% per year versus 1.1% per year; hazard ratio, 1.97; 95% CI, 1.41 to 2.71; ) [76].
Further, the combination of antiplatelet and anticoagulant therapy might be indicated for stroke prevention in a variety of conditions including AF, profound left ventricular dysfunction, and after prosthetic heart valve replacement. For this reason, the use of triple antithrombotic therapy (a dual antiplatelet regimen plus warfarin) is expected to increase along with an aging population. However, this approach carries an increased risk of bleeding complications [77].
Both the presence and degree of benefit associated with dual antiplatelet therapy are likely to depend on characteristics of the specific patient. The CHARISMA trial [71] demonstrated that dual antiplateltes (ASA plus clopidogrel) were more effective than ASA alone especially in patients with clinically evident cardiovascular disease (i.e., in secondary prevention CV death, MI, and stroke) rather than in patients with high risk profile but not established atherothrombotic disease (i.e., primary prevention). However, moderate bleeding events significantly increased (2% versus 1.3%, HR 1.6, CI 1.16 to 2.20; ) [77].
Several data indicate that the increased frequency of bleeding is influenced by both duration of therapy and ASA dosage: a dose <100 mg of ASA when given in combination with a thienopyridine seems to be associated with similar anti-ischemic efficacy but reduced bleeding event rates [78]. Further evidence is needed to define more complex antithrombotic treatment algorithm required in patients with different vascular comorbidities in whom more aggressive prevention approach is required.
6.2. Alternative Antiplatelet Treatments: Cilostazol, Sarpogrelate, and Triflusal
Cilostazol is a selective and potent selective type III phosphodiesterase (PDE 3A) inhibitor, leading to inhibition of platelet aggregation and vasodilation, with potential use in atherosclerotic conditions, including stroke [79, 80]. The Cilostazol Stroke Prevention Study (CSPS) [81, 82] was designed to evaluate safety and effectiveness of cilostazol in prevention recurrent stroke. In this double-blind, placebo-controlled trial, 1,095 patients (544 receiving cilostazol 100 mg twice daily and 548 receiving placebo) were enrolled. The recurrence rate of IS was significantly reduced by cilostazol with a 41.7% RRR (number needed to treat = 41). A subgroup analysis showed that patients with lacunar infarcts had a significant reduction in recurrence of IS, whereas no statistical significance was reached in patients with atherothrombotic or mixed-type infarctions. These results are in line with another PDE inhibitor, dipyridamole, currently used in stroke prevention, which caused withdrawal of therapy in 26% of patients in the ESPRIT trial [72].
Further potential use of cilostazol in clinical practice may derive from the benefits observed in diabetic patients. In patients with type 2 DM, cilostazol 100–200 mg/day significantly prevented IMT progression and reduced the number of silent brain infarctions which have been associated with increased risk of developing dementia [83]. Based on the results and the observed reduced efficacy of antiplatelets in diabetic patients [84, 85], cilostazol might represent an alternative to standard care at least in the subgroup of patients with lacunar stroke. This hypothesis was tested in the Cilostazol versus Aspirin for Secondary Ischemic Stroke Prevention (CASISP) trial [86], which was a pilot, multicentre, and double-blind trial, which randomly assigned 301 patients to cilostazol and 299 patients to aspirin treatment, with a follow-up period of 12 to18 months. The primary endpoint was any recurrence of stroke (IS, haemorrhagic stroke, or subarachnoid hemorrhage) during the trial period, as assessed by a follow-up MRI study. Cilostazol was associated with not-significant RRR of stroke (HR 0.62; 95% CI, 0.30–1.26; ) and significantly lower rates of symptomatic and asymptomatic cerebral hemorrhages (7 versus 1, ). The results suggested that cilostazol appeared to be a more effective and safer alternative to aspirin in secondary stroke prevention. Whether a reduced risk of intracranial bleedings is peculiar of PDE inhibitors, as suggested by the reduced occurrence of cerebral hemorrhage observed in patients treated with dipyridamole plus aspirin compared to aspirin alone [72], should be further investigated. Safety and effectiveness of cilostazol compared with aspirin in preventing recurrent stroke in patients with previous IS or TIA of arterial origin have been recently assessed. In this systematic review, two RCTs with 3,477 Asian participants were selected. Compared with aspirin, cilostazol was associated with a significantly lower risk of composite outcome of vascular events (6.77% versus 9.39%; RR 0.72, 95% CI, 0.57 to 0.91), haemorrhagic stroke (0.53% versus 2.01%, RR 0.26, 95% CI, 0.13 to 0.55), and minor adverse effects (8.22% versus 4.95%, RR 1.66, 95% CI, 1.51 to 1.83) [87].
Based on the available data, cilostazol seems to be more effective than aspirin in the prevention of vascular events in high-risk Asian patients. Further RCTs testing larger cohorts of vascular patients are needed in order to address the exact potential of cilostazol in the management of atherosclerosis and stroke prevention.
6.3. Sarpogrelate
The antiplatelet agent sarpogrelate is a selective inhibitor of 5-hydroxytryptamine receptors with dose-dependent inhibitory effect on platelet aggregation [88]. Similar to cilostazol, this drug has been used for years to treat patients with PAD in Japan. Based on this, its efficacy and safety in secondary stroke prevention has been tested in RCT versus aspirin in a Japanese ischemic stroke patients cohort [89]. In this study, 1,510 patients with recent infarction were enrolled which were randomly assigned to receive either sarpogrelate (100 mg × 2/day) or aspirin (81 mg/day). The study failed to demonstrate noninferiority of sarpogrelate to aspirin for prevention of stroke recurrence. However, bleeding events were significantly fewer with sarpogrelate than aspirin, although this might have been the result of the lower efficacy of sarpogrelate. A subgroup analysis of this study confirmed the superiority of aspirin in most patient subgroups, except in diabetics that could represent a specific target population of this drug, at least among Japanese stroke patients [90].
6.4. Triflusal
The efficacy of the antiplatelet agent triflusal (600 mg/d) versus aspirin (325 mg/d) for prevention of vascular events after stroke has been reported in a randomized, double-blind, and multicenter study. In this study, 2,113 patients with IS or TIA were randomly assigned to receive either triflusal (,058) or aspirin (,055). After a mean follow-up period of 30.1 months, the incidence of nonfatal stroke (HR, 1.09; 95% CI, 0.82 to 1.44), nonfatal acute myocardial infarction (HR, 0.95; 95% CI, 0.46 to 1.98,) and vascular death (HR, 1.22; 95% CI, 0.75 to 1.96) showed no differences between groups. A significantly higher incidence of major hemorrhages in the aspirin group was recorded (HR, 0.48; 95% CI, 0.28 to 0.82). The overall incidence of hemorrhage was significantly lower in the triflusal group (16.7% versus 25.2%) (OR, 0.76; 95% CI, 0.67 to 0.86; ). This study failed to show significantly superior efficacy of triflusal over aspirin in the long-term prevention of vascular events after stroke, but triflusal was associated with a significantly lower rate of hemorrhagic complications [91]. Further, a metaanalysis of 4 clinical trials comparing triflusal with aspirin including a total of 2,994 patients with IS or TIA who were followed from 6 to 47 months was performed. Of relevance, the authors reported no significant differences between aspirin and triflusal in the risk of serious vascular events (OR for aspirin versus triflusal, 1.02; 95% CI, 0.83–1.26). Aspirin was associated with a higher risk of hemorrhage, both major (OR, 2.42; 95% CI, 1.56–3.77) and minor (OR, 1.62; 95% CI, 1.31–2.01) [92]. Based on this metaanalysis, the European Stroke Organisation [93] recommends triflusal as an alternative to combined aspirin and dipyridamole, or to clopidogrel alone (Class I, Level A).
6.5. Carotid Endarterectomy (CEA) and Carotid Angioplasty with Stenting (CAS) in Secondary Stroke Prevention
The beneficial effect of CEA plus medical treatment versus medical treatment alone in prevention of stroke recurrence in symptomatic patients with high-grade carotid stenosis (>70%) has been extensively demonstrated over the last decades [9496]. These trials also demonstrated that CEA did not decrease the risk of stroke recurrence in patients with a symptomatic stenosis <50%. Conversely, patients with a moderate (50% to 69%) symptomatic stenosis may benefit from an intervention if this is performed by an experienced surgeon with a perioperative morbidity and mortality rate of <6%. Carotid angioplasty and stenting (CAS) has emerged as an alternative treatment for stroke prevention in patients deemed at high risk for conventional endarterectomy. However, its efficacy of in stroke prevention has not been clearly established yet. In this regard, two RCTs failed to establish noninferiority of CAS compared with CEA: the EVA 3S [97] and the SPACE [98] trials were both stopped prematurely for reasons of safety and futility because of a higher 30-day stroke and death rate in the CAS group. A more recent RCT in the International Carotid Stenting Study (ICSS) [99] which enrolled 1,713 patients has shown that the periprocedural risks of CAS are significantly higher than those of CEA. In particular, the risk of stroke and death are increased in CAS patients (as in the EVA 3s and SPACE trials), with similar low rates of myocardial infarction.
Few studies demonstrated noninferiority or superiority of CAS versus CEA. The Carotid and Vertebral Artery Transluminal Angioplasty Study (CAVATAS) [100] was a small randomized trial suggesting that CAS was equivalent to CEA or even superior in high-risk patients. The carotid Revascularization Endarterectomy versus Stenting Trial (CREST) [101] was a large RCT evaluating the two procedures in symptomatic patients with up to a 4-year followup. The study results were broadly consistent with those of previous trials. In particular, an equivalence between CAS and CEA regarding the primary composite end point of stroke, myocardial infarction, or death was observed. However, the rate of stroke or death in this trial was still significantly higher in the CAS group than in the CEA group, both during the periprocedural period and at 4 years.
Despite the evidence supporting CEA over CAS in secondary stroke prevention, it has been observed that the conclusions achieved by the above-mentioned trials might be faded by several methodological differences: the most important issue is the choice of primary end point, followed by timing of the primary end point (ranging from 30 to 120 days after randomization); further, the rate of periprocedural stroke in patients treated by CAS differed between studies, underlining the importance of training of proceduralists; inclusion or exclusion of a patient with preexisting coronary artery disease also contributed in different rates of cardiac complications after the procedure. Further, in the CREST trial, CAS tended to have greater efficacy below 70 years and CEA above 70 years, possibly reflecting increased technical challenge of stenting in older patients, such as the atherosclerotic burden in the internal carotid artery and aortic arch and increased arterial tortuosity [102]. This has been confirmed by a recent meta-analysis, suggesting that stenting for symptomatic carotid stenosis should be avoided in older patients (age ≥70 years), but might be as safe as endarterectomy in younger patients [103].
More recently, a systematic review and meta-analysis of 13 RCTs of CEA versus CAS enrolling 7,484 (80% with symptomatic carotid artery stenosis) has shown that, compared with CEA, CAS significantly increased the risk of any stroke (relative risk (RR), 1.45; 95% CI, 1.06–1.99) and decreased the risk of MI (RR, 0.43; 95% CI, 0.26–0.71). Of relevance, when analysis was restricted to the two most recent trials with the better methodology and more contemporary technique, we found stenting to be associated with a significant increase in the risk of any stroke (RR, 1.82; 95% CI, 1.35–2.45) and mortality (RR, 2.53; 95% CI, 1.27–5.08) and a nonsignificant reduction of the risk of MI (RR, 0.39; 95% CI, 0.12–1.23). The authors observed that, for every 1,000 patients opting for stenting rather than endarterectomy, 19 more patients would have strokes and 10 fewer would have MIs [104]. Another meta-analysis of 11 RCT performed through 2009 (not including CREST) has confirmed that CEA was superior to CAS with regard to short-term outcomes but the difference was not significant for intermediate term outcomes [105]. Further, the SPACE trial also reported a higher two-year rate of restenosis in the CAS than the CEA group, with similar 2-year rates of ipsilateral stroke [106]. Another single-center prospective randomized study of CEA versus CAS with a long followup of 5 years has shown a significantly higher incidence of relevant restenosis and neurologic symptoms after CAS [107].
Until more data are available, CEA remains the preferred treatment choice for symptomatic severe carotid artery stenosis. However, given the lack of significant difference in the rate of long-term outcomes, the individualization of treatment choices is appropriate. More long-term data are needed.
6.6. Early CEA/CAS
CEA has traditionally been delayed from 4 to 8 weeks because of fear of hemorrhagic transformation of the ischemic infarct. Pooled analysis from the European Carotid Surgery Trail (ECST) and the North American Symptomatic Carotid Endarterectomy Trial (NASCET) [108] has clearly shown that the benefit from CEA is maximal in symptomatic patients operated within 2 weeks of the index event. The analysis of long-term stroke prevention has shown that benefits from surgery decreased rapidly with time elapsed since the last neurological symptoms. Profit from CEA seems to depend not only on the degree of carotid stenosis, but also on delay in surgery after the presenting event, and the conclusion was that the procedure should ideally be done within 2 weeks of the patient’s last symptoms. Rothwell et al. [109] also showed that early treatment (either medical and/or surgical) of all patients presenting with a TIA or minor stroke can prevent up to 80% of early recurrent stroke. However, urgent CEA in preventing stroke recurrence proved effective in several observational single center studies. A prospective multicenter Italian study assessing safety and efficacy of early (1.5 days after the stroke) CEA after acute ischemic stroke has shown that if patients are strictly selected for early CEA after an acute stroke, early surgery patients have similar risks to elective surgery [110]. In the study of Ferrero et al. [111], cumulative rates of TIA/stroke and death after CEA were compared between patients undergoing to early (range, <48 hours) and delayed/deferred CEA (range, within 48 hours–24 weeks). The analysis demonstrated that early CEA in acute poststroke period for selected patients (not severe strokes and lesions <3 cm, no MCA occlusion) did not result in greater complication rate that performed delayed or deferred.
With growing experience in endovascular treatment, CAS has been proposed as an alternative to CEA, but data regarding the outcome of patients with acute stroke undergoing urgent endovascular surgery are still scarce. The main concern about CAS in urgent cases is that, while with CEA the plaque is completely removed, after stenting it is only remodeled and its stabilization is essential to avoid later embolic events. Safety and efficacy of early CAS after TIA (within 24–48 hours) has been demonstrated in single-center studies. Setacci et al. [112] reported the results of a small single-center study including 43 symptomatic patients who underwent to either early CAS (within 24 hours in TIA patient) or deferred CAS (between 1 and 30 days, in minor stroke patients) in selected patients (no major stroke, lesion <2.5 cm). The authors observed that early CAS is feasible and safe in selected patients with a first episode or recurrent TIA or minor stroke.
These studies suggest that safety and efficacy of emergency/urgent carotid endovascular revascularization depend on correct patient selection, and consequently on the reduction of the time loss between the index event and intervention, and on the specific skill of operators performing the procedure.
Recommendations
for patients with recent TIA or ischemic stroke within the past 6 months and ipsilateral severe (70% to 99%) carotid artery stenosis, CEA is recommended if the perioperative morbidity and mortality risk is estimated to be <6% (Class I; Level of Evidence A). For patients with recent TIA or ischemic stroke and ipsilateral moderate (50% to 69%) carotid stenosis, CEA is recommended depending on patient-specific factors, such as age, sex, and comorbidities, if the perioperative morbidity and mortality risk is estimated to be <6% (Class I; Level of Evidence B) [8]. When CEA is indicated for patients with TIA or stroke, surgery within 2 weeks is reasonable rather than delaying surgery if there are no contraindications to early revascularization (Class IIa; Level of Evidence B).
CAS is indicated as an alternative to CEA for symptomatic patients at average or low risk of complications associated with endovascular intervention when the diameter of the lumen of the internal carotid artery is reduced by >70% by noninvasive imaging or >50% by catheter angiography (Class I; Level of Evidence B). Among patients with symptomatic severe stenosis (>70%) in whom the stenosis is difficult to access surgically, medical conditions are present that greatly increase the risk for surgery, or when other specific circumstances exist, such as radiation-induced stenosis, or restenosis after CEA, CAS may be considered (Class IIb; Level of Evidence B). CAS in the above setting is reasonable when performed by operators with established periprocedural morbidity and mortality rates from 4% to 6%, similar to those observed in trials of CEA and CAS (Class IIa; Level of Evidence B) [8].
7. Conclusions and Future Directions
Despite the increased knowledge on the relevance of controlling modifiable vascular risk factors, to keep them under control remains a challenge. Earlier studies observed that patients with identified risk factors for stroke do not follow physicians’ suggestions regarding lifestyle changes or adherence to treatment prescribed to modify their risk [113, 114]. Several barriers can reduce the patient compliance to prescribed treatments: cultural gaps between physician and patients and socioeconomic factors, as well as the physician attitude [115]. In order to challenge this gap, few approaches have been attempted. Earlier studies have demonstrated that careful patient follow-up [116], intervention programs for coronary risk factor modification [117] and structured counseling provided by Stroke Prevention Clinics [118] were more effective than usual medical care in improving vascular risk factor control by increasing adherence to prescribed medications. More recently, a standardized electronic counseling (e-counseling) proved helpful in reducing BP in patients with HTN with greater reduction in SBP, pulse pressure, and total cholesterol as compared to general e-information [119]. Further, a structured risk factor modification program for secondary prevention of cardiovascular disease was found associated with a reduced risk of major vascular event, including nonfatal stroke [120], thus demonstrating that improving vascular risk factor control may translate into reduced cerebrovascular events. Based on this evidence, one of the objectives set by a panel of leaders in this field is to develop, implement, and evaluate a population approach for stroke prevention and public health communication strategies using traditional and novel (i.e., social media/marketing) techniques [7].
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114. J. Benson and N. Britten, “Patients' decisions about whether or not to take antihypertensive drugs: qualitative study,” British Medical Journal, vol. 325, no. 7369, pp. 873–876, 2002. View at Scopus
115. K. Gardner, A. Chappie, and A. Chapple, “Barriers to referral in patients with angina: qualitative study,” BMJ, vol. 319, no. 7207, pp. 418–421, 1999. View at Scopus
116. V. L. Burt, J. A. Cutler, M. Higgins et al., “Trends in the prevalence, awareness, treatment, and control of hypertension in the adult us population: data from the health examination surveys, 1960 to 1991,” Hypertension, vol. 26, no. 1, pp. 60–69, 1995. View at Scopus
117. R. F. DeBusk, “MULTIFIT: a new approach to risk factor modification,” Cardiology Clinics, vol. 14, no. 1, pp. 143–157, 1996. View at Scopus
118. M. S. Mouradian, S. R. Majumdar, A. Senthilselvan, K. Khan, and A. Shuaib, “How well are hypertension, hyperlipidemia, diabetes, and smoking managed after a stroke or transient ischemic attack?” Stroke, vol. 33, no. 6, pp. 1656–1659, 2002. View at Publisher · View at Google Scholar · View at Scopus
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Bibliography: Deep Walnuts [2]
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Title: Deep Walnuts [2]
Author: Dominic Harman
Year: 1999
Type: INTERIORART
ISFDB Record Number: 1282230
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Copyright (c) 1995-2011 Al von Ruff.
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News
Analyzed 8 days ago based on code collected 8 days ago.
Posted over 1 year ago by Joannes Vermorel
Evaluating the quality of a forecasting method can be dramatically complicated. In order to help the community finding out who's delivering the best forecasts out there :-), we have just produced a video concerning accuracy measurements of sales ... [More] forecasts. Not all metrics are born equal.
Special thanks to Ray Grover for the voice-over. [Less]
Posted almost 2 years ago by Joannes Vermorel
Our flagship inventory optimization product, Salescast, has benefited from a major documentation refresh about 3 months ago. Now it's the turn of the user interface (UI) to be extensively redesigned. Here below is a screenshot illustrating the new ... [More] look & feel (deployed today).
The previous UI was suffering from several problems:
not following UI standards, making it harder for everyone to figure out how Salescast was working. In this version, we have paid a lot more attention to make Salescast not only simpler, but closer to the de facto standards (Amazon, Facebook for example). People should not have to think about UI.
not focusing on changes, complicating the team usage of Salescast. In this version, the most notable UI addition is the Recent Activity listing that tells who has been doing what. Recent activity is a killer feature for teamwork. A lot of teamwork confusion happen job just because Bob did not tell Alice, he had just generated a new report. Now Alice will immediately notice what Bob has done, by simply looking at the recent activity.
confusion between queries and commands, triggering the generation of new reports (and causing extra charges!) while the intent was only to browse existing reports. Now, the commands are put on the right while the queries (read only) are gathered on the left. By establishing this simple distinction, we hope to siginificantly reduce the number of misclicks.
Since it's initial release 15 months ago, Salescast has steadily improved, and this release is certainly not our final word. More good stuff is coming. Stay tuned.
Side note: a new logo for Salescast is also under way :-) [Less]
Posted almost 2 years ago by Joannes Vermorel
Bitcoin is a nascent crypto-currency that has attracted a lot of attention lately. For those who've never heard of Bitcoin yet, you can check the good analysis of The Economist.
We believe that Bitcoin is a tremendous opportunity to ... [More] vastly reduce payment frictions for companies operating online such as Lokad.
Thus, in order to facilitate the bootstrap of the emerging Bitcoin economy, we have decided to accept Bitcoins as payment method.
Moreover, for the time being, Bitcoin payments come with a 10% discount as a favor made to early adopters. Contact us for up-to-date conversion rates.
We anticipate that the Bitcoin adoption will be faster among eCommerce compared to classical retail. Through Salescast, online merchants can forecast their sales and optimize their inventory levels based on those forecasts. We hope to make Bitcoin-powered eShops even more productive through advance inventory optimization. [Less]
Posted almost 2 years ago by Joannes Vermorel
A couple of weeks ago, we disclosed our plans concerning Shelfcheck, our future on-the-shelf availability optimizer targeting (physical) retailers. Since that time, we have been steadily moving forward, crushing a lot of point-of-sale ... [More] data.
Lokad isn't the only one company out there trying to tackle the OOS (out-of-shelf) issue, but there is very little literature about how to assess the respective merits of two OOS detectors. In this post, we review two fundamental metrics that define how good is a system at detecting OOS.
Intuitively, an indirect OOS detector (such as Shelfcheck) relies on the divergence between observed sales and the expected sales. Since random (aka unpredictable) fluctuation of the market can always happen, this approach, by construction, cannot be a perfect system (1), it's a tradeoff between sensibility and precision.
(1) Not being perfect does not imply being worthless.
The sensibility represents the percentage of OOS (aka the positives to be detected) that are captured by the system. This concept is already widely used in diverse areas ranging from medical diagnostics to airline security. The higher the sensibility the better the coverage of the system.
Yet, by increasing the sensibility, one also decreases the specificity of the system, that is to say, one decreases the percentage of non-OOS flagged as such (aka the negatives that should not be detected). In practice, it means that by pouring more and more alerts, the OOS detector gives more and more false alerts, wasting the time of the store teams looking for non-issues.
Although, specificity is not a very practical criteria in the case of retail. Indeed, OOS products only represent a small fraction of the non-OOS products. Several studies quote 8% OOS as being a relatively stable worldwide average. Hence, the specificity is typically very high, above 90%, even if the OOS detector happens to be producing pure random guesses. Hence, those high specificity percentages are somewhat misleading as they only reflect the imbalance that exists between OOS and non-OOS in the first place.
At Lokad, we prefer the precision that represents the percentage of accurately identified OOS within all alerts produced by the system. The precision directly translates into the amount of efforts that will not be wasted by the store staff checking for non-existent problems. For example, if the precision is at 50%, then one alert out of two is a false-alert.
Neither 100% sensibility nor 100% precision is possible, or rather if you have 100% sensibility then you have 0% precision (all products being classified as OOS all the time). The other way around, 100% precision indicates that you have 0% sensibility (no alert gets ever produced). The tradeoff sensibility vs precision cannot be escaped: if you want to detect anything, you need to accept that a some what you detect is incorrect.
In order to compare two OOS detectors, one needs to access their respective sensibility and precision. Then, in order to improve both the sensibility and precision, it remains possible to leverage a superior forecasting technology, as better forecasts will improve both the sensibility and precision.
Although, this raises another concern, how do you compare the following:
a detector A with 70% sensibility and 60% precision;
a detector B with 60% sensibility and 70% precision.
It turns out that this question cannot be addressed in a purely statistical manner: one needs to model the economic costs and benefits in order to assess the optimal choice.
Stay tuned for more. [Less]
Posted almost 2 years ago by Matthias Steinberg
Our growing development team in Russia has a new home: This week, Lokad’s new development center opened its doors in Ufa. The office opening is an important step in consolidating our development capabilities and expertise in Russia and ... [More] laying the foundations for further expansion. The office will be lead by Rinat Abdullin, Lokad’s Technology Leader.
Ufa is the capital of the Republic of Bashkortostan, Russia and situated near Eastern Europe's land boundary with Asia. Approximately 1 million people are currently enjoying average temperatures of up to 26C and endure up to -40C in Winter. Ufa's industry is dominated by oil refining and manufacturing.
The choice of location might puzzle, were it not for the Bashkir State University, which is among the top ten classical universities in Russia. It is especially strong in applied mathematics and information technologies. Many prominent applied mathematicians and physicists were among the faculty at different times, and the Department of Mathematics and the Ufa Institute of Mathematics including its Computing Center are among the best Russian schools in Applied Mathematics.
Our technology combines the triple mastery of statistics, enterprise software engineering and cloud computing in its most advanced forms. The only way to tackle this complexity is by working with the best people in its field worldwide, irrespective of where we find them. Ambitious new development initiatives such as Shelfcheck, an on-shelf availability monitoring tool for retail, require further investments in highly talented people. With Paris (France) and Ufa we now have two development locations that draw from extremely strong talent pools.
We are particularly pleased to see a red sofa appearing in the communal areas of the Ufa location. At the risk of giving away company secrets – a similar piece of furniture has been the breeding ground for some of the best ideas in Paris...
