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PMID:21473
|
Partial characterization of epididymal 5 alpha reductase in the rat.
|
Epididymal 5alpha reductase activity was found distitributed in the crude nuclear fraction (44 percent) and microsomal fraction (41 percent). Spermatozoa contaminating the nuclear preparation accounted for only 3 percent of its activity. There were no regional differences in the distribution of total 5alpha reductase activity. However, the nuclear enzyme was more active in caput than in other regions. Maximal activity was found at pH 6.2 and at 32 degrees C. Both enzymes had an absolute requirement of reduced dinucleotides. The microsomal preparation could only us NADPH while the nuclear enzyme could use NADPH and NADH. The apparent Km for the microsomal preparation was 0.62 +/- 0.05 X 10(-6)M and Vmax was 555 +/- 38 pmoles/mg protein/hour. The nuclear enzyme presented similar values. The reaction was not inhibited by accumulation of product in the medium, but other steroids such as progesterone, epitestosterone (17alpha-hydroxy-4-androsten-3-one) and 3-oxo-4-androstene-17beta-carboxylic acid were potent competitive inhibitors. The reaction was strongly inhibited by Hg, Zn and Cu. The properties of the epididymal reductase are similar to those of the prostatic enzyme.
|
Partial characterization of epididymal 5 alpha reductase in the rat. Epididymal 5alpha reductase activity was found distitributed in the crude nuclear fraction (44 percent) and microsomal fraction (41 percent). Spermatozoa contaminating the nuclear preparation accounted for only 3 percent of its activity. There were no regional differences in the distribution of total 5alpha reductase activity. However, the nuclear enzyme was more active in caput than in other regions. Maximal activity was found at pH 6.2 and at 32 degrees C. Both enzymes had an absolute requirement of reduced dinucleotides. The microsomal preparation could only us NADPH while the nuclear enzyme could use NADPH and NADH. The apparent Km for the microsomal preparation was 0.62 +/- 0.05 X 10(-6)M and Vmax was 555 +/- 38 pmoles/mg protein/hour. The nuclear enzyme presented similar values. The reaction was not inhibited by accumulation of product in the medium, but other steroids such as progesterone, epitestosterone (17alpha-hydroxy-4-androsten-3-one) and 3-oxo-4-androstene-17beta-carboxylic acid were potent competitive inhibitors. The reaction was strongly inhibited by Hg, Zn and Cu. The properties of the epididymal reductase are similar to those of the prostatic enzyme.
|
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] |
PMID:21474
|
Oxidative metabolism and de-ethynylation of 17alpha-ethynylestradiol by baboon liver microsomes.
|
Incubations of tritiated 17alpha-ethynylestradiol (EE2) with liver explants of baboon and mouse showed the primate species to be more efficient in the removal of the ethynyl group. Liver microsomes from sexually immature male and female baboons were then incubated with tritiated EE2 and estradiol (E2). Each hormone bound irreversibly to the microsomal pellet. Addition of glutathione reduced the irreversible or covalent association. Incubations with E2 demonstrated significant conversion to estrone (E1). The EE2 experiments demonstrated a conversion to estrone only in the presence of an NADPH-generating system, and the addition of SKF-525A reduced the conversion of EE2 to E1. The cleavage reaction appears to be an oxidative event.
|
Oxidative metabolism and de-ethynylation of 17alpha-ethynylestradiol by baboon liver microsomes. Incubations of tritiated 17alpha-ethynylestradiol (EE2) with liver explants of baboon and mouse showed the primate species to be more efficient in the removal of the ethynyl group. Liver microsomes from sexually immature male and female baboons were then incubated with tritiated EE2 and estradiol (E2). Each hormone bound irreversibly to the microsomal pellet. Addition of glutathione reduced the irreversible or covalent association. Incubations with E2 demonstrated significant conversion to estrone (E1). The EE2 experiments demonstrated a conversion to estrone only in the presence of an NADPH-generating system, and the addition of SKF-525A reduced the conversion of EE2 to E1. The cleavage reaction appears to be an oxidative event.
|
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] |
PMID:21478
|
[Triggering and regulatory mechanisms of ciliary motion in the ctenophore Bolinopsis. III. Electric excitability and inherent contractility of ciliated cells].
|
Rhythmical electrostimulation is able to produce in a tissue strip of Bolinopsis series of beats of combs-plates. However, the beat frequency, within one series, is only poorly controlled by electrical stimuli. Isolated comb-plate cells with a cilium cut off maintain rhythmical contractions stimulated both mechanically and electrically. A lot of microtubules has been found in the apical region of these cells. It is supposed that it is these microtubules, probably together with the associated root filaments, that are responsible for cell motility. Mutual mechanical stimulation of ciliary cells in these motions is, presumably, a signal providing intratissue propagation of metachronal wave.