[Less]
Posted almost 2 years ago by Joannes Vermorel
Last Wednesday, at the Strategies Logistique Event in Paris, we announced a new upcoming product named Shelfcheck. This product targets retailers and it will help them to improve on-shelf availability, a major issue for nearly all retail ... [More] segments.
In short, through an advance demand forecasting technology such as the one of Lokad, it becomes possible to accurately detect divergence between real-time sales and anticipated demand. If the observed sales for a given product drop to a level that is very improbable considering anticipated demand, then an alert can be issued to store staff for early corrective actions.
On-shelf availability has been challenging the software industry for decades, yet, as far we can observe, there is no solution even close of being satisfying available on the market at present day. For us, it represents a tough challenge that we are willing to tackle. We believe on-shelf availability is the sort of problem that is basically near-impossible to solve without cloud computing; at least near-impossible within reasonable costs for the retailer. Shelfcheck will be deployed on Windows Azure, a cloud computing platform that proves extremely satisfying for retail, as illustrated by the feedback we routinely get for Salescast.
A retail pain in the neck
Have you ever experienced frustration in a supermarket when facing a shelf where the product you were looking for was missing? Unless, you've been living under a rock, we seriously bet you face this situation more than once.
Recent studies (*) have shown that, on average, about 10% of the products offered in store are unavailable on average. Worse, the situation seems to have slightly degraded over the last couple of years.
(*) See materials published on ECR France.
In practice, there are many factors that cause on-shelf unavailability:
Product is not at the right place and customers can't find it.
Product has been stolen and incorrect electronic inventory level prevents reordering.
Packaging is damaged and customers put in back after having a close look.
...
Whenever a product is missing from a shelf, sales are lost, as studies have shown that clients go for a direct ersatz only 1/3 of the time. Then it generates client frustration too which turns into loss of loyalty. Missing products are putting an incentive on your clients to check if competitors are better.
With about 10% on-shelf unavailability, a back-of-the-envelope calculation indicates the losses for the retail industry amount for a rough 100 billion USD per year world-wide. Disclaimer: that's only an order of magnitude, not a precise estimate. Then, this estimate does not factor industry-wide cannibalizations, aka lost sales aren't always lost for everyone.
You can't improve what don't measure
On-shelf availability is especially painful because detecting the occurrence of the problem is so hard in the first place:
Direct shelf control is extremely labor intensive.
RFID remains too expensive for most retail segments.
Rule-based alerts are too inaccurate to be of practical use.
Thus, the primary goal of Shelfcheck is to deliver a technology that let retailers detect out-of-shelf (OOS) issues early on. Shelfcheck is based on an indirect measurement method: instead of trying to assess the physical state of the shelf itself, it looks at the consequences of an OOS within real-time sales. If sales for a given product drop to a level that is considered as extremely improbable, then the product is very likely to suffer an OOS issue, and an alert is issued.
By offering a reliable OOS metering system, Shelfcheck will help retailers to improve their on-shelf availability levels, a problem that proved to be extremely elusive to quantitative methods so far.
Store staff frustration vs. client frustration
The core technological challenge behind Shelfcheck is the quality of OOS alerts. Indeed, it's relatively easy to design a software that will keep pouring truckload of OOS alerts, but the time of the store staff is precious (expensive too), and they simply can't waste their energy chasing false-positive alerts, that is to say "phantom problems" advertized by the software, but having no real counterparts within the store.
Delivering accurate OOS, finely prioritized and mindful of the store geography (*) is the number one goal for Shelfcheck. Considering the amount of data involved when dealing with transaction data at the point of sale level, we believe that cloud computing is an obvious fit for the job. Fortunately, the Lokad team happens to somewhat experienced in this area.
(*) Store employees certainly don't want ending-up running to the other side of the hypermarket in order to check for the next OOS problem.
Product vision
The retail software industry is crippled by consultingware where multi-millions dollars solutions are sold and do not live up to expectations. We, at Lokad, do not share this vision. Shelfcheck will be following the vision behind Salescast, our safety stock optimizer.
Shelfcheck will be:
Plug & Play, with simple documented ways of feeding sales data into Shelfcheck.
Dead simple, no training required as Lokad manages all the OOS detection logic.
Robotized, no human intervention required to keep the solution up and running.
SaaS & cloud hosted, fitting any retailers from the family shop up to the largest retail networks.
The pricing will come as a monthly subscription. It will be strictly on demand, metered by the amount of data to be processed. We haven't taken any final decision on that matter, but we are shooting for something in the range of $100 per month per store. Cost will probably be lower for minimarkets and higher for hypermarkets. In any case, our goal is to make Shelfcheck vastly affordable.
Feature-wise, Shelfcheck is pre-alpha stage, hence, the actual feature set of the v1.0 is still heavily subject to change. Yet, here is the overview:
Real time OOS alerts available through web and mobile access.
Real time prioritization based on confidence in the alerts and financial impact of the OOS issues.
OOS dashboard feeding the management with key on-shelf performance indicators.
1 year free usage for early adopters
We are extremely excited by the sheer potential of this new technology. We are looking for volunteers to beta-test Shelfcheck. Lokad will offer:
Until Shelfcheck is released, early adopters will be provided with unlimited free access.
Upon release, early adopters will get 1 year of further free usage, within a limit of 10 points of sale.
In exchange, we will kindly ask for feedback, first about the best way for Lokad to acquire your sales data, and second about the usage of Shelfcheck itself.
Naturally, this feedback will drive our development efforts, bringing Shelfcheck closer to your specific needs. Want to grab the opportunity? Just mail us at contact@lokad.com. [Less]
Posted almost 2 years ago by Joannes Vermorel
Many people at Lokad are bloggers. We try to keep the official blog focused on our core forecasting business, but here is a sample of posts made by team members during the last week.
3 Low-Competition Niches In Retail ... [More] Software by Joannes Vermorel (@vermorel), Founder at Lokad. Software developers seem to be herd animals. They like to stay very close to each other. As a result, the marketplace ends up riddled with hundreds of ToDo lists while other segments are deserted, despite high financial stakes. During my routine browsing of software business forums, I have noticed that the most common answer to "Why the heck are you producing yet another ToDo list?" is the desperately annoying "Because I can’t find a better idea." Read more.
A simple, non-technical definition of cloud computing by Matthias Steinberg (@mfsteinberg), CEO at Lokad. Last year, I found myself researching the market for investment opportunities in ‘cloud computing companies’, driven by the insight that this was a rapidly growing market. Yet, it felt a bit like hunting the Yeti as I was having a hard time understanding what exactly cloud computing is, and what it is not. Realizing that I shared this confusion with a lot of my peers, I thought it is worth sharing some insights I gained in the last month. Read more.
Lokad.CQRS v2.0 Framework and Docs for Windows Azure by Rinat Abdullin (@abdullin), Technology Lead at Lokad.The project was started 1.5 years ago to provide simple light-weight service bus for Windows Azure Cloud. It did the job well and eventually got accepted into more projects inside Lokad and outside as well. This gave us more feedback from the production usages in cloud, along with new development experience and CQRS ideas. Second version builds upon that and also includes first decent documentation. Read more. [Less]
Posted almost 2 years ago by Joannes Vermorel
Our clients need better forecasts, but they also need better insights. Salescast, our webapp dedicated to retail, wholesale and manufacturing was having about 200 words of documentation, which, by any standard was thin.
Hence, over the ... [More] last couple of weeks, we gathered materials, and this morning, we released about 7000 words of documentation concerning Salescast ins and outs.
First, we clarify the scope of Salescast, with special focuses on large retail networks, eCommerce and manufacturers. In short, if your company has inventory, then Salescast do apply to your case.
Furthermore, we details pros and cons of statistics. There are limits in what can be achieved through statistics; and Lokad does not come with divination capabilities.
As a rather touchy subject, we also position Salescast against our competitors. Naturally, we have a rather biased viewpoint here. So in the end, it comes down to benchmark us and see for yourself.
Also, Salescast isn't even remotely intended as a silver-bullet for supply chain. Lokad is a rather social software company in the sense it plays well with buddies, aka other complementary apps.
Nonetheless, Salescast does have some nice features on its own. It delivers Excel reports with both forecasts and key inventory optimization metrics (but you probably know that already). Just in case, here is a sample Excel report.
At this point, people typically ask How does your forecasting engine works? and How much does it cost? We already had two pages, namely Technology and Pricing, addressing those two questions, but we have tried to adopt a more practical angle here.
Once all those items are cleared, it's time to get started with Lokad. From there, you should quickly end-up with your first forecast report delivered. Expect the unexpected, forecasting is a counter-intuitive science. [Less]
Posted about 2 years ago by Joannes Vermorel
Europe’s dedicated magazine for the supermarket sector ESM is featuring Lokad this month in a two page article titled Embracing the Cloud. The publication gives a great overview of Lokad to retailers and details why the cloud is such ... [More] a game changer for retail. ESM had taken notice of Lokad’s presentation at the EuroShop 2011 in Düsseldorf, which particularly impressed the Editor Kevin Kelly.
Also, we agreed to offer ESM’s readers a free proof of concept to benchmark the retailer’s current forecasting accuracy compared to the one delivered by Lokad. Consider a 3 weeks net execution time to get hard facts on your forecasting practice.
In this issue, ESM investigates the highly competitive European grocery market, including the growing prominence of private labels, covers areas of interest to both buyers and other senior management working in the retail and manufacturing sectors, such as supply chain and logistics management, technology (such as EPoS), packaging and design and environmental best practices. The magazine appears bimonthly. [Less]
Posted about 2 years ago by Joannes Vermorel
Joannes Vermorel, founder of Lokad, discusses with Bruno Aziza, Director, Worldwide Strategy, Analytics and Business Intelligence at Microsoft, how companies are using the cloud to turn hard data into sales forecasts that enable them to optimize inventory and staffing. The interview was shot in New York during NRF11.
Copyright © 2013 Black Duck Software, Inc. and its contributors, Some Rights Reserved. Unless otherwise marked, this work is licensed under a Creative Commons Attribution 3.0 Unported License . Ohloh ® and the Ohloh logo are trademarks of Black Duck Software, Inc. in the United States and/or other jurisdictions. All other trademarks are the property of their respective holders.
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Moderate Activity
Estimated Cost
Analyzed 8 days ago based on code collected 8 days ago.
Project Cost Calculator
$ .00
36,554 lines
9 person-years
$ 478,615 *
*Using the Basic COCOMO Model
Estimate seems way too high?
Ohloh scans all files at any given code location to calculate the cost estimate.
Ohloh lets you exclude files and direc-tories from this calculation on the Code Locations page. You can get a more realistic estimate by excluding:
• External dependencies or libraries
• Non-code files
About Cost Estimates
• Software cost estimation is tricky business even when all the variables are known -- knowlegdge which we certainly don't have.
• We calculate the estimated cost of the project using the Basic COCOMO model.
• For those familiar with the details, we are using coeffcients a=2.4 and b=1.05.
• Please note that COCOMO was created to model large institutional projects, which often don't compare well with distributed open-source projects.
• COCOMO is meant to include the design, specification drafting, reviewing and management overhead that goes along with producing quality software.
• This model seems to be most accurate with mature, large projects. Young projects with little activity are typically overvalued.
Copyright © 2013 Black Duck Software, Inc. and its contributors, Some Rights Reserved. Unless otherwise marked, this work is licensed under a Creative Commons Attribution 3.0 Unported License . Ohloh ® and the Ohloh logo are trademarks of Black Duck Software, Inc. in the United States and/or other jurisdictions. All other trademarks are the property of their respective holders.
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User:Emma Goddery
From OpenWetWare
Jump to: navigation, search
I am a new member of OpenWetWare!
Contents
Contact Info
Emma Goddery (an artistic interpretation)
I work in the Your Lab at Arizona State University. I learned about OpenWetWare from Dr. Karmella Haynes, and I've joined because I am a student in the 2012 BME103 course.
Education
• Dakota Ridge High School
• Arizona State University Biomedical Engineering
• Year, BS, Institute
Research interests
1. Interest 1
2. Interest 2
3. Interest 3
Publications
1. Goldbeter A and Koshland DE Jr. . pmid:6947258. PubMed HubMed [Paper1]
2. JACOB F and MONOD J. . pmid:13718526. PubMed HubMed [Paper2]
leave a comment about a paper here
3. Mark Ptashne. A genetic switch. Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory Press, 2004. isbn:0879697164. [Book1]
All Medline abstracts: PubMed HubMed
Useful links
Personal tools
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[17] that Christ may dwell in your hearts through faith; to the end that you, being rooted and grounded in love,
This work is licensed under a Creative Commons Attribution-ShareAlike 3.0 United States License.
An XML version of this text is available for download, with the additional restriction that you offer Perseus any modifications you make. Perseus provides credit for all accepted changes, storing new additions in a versioning system.
load focus Latin (Saint Jerome, Bible Foundation and On-Line Book Initiative)
load focus Greek (Brooke Foss Westcott, Fenton John Anthony Hort, 1885)
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Google Adds 'My Account' to its Homepage
Apr 22, 2005 • 2:17 pm | (4) by | Filed Under Google Search Engine
This week Barry blogged about Google My Search History and pointed to other sources from our industry that talked more in depth about this new direction. Today for the first time I noticed that Google has added "My Account" to its homepage as well as a link to "My Search History" and of course the ability to sign out. Here is a snapshot:
Remember that you can discuss about this topic at WebmasterWorld and SEW Forums.
Previous story: Yahoo! 360 Short URLs Weird
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Mahalo is Spam According to Google's Quality Guidelines
Mar 24, 2008 • 10:02 am | (2) by | Filed Under Other Search Topics
According to Aaron Wall, since Mahalo adds no value (if you remove the links which point to other pages on the Internet), it violates Google's spam guidelines.
While many people, particularly SEOs (who have been frustrated with Jason's commentary in the past -- though to me, he redeemed himself during the keynote at SESNY last week), lauded Aaron's statements, I have to disagree with Aaron's statement. Personally, do you think that this post on how to convert from a PC to Mac lacks any substance? I think it's a great piece.
Jason responds in the Sphinn thread:
Over time I think you’ll see our pages grow to be over 50% original content, 20% links, and 20% UGC (i.e. reviews, votes, comments). Most pages in the system are 50-70% complete.... over the next two to three years they will reach 80-90% complete thanks to the help of the community and they will be worthy of a top 30-50 ranking in 20-30% of the cases is my guess.
Furthermore, all pages with less than 400 unique words will be nofollowed.
Personally, I think that's a great step. And I have to continue to disagree with individuals who think it's appropriate to continue bashing Jason when he clearly rectified the situation last week -- since, well, I was there.
As one member puts it, "if an actual human being combs through that data to provide us a garbage free page, then it should not be considered scraping or spam."
Agreed on all fronts.
Forum discussion continues at Sphinn.
Previous story: Use "Directory" + [Local Area] to Find Local Directories
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Can Google Grow Forever?
May 13, 2010 • 8:37 am | (2) by | Filed Under Other Google Topics
A WebmasterWorld thread is discussion a recent BusinessInsider piece that says:
Google's share gains have flatlined.
A year ago, Google had 65% of the US search market. Last month, Google had 64.4% of the US search market. This after an amazing decade in which Google gained about a point of share per month.
Most Google bulls expected Google to eventually own considerably more than 65% share of the US search market. As Google's competitors collapsed, it seemed Google would eventually be able to gain the 80%-90% share that Google has in many other countries worldwide. Based on Google's flatlining in the past year, however, this incremental 15-25 point market share gain now seems like wishful thinking.
Honestly, I love Danny's When Losers Are Winners: How Google Can “Lose” Search Share & Yet Still Stomp Yahoo. Danny says at the end:
Today's news also highlight a long-standing issue that the industry has yet to address. Search share and search volume figures don't tell the entire story. Both can be "inflated." Or deflated, in a world where "blended" search might integrate information that previously required two searches to retrieve.
The thread gets into the topic of, can Google grow forever? BillyS said:
I've been hearing about the downfall of Google for at least five years now. They dominated search back then, they still dominate. All efforts to close the gap have failed up until now. There is no logical reason to expect that to change. I don't see a major threat on the horizon. In fact, I could argue that based on history Facebook's days are limited.
Another person said:
Agree. They can't go up forever. IE hit a peak and is now settling back into a lower market range which it has held for years. I expect G to stay in the 60% share range for many years to come.
It is a debatable topic, which is why we have forums.
Forum discussion at WebmasterWorld.
Previous story: Google's New "Short Answers" Are Not "New"
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Person:George Watson (14)
Browse
Capt. George Watson
b.est 1602
• HCapt. George Watsonest 1602 - 1688/9
• WPhebe Hicks1614/15 - 1663
m. 1635
1. Phebe WatsonAbt 1636 - Abt 1682
2. John Watson1638 -
3. Mary WatsonAbt 1642 - 1723
4. Elizabeth Watson1647 - 1723
5. Elizabeth WATSON1648/49 - 1723
6. Samuel Watson1648/49 - 1649
7. Jonathan Watson1651/52 -
8. Elkanah Watson1655/56 - 1689/90
Facts and Events
Name Capt. George Watson
Gender Male
Birth[4] est 1602
Marriage 1635 Plymouth, Plymouth, Massachusetts, United Statesto Phebe Hicks
Death[3] 31 Jan 1688/9 Plymouth, Plymouth, Massachusetts, United States
GEORGE WATSON, CAPTAIN, OWNER OF WATSON'S POND
Immigration: 1633 New England Note: Captain George Watson is purportedly born in Holme on Spaulding, co. Yorkshire, Grantham, co. Lincoln or in London according to various sources. Charles Edward Banks recorded his origin as Dedham, Essex, England. George became a Freeman March 1633/34 in Plymouth, although he did not appear on the Freeman List. It is possible that he emigrated from Dedham, Essex, England in 1631 and lived in Penobscot from 1631 to 1633, when he removed to Plymouth. He died at age 87, and an inventory of his estate is taken on 2nd February 1688/9.
11th Feb 1632: "Examinations of John Deacon, Henry Sampson, George Watson and Oliver Gallow before Captain Walter Neale in New England; of Thomas Willett and William Phipps before Captain Henry Keye; and of Edward Astley before Attorney - General Noye in the case of Edward Astley. (CSPC)."
[Note: Contrary to the long-held belief that George Watson is the son of Robert and Elizabeth Watson of London, there is no evidence that either person ever existed in the colonies. Therefore, the origins of George's birth and parents are yet to be identified. However, an unverified entry is found of a marriage 28th Jun 1602 for Robert and Elizabeth (Pye/Peye/Paye) Watson in Holbeach, Co. Lincoln, England. Their list of children included George, Samuel, Nathaniel, Frances, Thomas, John and Robert.]
"Mr. Watson was one of the most respectable and useful members of the early settlement at Plymouth, holding various offices of trust, and faithfully performing his public duties, while his prudence enabled him to become quite independent, owning large tracts of land. he reared up a family of four children - three having died in infancy - from whom have sprung many of the most useful and prominent men of the colony and state down to the present period." At the meeting of the Council of War at Plymouth, 12th May 1653, when public apprehension is aroused of armed conflict with the Dutch of New Amsterdam, two barques were pressed for the service, one the vessel in which George Watson sailed.
George is a captain in the Plymouth Militia. Also, he served in 1653 as Captain of the Bark Expedition under Captain Myles Standish, against the Dutch in New York.
George is deeded 1/2 share in the Taunton Iron Works on 29th March 1663/64 from James Leonard, Sr., father of his son-in-law Thomas Leonard who had married (1662) his daughter Mary Watson. He is deeded another 1/3 share in the Taunton Iron Works on 8th December 1676 from his son-in-law Thomas Leonard. George Watson accumulated wealth in Plymouth in other transactions as well. He received a house and land from Thomas Hope on 29th April 1670. He is deeded a house on 26th October 1670 from Nathaniel Masterson. He received meadow land in Mattapoiset from Joshua Tizdell (Tisdale) on 3rd June 1685. Also, George deeded land on 9th September 1681 to his son Elkanah.
George Watson bought about 40 acres of land and a pond in Taunton, Massachusetts, New England from John Gilbert in 1664. The pond and land became a Massachusetts State Park sometime in the 20th century, On the the property behind the bathrooms and guard station towards the pond, a large stone is placed there with a plaque that reads. "JOHN GILBERT BORN IN ENGLAND IN 1580 CAME TO AMERICA IN 1635, WAS ONE OF 46 COHANNET PURCHASERS, SETTLED NEAR THE GREAT WEIR IN 1638, WAS REPRESENTATIVE AT THE GENERAL COURT IN 1639 IN 1641, FOR HIS SERVICES TO THE TOWN OF TAUNTON, HE WAS GRANTED THE LAND NOW OCCUPIED BY THIS PARK, HE BUILT HIS FARM HOUSE AND NAMED IT "PONDSBROOKE" AND LIVED HERE UNTIL HIS DEATH IN 1657. HIS BODY IS BURIED NEAR THIS SPOT."
Interestingly enough I will note here that on the hill overlooking the pond under some pine trees near this plaque, there are small groups of cobblestones embedded in the ground separated by a few feet apart in between them. There are probably quite a few different relatives burie
References
1. P.O.B.: "Topographical Dictionary of 2885 English Emigrants to New England, Savage, p. 437.
2. D./P.O.D.: "Plymouth Church Records, 1620-1859", Vol. I, p. 262: "(1688).
3. Plymouth, Mass. Vital Records, in Massachusetts Society of Mayflower Descendants. Mayflower Descendant, 15:214.
George Watson deceased on the 31 day of January 168[worn] Being Entered into the 87th yeare of his age. [Follows an entry for May 1688, suggesting this is 1688/1689. If so, his birth would be about 1602.]
4. George Watson, in Anderson, Robert Charles. The Great Migration Begins: Immigrants to New England, 1620-1633. (Boston: New England Historic Genealogical Society, 1995).
ORIGIN: Unknown
MIGRATION: 1631
FIRST RESIDENCE: Penobscot
REMOVES: Plymouth by 1634
BIRTH: About 1602 based on age at death (but this is probably exaggerated by about five years).
DEATH: Plymouth 31 January 1688/9, "being 87 years of age" [PChR 1:262].
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Place:Nanpantan, Leicestershire, England
Watchers
NameNanpantan
TypeVillage
Located inLeicestershire, England
the text in this section is copied from an article in Wikipedia
Nanpantan is a small village in the Charnwood borough of Leicestershire, England. It is located in the south-west of the town of Loughborough, but the village is slightly separated from the main built-up area of Loughborough. It is also the site of the Nanpantan Reservoir.
The first edge rails used in a wagonway were on the Charnwood Forest Canal, in the section between Nanpantan and Loughborough.
Research Tips
This page uses content from the English Wikipedia. The original content was at Nanpantan. The list of authors can be seen in the page history. As with WeRelate, the content of Wikipedia is available under the Creative Commons Attribution/Share-Alike License.
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Place:Thaxted, Essex, England
Watchers
NameThaxted
Alt namesTachestedasource: Domesday Book (1985) p 105
TypeTown
Coordinates51.95°N 0.333°E
Located inEssex, England
source: Getty Thesaurus of Geographic Names
source: Family History Library Catalog
the text in this section is copied from an article in Wikipedia
Thaxted is a town in the Uttlesford district of Essex, England, with about 2,500 inhabitants.
Research Tips
This page uses content from the English Wikipedia. The original content was at Thaxted. The list of authors can be seen in the page history. As with WeRelate, the content of Wikipedia is available under the Creative Commons Attribution/Share-Alike License.
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Australian Bureau of Statistics
Celebrating the International Year of Statistics 2013
ABS Home > Statistics > By Catalogue Number
2901.0 - Census Dictionary, 2011
Latest ISSUE Released at 11:30 AM (CANBERRA TIME) 23/05/2011
Page tools: Print Page RSS Search this Product
2011 Census Dictionary >> Glossary >> Household mobility
Household mobility
See Internal migration.
Previous PageNext Page
© Commonwealth of Australia 2013
Unless otherwise noted, content on this website is licensed under a Creative Commons Attribution 2.5 Australia Licence together with any terms, conditions and exclusions as set out in the website Copyright notice. For permission to do anything beyond the scope of this licence and copyright terms contact us.
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User:CuriousMark
From NAS-Central Buffalo - The Linkstation Wiki
Revision as of 04:26, 6 January 2008 by CuriousMark (Talk | contribs)
Jump to: navigation, search
Contents
CuriousMark's Kurobox becomes a TiVo MediaServer Page
I have had a Kurobox for some time now and it has served well as a basic file server. I have been wanting to upgrade it to act as a media server for my TiVo, but didn't know enough Linux to feel comfortable attacking the task. Also, the thought of putting together all the pieces seemed a bit daunting. With the recent advent of nearly automatic upgrades to uboot and modern Debian distributions, plus the arrival of pyTiVo on the scene, it seems like now is the time to quit procrastinating and get started.
Resources
The primary resource is NAS-Central and the wonderful people who are behind it. I started by asking in the forums. Here is the thread that this page is based on. It shows the false starts and mis-steps that I will leave out here in order to keep things readable and usable. My thanks to Andre, mindbender, and davy_gravy for their help and patience with this Linux newbie.
This wiki is also a treasure trove of information and searches have often led me to a page with just the information I needed. It is good to see it growing and maturing.
Here are Mindbender's directions with annotations and hints based on my experience added.
how to install LNI uboot + 2.4.33.3-firmimg.bin + freelink to the original kurobox/LS1
Prerequisites:
Things to do to get the Kurobox ready and software needed.
This may be where to install a new bigger drive in the Kuro. It will wake up in EM-Mode and you should be able to work from there. I didn't do it that way, I had a working box, but planned on expanding the size of the boot partition. I didn't think about expanding it until I was well into the process and found that expanding it when I did, later on, may have been better than doing so before starting. Your results may differ, you could do it now. I will describe what I did when I get to that point in the upgrade process.
Next we need the uboot, flash kernel, and Freelink code. Mindbender shows getting it like this:
cd /mnt/<br>
wget http://downloads.nas-central.org/LS1_PPC/Bootloader/Uboot/Precompiled/u-boot-lsppchd-flash-1.2.0-r2.bin<br>
wget http://downloads.nas-central.org/ALL_LS_KB_PPC/Bootloader/UBoot/FirmimgBins/linux-2.4.33.3-list.mg.2-v3/firmimg.bin<br>
wget http://downloads.nas-central.org/LS1_PPC/Distributions/Freelink/TmpImages/tmpimage-FL-1-21-LS1.tgz
That is downloading the uboot binary + the 2.4.33.3 firmimg.bin and the freelink rootfs to /mnt/ which should be the point to which /dev/hda3 should be mounted.
Since my kurobox was only slightly updated, wget was not available to me. Instead I downloaded those files to my PC and then put them on the share drive of the kuro using windows networking. I then logged into the Kuro using telnet and moved them to the suggested directory. If you are working with a new drive, this may not be an option. The Kurobox supports ftp, and that is another way to get the files onto the box.
cd /mnt/<br>
mv /mnt/share/u-boot-lsppchd-flash-1.2.0-r2.bin u-boot-lsppchd-flash-1.2.0-r2.bin<br>
mv /mnt/share/firmimg.bin firmimg.bin<br>
mv/mnt/share/tmpimage-FL-1-21-LS1.tgz tmpimage-FL-1-21-LS1.tgz<br>
Now the code is where it belongs.
Here is a link to the flash map of the LS1/kurobox original. http://buffalo.nas-central.org/index.php/Information/PPCFlashROM
There you will see the bootloader-device is "/dev/fl2" and the firmimg.bin device is "/dev/fl1".
Updating Flash Memory and the boot partition
You can install the latest uboot without compiling by flashing the binary to /dev/fl2
see http://buffalo.nas-central.org/index.php?title=U-boot_bootloader#Flashing_U-Boot_from_Linux for the full up directions and warnings. This process was too daunting for me to try previously, so I am very glad that Mindbender provided the software and isntructions to just do it.
This is the critical part of this guide. in case anything goes wrong when flashing uboot you will need jtag to recover the box. Be warned! It worked for me on three boxes. It worked for me with no trouble either, but you should know you are doing this at your own risk, you really can brick your Kurobox.
installing uboot to /dev/fl2
dd if=u-boot-lsppchd-flash-1.2.0-r2.bin of=/dev/fl2 bs=1k
In case you have cmp on board of your currently installed execute the cmp command also to be completely sure that it worked.
I did not have cmp and did not compare. I ran into issues later and was able to compare from Freelink (as installed below) and found that Flashing worked flawlessly. If you do wish to compare, install it to your kurobox before beginning.
:# cmp u-boot-lsppchd-flash-1.2.0-r2.bin /dev/fl2<br>
:cmp: EOF on u-boot-lsppchd-flash-1.2.0-r2.bin
EOF on the file means it was a good comparison. The flash device doesn't end at the end of file, so this result is expected. Everything up to the end of file compared successfully.
installing the 2.4.33.3 firmimg.bin to /dev/fl1
If you do this uboot update, I also recommend you flash the 2.4.33.3/telnet/ftp-enabled firmimg.bin to flash. This has the functionality you are used to having on your kurobox. This is an updated flash kernel that has some nice features that really makes many tasks a breeze compared to the kuro's old EM-Mode
This is the kernel that will be used in EM-Mode and when you do a two flash uboot startup.
:dd if=firmimg.bin of=/dev/fl1 bs=1k
Again, if you have cmp onboard, use it to make sure everything was written correctly.
Do both of the above flash operations at the same time without rebooting. Then when you reboot you will see the box behaving differently. It should behave as shown in this youtube video by Mindbender:
http://www.youtube.com/watch?v=pTPrrUYd8O8
Watch the video closely keeping your eyes on the disk fault light. It will blink 1, 2 or 3 times to indicate which boot mode it will execute when the entry timer expires. Each time you press the power button, the number of flashes will increment. If you do nothing it will boot "single flash". At this point in the upgrade you effectively can only boot to 2 and 3 blink mode as you only have a bootable kernel in flash, but not on the hard drive in the /boot directory (thats why you are flashing the 2.4.33.3-firmimg.bin, btw. If you don`t install the flash kernel, and reboot uboot will have problems finding a kernel to boot. The 1-blink mode will work as well, but in reality it will be 2 blink mode as it defaults to the flash kernel if no kernel is found in /boot on the hard drive. The Kurobox software does not have a /boot directory, so this is exactly what happens. A blank disk would also do the same.
Installing freelink to /dev/hda1
Once you have booted into EM mode with uboot (three flashes of the Disk Fail light during the uboot portion of startup), you can install freelink to /dev/hda1. Login via telnet, seeing the above welcome screen, and then you can do several things. The first thing I did was to repartition the hard drive. I used fdisk and mke2fs to create a 5GiB root partition, a 256MiB swap partition and left the rest of th 200GB hard drive for the working partition. Good generic directions can be found at [Convert your PPC LinkStation into a full-blown Debian system].
mount_disk<br>
cd /mnt/hda1<br>
rm -r *<br>
tar xzvf ../hda3/tmpimage-FL-1-21-LS1.tgz<br>
That should be all, when you reboot again you will be running Freelink. This is now the beginning, Freelink needs configuring and addition of all the software you will want to make the box functional as a NAS and media server.
Installing U-Boot on a Kurobox HG
Here are Mindbender's directions.
how to install LNI uboot + 2.4.33.3-firmimg.bin to the kuroboxHG/LS HG/LS HS
This section is taken from Mindbender's post, but I have not tried it given that I only own an original Kurobox. It should work just as well as the standard update did for me.
Prerequisites:
cd /mnt/
wget http://downloads.nas-central.org/LSHG_PPC/Bootloader/Uboot/Precompiled/u-boot-lsppchg-flash-1.2.0-r2.bin
wget http://downloads.nas-central.org/ALL_LS_KB_PPC/Bootloader/UBoot/FirmimgBins/linux-2.4.33.3-list.mg.2-v3/firmimg.bin
(thats downloading the uboot binary + the 2.4.33.3 firmimg.bin to /mnt/ which should be the point where /dev/hda3 should be mounted to)
ok, here is the flash map of the LS HG/LS HS/kuroHG original. http://nas-central.org/index.php/Information/HGFlashROM there you see the bootloader-device /dev/mtd1 and the firmimg.bin device /dev/mtd0.
you can install the latest uboot without compiling by flashing the binary to /dev/mtd1 http://buffalo.nas-central.org/index.php?title=U-boot_bootloader#Flashing_U-Boot_from_Linux
this is the critical part of this guide. in case anything goes wrong when flashing uboot you ll need jtag. be warned. it worked for me on three boxes... installing uboot to /dev/mtd1
dd if=u-boot-lsppchd-flash-1.2.0-r2.bin of=/dev/mtd1 bs=1k
in case you have cmp on board of your currently installed execute the cmp command also to be completely sure that it worked.
installing the 2.4.33.3 firmimg.bin to /dev/mtd0
if you do this, i recommend to flash the 2.4.33.3/telnet/ftp-enabled firmimg.bin to flash....this has the functionality you are used to on your kurobox.
dd if=firmimg.bin of=/dev/mtd0 bs=1k
again, if you have cmp onboard then use it to make sure everything got written correctly.
do both at the same time without rebooting. then you can reboot the first time...you ll see the box behaving differently...hopefully that way: http://www.youtube.com/watch?v=pTPrrUYd8O8
then you effectively can only boot to 2 and 3 blink mode as you only have a bootable kernel in flash (thats why you are flashing the 2.4.33.3-firmimg.bin btw...if you don`t and reboot uboot will have problems to find a kernel to boot). the 1-blink mode will work as well, but in reality it will be 2 blink mode as it defaults to the flash kernel if no kernel is found in /boot
BOTH
Update to latest 2.6-kernel last desirable thing to do is to update to a late 2.6 by running andre`s webinstaller with variant-uboot mode: http://hvkls.dyndns.org/downloads/documentation/README-webinstaller.html
This did not go well for me at first. Here is my expanded set of directions that worked better.
Update Freelink before updating the kernel to 2.6
I found that the web installer failed and asked me to reinstall several items using apt-get when I ran it on a virgin Freelink installation. When I tried to do the re-installs, apt-get cried foul and failed saying I needed to run apt-get update first. Upon running that, it had a failure and recommended running it again. the second apt-get update worked with no error messages and then the re-installs worked fine. After all of that, the web-installer ran without a hitch. Here is a summary of the command lines and the order that should get things up and running quickest.
TBD
TBD
foonas is easier for testing you can between also test http://downloads.foonas.org/foonas/alpha/lsppchd/foonas-bootstrap-image-1.0-dev-lsppchd.rootfs-svn-1062.tar.gz it has the advantage that you do not need to provide a 2.6-kernel as it already has one included...and /etc/fstab is already modified the right way.....so you would only need to untar it to /dev/hda1 and then reboot to 1 blink mode.
Things left to do when upgrading to 2.6 remember that if you update to a 2.6 kernel using foonas you will have to modify /dev/fstab...in 2.4 all devices were called /dev/hdaX , in the current foonas 2.6-kernels they use the /dev/sdaX device. The 2.6 kernel installed by Andre's web installer uses hdaX and no modification to /def/fstab is required when using it.
Personal tools
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Error!
Success!
Visual Studio 2008 Samples for c#
0
kicks
Visual Studio 2008 Samples for c# (Unpublished)
LINQ Samples :We have created four sets of samples for you that are designed to help you learn LINQ quickly, and to act as a reference and guide for those with more advanced skills. A few handy utilities are also included.
Kicked By:
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Myanmar TaxationEdit This Page
From FamilySearch Wiki
Residents pay a progressive individual income tax of 3–30%.
The corporate tax rate is 30% with a 10% capital gains tax (40% for non-residents). Indirect taxes include a commercial tax on prescribed services, ranging from 5% to 30%, and on goods, ranging from 5% to 200%. There are also social security taxes, customs duties, royalties on natural resources, stamp tax, and property tax. The ratio of tax revenues to GDP is very low, estimated at between 2.3% to 3.6%, of which it is also estimated that 87% goes to the military.
Economy of Burma
From Wikipedia, the free encyclopedia
The Economy of Burma (Myanmar) is an emerging economy with an estimated nominal GDP of $51.93 billion[2] and a purchasing power adjusted GDP of $83.74 billion.[2] Real growth rate is estimated at 5.5% for the 2011 fiscal year.[3]
Historically, Burma was the main trade route between India and China since 100 BC. The Mon Kingdom of lower Burma served as important trading center in the Bay of Bengal. After Burma was conquered by British, it became the wealthiest country in Southeast Asia. It was also once the world's largest exporter of rice. It produced 75% of the world's teak and had a highly literate population.[4]
After a parliamentary government was formed in 1948, Prime Minister U Nu embarked upon a policy of nationalization. The government also tried to implement a poorly thought out Eight-Year plan. By the 1950s, rice exports had fallen by two thirds and mineral exports by over 96%. The 1962 coup d'état was followed by an economic scheme called the Burmese Way to Socialism, a plan to nationalize all industries. The catastrophic program turned Burma into one of the world's most impoverished countries.[5]
In 2011, when new President Thein Sein's government came to power, Burma embarked on a major policy of reforms including anti-corruption, currency exchange rate, foreign investment laws and taxation. Foreign investments increased from US$300 million in 2009-10 to a US$20 billion in 2010-11 by about 667%.[6] Large inflow of capital results in stronger Burmese currency, kyat by about 25%. In response, the government relaxed import restrictions and abolished export taxes. Despite current currency problems, Burmese economy is expected to grow by about 8.8% in 2011.[7] After the completion of 58-billion dollar Dawei deep seaport, Burma is expected be at the hub of trade connecting Southeast Asia and the South China Sea, via the Andaman Sea, to the Indian Ocean receiving goods from countries in the Middle East, Europe and Africa, and spurring growth in the ASEAN region.[8][9]
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• This page was last modified on 12 February 2013, at 20:21.
• This page has been accessed 574 times.
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Penitentiary Prisoner IndexEdit This Page
From FamilySearch Wiki
The Colorado State Archives has placed an Index to the Penitentiary Prisoners for the period 1871 to 1973 on its website. The Index gives the inmate's name and file number.
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143rd Regiment, Illinois Infantry (100 days, 1864)Edit This Page
From FamilySearch Wiki
Revision as of 01:54, 23 January 2013 by Ericcharlesperazzo (Talk | contribs)
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)
United States U.S. Military Illinois Illinois Military Illinois in the Civil War
143rd Regiment, Illinois Infantry (100 Days, 1864)
Contents
Brief History
The 143rd Regiment, Illinois Infantry (100 Days, 1864) was organized at Mattoon, Ill., and mustered in for 100 days June 11, 1864. It mustered out September 26, 1864.[1]
For more information on the history of this unit, see:
Companies in this Regiment with County of Origin
Men often enlisted in a company recruited in the counties where they lived though not always. After many battles, companies might be combined because so many men were killed or wounded. However if you are unsure which company your ancestor was in, try the company recruited in his county first.
The following counties of origin are taken from the Adjutant General's Report, found on the Illinois Civil War Rosters web site. Roster pages are from the same report found on the Internet Archives web site. The rosters show the men who served in each regiment, their residences, dates of enlistment and mustering out, and other remarks.
Company A - many men from Coles County - Coles County web site - Roster, vol. 7, page 192.
Company B - many men from Alexander County - Alexander County web site - Roster, vol. 7, page 193.
Company C - many men from Christian and Montgomery counties - Christian and Montgomery county web sites - Roster, vol. 7, page 195.
Company D - many men from Clay County - Clay County web site - Roster, vol. 7, page 196.
Company E - many men from Fayette County - Fayette County web site - Roster, vol. 7, page 198.
Company F - many men from Effingham and Madison counties - Effingham and Madison county web sites - Roster, vol. 7, page 199.
Company G - many men from Shelby County - Shelby County web site - Roster, vol. 7, page 200.
Company H - many men from Montgomery County - Montgomery County web site - Roster, vol. 7, page 202.
Company I - many men from Moultrie County - Moultrie County web site - Roster, vol. 7, page 203.
Company K - many men from Coles and Madison counties - Coles and Madison county web sites - Roster, vol. 7, page 205.
The Civil War Soldiers and Sailors database lists 1,127 men on its roster for this unit. Roster.
Other Sources
WEBSITES
Civil War Soldiers and Sailors System, (accessed 30 Dec. 2010) can be searched by soldier's name or by regiment; includes regimental rosters and additional history of the regiment.
BOOKS & FILMS
United States. War Department. Record and Pension Office. Compiled Records Showing Service of Military Units in Volunteer Union Organizations. (Washington, District of Columbia : The National Archives, c1964). 48th-53rd Infantry FHL US/CAN Film 1488474 . Other libraries with this microfilm.
References
1. National Park Service, The Civil War Soldiers and Sailors System (accessed 4 January 2011).
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For the half-year to 30 June 2013, the IPKat's regular team is supplemented by contributions from guest bloggers Stefano Barazza, Matthias Lamping and Jeff John Roberts.
Two of our regular Kats are currently on blogging sabbaticals. They are Birgit Clark and Catherine Lee.
Tuesday, 2 February 2010
Apple and Fujitsu in iPad trade mark dispute
Update: a wise blogger recently remarked that realising that someone else has already blogged about the same topic is one of the little quirks of a blogger's life. Not realising it and blogging anyway is another quirk and might explain the duplicity in the IPKat's reporting on the iPad dispute (see here for the Amerikat's take on the same story).
The N.Y. Times reports of a trade mark dispute between Apple and Japanese Fujitsu’s over the mark iPAD (in relation to class 9 goods). This dispute is reminiscent of Apple's trade mark dispute with Cisco, when it first introduced the iPhone several years ago.
While Apple uses the iPAD mark on a type of electronic device, which an uninformed lay person might describe as a type of laptop in the form of overgrown mobile phone (pictured to the right), Fujitsu's iPad (pictured above left) is meant for use in the retail industry, allowing retailers to check prices and inventories and has reportedly been on the market since 2002. A Fujitsu spokesperson is cited with the words "...it’s our understanding that the name is ours,” and Fujitsu appears to have applied for an US trade mark in 2003.
According to the report, Fujitsu's application was blocked by an earlier iPad mark for a handheld computer device (in the name of MAG-TEK, INC, US trade mark No. 3405828, this Kat assumes).
After initially abandoning its US application, Fujitsu decided to "revive" it last summer, one month before Apple filed its iPad application (under the name of a third party). After extending the opposition deadline several times, Apple now appears to have been set a deadline of 28 February 2010 to oppose Fujitsu's application No. 76497338 IPAD, which was published for opposition purposes on 1 September 2009.
While most observers predict that the current dispute will most likely settle, we also learn from the N.Y. Times report that German company Siemens and - perhaps more intriguingly - a Canadian lingerie company called Coconut Grove also own US trade marks for iPad.
A quick check of the USPTO trade mark register (right) reveals numerous IPAD marks in different permutations and that Siemens' mark covers "programmable electrical drives for controlling machine..." in class 9, whereas Coconut Grove's mark covers "pads for use in bras" in class 25.
We should know more once the opposition deadline has passed... in the meantime, Merpel thinks that this story will be helpful publicity for both sides's products.
Subscribe to the IPKat's posts by email here
Just pop your email address into the box and click 'Subscribe':
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Preferences
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[maemo-users] [maemo-users] Is it possible to boot from the MMC card?
From: Frantisek Dufka dufkaf at seznam.cz
Date: Mon Mar 27 18:52:19 EEST 2006
Well, there is some interesting stuff in /mnt/initfs/linuxrc which is a
script executed when device boots but you probably need access to serial
console (pins next to battery) to see the menu or use flasher to set
root device permanently. So the answer is - yes it is possible.
Also the script could be modified to check for the card and some file on
it and boot it only when it is found. And of course it can be easily
modified to stop booting at all ;-)
Frantisek
Disconnect wrote:
> On Mon, 27 Mar 2006, Jose Maria Rodriguez Millan did have cause to say:
>
>> <html><div style='background-color:'><DIV class=RTE>Hello,</DIV>
>> <DIV class=RTE> I'm taking my first steps in developing for this platform, and, although i've seen a developer system can be flashed on the device, i'm wondering if it's possible to somehow copy the developer system on the MMC card and then, boot from it.I guess that would make developing for the platform way easier.</DIV>
>> <DIV class=RTE>Any comments on this?</DIV></div><br clear=all><hr>Don't just search. Find. <a href="http://g.msn.com/8HMBEN/2749??PS=47575" target="_top">MSN Search</a> Check out the new MSN Search!
>
> Use text, mmkay?
>
> And no, right now there is no way to boot off mmc (or network, or..) but
> something could probably be hacked up. (See the multiboot-menu packages that
> were developed for the ipaq - the basic idea is that all versions share a
> kernel, but the initrd can 'chain' to other initrds and roots.)
> _______________________________________________
> maemo-users mailing list
> maemo-users at maemo.org
> https://maemo.org/mailman/listinfo/maemo-users
More information about the maemo-users mailing list
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Public water users could see flush tax quadruple while septic users would pay double
January 27, 2012 at 1:49 am
By Daniel Menefee
Dan@MarylandReporter.com
(Photo by ex.libris. Creative Commons license)
CORRECTION: Many homes on public water and sewer in Maryland could see their flush tax almost quadruple triple to over $117 a year for the Bay Restoration Fund — much higher than homes on septic systems, which would see the fee double to a $60 cap under Gov. Martin O’Malley’s proposed consumption-based tax. (Original calculation was incorrect.)
Howard County Deputy Chief of Utilities Jeff Welty said that under the proposal, homes on public water in the Baltimore-Washington suburbs would be the largest contributors to the Bay Restoration Fund through the flush tax. The proposal caps the flush tax on homes on septic at $5 per month, or $60 annually.
The O’Malley administration has said that a consumption-based tax would lower the monthly payment below the current $2.50 to $1.80 for a “low-end user” — a household using less than 2,000 gallons per month — such as a single person living at 1600 W. North Ave. in Baltimore, for instance, the governor has said.
The fee would increase to over $9 a month for “high-end users” — households using 8,000 gallons or more per month.
Usage levels called too conservative
Utility managers think these usage levels are too conservative and push average water users to the top of the scale.
Chestertown Utilities Manager Robert Sipes said someone who uses 8,000 gallons a month “is not necessarily a high-end user.” A family of three or four could easily exceed 8,000 gallons per month, with their flush taxes “exceed[ing] the $5 cap applied to septic users significantly. It would essentially triple the current flush tax for a couple with two kids on public water.”
The American Water Works Association reports that an average person uses 70 gallons of water each day. Using this formula, a household of four would use 8,400 gallons of water per month. CORRECTION: The governor’s proposal would increase this household’s tax from the current $30 to $117 $106.
Maryland municipal utility officials said their average home with two to three people uses about 5,000 gallons of water a month. Under O’Malley’s proposal, the flush tax from these homes would be higher, with those on city water paying $5.55 a month.
Residents of Maryland’s suburban counties would be paying the higher fees all around. Welty said average residential use in Howard County is 5,200 gallons. Harford County residents average 5,400 gallons, according to the county’s website. Officials in Elkton in Cecil County report nearly 60% of residential homes use more than 9,000 gallons per month. Under O’Malley’s proposal, this group of Elkton homes would pay $127 annually.
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Featured
From our Essentially Ellington photographer: Scene in New York City
Edmonds-Woodway High School parent Jon Howeiler accompanied the Edmonds-Woodway High School Jazz Ensemble 1 as they made their trip to New York City for the Essentially Elllington Festival at Lincoln Center last Thursday-Sunday. Here are some of his photos: »
Here’s your chance to name a park in Edmonds
The city is asking Edmonds’ residents for their ideas on a name for the SR 104 mini park located at 131 Sunset Ave. Located in the Ferry holding lanes, this park also serves as a stopping point for a restroom break for walkers that stroll around Edmonds. The park has also just had a minor face-lift. All suggested names will go to the Planning Board for consideration and a hearing. The Planning Boa... »
E-W Jazz Band honored at Essentially Ellington for outstanding clarinet section, soloist
Edmonds-Woodway High School’s Jazz Ensemble missed out on being named one of the top three bands Sunday in the Essentially Ellington Festival at the Lincoln Center in New York City, but the school did receive an award for outstanding clarinet section. In addition, E-W clarinet player Kyle Brooks received an award for outstanding soloist. The only Seattle-area band to make the final list was ... »
Reminder: Essentially Ellington continues today — top three bands announced at 1 p.m.
Edmonds-Woodway High School’s Jazz Ensemble performed Saturday, and more schools are performing today during the Essentially Ellington Jazz Festival in New York City. You can watch is all live here, including the announcement of the top three bands at 1 p.m. Pacific, followed by the finale concert and awards ceremony starting at 4:30 p.m. On Saturday and Sunday, Edmonds-Woodway High School&#... »
Edmonds scenic: Goose family out for a stroll
»
Seen Saturday in Edmonds: From soccer to fairies to whales to jazz, events galore
What do soccer players, fairies, jazz music and killer whales have in common? That was just a sampling of the events happening around Edmonds Saturday, as captured by My Edmonds News: »
Meet your SeaWolves at Edmonds TOP Food, 11 a.m. today!
Edmonds’ own premier development league soccer team, the North Sound SeaWolves, will be at Edmonds TOP Foods today — Saturday, May 11 — starting at 11 a.m. Here’s your chance to meet the players, get a poster and receive free tickets to the May 18 home opener! TOP Foods is located at 21900 Hwy 99 in Edmonds. »
Non-profit antique store Homestyle Mercantile focuses on joy of giving
Story, photos and video by Annie Wilson UW News Lab What started out as a handy way to relieve families of items from deceased family members while also raising money for charity soon grew into something much larger. About two years ago Reuben Sapien opened the Homestyle Mercantile, an antiques store in downtown Edmonds. The shop is one of several stores Sapien has started over the last 20 years f... »
Edmonds scenic: A fine day for a ferry ride
»
Edmonds scenic: Sporting that sunset glow
»
North Sound SeaWolves looking for host families for college players
Can you open your home to a college soccer player for two months? The North Sound SeaWolves – the Edmonds-based premier development league soccer team — is seeking host families for two college soccer players from out of the area who are looking for place to stay during the team’s two-month season, May 18-July 17. “These are college student-athletes that will be members of ... »
Reminder: Westgate UPS store to host grand opening, ribbon cutting Friday
Join our sponsor, the New UPS Store at Westgate Village, as they celebrate their grand opening and ribbon cutting with Mayor Dave Earling at 4:30 p.m. Friday, May 3. There will be prizes and contests, food, and the store will launch a book drive that will run through May 31. All books and money collected will help local children have the books they need to keep up their reading all summer. And rem... »
Great news for our readers: My Edmonds News partnering with Edmonds Deals!