|
[Triggering and regulatory mechanisms of ciliary motion in the ctenophore Bolinopsis. III. Electric excitability and inherent contractility of ciliated cells]. Rhythmical electrostimulation is able to produce in a tissue strip of Bolinopsis series of beats of combs-plates. However, the beat frequency, within one series, is only poorly controlled by electrical stimuli. Isolated comb-plate cells with a cilium cut off maintain rhythmical contractions stimulated both mechanically and electrically. A lot of microtubules has been found in the apical region of these cells. It is supposed that it is these microtubules, probably together with the associated root filaments, that are responsible for cell motility. Mutual mechanical stimulation of ciliary cells in these motions is, presumably, a signal providing intratissue propagation of metachronal wave.
|
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] |
PMID:21481
|
Splenic-gonadal fusion and adrenal cortical rest associated with bilateral cryptorchism.
|
A patient with bilateral cryptorchism had ectopic splenic tissue adherent to his left testicle and an adrenal cortical rest adherent to his right testicle. The embrylogic basis for these disorders is discussed and the literature reviewed.
|
Splenic-gonadal fusion and adrenal cortical rest associated with bilateral cryptorchism. A patient with bilateral cryptorchism had ectopic splenic tissue adherent to his left testicle and an adrenal cortical rest adherent to his right testicle. The embrylogic basis for these disorders is discussed and the literature reviewed.
|
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] |
PMID:21482
|
Uropharmacology: part VI. Parasympathetic depressants.
|
A discussion of the various parasympathetic depressants is presented. Belladonna alkaloids include atropine, the prototype, and scopolamine. Synthetic drugs include quaternary ammonium compounds with antimuscarinic activity such as methantheline, propantheline, and other drugs such as isopropamide, pipenzolate methylbromide, and diphemanil methylsulfate. A miscellaneous class of drugs such as hemicholinium, valethamate, and oxybutynin chloride also possesses parasympathetic depressant activity. The pharmacologic properties and clinical usage of these drugs are discussed.
|
Uropharmacology: part VI. Parasympathetic depressants. A discussion of the various parasympathetic depressants is presented. Belladonna alkaloids include atropine, the prototype, and scopolamine. Synthetic drugs include quaternary ammonium compounds with antimuscarinic activity such as methantheline, propantheline, and other drugs such as isopropamide, pipenzolate methylbromide, and diphemanil methylsulfate. A miscellaneous class of drugs such as hemicholinium, valethamate, and oxybutynin chloride also possesses parasympathetic depressant activity. The pharmacologic properties and clinical usage of these drugs are discussed.
|
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] |
PMID:21483
|
[Method of stabilizing ketamine anesthesia].
|
The authors eliminated the disavantages of intravenous ketamine anesthesia, interfering with its wide use (hypertension, tachicardia, hypersalivation, diplopia etc.) by means of preliminary ganglionic blockade. Clinical studies of ketamine anesthesis with ganglionic blockade and without the latter illustrated the advantages of the suggested method.
|
[Method of stabilizing ketamine anesthesia]. The authors eliminated the disavantages of intravenous ketamine anesthesia, interfering with its wide use (hypertension, tachicardia, hypersalivation, diplopia etc.) by means of preliminary ganglionic blockade. Clinical studies of ketamine anesthesis with ganglionic blockade and without the latter illustrated the advantages of the suggested method.
|
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] |
PMID:21480
|
[Effect of carbonic acid concentration in blood on content of keto-acids and redox state of nicotinamide coenzymes in rabbit tissues].
|
A rise in the level of carbonic acid in animal blood at practically unchanged pH causes an increase in the value of free [NAD+]/[NADH] and [NADP+]/[NADPH] ratios in the liver cells cytoplasm. The increase in the oxidizing power of the liver cells cytoplasm is observed against a background of a rise in both the total level of keto-acids in blood and concentration of pyruvate oxaloacetate and alpha-ketoglutarate in the liver and kidneys of animals. The level of lactate, glutamate and citrate in these tissues also grows. The level of ketonic bodies in blood of the animals with a higher concentration of carbonic acid is somewhat lower than that in the animals with a lower concentration of HCO3 and CO2 in blood.
|
[Effect of carbonic acid concentration in blood on content of keto-acids and redox state of nicotinamide coenzymes in rabbit tissues]. A rise in the level of carbonic acid in animal blood at practically unchanged pH causes an increase in the value of free [NAD+]/[NADH] and [NADP+]/[NADPH] ratios in the liver cells cytoplasm. The increase in the oxidizing power of the liver cells cytoplasm is observed against a background of a rise in both the total level of keto-acids in blood and concentration of pyruvate oxaloacetate and alpha-ketoglutarate in the liver and kidneys of animals. The level of lactate, glutamate and citrate in these tissues also grows. The level of ketonic bodies in blood of the animals with a higher concentration of carbonic acid is somewhat lower than that in the animals with a lower concentration of HCO3 and CO2 in blood.
|
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] |
PMID:21486
|
Acid elution of blood group antibodies from intact erythrocytes.