My Edmonds News has partnered with Edmonds Deals to bring great deals from Edmonds-area businesses and organizations to our readers. Edmonds Deals was launched March 1 of this year by Peter Wilson as a service of his online marketing firm, bizmktg.com. The service consists of the website (EdmondsDeals.com), a Facebook page and Twitter @EdmondsDeals. The deals on Edmonds Deals are local so you won&... »
Edmonds Beauty: Spring into fashion
By Aygin Ghorbani The most refreshing and certainly beautiful season is here — spring! While the sun tries to beam through the gray, rainy skies, we notice the days start to get longer, the birds start chirping and the flowers are blooming. It’s a sense a hope for most of us, a sense of happiness that the summer days that we are all waiting for are getting closer. Spring offers a lot, includ... »
Edmonds scenic: Cloud rainbow
»
Artfully Edmonds: Grab Bag
By Juliet Brewster Best of luck to the EWHS Jazz I! They have been selected as finalists in the Essentially Ellington jazz competition and festival at Lincoln Center in New York City. Fourteen other high school jazz bands from all over the country will compete, including our Seattle rivals, Garfield and Roosevelt. The band will be in NYC from May 9-12. Fed, White & Brew – Edmonds’ own A... »
That’s all, folks! Robin Hood Lanes says goodbye with special Dollar Day event
Story and photos by Larry Vogel Robin Hood Lanes, an Edmonds fixture since 1960, bid a heartfelt goodbye to the Edmonds community on Wednesday with a full day of dollar bowling, dollar beers, and dollar food items. The iconic bowling alley on Edmonds Way closes for good on Thursday, April 25. Robin Hood will missed by many. Eileen Jacoby, 92, has been a regular bowler here for decades. “It&#... »
Local Paralympic swimmer qualifies for World Championships in record time
Story and photos by Ruth Whyman UW News Lab Kayla Wheeler was born missing both her legs and her right arm. Sitting on the side of Shoreline Pool, waiting for a recent Sunday afternoon practice to start with Shadow Seals – a local swim team for disabled athletes – it is difficult to imagine how she might manage the physicality of swimming. This is what makes her athletic ability so much more impre... »
Edmonds scenics: Several looks at Sunday’s sunset
»
Modern Money: In search of groceries
By Denise Marinacci It’s been a little while since our friendly, neighborhood grocery store, Petosa’s, closed its doors. I’m sure I’m not the only one that misses the smiling faces, the delicious deli food, and for me, the convenience of running over when you’ve forgotten a key ingredient for dinner. Now that Edmonds is without a grocery, I set out to find the shops, stores, bakeries that sell foo... »
E-W grad stuck in MIT dorm as Boston police search for bombing suspect
With all eyes on Boston during a series of tragic events, Edmonds-Woodway High School graduate Alex Springer — now a freshman at MIT — has had quite a week. Springer, a 2012 E-W grad who also served as a student representative on the Edmonds City Council last year, called My Edmonds News Friday morning to describe his experiences during the days following the Boston Marathon bombing th... »
Edmonds scenic: Sundown on the dock
»
Edmonds scenic: Monday sunset
»
Edmonds scenic: Taking in the view
»
Edmonds scenic: Friday night sky
»
Artfully Edmonds: Saying hello…
By Juliet Brewster How rude of me, I never introduced myself in last week’s column. My name is Juliet Brewster, and if you spend much time in downtown Edmonds, you’ve probably seen me behind the counter in the bookshop, The Cheesemonger’s Table, or at Twist Yoga Studio. I’ve been running around Edmonds since I was bratty enough to be kicked out of an antique shop in the old Old Milltown, but left ... »
It’s official: Edmonds IGA Market will open in former Petosa’s space May 15
By Larry Vogel Less than three months after Petosa’s Family Grocer closed its doors, Edmonds will once again have a full-service grocery in the downtown core. “We plan to open the new Edmonds IGA Market on May 15,” said owner Mike Trask. A gala grand opening event will be held in June. No stranger to the grocery business, Trask has been at the helm of the Granite Falls IGA since ... »
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It's easy! Just pick the product you like and click-through to buy it from trusted partners of Quotations Book. We hope you like these personalized gifts as much as we do.
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212 - The Extra Degree
The one extra degree makes the difference. This simple analogy reflects the ultimate definition of excellence. Because it's the one extra degree of effort, in business and life, that can separate the good from the great. This powerful book by S.L. Parker and Mac Anderson gives great examples, great quotes and great stories to illustrate the 212° concept. A warning - once you read it, it will be hard to forget. Your company will have a target for everything you do ... 212°
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Quotes by Fabry, Joseph
We don't have a biography. Please consult wikipedia.
"The human spirit is your specifically human dimension and contains abilities other creatures do not have. Every human is spiritual; in fact, spirit is the essence of being human. You have a body that may become ill; you have a psyche that may become disturbed. But the spirit is what you are. It is your healthy core."
Fabry, Joseph on human spirit
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Project info for Sencience
Created 13 Aug 2003 at 04:10 UTC by raquib, last modified 19 Dec 2006 at 02:00 UTC by raquib.
Homepage: www.cs.rit.edu/~tak7130
Notes:
Project name is now being used to describe my Master's Thesis. Goal of the project: to allow a training paradigm over a self supervising learning system. The work is an extension of Michael H. Coen's thesis, hopefully to be complete May 2007.
Description: AI Software
This project has the following developers:
X
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RuneScape/Ores
From StrategyWiki, the video game walkthrough and strategy guide wiki
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This page contains unnecessary drivel. As our aim is to present helpful and complete guides for games, pages do not need to contain unnecessary information, such as the names of contributors, or mini guides for editing. If you are qualified, please edit it to remove the drivel, and then remove this template from the page.
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Drivel specifics: especially the last section on rune essence
Contents
[edit] Ore price listing
Rock Mining level required Exp received Selling price Notes
Rune Essence 1 5 To mine these rocks, players will need to complete the Rune Mysteries Quest. To learn more about rune essence, see the runecrafting skill guide. Rune essence does not have a respawn time, players only need to click once on the rock and they will mine until their inventory is full.
Clay 1 5 1-50gp Clay is used in crafting to make small items such as pots.
Copper 1 17.5 1-10gp One of the basic ores all mining beginners mine. Players can smelt a copper ore with a tin ore to make a bronze bar.
Tin 1 17.5 1-10gp One of the basic ores all mining beginners mine. It is smelted with Copper to make Bronze bars. Players can smelt a copper ore with a tin ore to make a bronze bar.
Limestone 1 26.5 This is used for building stuff in a Player-Owned House, such as a fireplace. It's also used for building temples in Shades of Mort'ton. You can mine it, or buy it from the Stone Mason in Keldagrim. Limestone is also needed in the members quest Shades of Mort'ton.
Blurite 10 17.5 Blurite is needed in the quest The Knight's Sword. The only non-member use occurs during the Knight's Sword quest.
Iron 15 35 5-100gp Common mining ore because it is also used to make steel. Has a 50% chance for success. Players can smelt an iron ore to make an iron bar.
Silver 20 40 100-400gp Silver is used to craft items, such as tiaras.
Coal 30 50 100-300gp An ore that is used in smelting all the high level ores except for gold. it is a valuable resource that is often fought over. Need level 30 mining and a good pickaxe or you'll be very slow. Coal can be smelted with iron ores, mithril ores, adamant ores, and runite ores to make steel, mithril, adamant, and rune bars.
Gold 40 65 120-200gp Gold is used to craft items, such as jewellery.
Gem 40 65 Gem rocks are only available to members who have completed the Shilo Village quest. Mining a gem rock gives a player a gem ranging from sapphire to diamond, which can be used for crafting.
Mithril 55 80 300-500gp Mithril is an uncommon rock to find. Mithril ores can be smelted with four coal ores to make a mithril bar.
Adamant 70 95 700-1000gp Rare rock that if found, there are few. Long respond time. Adamant ores can be smelted with six coal ores to make an adamant bar.
Runite/Rune 85 125 10000-14000gp Extermly Rare ore that is believed to be found in the wilderness. High respond time. Rune ores can be smelted with eight coal ores to make a rune bar.
[edit] How to turn a profit
Mining can be a very good skill to get money in for players who have patience. It might take a long time to get ores, but players can sell them for good prices. Players should sell their ores to other players, not to General Stores or the dwarf who owns the ore store in the Dwarven Mines. Iron and coal are the best ores to sell as many people want them to make steel bars.
So if a player has 1,000 coal ores and sells them all at 200 each, they've just made 200k. However, it takes time to mine coal, so it will take a long time to get rich off it. Also, please note one thing: you might be better off buying iron in the Dwarven Mine store, as they often sell it for 1gp each.
Copper, Tin, and Iron Ores have practically no value because so many people can mine them, so it is advised to bank your ores(if near a bank), or give it away(much faster than dropping one at a time).
[edit] Special ores
[edit] Elemental
For use in the Elemental Workshop quest.
[edit] Pure Essence
This type of essence is used for members to make the other higher level types of runes. You need to be a member and have 30 mining to be able to get it. The street price for it is about 100gp each.
[edit] Tips on Rune Essence
Tired of buying runes for such high prices? Well, upon completion of the Rune Mysteries Quest, you can talk to Sedridor in the Wizards Tower, Aubury in Varrock, Cromperty in Ardougne, Brimstail in the Gnome Stronghold, and someone in the Mage's Guild (66 Magic required) to go to the Rune Essence Mine! Once there, you can go back to the person you teleported from by use of a portal. If your mining level is 1-29 or you are not on a members-only world, you can mine the standard Rune Essence. This can be used to make Air, Mind, Water, Earth, Fire, and Body runes. They cannot be used on members' servers to make any other runes other than those six types!
If you are on a Members Server and have 30 mining, you can mine Pure Essence. Pure Essence can be used for rune crafting and can be traded on all servers, but can only be mined on members servers. You can make any rune with it (with the exception of Blood and Soul runes) and it sells better then standard Rune Essence.
Each Essence, Rune or Pure, gives 5 Mining XP. If you get the "Lost and found" Random Event, instead of receiving Law Runes, you get 9 noted essence. Ess is short for Essence in common RuneScape lingo.
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Pärnu
From Wikitravel
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Sand dunes in Pärnu
Pärnu [1] [2] is a resort seaside city (and estonia's summer capital) with a small harbour in the South-Western part of Estonia.
[edit] Understand
The city is famous for its spas, shallow white sandy beach and beautiful parks. It is also a popular place for conferences, theatre performances and concerts. In 1838 the first spa was opened and nowadays Pärnu is a health resort of international stature.
During the Great Northern War, the University of Tartu was relocated to Pärnu from 1699-1710. Tartu University still has a branch campus there today.
Since 1996 Pärnu has been known as Estonia's Summer Capital. Beach life, dozens of festivals, night clubs, big party crowds, concerts and funfairs - Pärnu has it all.
Pärnu is an ancient Hanseatic town where a medieval festival and an arts and crafts fair are held every summer. In 2010 Pärnu will host the 30th International Hanseatic Days.
[edit] Get in
[edit] By train
Pärnu has train connection to Tallinn. Trains depart from Tallinn and from Pärnu twice a day. Journey takes about 2,5 hours. Train schedules: Edelaraudtee
[edit] By car
Pärnu has road connections to Tallinn, Haapsalu, Tartu, Vändra, Paide and Riga in Latvia.
Distances table
1 km = 0.62 miles to Tallinn to Tartu to Viljandi to Narva to Paide to Haapsalu
from Pärnu 128 km 174 km 96 km 291 km 95 km 108 km
[edit] By bus
Buses arrive to Pärnu from Tallinn, Tartu, Haapsalu, Riga and several other places. Bus schedules and journey planner can be found at Bussireisid.
[edit] By boat
There is a ferry connection in from Kihnu (a small island in the Baltic Sea) to Pärnu and between Pärnu and Ruhnu. In winter there are also ice-roads between the islands of Pärnu bay.
Ticket Poland - Estonia
[edit] Get around
Pärnu downtown
[edit] By foot
Pärnu's old city is pretty small and navigateable by foot, but it's full of small boutiques. Lots of them are on Rüütli street.
[edit] By bus
Bus connection covers the whole city. There are 26 routes and their schedules can be found at Pärnu ATP-s website.
[edit][add listing] See
• Old town with buildings even from medieval times
• Beautiful wooden villas
• Beach park and promenade
• Pärnu, Lydia Koidula and Modern Art Museum and City gallery
• Pärnu Town Hall, Nikolai tn 3, Pärnu. The Town Hall is located in a house built in 1797 by the merchant P.R. Harder. In 1804, the Russian czar Alexander I stayed in this house during his visit to Pärnu. A new art nouveau building with neo-baroque details was completed in 1911 to the north side of the town hall. edit
• Church of Jekaterina, Vee tn 8, Pärnu. Completed in 1768, the Church of Jekaterina is the most style pure and ample baroque church in Estonia. The church has influenced the development of orthodox church architecture in whole Balticum. To this day Pärnu Russian congregation operates in the church. edit
• Seegi Maja (the Almshouse), Hospidali tn 1, Pärnu. Seegi Maja (the Almshouse) is known to be the oldest building in Pärnu. It was built in 1658 on the remains of the old almshouse of the Holy Spirit's Church as a shelter to the sick and the cripples.Based on research on the wooden raft under the basement of the building, the old almshouse dates back to 1250-1350. The building was restored to look the way it did in the 17th century. It houses the St. Peterburg Hotel and the restaurant Seegi Maja. edit
• Statue of Raimond Valgre, Mere puiestee 22, Pärnu. The musician Raimond Valgre who played in Pärnu in 1930s and brought the town a lot of fame, can today be found sitting in the park near Kuursaal, as a bronze statue. You too can sit down next to the beloved Estonian composer and enjoy his beautiful compositions. edit
• Eliisabet's Church, Nikolai tn 22, Pärnu. Eliisabet's Church, inaugurated in 1750, is the most outstanding sacral building of the Baroque period in Estonia. One of the best organs in Estonia is in Eliisabet's Church and the place is popular as a concert hall among music lovers. edit
[edit][add listing] Do
Pärnu has a long beach that opens to the south and has lovely, almost white sand. It is a major Baltic seaside resort and as previously stated, the summer capital of Estonia.
Completed in 2006, the attractive Beach Promenade has the feeling of a real resort and makes the Summer Capital's beaches inviting even in bad weather! Well thought-out lighting keeps the beach active even when the sun goes down, and the playing colours of the fountains are a sight in themselves in the darkness of the night.
It may rain in summer, when it does many Estonian holiday makers go indoors to the Vee Park, which is an indoor water park at the largest beachside hotel. As water parks go, this one is fantastic, and insures that your few days at the beach won't be wasted because of inclement weather. There are lot of interesting historical buildings in the city center. You may ask special brochure and discover all of them by your own
Just south of downtown is a modern art museum and art school. Exhibits change frequently, and are often edgy and provocative.
Outdoor activities:
• Horseback riding
• Sassi Talu, Kabriste küla, Audru vald (18 km west of Pärnu), +372 56 467 301 (), [3]. European-standards excellent service and reception; good both for novice and for experienced riders. edit
In winter time, consider one of the many spa hotels:
• Spa Estonia [4] Modern and moderate-priced; great variety of medical treatment and procedures.
• The Pärnu International Documentary and Anthropology Film Festival. edit
• David Oistrahh festival. edit
• Organ festival. edit
• Bacardi Feeling Beach festival. edit
• Hanseatic and Handicraft days. edit
• The Pärnu Jetty, Seedri tn 6, Pärnu. The jetty has gained a romantic aura as a place for lovers to take walks on; however, before taking a walk there, be sure to check the water level! The length of both jetties is a bit over two kilometres. edit
[edit] Learn
Pärnu College of Tartu University[5] is located in Pärnu.
[edit] Work
[edit][add listing] Buy
[edit][add listing] Eat
[edit] Budget
[edit] Mid-range
[edit] Splurge
[edit][add listing] Drink
Pärnu has several clubs and lounges, most of them located in the center near Ruutli street. There are also many cozy terraces and pubs around the numerous parks in the summer time.
[edit][add listing] Sleep
[edit] Budget
• Residence Konse, 44a Suur Joe. edit
[edit] Mid-range
• Spa Estonia [6]
• Victoria Hotel, [7]. Small historical hotel in the city centre edit
• Villa Andropoff, Valgeranna küla, Audru vald 88326, Pärnumaa, Estonia (9 km from Pärnu, 120 km from Tallinn), +372 4443 453 (, fax: +372 4442 004), [8]. checkin: 14:00; checkout: 12:00. The Resort and Conference Centre Villa Andropoff in Estonia nearby the historical baltic seaside resort Pärnu offers many-faceted possibilities for the whole family. Twin room 50EUR. (6471898.5,522843.4) edit
[edit] Splurge
[edit] Contact
[edit] Stay safe
[edit] Cope
[edit] Get out
If you dare leave urban pleasures behind, there is great countryside to explore. Many cultural and sports events offer entertainment all year round. The enthralling nature of Pärnu County, which offering ample, quality opportunities for active holidays, supplements the greenery of city parks and boulevards. Walks in the forest, kayak trips on rivers and the sea, riding, fishing, hunting trips, adventurous bog or canoe trips to Soomaa National Park and elsewhere in the county offer great escapes. In winter, you can partake in skiing, snowshoe and kick sledge trips, or a sledge safari and enjoy a sauna steam.
Kihnu Island is cosy and warm yet exotic – folk costumes are worn here every day and the handicraft of older generations remains highly valued. Several other romantic small islands and the Western Coast offer an opportunity to participate in local rites and customs.
• Valgeranna, unspoilt beach just 8 kilometres from Pärnu
• Kabli, quiet seaside village with bird-ringing centre and rural beaches
This article is an outline and needs more content. It has a template, but there is not enough information present. Please plunge forward and help it grow!
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Australian Bureau of Statistics
Celebrating the International Year of Statistics 2013
ABS Home > Statistics > By Catalogue Number
2016.2 - Census of Population and Housing: Selected Characteristics for Urban Centres and Localities, Victoria, 2001
Latest ISSUE Released at 11:30 AM (CANBERRA TIME) 26/03/2003
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INTRODUCTION
STATISTICS PRESENTED IN THIS PUBLICATION
This publication presents a range of social and housing statistics produced from the 2001 Census of Population and Housing for Victoria. For comparative purposes, it includes 1996 Census data based on 2001 Census geography.
The tables in this publication provide a selection of data for the population and their housing arrangements for Urban Centres/Localities (UC/L) and Sections of State (SOS). These geographic structures classify areas according to size of the population. Whilst the UC/L classification covers only part of each state and territory, the SOS classification covers each state and territory in its entirety. Refer to the Glossary for more information about the definitions of UC/L and SOS.
The data are presented on the basis of where people were counted on Census Night ('as enumerated'), and therefore data for each geographic area include visitors to that area, but exclude those people who were away on Census Night. All tables exclude overseas visitors unless otherwise stated.
Data released in Selected Social and Housing Characteristics, Victoria (cat.no. 2015.2) provide various characteristics for Statistical Divisions (SDs), Statistical Subdivisions (SSDs) and Statistical Local Areas (SLAs). Refer to Appendix 2-Census Products and Services for more information.
POPULATION MEASURES
Census counts should not be confused with the Australian Bureau of Statistics' (ABS) official population estimate, the Estimated Resident Population (ERP) which is used for electoral purposes and in assisting in the distribution of government funds to state and local governments. The ERP is the definitive population estimate and is derived from the census counts. For a fuller description of population measures and the derivation of ERP, please see paragraphs 6, 7 and 8 of the Explanatory Notes. Appendix 1-Estimated Resident Population includes a table showing census counts and ERP for each state and territory.
One of the important features of the Census is that it describes the characteristics of Australia's population and housing for small geographic areas and small population groups. While not available in this publication, data at the smallest geographic level (Collection District) are available in a range of census products. For more information on these products, please refer to Appendix 2-Census Products and Services. Concepts and definitions used in this publication are explained in the Glossary and more detailed information is available in the 2001 Census Dictionary (cat. no. 2901.0). The Explanatory Notes in this publication provide a discussion of the scope and coverage of the Census, the different measures of population, and the limitations of census data.
This publication is one of a series of publications which provide data for both UC/L and SOS geographic areas for each state and territory (cat. no's 2016.1-7). A similar publication is also available, providing data for urban centres across the whole of Australia (cat. no. 2016.0). See Appendix 2-Census Products and Services for more information.
SUMMARY OF FINDINGS
Person characteristics
Population distribution
The Census of Population and Housing conducted on 7 August 2001 counted 4,644,950 persons (including 32,853 overseas visitors) in Victoria, an increase of 6.2% or 271,430 persons since 1996 (4,373,520 persons).
There were 147 Urban Centres and 188 Localities in Victoria in 2001, two of which had population counts of over 100,000 persons: Melbourne (3,160,171) and Geelong (130,194). These Urban Centres accounted for 70.8% of the population.
Age
In 2001, the median age was 35 years, compared with 33 years in 1996. The proportion of persons aged 0-14 years decreased from 21.2% in 1996 to 20.4% in 2001, while the proportion of persons aged 65 years and over increased from 12.0% to 12.7% during the same period.
The highest percentages of people aged 0-14 years were recorded in the Localities of Moriac (36.0%), Merino (34.9%), Cabarita (34.1%) and Noorat (34.0%).
A high proportion of people aged 65 years and over was recorded in the Urban Centres of St Leonards (34.1%), Portarlington (32.4%) and Paynesville (31.2%), and in the Localities of Indented Head (33.3%), Edenhope (32.6%) and Minyip (31.9%).
Sex
In 2001, there were slightly more females (2,365,889) than males (2,279,061) with higher proportions of females recorded in urban areas (Major Urban (51.2%) and Other Urban (51.6%)) and Bounded Locality (50.1%). Proportionally more males were recorded in Rural Balance (51.9%).
Females outnumbered males in 215 Urban Centres and Localities. A higher proportion were recorded in the Urban Centres of Yarram (53.9%), Daylesford (53.9%) and Queenscliffe (53.8%), and in the Localities of Poowong (55.9%), Foster (55.4%) and San Remo (55.1%).
A higher proportion of males was recorded in the Urban Centres of Nagambie (53.3%), Barham-Koondrook (Koondrook Part) (52.1%), Kinglake (51.6%) and Yallourn North (51.4%), and in the Localities of Cann River (55.9%), Werribee South (55.8%), Dartmoor (55.7%) and Pyalong (55.5%).
Median individual income
The median weekly income grew from $290 in 1996 to $380 in 2001, an increase of 31.0%, with larger increases recorded in the Rural Balance (31.9% to $368) and Bounded Locality (34.1% to $303) sections of the state.
The highest median incomes were recorded in the Urban Centres of Mount Macedon ($547), New Gisborne ($517) and Hurstbridge ($463), while Localities with high median incomes were Dinner Plain ($603), Cabarita ($559) and Arcadia Downs ($534). The lowest median incomes were recorded in the Localities of Woomelang ($198), Loch Sport ($203) and Newlands Arm ($223).
Indigenous status
The number of persons who reported being of Indigenous origin increased by 16.8% to 25,079 persons in 2001, up from 21,474 persons in 1996, representing 0.5% of the total population (excluding overseas visitors).
Of the Urban Centres and Localities, 42.8% reported proportions of Indigenous persons equivalent to, or less than, the average for Victoria (0.5%). The highest proportions were recorded in the Localities of Cann River (10.6%), Halls Gap (5.0%) and Bruthen (4.9%), and in the Urban Centres of Heywood (7.4%) and Orbost (4.2%).
Birthplace
The number of persons born in Australia increased by 3.4% to 3,277,054 persons in 2001, up from 3,168,853 persons in 1996, representing 71.1% of the population of Victoria (excluding overseas visitors).
In 51 of the Urban Centres and Localities, 90.0% or more of the population were born in Australia. The highest proportions of Australian born were recorded in the Localities of Cabarita (96.3%), Noorat (95.9%) and Murrayville (94.4%).
Of the population counted (excluding overseas visitors), 23.4% (1,080,344 persons) stated that they had been born overseas, with the highest proportions recorded in Melbourne (29.4%), Robinvale (25.1%) and Venus Bay (L) (24.9%).
Internet and Personal computer use
Of the population counted (excluding overseas visitors), 43.4% (2,001,169 persons) used a personal computer at home in the week prior to Census Night. Of the same population, 38.6% (1,780,853 persons) used the Internet in the week prior to Census Night.
The highest proportions of persons who used a personal computer at home were recorded in the Localities of Arcadia Downs (64.6%), Baranduda (64.0%) and Menzies Creek (61.6%), while low proportions were recorded in the Localities of Gunbower (16.3%), Loch Sport (18.4%) and Nyah (18.4%).
Employed persons
In 2001, there were 2,082,216 employed persons (93.2% of the labour force ), an increase of 197,336 persons since 1996.
Urban Centres with the largest numbers of employed persons were Melbourne (1,447,136 or 93.3% of the labour force), Geelong (53,507 or 90.6%), Ballarat (29,970 or 90.2%) and Bendigo (28,254 or 90.8%).
Full-time employment
The number of full-time workers increased from 1,285,053 persons in 1996 to 1,354,647 in 2001. This represents 65.1% of the employed population in Victoria. As a proportion of persons in the labour force, the highest proportions of full-time workers were recorded in the Localities of Dartmoor (70.9%) and Waterford Park (68.8%). Urban Centres with the highest proportions of full-time workers were Wallan (65.7%) and Wandong-Heathcote Junction (64.2%).
Part-time employment
In 2001, 663,221 workers (31.9% of employed persons) were employed part-time, an increase of 106,799 persons since 1996. The highest proportions of part-time workers, as a proportion of persons in the labour force, were recorded in the Localities of Venus Bay (50.0%) and Loch Sport (42.5%). The Locality with the lowest proportion of part-time workers was Dartmoor (20.4%).
Unemployed persons
In 2001, there were 151,859 unemployed persons (6.8% of persons in the labour force), down from 196,189 persons (9.4%) in 1996. High unemployment rates were recorded in the Localities of Loch Sport (26.1%), Seaspray (24.6%) and Port Welshpool (21.6%).
For those aged 15-24 years, the unemployment rate fell to 12.8% from 16.1% in 1996. Localities which recorded a high level of youth unemployment were Quambatook (66.7%) and Talbot (50.0%).
The number of unemployed persons looking for full-time employment decreased by 31.5%, down from 155,021 persons in 1996 to 106,157 in 2001. However, those looking for part-time work increased by 11.0%, from 41,168 in 1996 up to 45,702 in 2001.
Occupation
In 2001, the largest occupation groups were Professionals and Associate Professionals (30.5% of employed persons), Intermediate Clerical, Sales and Service Workers (16.1%) and Tradespersons and Related Workers (12.2%). These proportions are similar to those in 1996.
The highest proportions of Professionals and Associate Professionals were recorded in Halls Gap (L) (52.0%), Shoreham (L) (48.4%) and Mount Macedon (45.6%).
The highest proportions of Intermediate Clerical, Sales and Service Workers were recorded in the Localities of Cranbourne South (21.6%), Bethanga (21.3%) and Goornong (20.9%).
The highest proportions of Tradespersons and Related Workers were recorded in the Localities of St Andrews Beach (25.3%), Hazeldene (23.4%) and Briagolong (23.3%).
Industry
In 2001, the three largest industries of employment were Manufacturing (15.3%), Retail Trade (14.8%), and Property and Business Services (11.4%). These were also the top three employing industries in 1996.
The highest proportions of persons employed in Manufacturing were recorded in the Localities of Strathmerton (53.1%), Dartmoor (50.0%) and Girgarre (46.8%).
The highest proportions of persons employed in Retail Trade were recorded in Loch (L) (28.8%), Marong (L) (23.6%) and Barham-Koondrook (Koondrook Part) (23.6%).
The highest proportions of persons employed in Property and Business Services were recorded in the Localities of Metung (15.7%), St Andrews (15.3%) and Dinner Plain (15.0%).
Dwelling Characteristics
Total number of dwellings
The number of dwellings increased by 8.2% from 1,773,220 in 1996 to 1,918,583 in 2001. Of these dwellings, 90.2% (1,731,343) were occupied private dwellings, 9.5% were unoccupied private dwellings and 0.2% were non-private dwellings.
Total number of persons
The number of persons in occupied private dwellings increased by 6.4% from 4,254,566 in 1996 to 4,525,065 in 2001. Of these, 71.0% (3,212,904 persons) were counted in Major Urban areas, similar to 1996 (68.6%).
Fully owned
Of the 1,731,343 occupied private dwellings in Victoria, 42.9% were fully owned, compared with 43.7% in 1996. The Rural Balance area recorded the highest proportion of fully owned dwellings (51.5%). In 1996, the highest proportion was Bounded Locality (52.0%), followed by the Rural Balance (51.9%).
Home ownership proportions were highest in the Localities of Underbool (78.2%), Indented Head (72.0%) and Woomelang (71.4%).
Being purchased
In Victoria, 27.8% of private dwellings were being purchased, similar to 27.1% in 1996. The Other Urban areas recorded the highest percentages in both 2001 (29.2%) and 1996 (29.3%).
High proportions of homes being purchased were recorded in Waterford Park (L) (64.6%), Seville East (L) (62.4%) and Miners Rest (61.3%).
Housing loan repayments
The median housing loan repayment increased by 14.7%, up from $741 in 1996 to $850 in 2001. High median loan repayments were recorded in Cranbourne South (L) ($1,325), Mount Macedon ($1,172) and Dinner Plain (L) ($1,166).
Rented dwellings
In 2001, 22.1% (382,981) of occupied private dwellings were being rented, compared with 23.2% in 1996. The Major Urban and Other Urban areas (both 23.4%) recorded the highest proportions, similar to 1996 (24.5% and 24.6% respectively).
The areas with the highest proportions of rented dwellings were Dinner Plain (L) (55.9%), Cann River (L) (43.8%) and Robinvale (40.3%).
Rent
The median weekly rent increased by 24.8%, from $121 in 1996 to $151 in 2001. The Major Urban areas ($165) recorded the highest median rent, compared with the Rural Balance ($87) and the Bounded Locality ($102).
A high median rent was recorded in the Urban Centres of Mount Macedon ($180), Beaconsfield Upper ($179), and Gisborne ($175), and in the Localities of Dinner Plain, Waterford Park (both $500) and Arcadia Downs ($237).
Family Characteristics
The number of families grew from 1,152,887 in 1996 to 1,222,690 in 2001, an increase of 6.1%. Of these families, 48.8% were couples with dependent and non-dependent children, 34.4% were couples without children, 14.8% were one parent families and 1.9% were of other family type. In the Rural Balance area, 47.6% of couple families had dependent children.
The highest proportions of couple families with dependent children were recorded in the Localities of Arcadia Downs (66.3%), Cabarita (66.2%), Moriac (64.8%) and Miners Rest (63.4%).
The highest proportions of couple families without children were recorded in the Localities of Loch Sport (76.4%), Metung (71.2%) and Newlands Arm (70.1%).
The highest proportions of one parent families were recorded in the areas of Seaspray (L) (27.5%), Millgrove (L) (25.6%) and Woori Yallock (24.2%).
Households Characteristics
The number of households grew from 1,554,456 in 1996 to 1,667,687 in 2001, an increase of 7.3%. Of these households, 72.3% were family households, 23.8% were lone person households and 3.8% were group households. These proportions are similar to those in 1996 (73.0%, 22.9% and 4.0% respectively).
The highest proportions of lone person households were recorded in the Localities of Jeparit (43.6%), Gunbower (43.4%) and Seaspray (42.6%).
Localities recording the highest proportions of group households were Dinner Plain (12.0%) and Buxton (6.5%).
© Commonwealth of Australia 2013
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Australian Bureau of Statistics
Celebrating the International Year of Statistics 2013
ABS Home > Statistics > By Release Date
6427.0 - Producer Price Indexes, Australia, Jun 2009
Previous ISSUE Released at 11:30 AM (CANBERRA TIME) 20/07/2009
Page tools: Print Page Print All RSS Search this Product
JUNE KEY FIGURES
Mar Qtr 09 to Jun Qtr 09
Jun Qtr 08 to Jun Qtr 09
STAGE OF PRODUCTION
% change
% change
Final (Stage 3) commodities (excl. exports)
-0.8
2.1
Domestic
0.0
0.3
Imports
-5.9
14.3
Intermediate (Stage 2) commodities
-1.9
-0.8
Domestic
-1.1
-0.4
Imports
-7.9
-3.2
Preliminary (Stage 1) commodities
-2.7
-1.9
Domestic
-2.0
0.0
Imports
-7.5
-13.4
Final Stage, Base 1998-99 = 100.0
Final Stage, Quarterly % change
JUNE KEY POINTS
FINAL (STAGE 3) COMMODITIES
• decreased by 0.8% in the June quarter 2009.
• mainly due to price decreases in industrial machinery and equipment manufacturing (-5.6%), building construction (-0.5%), and electronic equipment manufacturing (-10.5%).
• partially offset by price increases in motor vehicle and part manufacturing (+2.0%) and petroleum refining (+5.0%).
• increased by 2.1% through the year to June quarter 2009.
INTERMEDIATE (STAGE 2) COMMODITIES
• decreased by 1.9% in the June quarter 2009.
• mainly due to price decreases in iron and steel manufacturing (-11.8%), coal mining (-17.5%), basic chemical manufacturing (-8.7%) and property operators and developers (-2.2%).
• partially offset by price increases in electricity, gas and water supply (+2.9%), basic non-ferrous metal manufacturing (+8.0%) and grain, sheep, beef and dairy cattle farming (+2.0%).
• decreased by 0.8% through the year to June quarter 2009.
PRELIMINARY (STAGE 1) COMMODITIES
• decreased by 2.7% in the June quarter 2009.
• mainly due to price decreases in coal mining (-17.5%), iron and steel manufacturing (-11.4%) and basic chemical manufacturing (-8.6%).
• partially offset by price rises in basic non-ferrous metal manufacturing (+8.1%) and electricity, gas and water supply (+2.7%).
• decreased by 1.9% through the year to June quarter 2009.
NOTES
FORTHCOMING ISSUES
ISSUE (QUARTER) Release Date
September 2009 26 October 2009
December 2009 25 January 2010
March 2010 27 April 2010
June 2010 26 July 2010
FORTHCOMING CHANGES
As foreshadowed in the March quarter 2009 issue, the content and format of this publication will change from September quarter 2009, to reflect an updated weighting pattern and the new industrial classification (ANZSIC 2006) for some of the producer price index series. Further details regarding these changes can be found in Information Paper: Update on ANZSIC 2006 Implementation for Producer and International Trade Price Indexes (cat. no. 6427.0.55.002).
ROUNDING
Any discrepancies between totals and sums of components in this publication are due to rounding
DATA REFERENCES
Data referenced in the Key points and Commentary are available from the tables shown in this publication, or in the corresponding tables of this publication on the ABS website <http://www.abs.gov.au>.
INQUIRIES
For further information about these and related statistics, contact the National Information and Referral Service on 1300 135 070 or Lee Taylor on Canberra (02) 6252 6251.
SUMMARY COMMENTARY
COMMENTARY
Links to Other Parts of This Release
To access the "Commentary" page use this link
Commentary
To access the "Price Indexes and Contract Price Indexation" page use this link
Price Indexes and Contract Price Indexation
© Commonwealth of Australia 2013
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Australian Bureau of Statistics
Celebrating the International Year of Statistics 2013
ABS Home > Statistics > By Release Date
6227.0 - Education and Work, Australia, May 2010 Quality Declaration
Previous ISSUE Released at 11:30 AM (CANBERRA TIME) 31/10/2011
Page tools: Print Page Print All RSS Search this Product
This data cube reports on an indicator from the National Agreement for Skills and Workforce Development, and encompasses date for 2001-2010.
© Commonwealth of Australia 2013
Unless otherwise noted, content on this website is licensed under a Creative Commons Attribution 2.5 Australia Licence together with any terms, conditions and exclusions as set out in the website Copyright notice. For permission to do anything beyond the scope of this licence and copyright terms contact us.
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Australian Bureau of Statistics
Celebrating the International Year of Statistics 2013
ABS Home > Statistics
Statistics by Title
A B C D E F G H I J K L M N O P Q R S T U V W X Y Z 0-9
Y
Year 2000 Problem, Australia, Jun 1999 (cat no. 8152.0)
Year 2000 Problem, Australia, Preliminary, Jun 1999 (cat no. 8151.0)
Year Book Australia, 2012 (cat no. 1301.0)
Year Book Australia on CD-ROM, 2006 (cat no. 1301.0.30.001)
Youth, Australia: A Social Report, 1997 (cat no. 4111.0)
© Commonwealth of Australia 2013
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Research article
AGE-BSA down-regulates endothelial connexin43 gap junctions
Chi-Young Wang1, Hung-Jen Liu2, Heng-Ju Chen3, Yi-Chun Lin4,5, Hsueh-Hsiao Wang4,6, Ta-Chuan Hung4,5 and Hung-I Yeh4,6*
Author Affiliations
1 Department of Veterinary Medicine, College of Veterinary Medicine, National Chung Hsing University, 250 Road Kuo Kuang, Taichung 402, TAIWAN
2 Institute of Molecular Biology, College of Life Sciences, National Chung Hsing University, 250 Road Kuo Kuang, Taichung 402, TAIWAN
3 Graduate Institute of Biotechnology, National Ping-Tung University of Science and Technology, 1 Road Shuefu, Pingtung 912, TAIWAN
4 Departments of Internal Medicine and Medical Research, Mackay Memorial Hospital, 45 Road Minsheng, New Taipei City 251, TAIWAN
5 Department of Nursing, Mackay Medicine, Nursing and Management College, 92 Road Shengjing, Taipei 112, TAIWAN
6 Department of Medicine, Mackay Medical College, 46 Road Zhongzheng, New Taipei City 252, TAIWAN
For all author emails, please log on.
BMC Cell Biology 2011, 12:19 doi:10.1186/1471-2121-12-19
The electronic version of this article is the complete one and can be found online at: http://www.biomedcentral.com/1471-2121/12/19
Received:13 December 2010
Accepted:16 May 2011
Published:16 May 2011
© 2011 Wang et al; licensee BioMed Central Ltd.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Background
Advanced glycation end products generated in the circulation of diabetic patients were reported to affect the function of vascular wall. We examined the effects of advanced glycation end products-bovine serum albumin (AGE-BSA) on endothelial connexin43 (Cx43) expression and gap-junction communication.
Results
In human aortic endothelial cells (HAEC) treated with a series concentrations of AGE-BSA (0-500 μg/ml) for 24 and 48 hours, Cx43 transcript and Cx43 protein were reduced in a dose dependent manner. In addition, gap-junction communication was reduced. To clarify the mechanisms underlying the down-regulation, MAPKs pathways in HAEC were examined. Both a MEK1 inhibitor (PD98059) and a p38 MAPK inhibitor (SB203580) significantly reversed the reductions of Cx43 mRNA and protein induced by AGE-BSA. Consistently, phosphorylation of ERK and p38 MAPK was enhanced in response to exposure to AGE-BSA. However, all reversions of down-regulated Cx43 by inhibitors did not restore the functional gap-junction communication.
Conclusions
AGE-BSA down-regulated Cx43 expression in HAEC, mainly through reduced Cx43 transcription, and the process involved activation of ERK and p38 MAPK.
Background
Diabetes is known to accelerate the process of atherosclerosis, in which endothelial dysfunction plays a key role [1]. Previous studies have shown that diabetes alters the expression of a variety of molecules involved in maintenance of endothelial function [2]. Animal experiments also demonstrated that induction of diabetes, either alone or on top of hyperlipidemia, suppress the expression of endothelial gap junctions [3,4]. Gap junctions are cell membrane channels made of paired hexamers of connexins, which allow exchange of ions and small signaling molecules between the cytoplasmic compartments of adjacent cells. In mammals endothelial cells mainly express connexin43 (Cx43), Cx40, and Cx37 [5], of which Cx43 is by far the predominant in the cultured endothelial cells [6,7]. Several reports have shown that endothelial Cx43 gap junctions are down-regulated by factors causing endothelial dysfunction [8], such as aging [9], hypertension [10], and arsenic trioxide [11], the last of which was even reported to induce endothelial lesion. In contrast, other factors causing endothelial dysfunction, such as oscillatory shear stress, were shown to enhance Cx43 expression [12].
One key offending factor underlying the toxic effects of diabetes is glucose, the high level of which had been proved to affect the activities of endothelial cells in many aspects, including suppression of gap junctions [13,14]. Since diabetes is associated with a more severe form of vascular disease, we suspected that molecules underlying the detrimental effects of diabetes other than glucose also affect endothelial gap junctions.
During chronic exposure to elevated blood glucose advanced glycation end products (AGE) are generated in the circulation. AGE are also versatile molecules and have been reported to possess multiple actions in the vascular wall, such as changes of release of cytokines, induction of expression of cell adhesion molecules, impairment of endothelial vasodilatation, and triggering of chronic inflammation [15]. However, the effect of AGE on endothelial gap junctions remained unclear. To this end, we examined the expression of Cx43 as well as the gap-junction communication in human aortic endothelial cells (HAEC) treated with AGE and explore underlying mechanisms. A previous study had shown that the average serum AGE levels in diabetic patients ranged between 28.8 and 87.2 μg/ml and may reach 160-500 μg/ml in severe cases [16].
Methods
Generation of AGE-BSA
AGE-BSA was prepared by incubation of BSA (Fraction V, Sigma Chemical Co., St. Louis, MO, USA) at a concentration of 50 mg/ml with 0.5 M glucose in 10 mM phosphate-buffered saline (PBS) containing 0.5 mM EDTA, pH 7.4, at 37°C for 12 weeks. For a control group, the same concentration of BSA was incubated with PBS containing EDTA without glucose for 12 weeks. Free glucose was removed by extensive dialysis against PBS. The brown color of AGE-BSA showed the typical appearance of AGE. SDS-PAGE analysis showed the formation of AGE-BSA monomer and dimer with molecular weights approximately equaling to 78 and 157 kDa, respectively. However, unglycated BSA was about 69 kDa. The ratio of relative fluorescence intensities of AGE-BSA to unglycated BSA was approximately 64 folds in our preparations. Results were consistent with other's report [17]. For a control group, BSA was processed as above procedures. BSA and AGE-BSA were stored at -70°C until use.
Cell culture
HAECs (Cascade Biologics) were seeded in 1% gelatin-coated plasticware and incubated at 37°C under a humidified 95% air and 5% CO2 atmosphere. Cells grown to confluence were dissociated with 3 ml of 0.25% trypsin-EDTA (GIBCO) at 37°C for 3 min. The suspension was diluted with 7 ml medium 200 supplemented with LSGS, centrifuged at 1200 rpm for 8 min, and resuspended in the culture medium. Cells were then replated in 35-mm Petri dishes (5 × 104 cells per cm2) and allowed to grow to confluence or seeded at the same density onto 12-mm glass coverslips coated with 1% gelatin (Sigma). Cells of passage 4 to 6 were used in the subsequent experiments. To evaluate the relative mRNA level of Cx37, Cx40, and Cx43, human umbilical vein endothelial cells (HUVEC, from PromoCell), maintained in endothelial cell growth medium (EGM, from PromoCell) were used as a reference. The confluent cells were treated with AGE-BSA at series concentrations of 25, 50, 100, 250, and 500 μg/ml for 24 and 48 hours. For a control group, cells were treated with BSA at series concentrations of 25, 50, 100, 250, and 500 μg/ml for 24 hours. For examination of activation of MAPK signaling pathways, cells were incubated with AGE-BSA at concentrations of 25, 50, 100, 250, and 500 μg/ml for 2 and 6 hours. For time course experiments, cells were incubated with AGE-BSA at 500 μg/ml for 0, 0.5, 1, 2, 4, and 6 hours. For drug treatments, cells were treated with 40 μM of PD98059 (Sigma), 15 μM of SB203580 (Sigma), and 10 μM of SP600125 (Sigma) for 30 min and then incubated with 500 μg/ml AGE-BSA for 24 hours.
Western blotting
Cells were collected in RIPA buffer containing 150 mM NaCl, 5 mM EDTA, 1% NP40, 2 mM PMSF, and 50 mM Tris-HCl, pH 7.4 or SB buffer containing 20% SDS, 0.1 M Tris-HCl, pH 6.8, and 10 mM EDTA followed by sonication for 30 sec. Thirty microgram of sample was loaded in each lane, resolved by 12% SDS-PAGE, and transferred onto a PVDF membrane (Amersham, UK). The membrane was incubated with the monoclonal anti-Cx43 antibody (1:1000; Chemicon), anti-Cx43 antibody (1:1000; BD Biosciences), anti-β actin antibody (1:2000; Chemicon), anti-ERK antibody (1:1000; Cell signaling), anti-p38 MAPK antibody (1:1000; Biosource), anti-JNK antibody (1:2000; Cell signaling), anti-phosphorylated ERK antibody (1:1000; Cell signaling), anti-phosphorylated p38 MAPK antibody (1:1000; Biosource), and anti-phosphorylated JNK antibody (1:1000; Cell signaling) at room temperature for 1 hour. After three washes with TBST (20 mM Tris pH 7.6, 150 mM NaCl, 0.1% Tween 20), a horseradish peroxidase-conjugated mouse anti-rabbit or donkey anti-mouse IgG (1:3000 in TBST plus 10% BSA) was added, and an enzyme-linked chemiluminescence system (ECL; Amersham, UK) was applied to check the bound antibody.
Real-time PCR and Semi-quantitative RT-PCR
Total RNA was extracted using Trizol reagent (Invitrogen Life Technologies, Carlsbad, USA) according to the manufacturer's instructions. After phenol treatment and drying, RNA was dissolved in RNase-free water for determination of concentration using spectrophotometer. RNA quality was checked on agarose electrophoresis. For real-time PCR analysis, endothelial cDNA were amplified with primers specific for Cx43 (sense: 5'-CCT CTC GCC TAT GTC TCC TC-3') (antisense: 5'-GCT CAC TTG CTT GCT TGT TG-3'), Cx37 (sense: 5'-CGT AGA GCG TCA GAT GGC-3') (antisense: 5'-GCA CAC TGG CGA CAT AGG-3'), Cx40 (sense: 5'-TCA ATC CCT TCA GCA ATA ATA TG-3') (antisense: 5'-GTG ACC TGG TGA GAC TCC-3'), and β-actin (sense: 5'-TCC TGT GGC ATC CAC GAA ACT-3') (antisense: 5'-GAA GCA TTT GCG GTG GAC GAT-3') using iQTM SYBR Green Supermix reagent and detected with iQTM Single Color Real-Time PCR Detector System (all from Bio-Rad, California, USA). Relative mRNA levels were normalized with the corresponding levels of β-actin.
For Semi-quantitative RT-PCR, total RNA (2 μg) was reverse-transcribed into complementary DNA (cDNA) using the SuperScrip™ II reverse transcriptase and random primers (Invitrogen Life Technologies) according to the manufacturer's recommendations and diluted with DDW to a final volume of 20 μl. The optimal number of PCR cycles was determined for Cx43 (Cx-1: 5'-TCTGAGTGCCTGAACTTGC-3' and Cx-2: 5'-ACTGACAGCCACACCTTCC-3') and GADPH (GA-1: 5'-CATTGACCTCAACTACATGG-3' and GA-2: 5'-TTGCCCACAGCCTTGGCAGC-3') so that the amplification process was conducted during the exponential phase of amplification. Amplification of the GADPH gene transcript was used as the internal control to stringently control for any variability in RNA degradation and RT efficiency. Reactions were carried out in a 50 μl final reaction volume.
Immunofluorescence detection
Confluent cells grown on coverslips were fixed with methanol at -20°C for 5 min. After blocking with 0.5% BSA, the cells were incubated with the anti-Cx43 antibody (1:1000; Chemicon) at 37°C for 1 hour, followed by incubation with a CY3 conjugated donkey anti-mouse antibody (Chemicon). The cells were then incubated with FITC-conjugated lectin Ulex europaeus agglutinin-1 (UEA-1; 10 μg/ml; Sigma) for 1 hour to confirm the identity of endothelial cells [18] or bisbenzamide (1 μg/ml; Sigma) for 15 min. Finally, the cells were mounted and examined using a Leica TCS SP confocal laser scanning microscope.
Scrape loading assay
Gap-junction communication in HAECs treated with 250 and 500 μg/ml of AGE-BSA for 24 and 48 hours was evaluated by scrape loading assay. For drug experiments, HAEC were respectively pretreated with i) 40 μM of PD98059 and ii) 15 μM of SB203580 prior to addition of the AGE-BSA (500 μg/ml) for 24 hours. The medium of confluent cells were removed and rinsed with Hank's balanced salt solution (GIBCO). A 27-gauage needle was used to create multiple scrapes through the cell monolayer in the presence of PBS containing 0.5% rhodamine-dextran and 0.5% Lucifer yellow. After 3 min of incubation at room temperature, the culture was rinsed three times and then incubated for 5 min in medium 200 supplemented with LSGS to allow the loaded dye to transfer to adjoining cells. The cells were viewed and recorded using a fluorescence microscope.
MTT assay
Cells were incubated in medium 200 supplemented with LSGS containing 0.5 mg/ml MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide]. After 1 hour, the MTT solution was removed and dimethylsulfoxide (100 μl/well) was added.
Absorbances (550 nm) of the supernatant were read using a microplate spectrophotometer (Spectra Max 340; Molecular Devices, Sunnyvale, CA). Cell viability was expressed as the percentage of the absorbance values of treated cells to controls.
Analysis
Densitometric scanning and analysis were performed on immunoblots and gel images using Imagemaster (Amersham Pharmacia Biotec, NJ, USA). Within each lane, total amounts of bands of Cx43 were divided by those of actins as loading controls. For RT-PCR, amounts of Cx43 were divided by those of GADPH. For the scrape loading assay, the area between the bilateral edges of lucifer yellow transfer and the scrape line was measured. The value of total amount of each sample was presented as mean (%) ± S.E. The significant differences are analyzed by t-test.
Results
Real-time PCR showed that Cx37, Cx40, and Cx43 existed in both HAEC and HUVEC. However, the expression levels differed markedly (Figure 1). Regarding HAEC, Cx43 transcripts were more than 10-fold abundant, compared to Cx37 or Cx40 (Cx43 vs. Cx37 or Cx40, both P <0.01), in contrast to HUVEC, in which Cx37 was the most abundant. In addition, the relative mRNA expression levels of individual connexins in HAEC were lower than those in HUVEC (all P <0.05, see Figure 1). Consistently, immunoconfocal examination of the untreated HAEC showed that Cx43 was abundantly expressed at the cell borders, typical for gap junctions, but Cx37 and Cx40 were rarely seen (Figure 2). Treatment with AGE-BSA for 24 hours at doses of 100 and 500 μg/ml lead to a marked reduction of Cx43 and Cx37 and Cx40 remained rarely seen (Figure 2). Therefore, the following experiments in HAEC were focused on Cx43.
Figure 1. Analysis of the mRNA profile of Cx37, Cx40, and Cx43 in HAEC and HUVEC showed distinct expression patterns in each type of cells and in HAEC Cx43 was the most abundant transcripts. Real-time PCR showed the existence of Cx37, Cx40, and Cx43 transcripts. Cx43 and Cx37 were the most abundant connexin transcripts expressed in HAEC and HUVEC, respectively. In HAEC, compared to Cx37 and Cx40, Cx43 was more than 10-fold abundant. *, p < 0.05; #, p < 0.005, HAEC vs. HUVEC. **, p < 0.01 for Cx43 vs. each of Cx37 and Cx40 in HAEC.
Figure 2. Expression of Cx37, Cx40, and Cx43 gap junctions in HAEC evaluated by immunoconfocal microscopy showed that in untreated cells Cx43 was abundantly expressed at the cell borders but Cx37 and Cx40 were rarely observed. AGE-BSA reduced Cx43 signals in a dose-dependent manner. Images were respectively obtained from cells treated with 100 μg/ml and 500 μg/ml of AGE-BSA for 24 hours. Compared with the control group (A), Cx43 gap junctions (red spots) are reduced as the dose of AGE-BSA increases (B and C). Note no conceivable signals of Cx37 and Cx40 were found in all cells. Blue label, nucleus. Green label, lectin UEA1. The concentration of AGE-BSA and the incubation time are indicated in the upper left of each image. h, hours. All images are of the same magnification. Bar, 50 μm.