|
Human IgM and IgG antibodies against blood group antigens (A, B, D, C, c, E, e, Fya, K), autoantibodies and mouse IgM and IgG antibodies against sheep erythrocytes have been eluted from intact human and sheep red cells by glycine-HCl buffer, pH 3.0. The yield of human antibodies was higher with acid than with heat and ether elution, and the contamination of hemoglobin in the eluate was negligible. The acid elution method is very simple and rapid and, therefore, highly suitable for experimental as well as routine immunohematological work.
|
Acid elution of blood group antibodies from intact erythrocytes. Human IgM and IgG antibodies against blood group antigens (A, B, D, C, c, E, e, Fya, K), autoantibodies and mouse IgM and IgG antibodies against sheep erythrocytes have been eluted from intact human and sheep red cells by glycine-HCl buffer, pH 3.0. The yield of human antibodies was higher with acid than with heat and ether elution, and the contamination of hemoglobin in the eluate was negligible. The acid elution method is very simple and rapid and, therefore, highly suitable for experimental as well as routine immunohematological work.
|
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] |
PMID:21484
|
[Relationship between glycolysis and pentose cycle intensity and thrombocyte aggregation induced by ADP in man].
|
Aggregation of thrombocytes, caused by ADP, affected the rate of anaerobic breakdown of carbohydrates due to inhibition of the glucose degradation, while the rate of lactate formation was not distinctly altered. Content of NAD was decreased simultaneously with an increase in content of NADP; the activity of glucose-6-phosphate dehydrogenase was distinctly increased but the unaltered activity of 6-phosphogluconate dehydrogenase was observed; content of reduced glutathione was increased 15-fold. The activity of NADP-dependent glutathione reductase was increased under effect of ADP; at the same time, the activity of NAD-dependent glutathione reductase did not depend on the diphosphate effect.
|
[Relationship between glycolysis and pentose cycle intensity and thrombocyte aggregation induced by ADP in man]. Aggregation of thrombocytes, caused by ADP, affected the rate of anaerobic breakdown of carbohydrates due to inhibition of the glucose degradation, while the rate of lactate formation was not distinctly altered. Content of NAD was decreased simultaneously with an increase in content of NADP; the activity of glucose-6-phosphate dehydrogenase was distinctly increased but the unaltered activity of 6-phosphogluconate dehydrogenase was observed; content of reduced glutathione was increased 15-fold. The activity of NADP-dependent glutathione reductase was increased under effect of ADP; at the same time, the activity of NAD-dependent glutathione reductase did not depend on the diphosphate effect.
|
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] |
PMID:21489
|
[Pathophysiology of lipolysis in human adipose tissue (author's transl)].
|
Lipolysis in human adipose tissue was measured as glycerol release in isolated fat cells and in adipose tissue homogenates. In isolated fat cells lipolysis proceeded optimally at pH 7.4, was stimulated 3.5 fold by noradrenaline and was not influenced by serum or protamine. In adipose tissue homogenates lipolysis was stimulated 4 fold by serum. Serum-stimulated lipolytic activity was optimal at pH 8.0, was inhibited by 1 M sodium chloride and protamine and was not influenced by noradrenaline. Lipolytic activity in isolated fat cells is ascribed on the basis of these observations mainly to the action of hormone-sensitive lipase. whereas lipolysis in adipose tissue homogenates in the presence of serum seems to be regulated by lipoprotein lipase. Thus, the lipolytic processes involved in the mobilization of triglycerides from adipose tissue and in the uptake or triglycerides into adipose tissue can be assessed separately, using the two described methods. The re-esterification of FFA, the second pathway in the mobilization of triglycerides, has also been investigated.
|
[Pathophysiology of lipolysis in human adipose tissue (author's transl)]. Lipolysis in human adipose tissue was measured as glycerol release in isolated fat cells and in adipose tissue homogenates. In isolated fat cells lipolysis proceeded optimally at pH 7.4, was stimulated 3.5 fold by noradrenaline and was not influenced by serum or protamine. In adipose tissue homogenates lipolysis was stimulated 4 fold by serum. Serum-stimulated lipolytic activity was optimal at pH 8.0, was inhibited by 1 M sodium chloride and protamine and was not influenced by noradrenaline. Lipolytic activity in isolated fat cells is ascribed on the basis of these observations mainly to the action of hormone-sensitive lipase. whereas lipolysis in adipose tissue homogenates in the presence of serum seems to be regulated by lipoprotein lipase. Thus, the lipolytic processes involved in the mobilization of triglycerides from adipose tissue and in the uptake or triglycerides into adipose tissue can be assessed separately, using the two described methods. The re-esterification of FFA, the second pathway in the mobilization of triglycerides, has also been investigated.
|
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] |
PMID:21485
|
[Specificity of glutamine metabolism in pre-tumorous diseases and cancer of the human stomach].