In cells treated with AGE-BSA at doses from 25 to 250 μg/ml for 24-48 hours, no changes of the cell density and morphology were observed (Figure 3). However, cells became slightly retracted and reduced in density after treated with 500 μg/ml of AGE-BSA for more than 24 hours (Figure 3L). For Cx43, regardless of the 24 or 48 hours treatment, the levels of expression gradually decreased as the dose of AGE-BSA increased (Figure 3). Western blotting also verified the dose-dependent effect of AGE-BSA on Cx43 expression (Figure 4). After exposure to 500 μg/ml of AGE-BSA for 24 hours (Figure 4A) and 48 hours (Figure 4B), the relative expression levels of Cx43 protein were respectively reduced to 61.2 ± 7.3% and 43.9 ± 8.8% (both P <0.05, compared to the control groups). A control group of cells treated with dialyzed, long term stored BSA for 24 hours showed no changes of Cx43 proteins (Additional file 1 Figure S1). There was no difference in decreasing trends or patterns of Cx43 proteins extracted from AGE-BSA-treated cells with lysis buffers containing NP40 or SDS as well as no difference in Cx43 expression when the samples were detected using anti-Cx43 antibodies from various sources (Additional file 2 Figure 2S and Additional file 3 Figure 3S). The function of gap-junction communication was checked using the method of scrape-loading/dye transfer (Figure 5A). The areas of dye transfer were significantly reduced in cells treated with 500 μg/ml of AGE-BSA for 24 hours and 250 μg/ml of AGE-BSA for 48 hours (Figure 5B; both P <0.05). Moreover, because cells were not well contacted after exposure to 500 μg/ml of AGE-BSA for 48 hours, the inhibition of gap-junction communication was unable to quantify (Figure 5B). The effects of AGE-BSA on viability of cells after exposure for 24 hours were assessed using MTT assay. The relative viability of cells was 94.2 ± 0.2% (100 μg/ml AGE-BSA), 90.2 ± 1.4% (250 μg/ml AGE-BSA), and 81.6 ± 0.8% (500 μg/ml AGE-BSA), respectively (all P <0.05, compared to the control groups) (Figure 6).
Figure 3. Reduced expression of endothelial Cx43 gap junctions by AGE-BSA, as evaluated by immunoconfocal microscopy. The levels of Cx43 decreased as the dose of AGE-BSA increased regardless of 24 or 48-hour treatment. Images A-F and G-L were respectively obtained from cells treated with AGE-BSA for 24 and 48 hours. Compared with the control group (A and G), Cx43 gap junctions (red spots) are reduced as the dose of AGE-BSA increases (B-F and H-L). Blue label, nucleus. The concentration of AGE-BSA and the incubation time are indicated in the upper left of each image. h, hours. All images are of the same magnification. Bar, 50 μm.
Figure 4. Dose-dependent reduction of expression of Cx43 protein by AGE-BSA, as detected by Western blotting. Note that a dose-dependent reduction is seen in cells treated with AGE-BSA for both 24 (A) and 48 hours (B). For each dose of treatment, the relative level of the total amount of Cx43 protein is shown in the histogram at the top of the blot. The relative expression levels of Cx43 protein significantly reduced to 61.2 ± 7.3% and 43.9 ± 8.8% after cells exposure to 500 μg/ml of AGE-BSA for 24 and 48 hours, respectively. *, p < 0.05 compared to the untreated cells.
Additional file 1. Figure S1 -Comparison of the effects of AGE-BSA and BSA on expression of Cx43 proteins, as detected by Western blotting. No effects on the expression levels of Cx43 protein by BSA were detected. Note that a dose-dependent reduction is only seen in cells treated with AGE-BSA (A) but not with dialyzed, long-term stored BSA (B). Cells were treated for 24 hours.
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Additional file 2. Figure S2 -Comparison of different lysis buffers in extraction of Cx43 proteins from HAEC treated with a serious concentrations of AGE-BSA, as detected by Western blotting. No differences on the expression levels of Cx43 protein were detected using both extraction buffers. Note that no difference in decreasing trends or patterns of Cx43 proteins extracted from cells using either NP40 (A) or SDS (B) buffers.
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Additional file 3. Figure S3 -Comparison of anti-Cx43 antibodies of various sources in detecting Cx43 proteins from cells treated with a serious concentrations of AGE-BSA, as examined by Western blotting. No differences on the expression levels of Cx43 proteins were detected using both anti-Cx43 antibodies. Note the same pattern of a dose-dependent reduction was seen using antibodies of different sources (antibody used in A, from Chemicon; in B from, BD Biosciences).
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Figure 5. Reduction of gap-junction communication by AGE-BSA, as evaluated using scrape loading assay. A. Bottom images are phase-contrast micrographs of the upper fluorescent images. The concentration of AGE-BSA (in μg/ml) is indicated in the upper left of each paired images. B. Analysis of the area between the bilateral edges of lucifer yellow transfer and the scrape line is shown in the histogram. Bar, 150 μm. Note that significant decrements were seen in cells treated with 250 μg/ml for 24 and 48 hours, and 500 μg/ml for 24 hours, respectively. *, p < 0.05 compared to the untreated cells. h, hours. The inhibition of gap-junction communication at 500 μg/ml of AGE-BSA for 48 hours was not quantified because cells were not well contacted.
Figure 6. Dose-dependent reduction in viability of cells treated with AGE-BSA, as detected by MTT assay. Note a dose-dependent reduction is seen in cells treated with AGE-BSA for 24 hours. For each dose of treatment, the relative level of the total amount of cells is shown in the histogram. The relative viability of cells significantly reduced to 94.2 ± 0.2%, 90.2 ± 1.4%, and 81.6 ± 0.8% post exposure to 100 μg/ml, 250 μg/ml, and 500 μg/ml, respectively. *, p < 0.05 compared to the untreated cells.
To explore whether alteration of Cx43 expression by AGE-BSA was regulated through MAPK cascades, three signal pathways, ERK, JNK, and p38 MAPK were separately blocked before addition of AGE-BSA. PD98059, an MEK1 inhibitor which blocks ERK pathway, significantly reversed the reduction of Cx43 proteins in cells treated with AGE-BSA for 24 hours (relative expression levels, without PD98059, 62.3 ± 4.8%; with PD98059, 84.1 ± 4.4%, p < 0.05 see Figure 7A). Similarly, SB203580, a p38 MAPK inhibitor, also significantly reversed the reductions (without SB203580, 60.5 ± 1.6%; with SB203580, 79.6 ± 2.9%, p < 0.05 see Figure 7B). The basal levels of Cx43 proteins were not affected after the addition of these inhibitors. In contrast, no such effects were seen in cells treated with SP600125, a JNK inhibitor (data not shown). Immunoconfocal microscopy also confirmed that AGE-BSA induced reduction of Cx43 was reversed by PD98059 or SB203580 to the levels comparable to the control group and the reversed Cx43 mainly located at cell borders (Figure 7C). The gap-junction communication was measured when cells were treated with 500 μg/ml of AGE-BSA for 24 hours in the presence of either PD98059 (40 μM) or SB203580 (15 μM). Whereas Cx43 proteins returned to approximately the control levels (Figure 7A-C), the areas of dye transfer remained close to those of cells treated with 500 μg/ml of AGE-BSA for 24 hours (all P <0.05, compared to the control groups; see Figure 7D). Taken together, our results indicated the reversed Cx43 by MEK1 or p38 MAPK inhibitor remained at the cell borders but exhibited impaired communication function.
Figure 7. PD98059 and SB203580 respectively blocked the down-regulating effect of AGE-BSA on Cx43 protein (as detected by Western blotting and immunoconfocal microscopy) but not the effect on the gap-junction communication (by scrape loading). Note the relative expression levels of Cx43 in cells treated with AGE-BSA (500 μg/ml; 24 hours) were reversed from 62.3 ± 4.8% to 84.1 ± 4.4% by PD98059 (A) and from 60.5 ± 1.6% to 79.6 ± 2.9% by SB203580 (B), respectively. The results were further confirmed by immunoconfocal microscopy (C). The inhibition of gap-junction communication by AGE-BSA (500 μg/ml; 24 hours) was not rescued in the presence of PD98059 and SB203580 (D). *, p < 0.05 compared to the untreated cells. Images in (C) were obtained from the control group, cells treated with AGE-BSA, cells treated with AGE-BSA and PD98059, and cells treated with AGE-BSA and SB203580, respectively. Note that Cx43 gap junctions (red spots) were reversed near the level of the control group by PD98059 and SB203580. Blue label, nucleus. The concentrations of AGE-BSA and inhibitor used are indicated in the upper left of each image. All images are of the same magnification. Bar, 50 μm.
In addition, to understand the effect of AGE-BSA on protein synthesis at the transcription level, Cx43 transcripts was measured using semi-quantitative RT-PCR, which showed a dose-dependent decrease of the transcripts in cells treated with AGE-BSA for 24 hours (relative expression levels, 500 μg/ml of AGE-BSA, 57.1 ± 8.9%; p < 0.05 compared to the control group; see Figure 8A). This effect was more obvious after treatment for 48 hours (500 μg/ml of AGE-BSA, 38.3 ± 5.5%; p < 0.05; see Figure 8B). Moreover, the decrement of Cx43 transcripts at exposure of 500 μg/ml of AGE-BSA followed a time-dependent manner, as early as 6 hours, a significant decrease was noticed (77.5 ± 5.1%; p < 0.05; see Figure 8C). The down-regulation of Cx43 transcripts by AGE-BSA (500 μg/ml) seen at 24 hours was significantly reversed in cells pre-treated with PD98059 or SB203580 (without PD98059 or SB203580, 65.1 ± 2.1%; with PD98059, 95.1 ± 5.3%; with SB203580, 98.8 ± 9.4%, all p < 0.05; see Figure 8D). No affections on the basal levels of Cx43 transcripts were seen after the addition of these inhibitors. This indicated that the AGE-BSA-induced reduction in Cx43 transcription was mediated by ERK and p38 MAPK. Phosphorylation of ERK and p38 MAPK was activated dose-dependently in cells treated with AGE-BSA for 2 hours (Figure 9A) and 6 hours (Figure 9B), while no such change was seen in JNK. Taken together, these data indicated that the down-regulation of Cx43 expression was through ERK and p38 MAPK signaling pathways.
Figure 8. Reduced transcription of Cx43 by AGE-BSA and partial recovery of reduced Cx43 transcript by each of PD98059 and SB203580, as detected by semi-quantitative RT-PCR. Note that a dose-dependent reduction is seen in cells treated with AGE-BSA for both 24 (A) and 48 hours (B); a time-dependent reduction is seen in cells treated with 500 μg/ml of AGE-BSA for different time periods (C). The relative transcription levels of Cx43 of AGE-BSA-treated cells were significantly reversed by PD98059 from 65.1 ± 2.1% to 95.1 ± 5.3% or by SB203580 from 65.1 ± 2.1% to 98.8 ± 9.4% (D). For each dose of treatment, the relative level of Cx43 transcripts is shown in the histogram at the top of the blot. h, hours. *, p < 0.05 compared to the untreated cells.
Figure 9. Activation of ERK and p38 MAPK but not JNK signaling pathways by AGE-BSA. Representative western blots of ERK, phosphorylated ERK, p38 MAPK, phosphorylated p38 MAPK, JNK, and phosphorylated JNK in cells treated with AGE-BSA for 2 (A) and 6 hours (B). Phosphorylation of ERK and p38 MAPK was activated dose-dependently but no such effect was seen for JNK.
Discussion
This study demonstrates that, in cultured HAEC, i) Cx43 is the predominant gap junction component protein, unlike HUVEC; ii) AGE-BSA reduced Cx43 transcript expression in a time-dependent manner; and iii) AGE-BSA reduced Cx43 protein expression and attenuated gap-junction intercellular communication in a dose-dependent manner; Although a substantial portion of HAEC died when the cells were treated with high dose of AGE-BSA (500 μg/ml) for more than 24 hours, the down-regulating effects of AGE-BSA on Cx43 expression should not be mainly attributed to a general reduction in protein expression during cell dying, since the expression level of actin protein along the reduction of Cx43 was relatively stable (see Figure 4). In addition, inhibition of gap-junction communication activity may cause cell death, as seen in retinal capillary cells, in which attenuation of gap-junction communication with high glucose treatment disturbed normal transport of small molecules and calcium ions, leading to cell death [14,19]. The reduction of Cx43 transcripts by AGE-BSA was partially recovered by inhibitors of each of the ERK1 and p38 MAPK. Moreover, the activation of the MAPK cascades was further confirmed by the increase of phosphorylated ERK and p38 after a short time exposure to AGE-BSA indicating that activation of both the ERK and p38 MAPK cascades is involved in this process. Despite the down-regulation of Cx43 expression by AGE-BSA can be abrogated by the inhibitors and the reversed Cx43 mainly stayed at the cell borders, the gap-junction communication was impaired. These novel findings are complementary to our current knowledge regarding the effects of diabetes on endothelial gap junctions.
The results of the present study, in conjunction with those from other in vitro experiments investigating the effect of high glucose on endothelial gap junctions [13,14], indicate that in the circulation of diabetic individuals high glucose per se as well as the glycated albumin, which is generated in high glucose milieu, affects endothelial gap junction. Interestingly, both the actions of high glucose and the glycated albumin on endothelial gap junctions may share similar mechanisms. In cultured microvasular endothelial cells treated with high glucose, the Cx43 mRNA was reduced [13]. The same effect was observed for the AGE-BSA-induced down-regulation of Cx43 in this study [14].
Phosphorylation of the members of connexin, including Cx43, is well known to play a key role in regulation of gap-junction communication [20]. The effects of phosphorylation on the communication can be either enhancing or inhibiting. The phosphorylation status of Cx43 is believed to affect the oligomerization of Cx43 into connexons and delivery of integral connexins to justified locations [21-23]. Of the kinases involved in phosphorylation of Cx43, ERK and p38 MAPK were reported to attenuate the gap-junction communication [24]. These data mainly came from non-endothelial cells. On the other hand, activation of the ERK and p38 MAPK cascades was also reported to affect the expression level of Cx43 protein. In cultured smooth muscle cells isolated from human saphenous vein, inhibition of ERK and p38 MAPK by using siRNA specific to the kinases were shown to attenuate the enhanced expression of Cx43 induced by angiotensin II [24]. Considering in the present study that i) a close association between the AGE-BSA-induced reduction in the Cx43 mRNA level and the reduction of Cx43 protein is appeared; ii) the down-regulation of endothelial Cx43 transcript and protein by AGE-BSA is blocked by inhibitors of kinase (ERK and p38 MAPK) cascades; and iii) phosphorylations of ERK and p38 MAPK were activated dose-dependently, one may question whether in endothelial cells after exposure to AGE-BSA there is a link between activation of the MAPK and enhanced proteolysis of Cx43, as reported in lens epithelial cells that phosphorylation is a signal for Cx43 proteins to enter a proteasome-dependent degradation [25]. Moreover, activation of the ERK and p38 MAPK cascades in endothelial cells exposed to AGE-BSA results in regulation of Cx43 protein in a direction opposite to that seen for smooth muscle cells [24]. Recent findings indicated that AGE-HSA (human serum albumin) could promote endothelial progenitor cells (EPCs) senescence and apoptosis via ERK and p38 MAPK pathways, leading to the down-regulation of the number of EPCs [26,27]. This decreases the protective capacity of EPCs on atherosclerosis in diabetic patients and increases their risks for cardiovascular diseases [28]. Similarly, glucose, AGE, and methylglyoxal, the precursor of AGE, could enhance apoptosis of human cardiac myocytes and collagen deposition, an important event of diabetic cardiomyopathy, in rat cardiac fibroblasts accompanied by temporal activation of ERK1/2, p38 MAPK, and nuclear O-GlcAcylation [13,29,30]. Taken together, our findings in the present study suggested that reduced Cx43 transcription is the major mechanism underlying the down-regulation of Cx43 protein in HAEC treated with AGE-BSA. More studies are required to determine whether an increased turnover rate of Cx43 plays a significant role in this process. Although the down-regulation of Cx43 expression was abolished by inhibitors, these rescued Cx43 proteins may already lose their membrane-association and consequently be unable to assemble properly to exert their activity. It is also possible that gap junctions were induced to partially close during the blockage of those signaling pathways. These two phenomenons were already reported in cells under certain physical and chemical offenses [31,32].
Down-regulation of Cx43 gap junctions in endothelial cells by AGE-BSA has implications in the pathogenesis of diabetic vasculopathy, which, as mentioned before, starts with endothelial dysfunction [1]. Our recent findings indicated that in endothelial cells down-regulation of Cx43 per se activates the cells to a pathological status, in which pro-coagulatory molecules, such as plasminogen activation inhibitor 1 and von Willebrand factor, were up-regulated and the viability, angiogenic capacity, and proliferation of the cells were reduced [33]. These changes of endothelial cells due to insufficient expression of Cx43 may further aggravate the harmful effects of AGE-BSA on the vascular cells.
Conclusions
This study provided the evidence that AGE-BSA down-regulated Cx43 expression in HAEC, mainly through reduced Cx43 transcription, and this process involved activation of ERK and p38 MAPK.
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
WC-Y, LH-J, CH-J designed, carried out the experiments, analyzed data and drafted the manuscript. LY-C performed the gap junction communication assay, analyzed data and commented on the manuscript. WH-H performed the real-time PCR, analyzed data and commented on the manuscript. HT-C designed experiments, analyzed data and commented on the manuscript. YH-I supervised the project and corrected the final manuscript. All authors read and approved the final manuscript.
Acknowledgements
Supported by grants NSC-94-2312-B-020-001 and NSC99-2321-B-020-004-MY3 from the National Science Council, Taiwan and MMH-E 95003 from the Medical Research Department of the Mackay Memorial Hospital, Taiwan.
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Decreased expression of extracellular matrix proteins and trophic factors in the amygdala complex of depressed mice after chronic immobilization stress
Soonwoong Jung, Younghyurk Lee, Gyeongwha Kim, Hyeonwi Son, Dong H Lee, Gu S Roh, Sang S Kang, Gyeong J Cho, Wan S Choi and Hyun J Kim*
BMC Neuroscience 2012, 13:58 doi:10.1186/1471-2202-13-58
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Research article
Proximity of food retailers to schools and rates of overweight ninth grade students: an ecological study in California
Philip H Howard1*, Margaret Fitzpatrick1 and Brian Fulfrost2
Author affiliations
1 Department of Community, Agriculture, Recreation and Resource Studies, Michigan State University, 316 Natural Resources, East Lansing, MI, 48824, USA
2 Design, Community & Environment, 1625 Shattuck Ave, Suite 300, Berkeley, CA, 94709, USA
For all author emails, please log on.
Citation and License
BMC Public Health 2011, 11:68 doi:10.1186/1471-2458-11-68
Published: 31 January 2011
Abstract
Background
The prevalence of obesity and overweight in youth has increased dramatically since the 1980s, and some researchers hypothesize that increased consumption of low-nutrient, energy-dense foods is a key contributor. The potential importance of food retailers near schools has received increasing attention, but public health research and policy has focused primarily on fast food restaurants. Less is known about the relationship between overweight/obesity and other types of retailers. This study aims to investigate the potential associations between nearby 1) fast food restaurants, 2) convenience stores, and 3) supermarkets, and rates of overweight students in California schools.
Methods
We examined the rate of overweight ninth grade students in public schools in 2007 using linear regression. The percentage of overweight students per school was determined by a state required physical fitness test, with three different options for measuring individual body composition. Our key independent variables were the presence of three different types of retailers within 800 m network buffers of the schools. Additional independent variables included school ethnic, gender and socioeconomic composition, as well as urban/non-urban location. We obtained the data from the California Department of Education and ESRI, Inc.
Results
The presence of a convenience store within a 10-minute walking distance of a school was associated with a higher rate of overweight students than schools without nearby convenience stores, after controlling for all school-level variables in the regression (1.2%, 95% confidence interval 0.03, 2.36). Nearby fast food restaurants and supermarkets, however, were not associated with school rates of overweight students.
Conclusions
Public health researchers and policy-makers interested in the food environments outside schools should expand their recent focus on nearby fast food restaurants to include convenience stores, which may also be important sources of low-nutrient, energy-dense foods for students.
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This article is part of the series Data standardization, sharing and publication.
Correspondence
Adventures in data citation: sorghum genome data exemplifies the new gold standard
Scott C Edmunds1*, Tom J Pollard2,3,4, Brian Hole2,5 and Alexandra T Basford1
Author Affiliations
1 GigaScience, BGI-Hong Kong Ltd., 16 Dai Fu Street, Tai Po Industrial Estate, NT, Hong Kong
2 Ubiquity Press, Gordon House, 29 Gordon Square, London, WC1H 0PP, United Kingdom
3 Department of Space & Climate Physics, Mullard Space Science Laboratory, University College London, Surrey, RH5 6NT, United Kingdom
4 University College Hospital, 235 Euston Road, London, NW1 2BU, United Kingdom
5 Institute of Archeology, University College London, 31–34 Gordon Square, London, WC1H 0PY, United Kingdom
For all author emails, please log on.
BMC Research Notes 2012, 5:223 doi:10.1186/1756-0500-5-223
The electronic version of this article is the complete one and can be found online at: http://www.biomedcentral.com/1756-0500/5/223
Received:16 December 2011
Accepted:9 May 2012
Published:2 July 2012
© 2012 Edmunds et al.; licensee BioMed Central Ltd.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Scientific progress is driven by the availability of information, which makes it essential that data be broadly, easily and rapidly accessible to researchers in every field. In addition to being good scientific practice, provision of supporting data in a convenient way increases experimental transparency and improves research efficiency by reducing unnecessary duplication of experiments. There are, however, serious constraints that limit extensive data dissemination. One such constraint is that, despite providing a major foundation of data to the advantage of entire community, data producers rarely receive the credit they deserve for the substantial amount of time and effort they spend creating these resources. In this regard, a formal system that provides recognition for data producers would serve to incentivize them to share more of their data.
The process of data citation, in which the data themselves are cited and referenced in journal articles as persistently identifiable bibliographic entities, is a potential way to properly acknowledge data output. The recent publication of several sorghum genomes in Genome Biology is a notable first example of good data citation practice in the field of genomics and demonstrates the practicalities and formatting required for doing so. It also illustrates how effective use of persistent identifiers can augment the submission of data to the current standard scientific repositories.
Discussion
One of the key lessons learned from the Human Genome Project, taking a page from the C. elegans community [1], was that making data broadly and freely available prior to publication was profoundly valuable to the field of genomics [2]. Subsequent genomics projects have tried to follow this practice as laid out in the Bermuda Rules [3] and ultimately enshrined in the Fort Lauderdale agreement [4]. The wider biological science community has also attempted to follow similar practices, as outlined in the guidelines published from the Toronto International Data Release Workshop [5], but adoption has been held back by a lack of easy-to-access repository infrastructure for many fields as well as an absence of incentives for authors to go through the time and effort necessary to make their work openly and easily available to others.
The benefits of making data available to the research community as a whole can be calculated [6]: there is a measurable trend towards an author’s work accumulating additional citations as a result of the supporting data being publically accessible [7,8]. Again, however, the lack of a universally recognized tagging system linking investigators to their deposited data has hindered authors from receiving due credit [9]. Recent scandals relating to falsified data that went long undetected in medicine [10] and psychology [11] also highlight the need to make data easily accessible for purposes of validation and to maintain public trust in science. One notable attempt to address the issues of gaining complete access to data is the Dryad repository, which serves as a storehouse for smaller datasets directly affiliated with publications in the biosciences [12].
The next step forward in this regard is the recent publication of the genomes of three strains of the important food crop Sorghum bicolor published in Genome Biology[13]. This work follows the best practices of the genomics community by having the supporting raw and useful processed data available in the relevant and available data repositories. However, for the first time in the long-established data-sharing practices of the community, this process has been supplemented specifically by integrating into the reference section a citation for the collective dataset. Thus, in addition to having the raw data [SRA046843], assemblies [GenBank: AHAO00000000-AHAQ00000000], mutation [1056306], and structural variation data [nstd63] available in a number of NCBI databases, the data citation included in the reference section [14] makes available the same data, along with additional information, from a single point of access. In addition, and perhaps of even greater value, these data are persistently linked via a citable DataCite Digital Object Identifier (DOI), and are hosted on the GigaScience[15]GigaDB database [16].
GigaScience and GigaDB
GigaScience is a journal and data publishing project set up in conjunction with BGI (formally the Beijing Genomics Institute) [17], one of the world’s largest genomics data producers, and BioMed Central [18]. Using BGI’s large data storage and cloud computing infrastructure, GigaScience has created a novel publication format that integrates manuscript publication with data hosting. The data associated with articles are hosted in the connected GigaDB database and given DOIs to make them more searchable and trackable as well as independently citable.
To demonstrate the utility of data citation and to promote extremely rapid data release and dissemination of unpublished datasets independently of the journal, GigaDB has recently released large-scale data resources created by the BGI and its many external collaborations. Importantly, these data are the first cases in which whole genome-type data have been released with DOIs. GigaScience has been working with DataCite through the British Library to enable these and future datasets to receive DOIs.
DataCite
Founded in December 2009, DataCite is an international partnership working towards a global citation framework for research data, with the aim of enabling researchers to find, access, and reuse datasets with confidence. Since its foundation, DataCite has been building a community to collaboratively develop services and good practices for data citation [19,20].
DataCite is initially leveraging the DOI system for research data. DOIs identify a resource, rather than the location of the resource, allowing the creation of persistent and stable references and offering an easy way to connect articles with their underlying data. The DOI system is governed by the International DOI Foundation (IDF), a non-profit organisation, with DataCite operating as a Registration Agency. This relationship confers DataCite and its member organisations [21] with rights and infrastructure to register DOIs. The crucial advantage of the DOI system over alternatives is that it is already familiar to researchers, publishers, and libraries.
DataCite provides a service for trusted repositories to mint DOIs for datasets and, since August 2011, collects mandatory metadata on every DOI minted. Services are being developed around this open ‘metadata store’ to promote discovery and to facilitate access to original cited datasets. DataCite DOIs resolve to public pages describing the datasets and providing a route for access. DataCite is also developing a content negotiation service that will allow metadata, and possibly even the datasets themselves, to be requested directly via DOIs [22].
A cost is associated with managing persistence and with assigning identifiers, and so there is a cost associated with DOIs. In April 2011 the International DOI Foundation changed the charging model from a per-DOI cost to a cost-sharing model [23]. Essentially this means that repositories can pay a flat-fee to a Registration Agency for the right to mint virtually unlimited DOIs.