|
Content of free glutamine and the activity of glutamine synthetase and glutamine transaminase were studied in practically healthy persons and in patients with chronic atrophic gastritis, ulcerous disease, polyposis and with gastric carcinoma. The enzymatic activity was estimated in the areas of ulcerous impairment, of polypous vegetation of malignant neoplasm as well as in mucous membrane out of the impaired zone and in whole blood of all the patients studied. The tissues for biochemical tests were obtained by the directed gastrobiopsy. Content of glutamine was decreased in blood of patients with gastric carcinoma and increased in mucous membrane adjacent to the malignant tissue. Occurrence of the glutamine transaminase activity in the tissue areas studied was due to the specific glutamine metabolism during pretumoral diseases and gastric carcinoma, whereas the unimpaired gastric mucosa did not have the distinct enzymatic activity.
|
[Specificity of glutamine metabolism in pre-tumorous diseases and cancer of the human stomach]. Content of free glutamine and the activity of glutamine synthetase and glutamine transaminase were studied in practically healthy persons and in patients with chronic atrophic gastritis, ulcerous disease, polyposis and with gastric carcinoma. The enzymatic activity was estimated in the areas of ulcerous impairment, of polypous vegetation of malignant neoplasm as well as in mucous membrane out of the impaired zone and in whole blood of all the patients studied. The tissues for biochemical tests were obtained by the directed gastrobiopsy. Content of glutamine was decreased in blood of patients with gastric carcinoma and increased in mucous membrane adjacent to the malignant tissue. Occurrence of the glutamine transaminase activity in the tissue areas studied was due to the specific glutamine metabolism during pretumoral diseases and gastric carcinoma, whereas the unimpaired gastric mucosa did not have the distinct enzymatic activity.
|
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] |
PMID:21494
|
[On characterization of lipoxygenase from barley (author's transl)].
|
Lipoxygenase from brewing barley of Nordbaden has optimum activity at alkaline pH and predominantly forms 9-hydroperoxy-10-trans, 12-cis-octadecadienoic acid (9-LHPO) from linoleic acid. While at pH 7 90% 9-LHPO is produced, its proportions drops during incubation at more alkaline pH (pH 7,75) to 70%. This positional specifity is not caused by isoenzymes. While linoleic acid methylester is converted only to a lesser degree, trilinolein is not accepted as substrate.
|
[On characterization of lipoxygenase from barley (author's transl)]. Lipoxygenase from brewing barley of Nordbaden has optimum activity at alkaline pH and predominantly forms 9-hydroperoxy-10-trans, 12-cis-octadecadienoic acid (9-LHPO) from linoleic acid. While at pH 7 90% 9-LHPO is produced, its proportions drops during incubation at more alkaline pH (pH 7,75) to 70%. This positional specifity is not caused by isoenzymes. While linoleic acid methylester is converted only to a lesser degree, trilinolein is not accepted as substrate.
|
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] |
PMID:21495
|
[The idiopathic respiratory distress syndrome from the obstetrical point of view].
|
Frequency of idiopathic respiratory distress syndrome (IRDS) of low birth weight children was examined over 15 years (1960 to 1975) in the Frauenklinik of the Ernst-Moritz-Arndt-University of Greifswald. Possible pathogenetic risk factors were proofed. The validity of the Apgar-score and blood gas analyses for the development of IRDS was investigated. Finally, the coincidence between mode of delivery, early rupture of membrane and imminent intrauterine infection in relation to the frequency of IRDS was shown.
|
[The idiopathic respiratory distress syndrome from the obstetrical point of view]. Frequency of idiopathic respiratory distress syndrome (IRDS) of low birth weight children was examined over 15 years (1960 to 1975) in the Frauenklinik of the Ernst-Moritz-Arndt-University of Greifswald. Possible pathogenetic risk factors were proofed. The validity of the Apgar-score and blood gas analyses for the development of IRDS was investigated. Finally, the coincidence between mode of delivery, early rupture of membrane and imminent intrauterine infection in relation to the frequency of IRDS was shown.
|
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] |
PMID:21496
|
[Induction of labor with prostaglandin F 2 alpha (a comparative study using oxytocin and prostaglandin)].
|
The authors investigated the effectiveness of prostaglandin F2alpha administered by intravenous drop infusion to induce labour. They compared their results with those obtained in oxytocin-induced labour. They determined that prostaglandin F2alpha can without any danger be administered in the induction of labour. The process of labour and the ensuing results indicate that prostaglandin is less of a burden to the uteroplacental circulation. In administering either of the two drugs, the authors deem the principle of applying the "smallest" dose important.
|
[Induction of labor with prostaglandin F 2 alpha (a comparative study using oxytocin and prostaglandin)]. The authors investigated the effectiveness of prostaglandin F2alpha administered by intravenous drop infusion to induce labour. They compared their results with those obtained in oxytocin-induced labour. They determined that prostaglandin F2alpha can without any danger be administered in the induction of labour. The process of labour and the ensuing results indicate that prostaglandin is less of a burden to the uteroplacental circulation. In administering either of the two drugs, the authors deem the principle of applying the "smallest" dose important.
|
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] |
PMID:21497
|
[Behavior of various serum and urinary peptidases in pregnancy].
|
The investigations were carried out on activities of leucinearylamidase (LAP) and (GGTP) gammaglutamyltranspeptidase in serum and urine of women with physiological and prolonged pregnancy. The results shown on pathological increase of LAP-activity in serum and increase excretion of both enzymes in to the urine during prolonged pregnancy. These findings may appear very important for the explain of pathomechanism of prolonged pregnancy and diagnostic purpose.
|
[Behavior of various serum and urinary peptidases in pregnancy]. The investigations were carried out on activities of leucinearylamidase (LAP) and (GGTP) gammaglutamyltranspeptidase in serum and urine of women with physiological and prolonged pregnancy. The results shown on pathological increase of LAP-activity in serum and increase excretion of both enzymes in to the urine during prolonged pregnancy. These findings may appear very important for the explain of pathomechanism of prolonged pregnancy and diagnostic purpose.
|
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] |
PMID:21499
|
[Relationship between the incidence of neurologic complications following smallpox vaccination and the generation and infectious activity of vaccine from strain L-IVP].
|
A strictly controlled epidemiological trial was applied to the study of the incidence of neurological complications in children inoculated with vaccine from the L-IVP strain of the 2nd, 3rd, and 5th passages (generations). To ascertain a possible connection between the neuropathogenicity with the infectious activity the persons vaccinated were divided into 3 groups in accordance with the infectiousness level of the preparation obtained: the I group--3--5-10(8)OOU/ml; the II group--6--8-9-10(8) OOU/ml; the III group over 9-10(8) OOU/ml. The least number of neurological complications was recorded among children primarily inoculated with the vaccine of the 3rd passage with the infectiousness titre of over 9-10(8) OOU/ml.
|
[Relationship between the incidence of neurologic complications following smallpox vaccination and the generation and infectious activity of vaccine from strain L-IVP]. A strictly controlled epidemiological trial was applied to the study of the incidence of neurological complications in children inoculated with vaccine from the L-IVP strain of the 2nd, 3rd, and 5th passages (generations). To ascertain a possible connection between the neuropathogenicity with the infectious activity the persons vaccinated were divided into 3 groups in accordance with the infectiousness level of the preparation obtained: the I group--3--5-10(8)OOU/ml; the II group--6--8-9-10(8) OOU/ml; the III group over 9-10(8) OOU/ml. The least number of neurological complications was recorded among children primarily inoculated with the vaccine of the 3rd passage with the infectiousness titre of over 9-10(8) OOU/ml.
|
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] |
PMID:21500
|
[Role of autoimmune processes in the pathogenesis of post-vaccinal lesions of the nervous system].
|
Subcutaneous sensitization of guinea pigs with -vaccine, and also an intracardiac injection of smallpox or measles vaccine induced production of brain autoantibodies, whereas subcutaneous or intradermal immunization of the animals with liver viral vaccines was not accompanied by the formation of autoantibodies and development of the pathological processes in the nervous system tissue. tthe greatest changes in the brain tissue in the form of circulatory disturbances and inflammatory-dystrophic phenomena in combination with high autoantibody titres in the blood were noted in intracardiac injection of smallpox vaccine and in subturbancessensitization with AK-Vaccine and the least (short-term circulatory disturbances, transitory signs of serous meningitis and neuron dystrophy against the background of low blood auto anitbody content)--in intracardiac injection of measles vaccine. Administration of live viral vaccines into the circulation of animals against the background of their sensitization with -vaccine led to reduction of blood autoantibody level coinciding in time with the periods of marked pathomorphological changes in the brain tissue.
|
[Role of autoimmune processes in the pathogenesis of post-vaccinal lesions of the nervous system]. Subcutaneous sensitization of guinea pigs with -vaccine, and also an intracardiac injection of smallpox or measles vaccine induced production of brain autoantibodies, whereas subcutaneous or intradermal immunization of the animals with liver viral vaccines was not accompanied by the formation of autoantibodies and development of the pathological processes in the nervous system tissue. tthe greatest changes in the brain tissue in the form of circulatory disturbances and inflammatory-dystrophic phenomena in combination with high autoantibody titres in the blood were noted in intracardiac injection of smallpox vaccine and in subturbancessensitization with AK-Vaccine and the least (short-term circulatory disturbances, transitory signs of serous meningitis and neuron dystrophy against the background of low blood auto anitbody content)--in intracardiac injection of measles vaccine. Administration of live viral vaccines into the circulation of animals against the background of their sensitization with -vaccine led to reduction of blood autoantibody level coinciding in time with the periods of marked pathomorphological changes in the brain tissue.
|
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-0.05061998963356018,
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0.01990891620516777
] |
PMID:21501
|
Pseudomonas cytochrome c peroxidase. XIII. pH-denaturation of the enzyme.