A brief history of data citation
Environmental sciences researchers have been using the Pangaea [24] database to host data associated with their manuscripts for many years (for an example from 2005 see [25]), and Dryad has more recently created a similar model with biomedical data [26]. These have been notable successes in the movement for better data access. However, in cases where journals have included such datasets in their references, the datasets are often treated and formatted the same way as links on the web and, thus, are not listed by citation indices (such as article [25] and its associated dataset [27]). The long-established Protein Data Bank (PDB) biological macromolecular structure data archive [28] also uses DOIs, but other than rare exceptions [29,30], very few research articles have used DOIs to reference structures. Publishing data by wrapping and integrating it into the established journal infrastructure is underway in a number of research areas such as the Earth Systems Science Data journal [31], and there have been attempts to semantically enhance and integrate links with data via DOIs in biodiversity [32] and infectious disease research [33]. This has facilitated discovery and access to the underlying data, but these examples have not utilized or been citable using current journal indexing services.
As DOIs issued for GigaDB datasets have been associated with and published alongside journal manuscripts, the GigaDB project appears to be the first time that genomic datasets have been released prior to manuscript publication in this citable DOI form. Although there have been public calls [2] and journal editorials [9] encouraging such a system, the practicalities and consequences of releasing data in a citable form before the publication of their associated manuscripts have been unclear, especially with widely varying journal editorial policies regarding pre-publication dissemination of results. Relevant to this is a commonly acknowledged editorial guideline from the New England Journal of Medicine that outlines limitations on prepublication release of information known as the “Ingelfinger rule” [34]. It effectively states that a manuscript may not be considered for publication if its substance has been submitted or reported elsewhere and it has made many researchers wary of publicizing preliminary data. However, there are a number of ambiguities as to how this restriction is reconciled with the biological, and in particular the genomic, community’s code of practice regarding pre-publication data deposition in public databases.
An interesting test case was the first dataset to be given a data citation by BGI [35]. This high-profile dataset, the first publicly available genome of the E. coli 0104:H4 pathogen responsible for the 2011 European outbreak, was released prior to the publication of an associated article [36]. Researchers at the BGI collaborated with the University Medical Centre Hamburg-Eppendorf to rapidly sequence the genome of the pathogen. Due to the seriousness of the situation (with 50 human deaths and over 4000 people infected), it was clear that it was not in the public’s best interest to hold back that information in order to follow standard research practices of first analysing the data and then waiting to release them until after acceptance of the resulting manuscript. Instead, the decision was made to immediately release the dataset under the most open public domain waiver, CC0 [37], to maximize its use by the community. By giving the dataset a DOI it was possible to not only enable the research community to cite and credit the authors, but also to mark the time of data release, making this less commonly used route of data release more attractive to the authors.
Of greatest interest, perhaps, was that the rapid release of the E. coli genome data enabled an international community of “crowdsourced” researchers to pool resources and carry out expeditious “open-source” analysis of the organism, a level of instantaneous collaboration that has not been seen before. This high-profile distributed problem-solving approach substantially aided in limiting the health crisis, with strain-specific diagnostic primers disseminated within five days of the release of the sequence data (sequence available from the DOI landing page [35]), and the draft unassembled genome sequence data subsequently enabled the development of a targeted bactericidal agent to kill the pathogen [38]. It also brought to light a potentially useful way of scientifically addressing similar outbreaks in the future. Additionally, results of these analyses were published in the New England Journal of Medicine a few months later, showing that data citation can complement the traditional forms of academic credit [36].
Many other unpublished datasets have been released in the GigaDB database with DataCite DOIs, and a number of these have subsequently been used in scholarly journal articles. DOIs for two BGI datasets [39,40] used in a Nature Biotechnology[41] article were listed in the article’s Accession Codes section, but were not included in the reference section due to citation limits and policies treating them as non-refereed sources of information such as websites. The authors of the Sorghum bicolor genome paper worked very closely with the editors of Genome Biology to ensure that it followed the best practice guidelines and current recommendations regarding how best to cite data, and included the data citation in the reference section [42]. Since the publication of this paper, there have been positive recent developments from publishers such as Springer and Nature providing examples of data cited in this way [43,44].
Discoverability, accessibility, and preservation
Discoverability and accessibility of data are separate issues and should be treated as such. There are times when it may be necessary to limit accessibility to a dataset, but this should not prevent it from being archived and made discoverable via open metadata and a persistent identifier such as a DOI. Equally, discoverability and long-term preservation of data can be dealt with as separate issues. Repositories should be carefully selected to ensure a preservation plan is in place, but we must accept that some datasets will be lost over time, for example due to limited storage capacity. By ensuring that persistent identifier organizations are provided with open metadata, it should at least be possible to keep a record of a dataset’s existence and provenance however. DataCite, for example, collects metadata for all datasets that are allocated DataCite DOIs. In the event of a dataset becoming unavailable, the appropriate DOI can be updated to resolve to the associated metadata record.
How to cite data
Given the importance of data in promoting research, and the needs of data producers to gain credit for their work in the same manner that researchers using these data are recognized, datasets should be cited as research articles are cited. Further to this, even though many journals currently tend to remove URLs from the reference list, both DataCite and CrossRef recommend displaying DOIs within references as full URLS. They consider this best practice because it emphasises the actionable link and allows readers to readily access the underlying data. The DOI in URL form not only serves the same function as a journal volume, issue and page number do for a printed article, but also gives the combined advantages of linked access and the assurance of persistence, the lack of which in the past being part of the reason many journals have been reluctant to cite plain vanilla URLs. An example of what can be considered a new gold standard for data citation is the way in which the data that underpin the recently published sorghum paper [14] were cited in the reference section, as follows:
Zheng, L-Y; Guo, X-S; He, B; Sun, L-J; Peng, Y; Dong, S-S; Liu, T-F; Jiang, S; Ramachandran, S; Liu, C-M; Jing, H-C (2011): Genome data from sweet and grain sorghum (Sorghum bicolor). GigaScience. http://dx.doi.org/10.5524/100012 webcite
It is important to note that the DOI will always point to the version of the data that were used for the study in which they were cited, enabling other researchers to use them for validation and comparative studies with confidence.
Formally citing the dataset in this manner not only clearly identifies the dataset, but also paves the way for data discovery and citation tracking via existing bibliometric services. These services have traditionally focused on DOIs in the reference section only, so including data citations here can help such services to start using them more quickly as existing systems will require less modification.
Aiding the adoption of data citation
Many journals now include ‘Data Accessibility’ sections that provide information about the data used in the paper and explain where these data can be accessed. Journal editors are also starting to draw up new guidelines for data citation, but these approaches still remain inconsistent. In this regard, it is crucial that high-profile journals take the lead in citing data in a manner that drives adoption of good practices and raises awareness of this issue to the broader research community.
Journal editors and publishers who promote consistent and equitable means of citing data, as exemplified by the handlers of the sorghum paper [13], should be commended. Defining formal mechanisms for dataset citation is essential for making datasets more readily tracked and easily accessed. Furthermore, it provides the only real means for data producers to obtain appropriate recognition for their work, promoting more rapid data release potentially prior to the much more time-consuming process of manuscript publication. It also gives recognition and makes clear the role of the researchers investing the most effort in producing the dataset, who may not have received similar credit in an eventual more analysis-focused publication. As research is being carried out with ever increasing amounts of data, widespread data availability will serve to enhance scientific progress and provide greater public benefit from the investments made to create these sharable data resources.
Competing interests
SCE and AB are employees of the BGI and work on the GigaScience project. TP is a previous employee of the British Library and DataCite. BH is a previous employee of the British Library and Dryad-UK.
Authors' contributions
SE, TP and AB assigned DOIs to the BGI datasets highlighted here. All authors contributed to the writing, and read and approved the final manuscript.
Acknowledgements
SCE and AB are supported by the BGI. The authors would like to thank the authors of the Genome Biology sorghum paper for depositing their work in this way, and Shaoguang Liang for helping set up the GigaScience database. We would also like to thank Laurie Goodman for feedback and support.
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On Grammatical Differences between Daily British and American English
Bin Zhang, Zhaofeng Jiang
Abstract
The grammatical differences between daily British English and American English are mainly in terms of the usage of prepositions, auxiliary verbs, articles, pronouns, adjectives and adverbs, and tense and subjunctive mood. These differences exert influence on English learning and interpersonal communication. This paper tries to illustrate the differences of grammar between British and American daily English based on the authors’ teaching experience and learning experience in the UK. By comparison, English learners would understand more about the differences and be more effective while communicating with the people from the US and the UK.
Full Text: PDF
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Asian Social Science ISSN 1911-2017 (Print) ISSN 1911-2025 (Online)
Copyright © Canadian Center of Science and Education
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Building the Capacity of Farmer Based Organisation for Sustainable Rice Farming in Northern Ghana
Quaye Wilhemina, Yawson Ivy, Manful John Tawiah, Gayin Joseph
Abstract
This study assessed the perceptions of stakeholders concerning implementation activities of a Food Security and Rice
Producers Organisation Project (FSRPOP) in Northern Ghana. The project aimed at building the capacities of farmer
based organisations (FBOs) to assist rice farmers access credit, organise production inputs and improve market access.
The study results showed that although access to input supply and production credit improved, enhancing farmers’
marketing capacity was not successful. The management capacity of the FBOs was weak in performing more complex
administrative issues and market facilitating roles. Timely provision of production inputs, use of custom based
processing and credit inventory system for maximum profit were some of the lessons learnt. Facilitation of farmer -
medium scale buyer linkages and the development of lessons-based action plan for change with beneficiaries were
recommended
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Journal of Agricultural Science ISSN 1916-9752 (Print) ISSN 1916-9760 (Online)
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<D <M <Y
Y> M> D>
: C'mon! Ask the tough question!
(The tough question being "If every time particular copyrights are about to expire Congress extends copyright terms, as happened throughout the 20th century, doesn't that violate the spirit of the 'limited time' clause?" The corollary question being: "Will there be a point at which your clients will say 'OK, that's enough' and stop lobbying for another extension?")
I thought of that question as soon as I saw the headline of the article, and read the whole article hoping it would be asked, and it wasn't. Bah!
: I wasn't planning on including this photo in a roundup, because it seemed like a really cheap shot, but after I found it, Mike Popovic also found it and told me about it, so why not?
: Well, Uncle Sam and Uncle Gray have cashed my tax checks, so I'm the poorest I've been in a while. I don't understand why my witholdings were so far off from what I actually owed in taxes.
[Main]
Unless otherwise noted, all content licensed by Leonard Richardson
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< Alternate ETag Validation Functions
Tiny Youtube Party >
: More REST stuff, this time outsourced to Martin Fowler. In the embarassingly-titled "Richardson Maturity Model: steps toward the glory of REST", Fowler talks about the maturity model I set out at QCon in 2008, and shows examples of services at the different maturity levels.
Filed under:
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Rate This Article
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Ecoregions of North Carolina and South Carolina (EPA)
Ecoregions of North Carolina and South Carolina (EPA)
This article has been reviewed by the following Topic Editor: Mark McGinley
Ecoregions of North Carolina and South Carolina.
Ecoregions denote areas of general similarity in ecosystems and in the type, quality, and quantity of environmental resources. They are designed to serve as a spatial framework for the research, assessment, management, and monitoring of ecosystems and ecosystem components. By recognizing the spatial differences in the capacities and potentials of ecosystems, ecoregions stratify the environment by its probable response to disturbance (Bryce and others, 1999). These general purpose regions are critical for structuring and implementing ecosystem management strategies across federal agencies, state agencies, and nongovernment organizations that are responsible for different types of resources within the same geographical areas (Omernik and others, 2000).
The approach used to compile this map is based on the premise that ecological regions are hierarchical and can be identified through the analysis of the spatial patterns and the composition of biotic and abiotic phenomena that affect or reflect differences in ecosystem quality and integrity (Wiken 1986; Omernik 1987, 1995). These phenomena include geology, physiography, vegetation, climate, soils, land use, wildlife, and hydrology. The relative importance of each characteristic varies from one ecological region to another regardless of the hierarchical level. A Roman numeral hierarchical scheme has been adopted for different levels of ecological regions. Level I is the coarsest level, dividing North America into 15 ecological regions. Level II divides the continent into 52 regions (Commission for Environmental Cooperation Working Group 1997). At level III, the continental United States contains 104 ecoregions and the conterminous United States has 84 ecoregions (United States Environmental Protection Agency [USEPA] 2002). Level IV is a further subdivision of level III ecoregions. Explanations of the methods used to define the USEPA’s ecoregions are given in Omernik (1995), Omernik and others (2000), and Gallant and others (1989).
Ecological and biological diversity of the Carolinas is enormous. The two states contain barrier islands and coastal lowlands, large river floodplain forests, rolling plains and plateaus, forested mountains, and a variety of aquatic habitats. There are 5 level III ecoregions and 29 level IV ecoregions in North Carolina and South Carolina and most continue into ecologically similar parts of adjacent states.
Level III Ecoregions of the Conterminous United States. (Map source: USEPA, 2000)
The level III and IV ecoregions on this poster were compiled at a scale of 1:250,000 and depict revisions and subdivisions of earlier level III ecoregions that were originally compiled at a smaller scale (USEPA 2002; Omernik 1987). This poster is part of a collaborative project primarily between USEPA Region IV, USEPA National Health and Environmental Effects Research Laboratory (Corvallis, Oregon), North Carolina Department of Environment and Natural Resources (NCDENR), South Carolina Department of Health and Environmental Control (SCDHEC), and the United States Department of Agriculture-Natural Resources Conservation Service (NRCS). Collaboration and consultation also occurred with the United States Department of Agriculture-Forest Service (USFS), United States Department of the Interior-Geological Survey (USGS)-Earth Resources Observation Systems (EROS) Data Center, and with other State of North Carolina and State of South Carolina agencies.
The project is associated with an interagency effort to develop a common framework of ecological regions (McMahon and others, 2001). Reaching that objective requires recognition of the differences in the conceptual approaches and mapping methodologies applied to develop the most common ecoregion-type frameworks, including those developed by the USFS (Bailey and others, 1994), the USEPA (Omernik 1987, 1995), and the NRCS (U.S. Department of Agriculture-Soil Conservation Service, 1981). As each of these frameworks is further refined, their differences are becoming less discernible. Regional collaborative projects such as these in North Carolina and South Carolina, where some agreement has been reached among multiple resource management agencies, are a step toward attaining consensus and consistency in ecoregion frameworks for the entire nation.
45. Piedmont
Considered the non-mountainous portion of the old Appalachians Highland by physiographers, the northeast-southwest trending Piedmont ecoregion comprises a transitional area between the mostly mountainous ecoregions of the Appalachians to the northwest and the relatively flat coastal plain to the southeast. It is a complex mosaic of Precambrian and Paleozoic metamorphic and igneous rocks with moderately dissected irregular plains and some hills. Once largely cultivated, much of this region is in planted pine or has reverted to successional pine and hardwood woodlands. The historic oak-hickory-pine forest was dominated by white oak (Quercus alba), southern red oak (Quercus falcata), post oak (Quercus stellata), and hickory (Carya spp.), with shortleaf pine (Pinus echinata), loblolly pine (Pinus taeda), and to the north and west, Virginia pine (Pinus virginiana). The soils tend to be finer-textured than in coastal plain regions.
45a. The Southern Inner Piedmont is generally higher in elevation with more relief than 45b. As a transitional region from the Blue Ridge (66) to the Piedmont, it contains some mountain outliers, and it receives more rainfall than 45b and 45c. The rolling to hilly well-dissected upland contains mostly gneiss and schist bedrock that is covered with clayey and micaceous saprolite. It is warmer than 45e to the north, and contains thermic soils rather than 45e’s mesic soils. The region is now mostly forested, with major forest types of oak-pine and oak-hickory, and less loblolly-shortleaf pine forest than 45b. Open areas are mostly in pasture, although there are some small areas of cropland.
45b. The Southern Outer Piedmont ecoregion has lower elevations, less relief, and less precipitation than 45a. The landform class is mostly irregular plains rather than the plains with hills of 45a and 45e. Pine (mostly loblolly and shortleaf) dominates on old field sites and pine plantations, while mixed oak forest is found in less heavily altered areas. Gneiss, schist, and granite are typical rock types, covered with deep saprolite and mostly red, clayey subsoils. Kanhapludults are common soils, such as the Cecil, Appling, and Madison series. Some areas within this region have more alkaline soils, such as the Iredell series, formed over diabase, diorite, or gabbro, and may be associated with areas once known as blackjack oak prairies.
45c. The Carolina Slate Belt extends from southern Virginia, across the Carolinas, and into Georgia. The mineral-rich metavolcanic and metasedimentary rocks with slatey cleavage are finer-grained and less metamorphosed than most Piedmont regions. Some parts are rugged, such as the Uwharrie Mountains, and many areas are distinguished by trellised drainage patterns. Silty and silty clay soils, such as the Georgeville and Herndon series, are typical. Streams tend to dry up and water yields to wells are low as this region contains some of the lowest water-yielding rock units in the Carolinas.
45e. Similar to 45a, the rolling to hilly Northern Inner Piedmont has higher elevations, more rugged topography, and more monadnocks or mountain outliers than other areas of the Piedmont. It has colder temperatures, more snowfall, and a shorter growing season than in 45a, b, c, and f, and it has mostly mesic soils rather than the thermic soils that cover other regions of the Carolina Piedmont. The region contains more Virginia pine and less shortleaf pine than 45b and 45c, more chestnut oak, and many mountain disjunct plant species. Streams tend to have higher gradients than in the Outer Piedmont regions, and contain many mountain-type macroinvertebrate species.
45f. The Northern Outer Piedmont is composed of mostly gneiss and schist rock intruded by granitic plutons, and veneered with saprolite. It is lithologically distinct from the adjacent Piedmont regions 45c and 45g, as well as from the younger unconsolidated sediments of 65m. Rocks and soils are similar to 45b, but 45f is cooler with a shorter growing season. The region contains more loblolly pine compared to the Virginia pine and shortleaf pine found in the Piedmont to the west, but it also contains local concentrations of mountain disjunct plant species. At the eastern boundary, the Fall Line is a broad transition zone where Piedmont rocks occur on the same landscape with Coastal Plain sediments. Some areas near this boundary have metavolcanic and metasedimentary rocks similar to 45c.
45g. The Triassic Basins of the Carolinas occur in four narrow bands and have unusual Piedmont geology of unmetamorphosed shales, sandstones, mudstones, siltstones, and conglomerates. Local relief and elevations are often less than in surrounding regions, and, with rocks that are easier to erode, stream valleys that cross the region tend to widen. Soils tend to be clayey with low permeability, and streams have low base flows. The clay has a high shrink-swell potential that can hinder construction; it is also utilized by many brick makers in the region. A mosaic of mixed and deciduous forest, pasture, cropland, and urban land cover occurs here.
45i. The Kings Mountain ecoregion is a hilly, somewhat rugged area with some northeast- to southwest-trending ridges and distinctive metasedimentary and metavolcanic rocks. Aluminum-rich quartz-sericite schist is common. The metamorphic grade is generally lower than adjacent geologic belts and the rocks contain an unusual variety of mineral deposits. Mining strongly influenced the early development of the region, including an iron industry in the late 1700’s to late 1800’s, and later production of marble, lime, gold, lead, silver, pyrite, lithium, mica, feldspar, silica, and clay. Soils are often a very fine sandy to silty texture, similar to 45c. The region is covered with oak-hickory-pine forest, and Virginia pine is common.
Several major land cover transformations have occurred in the Piedmont over the past 200 years, from forest to farm, back to forest, and now in many areas, spreading urban- and suburbanization. The Piedmont contains most of the largest urban areas of the Carolinas, with relatively high regional population densities and rates of growth.
Most rocks of the Piedmont are covered by a thick mantle of saprolite, except along some major stream valley bluffs and on a few scattered granitic domes and flatrocks. Rare plants and animals are found on rock outcrops, such as at Forty Acre Rock in Lancaster County, South Carolina. (Photo: Mike Creel, SCDNR)
Beaver (Castor canadensis) populations have increased in the Piedmont. They can create wetland habitat for a variety of plant and animal species, reduce downstream sedimentation, and improve water quality.
The Carolina wren (Thryothorus ludovicianus), the state bird of South Carolina, nests from the mountains, across the Piedmont and into the Coastal Plain. (Photo:SCDNR)
63. Middle Atlantic Coastal Plain
Ecoregion 63 is found primarily in the Carolinas and other states to the north, and has a broad transitional boundary with Ecoregion 75 to the south. It consists of low elevation, flat plains, with many swamps, marshes, and estuaries. Forest cover in the region, once dominated by longleaf pine in the Carolinas, is now mostly loblolly and some shortleaf pine, with patches of oak, gum, and cypress near major streams, as compared to the mainly longleaf-slash pine forests of the warmer Southern Coastal Plain (75). Its low terraces, marshes, dunes, barrier islands, and beaches are underlain by unconsolidated sediments. Poorly drained soils are common, and the region has a mix of coarse and finer textured soils compared to the mostly coarse soils in the majority of Ecoregion 75. Ecoregion 63 is typically lower, flatter, more poorly drained, and more marshy than Ecoregion 65. Pine plantations for pulpwood and lumber are typical, with some areas of cropland.
63b. The Chesapeake-Pamlico Lowlands and Tidal Marshes occur on the lowest marine terrace with elevations ranging from sea level to about 25 feet. The western boundary of 63b generally occurs at the Suffolk Scarp. The region is characterized by nearly level plains with some broad shallow valleys, seasonally wet soils (Aquults), brackish and fresh streams, and broad estuaries affected by wind tides. It is flatter and lower in elevation than 63e, with a slightly longer growing season than 63e and 65m. Some major areas of cropland are found in the region, growing corn, wheat, soybeans, and potatoes. Lake Mattamuskeet, the largest natural lake in North Carolina, provides valuable wintering areas for geese, swans, ducks, and other birds.
63c. Nonriverine Swamps and Peatlands are flat, poorly drained areas containing organic soils of peat and muck. The dark reddish-brown to black soils, acidic and nutrient-poor, often contain logs, stumps, and other woody matter from bald cypress and Atlantic white cedar trees. Pocosin lakes occur in some areas. The vegetation of the high and low pocosins contains a dense shrub layer, along with stunted pond pine, swamp red bay, and sweet bay. Swamp forests are dominated by swamp tupelo, bald cypress, and Atlantic white cedar. Fire during drought periods, logging, and construction of drainage ditches have affected natural vegetation patterns. Several areas of mineral and shallow organic soils have been drained and cultivated for crops of corn, soybeans, and wheat.
63d. Virginian Barrier Islands and Coastal Marshes occur in the northeast corner of North Carolina and contain salt, brackish, and freshwater marshes, dunes, beaches, and barrier islands that enclose Currituck Sound. The Quaternary-age deposits of unconsolidated sand, silt, and clay form dynamic landscapes affected by ocean wave, tide, wind, and river energy. The nearshore ocean water, influenced by the longshore Virginia Current, tends to be colder than in most of 63g, especially south of Cape Hatteras, where warmer Gulf Stream waters occur. On the barrier islands, northern beach grass and deciduous oaks are typical, compared to the sea oats and evergreen live oak more commonly found to the south in 63g. Salt marshes are dominated by saltmarsh and saltmeadow cordgrasses and black needlerush, while the freshwater marshes of upper Currituck Sound contain bulrush, cattail, sawgrass, and big cordgrass. The marshes provide wintering habitat for geese, ducks, and wading birds. Piping plover and loggerhead sea turtles occasionally nest along the beaches.
63e. The Mid-Atlantic Flatwoods occupies the middle portion of the coastal plain in northern North Carolina and southern Virginia. Upland surfaces are wider, lower in elevation, with less local relief, and have more poorly drained soils compared to Ecoregion 65m. Soils such as Aquults and some Udults formed in the mostly Pleistocene-age clays and sands. With slow natural subsurface drainage, except near streams, artificial drainage is common for agriculture and forestry operations. Corn, peanuts, and cotton are typical crops. Although similar to 63h, the Mid-Atlantic Flatwoods historically had a lower frequency of fire, less longleaf pine, and a different mix of grasses than in 63h. There are fewer Carolina Bays, and the region tends to be biologically less diverse than 63h in terms of plants and aquatic macroinvertebrates.
63g. The Carolinian Barrier Islands and Coastal Marshes covers most of the North Carolina coast, extending from Bodie Island in the north to North Myrtle Beach, South Carolina in the south. Similar to 63d, the region contains marshes, dunes, beaches, and barrier islands, but it tends to be slightly warmer and wetter. In the north, the boundary with 63d is transitional, and there is a high diversity of vegetation in the maritime forests in the boundary area where northern and southern maritime forests overlap, such as at Nags Head Woods. The maritime forests include live oak, laurel oak, loblolly pine, red cedar, yaupon holly, wax myrtle, dwarf palmetto, with cabbage palm (Sabal palmetto) in the south. Pamlico Sound is a shallow estuary supporting an important nursery for 90 percent of all the commercial seafood species caught in North Carolina, as well as vast recreational fisheries.
63h. The nearly level coastal plain of the Carolina Flatwoods has less relief, wider upland surfaces, and larger areas of poorly drained soils than the adjacent, higher elevation Ecoregion 65l. Covered by shallow coastal waters during the Pleistocene, the resultant terraces and shoreline-related landforms are covered typically by fine-loamy and coarse-loamy soils, with periodically high water tables. Other areas have clayey, sandy, or organic soils, contributing to the region’s plant diversity. Carolina bays and pocosins are abundant in some areas. The region is a significant center of endemic biota, with more biological diversity and rare species compared to 63e. Pine flatwoods, pine savannas, freshwater marshes, pond pine woodlands, pocosins, and some sandhill communities were once common. Loblolly pine plantations are now widespread with an active forest industry. Artificial drainage for forestry and agriculture is common. North Carolina’s blueberry industry is concentrated on some of the sandy, acidic soils of the region.