|
The effect of pH on the oxidized Pseudomonas cytochrome c peroxidase molecule was studied by measuring the peroxidatic activity, the sedimentation velocity, the circular dichroic spectra in the far UV and Soret regions, and the optical absorption spectra of the enzyme in the pH range 2.5-13.0 at a constant ionic strength (micron = 0.1). The enzyme was stable in a narrow pH region, pH 6.0 - 7.4. In the low pH range the gross tertiary structure was observed to change quite simultaneously with the enzymatic activity and secondary structure. The optical absorption spectra indicated that there were no coordinated internal protein liqands in the 6th coordination positions of the heme prosthetic groups at the lowest pH studied. In the high pH range the secondary structure and the protein environment of hemes were observed to remain stable after the tertiary structure had changed and the activity had decreased. According to the optical absorption spectra the 6th internal protein ligands of hemes were retained at the highest pH studied.
|
Pseudomonas cytochrome c peroxidase. XIII. pH-denaturation of the enzyme. The effect of pH on the oxidized Pseudomonas cytochrome c peroxidase molecule was studied by measuring the peroxidatic activity, the sedimentation velocity, the circular dichroic spectra in the far UV and Soret regions, and the optical absorption spectra of the enzyme in the pH range 2.5-13.0 at a constant ionic strength (micron = 0.1). The enzyme was stable in a narrow pH region, pH 6.0 - 7.4. In the low pH range the gross tertiary structure was observed to change quite simultaneously with the enzymatic activity and secondary structure. The optical absorption spectra indicated that there were no coordinated internal protein liqands in the 6th coordination positions of the heme prosthetic groups at the lowest pH studied. In the high pH range the secondary structure and the protein environment of hemes were observed to remain stable after the tertiary structure had changed and the activity had decreased. According to the optical absorption spectra the 6th internal protein ligands of hemes were retained at the highest pH studied.
|
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] |
PMID:21498
|
[Interneuronal synapses with electrical and chemical mechanisms of transmission and the evolution of the central nervous system].
|
It becomes increasingly evident that not only chemical but also electrotonic synapses are characteristic feature of the vertebrate brain. Experiments carried out on the isolated perfused central nervous system of lampreys and frogs have shown that in cyclostomes spinal motoneurons receive at least 4 different electrotonic inputs: from giant reticulospinal axons, from smaller descending fibers intrinsic to the cord, from primary afferents and ventral roots. In amphibian motoneurons it is possible to identify 3 electrotonically mediated monosynaptic actions: from primary afferents, recurrent, and reticulospinal. The exact mode of transmission of monosynaptic excitation 1a and supraspinal actions in mammals remains to be elucidated. The trends of evolutionary development of synaptic transmissional processes and synaptic organization in vertebrate nervous system are discussed.
|
[Interneuronal synapses with electrical and chemical mechanisms of transmission and the evolution of the central nervous system]. It becomes increasingly evident that not only chemical but also electrotonic synapses are characteristic feature of the vertebrate brain. Experiments carried out on the isolated perfused central nervous system of lampreys and frogs have shown that in cyclostomes spinal motoneurons receive at least 4 different electrotonic inputs: from giant reticulospinal axons, from smaller descending fibers intrinsic to the cord, from primary afferents and ventral roots. In amphibian motoneurons it is possible to identify 3 electrotonically mediated monosynaptic actions: from primary afferents, recurrent, and reticulospinal. The exact mode of transmission of monosynaptic excitation 1a and supraspinal actions in mammals remains to be elucidated. The trends of evolutionary development of synaptic transmissional processes and synaptic organization in vertebrate nervous system are discussed.
|
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] |
PMID:21503
|
Correlation of pituitary and testicular responses to stimulation tests in cryptorchid children.
|
LH-RH test and HCG stimulation test were performed in 154 cryptorchid boys aged 1 month to 15 years (64 unilateral and 90 bilateral). Basal plasma LH levels and LH response to LH-RH were significantly lower from infancy to early puberty in cryptorchids compared with controls. Basal FSH levels and FSH response to LH-RH were normal. The post-HCG rise of plasma testosterone was reduced until mid-puberty. A significant positive correlation was found between post-HCG testosterone levels and pre- and post-LH-RH levels of LH. This correlation suggests that testicular maldescent and the decreased ability of Leydig cells to respond to a short course of HCG may result from an early defect or a delay of pituitary LH secretion.
|
Correlation of pituitary and testicular responses to stimulation tests in cryptorchid children. LH-RH test and HCG stimulation test were performed in 154 cryptorchid boys aged 1 month to 15 years (64 unilateral and 90 bilateral). Basal plasma LH levels and LH response to LH-RH were significantly lower from infancy to early puberty in cryptorchids compared with controls. Basal FSH levels and FSH response to LH-RH were normal. The post-HCG rise of plasma testosterone was reduced until mid-puberty. A significant positive correlation was found between post-HCG testosterone levels and pre- and post-LH-RH levels of LH. This correlation suggests that testicular maldescent and the decreased ability of Leydig cells to respond to a short course of HCG may result from an early defect or a delay of pituitary LH secretion.
|
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0.06495856493711472,
-0.0029615277890115976
] |
PMID:21504
|
Comparison of a new antihistamine HC 20-511 with cyproheptadine (Periactin) in chronic urticaria.
|
The effect and tolerance of a new antihistamine, HC 20-511, in daily doses of 2 and 4 mg was compared with cyproheptadine (Periactin) 8 mg daily in 50 cases of chronic urticaria. HC 20-511 4 mg daily proved to be the most effective, rapid and long-lasting. HC 20-511 had less sedative effect than cyproheptadine, but both possessed the same appetite-stimulating effect. However, this was not noted with the smaller dosage of HC 20-511.
|
Comparison of a new antihistamine HC 20-511 with cyproheptadine (Periactin) in chronic urticaria. The effect and tolerance of a new antihistamine, HC 20-511, in daily doses of 2 and 4 mg was compared with cyproheptadine (Periactin) 8 mg daily in 50 cases of chronic urticaria. HC 20-511 4 mg daily proved to be the most effective, rapid and long-lasting. HC 20-511 had less sedative effect than cyproheptadine, but both possessed the same appetite-stimulating effect. However, this was not noted with the smaller dosage of HC 20-511.
|
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0.03449137136340141,
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] |
PMID:21505
|
Action of enflurane (Ethrane) on the neuromusclar block induced by AH 8165 D.
|
The neuromuscular block induced by AH 8165 D, a new non-depolarizing muscle relaxant, has been studied in two series of anesthetised patients. All the anesthetics were induced by Althesin, and intubations performed under local anesthesia. The first series of 15 patients received only 75% nitrous oxide and 25% oxygen for the first hour of anesthesia. The second series received in addition 1 to 1.5% of enflurane. After an hour all the patients received a single dose of AH 8165 D. Dose-response curves were established in both series and it was found that without enflurane the ED50 of AH 8165 D was 8.29 mg/m2 and with enflurane 4.07 mg/m2. Therefore enflurane potentiates the action of AH 8165 D by 2.03 times, although it is impossible from this study to indicate precisely how and where enflurane can affect neuromuscular transmission.
|
Action of enflurane (Ethrane) on the neuromusclar block induced by AH 8165 D. The neuromuscular block induced by AH 8165 D, a new non-depolarizing muscle relaxant, has been studied in two series of anesthetised patients. All the anesthetics were induced by Althesin, and intubations performed under local anesthesia. The first series of 15 patients received only 75% nitrous oxide and 25% oxygen for the first hour of anesthesia. The second series received in addition 1 to 1.5% of enflurane. After an hour all the patients received a single dose of AH 8165 D. Dose-response curves were established in both series and it was found that without enflurane the ED50 of AH 8165 D was 8.29 mg/m2 and with enflurane 4.07 mg/m2. Therefore enflurane potentiates the action of AH 8165 D by 2.03 times, although it is impossible from this study to indicate precisely how and where enflurane can affect neuromuscular transmission.
|
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] |
PMID:21506
|
Duration of action of analgesic supplements to anesthesia. A double-blind comparison between morphine, fentanyl and sulfentanil.
|
In a double-blind trial, in a total of 45 patients, sulfentanil was compared with fentanyl and morphine in equipotent doses, as a narcotic supplement to anesthesia. Initially, morphine was shown to have a significantly longer duration of effect than fentanyl and sulfentanil, which for the first 3 doses had similar durations of action to each other. Later doses of fentanyl, however, had an extended effect, presumably because of cumulation. This was not seen with sulfentanil.
|
Duration of action of analgesic supplements to anesthesia. A double-blind comparison between morphine, fentanyl and sulfentanil. In a double-blind trial, in a total of 45 patients, sulfentanil was compared with fentanyl and morphine in equipotent doses, as a narcotic supplement to anesthesia. Initially, morphine was shown to have a significantly longer duration of effect than fentanyl and sulfentanil, which for the first 3 doses had similar durations of action to each other. Later doses of fentanyl, however, had an extended effect, presumably because of cumulation. This was not seen with sulfentanil.
|
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] |
PMID:21507
|
Possible pathogenesis of Huntington's chorea and a new approach to treatment.
|
Recent identification of glutamate as a transmitter in neocortical efferents and discovery of toxic effects of excessive amounts of extra-cellular glutamate or its analogues on some brain neurons offer an explanation of the pathogenesis of Huntington's chorea. Cell death as well as consequent biochemical changes and clinical symptoms in this disease possibly result from excessive excitation of the neostriatal, and of some other neurons by glutamate released from neocortical axon terminals. If so, the development of Huntington's chorea could be prevented or arrested by blockers of glutamate transmissions, and folate should be reduced to the minimum in the diet of the genetically marked families.
|
Possible pathogenesis of Huntington's chorea and a new approach to treatment. Recent identification of glutamate as a transmitter in neocortical efferents and discovery of toxic effects of excessive amounts of extra-cellular glutamate or its analogues on some brain neurons offer an explanation of the pathogenesis of Huntington's chorea. Cell death as well as consequent biochemical changes and clinical symptoms in this disease possibly result from excessive excitation of the neostriatal, and of some other neurons by glutamate released from neocortical axon terminals. If so, the development of Huntington's chorea could be prevented or arrested by blockers of glutamate transmissions, and folate should be reduced to the minimum in the diet of the genetically marked families.