63n. The Mid-Atlantic Floodplains and Low Terraces are mostly a continuation of the riverine Ecoregion 65p, although a few floodplains mapped in this region originate within Ecoregion 63. Large, sluggish rivers, deep-water swamps, oxbow lakes, and alluvial deposits with abrupt textural changes characterize 63n. Brownwater floodplains originate in or cross the Piedmont (45) and the sediments contain more weatherable minerals than the blackwater floodplains that have their watersheds entirely within the coastal plain. Cypress-gum swamps are common, along with bottomland hardwoods of wetland oaks, green ash, red maple, and hickories.
Saltmarsh cordgrass (Spartina alterniflora) in the coastal marshes of Ecoregion 63g. The roots of spartina grass help hold the marsh soil and provide a footing for oysters and ribbed mussels. When the grass dies back in the fall, bacteria and fungi break it down into detritus, the base of the marsh food web. The detritus is eaten by crabs, snails, mussels, oysters, clams, and worms, which in turn are food for fish, shellfish, birds, and mammals.
Ghost crabs (Ocypode quadrata) inhabit the dry upper portions of beaches, although several times a day they must return to ocean waters to wet their gills.
Longleaf pine forests are fire-dependent. In the absence of fire, longleaf pine and its associated plants and animals, some of which are rare and occur only in longleaf pine ecosystems, are replaced by other species. (Photo: Ted Borg, SCDNR)
The carnivorous Venus fly traps (Dionaea muscipula) are native to some of the pocosins and peat bogs of Ecoregion 63 in the Carolinas.
The alligator (Alligator mississippiensis) is an integral component of many wetland ecosystems in the southern part of Ecoregion 63. North Carolina is usually considered the northern extent of its habitat range. (Photo: SCDNR)
65. Southeastern Plains
These irregular plains with broad interstream areas have a mosaic of cropland, pasture, woodland, and forest. Natural vegetation was predominantly longleaf pine, with smaller areas of oak-hickory-pine. On some moist sites, especially in the far south near Florida, Southern mixed forest occurred with beech, sweetgum, southern magnolia, laurel and live oaks, and various pines. The Cretaceous or Tertiary-age sands, silts, and clays of the region contrast geologically with the older metamorphic and igneous rocks of the Blue Ridge (66) and Piedmont (45). Elevations and relief are greater than in the Southern Coastal Plain (75), but generally less than in much of the Piedmont. Streams in this area are relatively low-gradient and sandy-bottomed.
65c. The Sand Hills are a rolling to hilly region composed primarily of Cretaceous-age marine sands and clays, capped in places with Tertiary sands, deposited over the crystalline and metamorphic rocks of the Piedmont (45). Many of the droughty, low-nutrient soils formed in thick beds of sand, although some soils contain more loamy and clayey horizons. Some upland areas are underlain by plinthite, and sideslopes tend to have fragipans that perch water and cause lateral flow and seepage. Stream flow is consistent; streams seldom flood or dry up because of the large infiltration capacity of the sandy soil and the great groundwater storage capability of the sand aquifer. On drier sites, turkey oak and blackjack oak grow with longleaf pine and a wiregrass ground cover. Shortleaf-loblolly pine forests and other oak-pine forests are now more widespread due to fire suppression and logging. The Sand Hills are a center of rare plant diversity in the Carolinas. The region is also known for its peach orchards, golf courses, and horse farms.
65l. The Atlantic Southern Loam Plains ecoregion is lower, flatter, more gently rolling, with finer-textured soils than 65c. It is a major agricultural zone, with deep, well-drained soils, and more cropland than 65c or 63h. The flora is varied due to the variety of edaphic conditions, but is generally more mesic than found in 65c, and more xeric than in 63h. The region has the highest concentration of Carolina bays. These are shallow, elliptical depressions, often swampy or wet in the middle with dry sandy rims. Carolina bays not drained for agriculture often contain rare or endangered plant and animal species.
65m. The dissected Rolling Coastal Plain extends south from Virginia and covers much of the northern upper coastal plain of North Carolina. Relief, elevation, and stream gradients are generally greater than in Ecoregion 63 to the east, and soils tend to be better drained. It has a slightly cooler and shorter growing season than 65l, but is a productive agricultural region with typical crops of corn, soybeans, tobacco, cotton, sweet potatoes, peanuts, and wheat. The region appears to be biologically less diverse than the coastal plain regions 65l and 63h to the south.
65p. Southeastern Floodplains and Low Terraces comprise a riverine ecoregion that provides important wildlife corridors and habitat. Composed of alluvium and terrace deposits of sand, clay, and gravel, the region includes large sluggish rivers and backwaters with ponds, swamps, and oxbow lakes. It includes oak-dominated bottomland hardwood forests, and some river swamp forests of bald cypress and water tupelo. Similar to 63n, the flood-prone region includes brownwater floodplains and blackwater floodplains. The brownwater floodplains originate in or cross the Piedmont (45) and the sediments contain more weatherable and mixed minerals than the blackwater floodplains that have their watersheds entirely within the coastal plain. The low terraces are mostly forested, although some cropland or pasture occurs in some areas that are better drained.
Longleaf pine forests once covered many portions of the Carolina coastal plain, along with smaller, scattered areas of mixed pine and hardwood forests. Over the past three centuries, naval stores or pine tar production, logging, open range cattle and feral hog grazing, agriculture, and fire suppression removed almost all of the longleaf pine forests.
Kudzu is an introduced fast-growing vine that was often planted in the 1930's and 1940's to control soil erosion. The vines can grow a foot per day, climbing and covering trees, poles, and buildings. It can kill trees and other native vegetation by blocking sunlight.
66. Blue Ridge
The Blue Ridge extends from southern Pennsylvania to northern Georgia, varying from narrow ridges to hilly plateaus to more massive mountainous areas with high peaks. The mostly forested slopes, high-gradient, cool, clear streams, and rugged terrain occur primarily on metamorphic rocks with minor areas of igneous and sedimentary geology. Annual precipitation of over 100 inches can occur in the wettest areas, while dry basins can average as little as 40 inches. The southern Blue Ridge is one of the richest centers of biodiversity in the eastern U.S. It is one of the most floristically diverse ecoregions, and includes Appalachian oak forests, northern hardwoods, and, at the highest elevations in Tennessee and North Carolina, Southeastern spruce-fir forests. Shrub, grass, and heath balds, hemlock, cove hardwoods, and oak-pine communities are also significant.
66c. The New River Plateau is a high, hilly plateau with less relief and a different land cover mosaic than surrounding Blue Ridge ecoregions. It has less dense woodland and forest cover, and more land devoted to pasture, orchards, cropland, livestock and dairy farms, and Christmas tree production. Elevations are generally between 2500-3500 feet, with a few higher peaks. Oak dominates most of the forests, with beech, birch, hemlock, and poplar on more moist sites and pines on drier areas.
66d. The Southern Crystalline Ridges and Mountains occur primarily on Precambrian-age igneous and high-grade metamorphic rocks, in contrast to the sedimentary and metasedimentary rocks of 66e and 66g. The crystalline rock types are mostly gneiss and schist, covered by well-drained, acidic, loamy soils. Some small areas of mafic and ultramafic rocks also occur, producing more basic soils. The region has greater relief and higher elevations than 66l, 66c, and 66j. Elevations of this rough, dissected region are generally 1200-4500 feet. The southern part of the region is wetter than the north. It is mostly forested, with chestnut oak (and formerly American chestnut) dominating on most slopes and ridges. There are a few small areas of pasture, apple orchards, Fraser fir Christmas tree farms, or minor cropland.
66e. The Southern Sedimentary Ridges in North Carolina consist of small areas near the Tennessee border in western Ashe, Watauga, Mitchell, Yancy, and Madison counties. The disjunct areas contain Cambrian-age sedimentary rocks of shale, sandstone, siltstone, conglomerate, and dolomite. Some metasiltstone or metasandstone occurs, but it is material of very low-grade metamorphism. One of the larger areas, in Madison County, is associated with the Hot Springs Window, an opening where the major thrust sheet was eroded to expose younger, underlying rocks such as the Shady Dolomite and Rome Formation shale and siltstone. Slopes of the region are typically steep and forested, with elevations ranging from 1500-4900 feet.
66g. The Southern Metasedimentary Mountains in North Carolina contain rocks that are not as strongly metamorphosed as the gneisses and schists of 66d. The geologic materials are mostly late Pre-Cambrian and include metagraywacke, metasiltstone, metasandstone, metaconglomerate, slate, schist, phyllite, and quartzite. These are steep, dissected, biologically diverse mountains that are densely forested. The Appalachian oak forests and, at higher elevations, the northern hardwoods forests include a variety of oaks and pines, as well as silverbell, hemlock, yellow poplar, basswood, buckeye, yellow birch, and beech. Much of the region is public land managed by the National Park Service or U.S. Forest Service.
66i. The High Mountains ecoregion includes several disjunct high-elevation areas generally above 4500 feet. The region has a more severe, boreal-like climate than surrounding regions, with wind and ice affecting vegetation, and it has frigid soils rather than [[mesic soi. Evergreen red spruce and Fraser fir forests are found at the higher elevations, and red oak forests and northern hardwood forests with beech, yellow birch, yellow buckeye, and sugar maple are common. The spruce-fir forests have been affected by the balsam wooly adelgid, a non-native insect that kills mature Fraser firs, and some forest growth declines are possibly linked to air pollutants. Heath balds dominated by evergreen rhododendron and mountain laurel, and grassy balds are found on some slopes and ridgetops. Northern flying squirrels, Blackburnian warblers, black-capped chickadees, and common ravens are seen in this region.
66j. The Broad Basins ecoregion is drier, has lower elevations and less relief than the more mountainous Blue Ridge regions (66g, 66d). It also has less bouldery colluvium than those two surrounding regions and more saprolite. The soils are mostly deep, well-drained, loamy to clayey Ultisols, although there are variations between the uplands, the high and low terraces, and the floodplains. The Asheville basin has the lowest annual precipitation amounts in North Carolina, receiving less than 42 inches. Compared to the higher mountainous ecoregions of 66, the Broad Basins have a mix of oaks and pines more similar to the Piedmont (45), with more shortleaf and Virginia pine, and white, southern red, black, and scarlet oaks. Although some areas of this rolling foothills region are mostly forested, overall it has more pasture, cropland, industrial land uses, and human settlement than other Blue Ridge ecoregions. Outlines of abandoned fields with pine-hardwood succession are apparent on many lower slopes.
66k. Similar to some parts of 66d, the Amphibolite Mountains are a botanically diverse area with many rare species, including some relict and disjunct species from areas much further north. The rugged, steeply sloping mountains are composed of Precambrian amphibolite and gneiss. The amphibolite, a metamorphosed black volcanic rock, formed from lavas that spilled on the floor of a shallow sea, mixing with layers of mud, sand, and volcanic ash. In some areas this rock weathers to produce shallow soils high in calcium and magnesium, and less acidic than most Appalachian soils. Oak forests (formerly American chestnut forests) dominate on south, east, and west facing slopes with an understory of Catawba rhododendron, mountain laurel, flame azalea, and dogwood. Cove forests and northern hardwood forests are found on north slopes, and include sugar maple, ash, yellow birch, tulip tree, and basswood.
66l. The open low mountains of the Eastern Blue Ridge Foothills are lower in elevation (1000-2800 feet) than most Blue Ridge regions and have more Piedmont influences. The region includes the Brushy Mountains to the north and the South Mountains to the south. Covered with mixed oak and oak-hickory-pine forests, these mountains tend to be slightly drier and warmer than most of Ecoregion 66. The South Mountains contain forested areas that harbor many uncommon or rare plant species, including turkey beard (Xerophyllum asphodeloides) on xeric ridges and one of North America’s rarest orchids, the small whorled pogonia (Isotria medeoloides).
66m. The prominent ridges and knobs of the Sauratown Mountains rise more than 1000 feet above the rolling Piedmont (45) surface. Sometimes called monadnocks or inselbergs, these isolated mountain outliers are formed in part by their caps of erosion-resistant quartzite. The region has both Piedmont and Blue Ridge vegetation communities: mostly oak and oak-pine forests with some Canadian and Carolina hemlock in moist areas. Other mountain flora found here include rhododendron, azalea, galax, mountain laurel, pitch pine, table mountain pine, and various ferns.
The Blue Ridge is part of one of the richest temperate broadleaf forests in the world, with a high diversity of flora and fauna. Black bear, deer, wild boar, turkey, grouse, songbirds, reptiles, many species of amphibians, thousands of species of invertebrates, and a variety of small mammals are found here. (Photo: SCDNR)
The saw-whet owl (Aegolius acadicus), a species of special concern in North Carolina, often roosts during the day in red spruce trees of 66i and nests in cavity trees such as yellow birch or in nest boxes set out for flying squirrels. (Photo: John and Karen Hollingsworth)
The high-elevation spruce-fir forests of 66i are buffeted by higher winds, colder temperatures, and greater precipitation than lower elevations. Growth declines, insect damage, and air pollution are continuing concerns.
Black bears are found in many parts of the Blue Ridge, but tend to be reclusive by nature. (Photo: John Lucas, SCDNR)
Catawba rhododendron grow at various elevations in the Blue Ridge, often densely on balds at higher elevations. (Photo: Joe and Monica Cook)
The red squirrel (Tamiasciurus hudsonicus) is one of the smaller but louder squirrels of the Blue Ridge. It collects nuts, seeds, birds' eggs, and mushrooms from forests generally higher in elevation than occupied by the gray squirrel.
75. Southern Coastal Plain
The Southern Coastal Plain extends from South Carolina and Georgia, through much of central Florida, and along the Gulf coast lowlands of the Florida Panhandle, Alabama, and Mississippi. From a national perspective, it appears to be mostly flat plains, but it is a heterogeneous region also containing barrier islands, coastal lagoons, marshes, and swampy lowlands along the Gulf and Atlantic coasts. In Florida, an area of discontinuous highlands contains numerous lakes. This ecoregion is generally lower in elevation with less relief and wetter soils than Ecoregion 65. It is warmer and has a different mix of vegetation than Ecoregion 63. Once covered by a variety of forest communities that included trees of longleaf pine, slash pine, pond pine, beech, sweetgum, southern magnolia, white oak, and laurel oak, land cover in the ecoregion as a whole is now mostly slash and loblolly pine with oak-gum-cypress forest in some low lying areas, citrus groves in Florida, pasture for beef cattle, and urban.
75i. Floodplains and Low Terraces are a continuation of the riverine 65p ecoregion across the Southern Coastal Plain. Similar to 63n, the broad floodplains and terraces of major rivers, such as the Savannah in South Carolina, comprise the region. Composed of stream alluvium and terrace deposits of sand, silt, clay, and gravel, along with some organic muck and swamp deposits, the region includes large sluggish rivers and backwaters with ponds, swamps, and oxbow lakes. River swamp forests of bald cypress and water tupelo and oak-dominated bottomland hardwood forests provide important wildlife habitat.
75j. The Sea Islands/Coastal Marsh region contains the lowest elevations in South Carolina and is a highly dynamic environment affected by ocean wave, wind, and river action. Mostly sandy soils are found on the barrier islands, while organic and clayey soils often occur in the freshwater, brackish, and salt marshes. Maritime forests of live oak, red cedar, slash pine, and cabbage palmetto grow on parts of the sea islands, and various species of cordgrass, saltgrass, and rushes are dominant in the marshes. The coastal marshes are important nursery areas for fish, crabs, shrimp, and other marine species. During the colonial and antebellum periods in the 1700's and 1800's, a plantation agriculture economy dominated the region, producing rice, indigo, and Sea Island cotton.
Tolerant of salt spray, dry sandy soils, or saturated conditions, cabbage palms (Sabal palmetto) are uniquely adapted to the coastal environment. Flexible trunks and shredded, divided leaves bend but tend not to break in hurricanes. (Photo: Allen Sharpe, SCETV)
Notes
• The full, original version of this entry is located here: http://www.epa.gov/wed/pages/ecoregions/ncsc_eco.htm. That description contains additional maps, as well as information on the physiography, geology, soil, potential natural vegetation, and the land use and land cover of the ecoregion.
• PRINCIPAL AUTHORS: Glenn E. Griffith (NRCS), James M. Omernik (USEPA), Jeffrey A. Comstock (Indus Corporation), Michael P. Schafale (NCDENR), W. Henry McNab (USFS), David R. Lenat (NCDENR), Trish F. MacPherson (NCDENR), James B. Glover (SCDHEC), and Victor B. Shelburne (Clemson University).
• COLLABORATORS AND CONTRIBUTORS: James E. Harrison (USEPA), David L. Penrose (NCDENR), Ronald C. Ahle (South Carolina Department of Natural Resources [SCDNR]), Roy L.Vick, Jr. (NRCS), Ben Stuckey, Jr. (NRCS), Dennis L. Law (USFS), Robert K. Peet (University of North Carolina), Richard T. Renfrow (SCDHEC), Paul G. Nystrom (SCDNR), Richard L. Scharf (SCDNR), Chip Smith (NRCS), Alan J. Woods (Dynamac Corporation), and Thomas R. Loveland (USGS).
• REVIEWERS: Stanley W. Buol (North Carolina State University), Berman D. Hudson (NRCS), Charles F. Kovacik (University of South Carolina), Rudy E. Mancke (University of South Carolina), and Gerard McMahon (USGS).
• CITING THIS POSTER: Griffith, G.E., Omernik, J.M., Comstock, J.A., Schafale, M.P., McNab, W.H., Lenat, D.R., MacPherson, T.F., Glover, J.B., and Shelburne, V.B., 2002, Ecoregions of North Carolina and South Carolina, (color poster with map, descriptive text, summary tables, and photographs): Reston, Virginia, U.S. Geological Survey (map scale 1:1,500,000).
• This project was partially supported by funds from the North Carolina Department of Environment and Natural Resources and the South Carolina Department of Health and Environmental Control through grants provided by the U.S. Environmental Protection Agency Region IV under the provisions of Sections 104(b) and 319(h) of the Federal Water Pollution Control Act.
Literature Cited
• Bailey, R.G., Avers, P.E., King, T., and McNab, W.H., eds., 1994, Ecoregions and subregions of the United States (map) (supplementary table of map unit descriptions compiled and edited by McNab, W.H. and Bailey, R.G.): Washington, D.C., U.S. Department of Agriculture-Forest Service, scale 1:7,500,000.
• Bryce, S.A., Omernik, J.M., and Larsen, D.P., 1999, Ecoregions - a geographic framework to guide risk characterization and ecosystem management: Environmental Practice, v. 1, no. 3, p. 141-155.
• Commission for Environmental Cooperation Working Group, 1997, Ecological regions of North America - toward a common perspective: Montreal, Quebec, Commission for Environmental Cooperation, 71 p.
• Gallant, A.L., Whittier, T.R., Larsen, D.P., Omernik, J.M., and Hughes, R.M., 1989, Regionalization as a tool for managing environmental resources: Corvallis, Oregon, U.S. Environmental Protection Agency, EPA/600/3-89/060, 152 p.
• McMahon, G., Gregonis, S.M., Waltman, S.W., Omernik, J.M., Thorson, T.D., Freeouf, J.A., Rorick, A.H., and Keys, J.E., 2001, Developing a spatial framework of common ecological regions for the conterminous United States: Environmental Management, v. 28, no. 3, p. 293-316.
• Omernik, J.M., 1987, Ecoregions of the conterminous United States (map supplement): Annals of the Association of American Geographers, v. 77, no. 1, p. 118-125, scale 1:7,500,000.
• Omernik, J.M., 1995, Ecoregions - a spatial framework for environmental management, in Davis, W.S., and Simon, T.P., eds., Biological assessment and criteria-tools for water resource planning and decision making: Boca Raton, Florida, Lewis Publishers, p. 49-62. ISBN: 0873718941.
• Omernik, J.M., Chapman, S.S., Lillie, R.A., and Dumke, R.T., 2000, Ecoregions of Wisconsin: Transactions of the Wisconsin Academy of Sciences, Arts and Letters, v. 88, no. 2000, p. 77-103.
• U.S. Department of Agriculture-Soil Conservation Service, 1981, Land resource regions and major land resource areas of the United States: Agriculture Handbook 296, 156 p.
• U.S. Environmental Protection Agency, 2002, Level III ecoregions of the continental United States (revision of Omernik, 1987): Corvallis, Oregon, U.S. Environmental Protection Agency-National Health and Environmental Effects Research Laboratory, Map M-1, various scales.
• Wiken, E., 1986, Terrestrial ecozones of Canada: Ottawa, Environment Canada, Ecological Land Classification Series no. 19, 26 p. ISBN: 0662147618.
Disclaimer: This article is taken wholly from, or contains information that was originally published by, the Environmental Protection Agency). Topic editors and authors for the Encyclopedia of Earth may have edited its content or added new information. The use of information from the Environmental Protection Agency) should not be construed as support for or endorsement by that organization for any new information added by EoE personnel, or for any editing of the original content.
Citation
Glenn E. Griffith, James M. Omernik (Lead Author);Mark McGinley (Topic Editor) "Ecoregions of North Carolina and South Carolina (EPA)". In: Encyclopedia of Earth. Eds. Cutler J. Cleveland (Washington, D.C.: Environmental Information Coalition, National Council for Science and the Environment). [First published in the Encyclopedia of Earth July 6, 2008; Last revised Date July 6, 2008; Retrieved May 18, 2013 <http://www.eoearth.org/article/Ecoregions_of_North_Carolina_and_South_Carolina_(EPA)>
The Authors
A biography for this person is not yet available. We encourage authors to submit biographical information, if you have not done so and would like to, contact Arielle Conti. ... (Full Bio)
James M. Omernik is a retired geographer formerly with the U.S. Environmental Protection Agency's National Health and Environmental Effects Research Laboratory is Corvallis, OR. He currently works parttime for the U.S. Geological Survey. Omernik began his career in geography with the Defense Intelligence Agency in the early 1960's and was with the EPA from 1972 to 2002. At EPA he has led in the design and development of national and regional maps of stream nutrient concentrations attri ... (Full Bio)
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New Hampshire Births and Christenings (FamilySearch Historical Records)Edit This Page
From FamilySearch Wiki
Revision as of 13:49, 2 May 2011 by TimothyNB (Talk | contribs)
This article describes a collection of historical records available at FamilySearch.org.
Contents
Collection Time Period
This index covers events from 1714 through 1904.
Record Description
This index is an electronic database of information compiled from a variety of sources including the following:
• Family Records
• Church Records
• Civil Registration
This index is not complete for any particular place or region.
Record Content
The key genealogical facts found in birth index entries may include:
• Name of the child
• Gender
• Names of the parents
• Birth date
• Birthplace
• Christening date (if the source is a church record)
• Family History Library Microfilm and item numbers for the source materials
Coverage Table
This collection is a partial index of records for the localities listed below as of April 2010. The table below shows the number of records by locality. Localities not listed may not have any records in this collection.
Most of these records date from the time period indicated in the columns below; however, there may be records before and after these dates. Record counts where the locality information was incomplete are included below under the country name. Because of this, you may wish to search only by state.
As this is an index of records compiled from various sources, it is strongly recommended that you verify the information in original records. Due to privacy laws not all records may be displayed.
Locality Births and Christenings, 1733-1900 Marriages, 1783-1920 Deaths and Burials, 1784-1949
Belknap 0 11,711 3,572
Carroll 6,542 11,324 2,142
Cheshire 886 24,286 7,504
Coos 0 8,074 4,584
Grafton 1,729 18,149 4,926
Hillsborough 1,553 113,964 61,721
Merrimack 1,823 27,478 15,405
New Hampshire 379,246 336,160 112,685
Rockingham 12,365 57,836 40,075
Strafford 0 22,727 5,315
Sullivan 315 13,337 4,731
Total 404,459 645,046 262,660
How to Use the Record
Use this index to help you learn more about your ancestors. The information could help you identify family relationships and lineages as well as direct you to original records of your ancestors, which may contain additional information.
In birth or christening records, if a surname is not listed for the child, the indexer often assigns the father’s surname to the child. This surname may not be correct. So if you are looking for a birth or christening, search by the given name of the child, adding parents' names and as much locality information as is permitted.
Finding the Original Source for an Entry in This Collection
Each entry in this index has a source listed which includes a batch number. You will need to trace the batch number for the individual entry to learn its source. Please see the following wiki articles for more information on batch numbers:
If an FHL film number is given in the entry for your ancestor, search for it in the Family History Library Catalog.
Record History
For over 30 years, volunteer indexers extracted this information from microfilm copies of the original records. In 1998, a few of the entries were published on 7 CDs by the Family History Department of The Church of Jesus Christ of Latter-day Saints as the "North America Vital Records Index". This is an index of some births and christenings throughout New Hampshire. The index is not necessarily complete for any particular place or region.
Why This Record Was Created
The Church of Jesus Christ of Latter-day Saints sponsored this index to help individuals find information about their ancestors.
Record Reliability
Church records and civil registration were official records and are some of the most reliable sources of information available for those who were born, married, or died in New Hampshire.
Related Web Sites
New Hampshire Birth Record Search
New Hampshire Genealogy and History
Related Wiki Articles
New Hampshire Vital Records
Contributions to This Article
We welcome user additions to FamilySearch Historical Records wiki articles. Guidelines are available to help you make changes. Thank you for any contributions you may provide. If you would like to get more involved join the WikiProject FamilySearch Records.
Citing FamilySearch Historical Collections
When you copy information from a record, you should also list where you found the information. This will help you or others to find the record again. It is also good to keep track of records where you did not find information, including the names of the people you looked for in the records.
A suggested format for keeping track of records that you have searched is found in the Wiki Article: How to Cite FamilySearch Collections.
Examples of Source Citations for a Record in This Collection
"New Hampshire Births and Christenings, 1714-1904." index, FamilySearch (https://www.familysearch.org): accessed 31 March 2011. entry for Martha Cole, 12 July 1899; citing Birth Records, FHL microfilm 2,156,071; Index entries derived from digital copies of original and compiled records.
Sources of Information for This Collection
"New Hampshire Births and Christenings, 1714-1904," index, FamilySearch (https://www.familysearch.org). Index entries derived from digital copies of original and compiled records.
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