|
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] |
PMID:21508
|
Acid production from Lycasin, maltitol, sorbitol and xylitol by oral streptococci and lactobacilli.
|
The acid production from maltitol was compared with the acid production from hydrogenated starch hydrolysate (Lycasin), sorbitol and xylitol by a number of oral strains and reference strains of Streptococcus mutans, S. sanguis, S. salivarius, S. mitior, S. milleri, S. faecalis, S. faecium, S. avium, Lactobacillus casei and L. salivarius. The polyols were added to a final concentration of 1.0% to two different basal media. Incubation was performed at 37 degrees C for 7 days after which the pH was recorded. Maltitol was fermented only by the lactobacilli (about two thirds of the strains). Lycasin was fermented by all strains of S. faecalis, more than 90% of the lactobacilli, about half of the S. sanguis strains, about one third of the S. mutans strains, and by a few other streptococcal strains. Acid production from sorbitol was observed among more than 80% of the S. mutans strains and the S. faecalis strains and most of the lactobacilli strains. Sorbitol-fermenting strains of S. sanguis and of S. mitior, all isolated from sorbitol-consumers, were observed. No other sorbitol-fermenting streptococci were found. Only the reference strains L. salivarius subsp. salivarius ATCC 11741 and S. avium ATCC 14025 fermented xylitol.
|
Acid production from Lycasin, maltitol, sorbitol and xylitol by oral streptococci and lactobacilli. The acid production from maltitol was compared with the acid production from hydrogenated starch hydrolysate (Lycasin), sorbitol and xylitol by a number of oral strains and reference strains of Streptococcus mutans, S. sanguis, S. salivarius, S. mitior, S. milleri, S. faecalis, S. faecium, S. avium, Lactobacillus casei and L. salivarius. The polyols were added to a final concentration of 1.0% to two different basal media. Incubation was performed at 37 degrees C for 7 days after which the pH was recorded. Maltitol was fermented only by the lactobacilli (about two thirds of the strains). Lycasin was fermented by all strains of S. faecalis, more than 90% of the lactobacilli, about half of the S. sanguis strains, about one third of the S. mutans strains, and by a few other streptococcal strains. Acid production from sorbitol was observed among more than 80% of the S. mutans strains and the S. faecalis strains and most of the lactobacilli strains. Sorbitol-fermenting strains of S. sanguis and of S. mitior, all isolated from sorbitol-consumers, were observed. No other sorbitol-fermenting streptococci were found. Only the reference strains L. salivarius subsp. salivarius ATCC 11741 and S. avium ATCC 14025 fermented xylitol.
|
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] |
PMID:21511
|
[Cogan's syndrome].
|
Since Cogan described his first case in 1945, about 50 cases have been published in the world literature, A 23 year old patient who suffered from this disease led us to make a detailed study of previous publications. The cochleo-vestibular, the etiological and the ophtalmical aspects of the case were studied, and, also the differential diagnosis and the therapeutical possibilities.
|
[Cogan's syndrome]. Since Cogan described his first case in 1945, about 50 cases have been published in the world literature, A 23 year old patient who suffered from this disease led us to make a detailed study of previous publications. The cochleo-vestibular, the etiological and the ophtalmical aspects of the case were studied, and, also the differential diagnosis and the therapeutical possibilities.
|
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] |
PMID:21512
|
Relapsing polychondritis. A clinical, pathologic-anatomic and histochemical study of 2 cases.
|
A 57-year-old man and a 70-year-old woman with relapsing polychondritis are reported. The man, suffering from arthralgias, respiratory obstruction, external ear and sanddle-nose deformities, conjunctivitis and irido-cyclitis, died after 4 years from airway obstruction because of tracheal and bronchial collapse. The woman is alive 8 months after the development of respiratory obstruction, probably caused by radiographically demonstrated tracheal obstruction, a saddle-nose deformity and hearing impairment. Microscopically, the involved cartilages showed degenerative and slight inflammatory changes and were eventually replaced by fibrous tissue. Histochemical studies, utilizing staining with Alcian blue at controlled electrolyte concentrations (Scott technique) and at controlled pH:s, with or without digestion with bacterial chondroitinase ABC; and staining with the PAS-method, with or without diastase digestion, revealed a complete or relative loss of glucosaminoglycans and glycogen. A biosynthetic defect is considered unlikely to be the primary pathogenetic mechanism of relapsing polychondritis. Histological and histochemical examination of biopsies from involved cartilages contribute to a definite diagnosis.
|
Relapsing polychondritis. A clinical, pathologic-anatomic and histochemical study of 2 cases. A 57-year-old man and a 70-year-old woman with relapsing polychondritis are reported. The man, suffering from arthralgias, respiratory obstruction, external ear and sanddle-nose deformities, conjunctivitis and irido-cyclitis, died after 4 years from airway obstruction because of tracheal and bronchial collapse. The woman is alive 8 months after the development of respiratory obstruction, probably caused by radiographically demonstrated tracheal obstruction, a saddle-nose deformity and hearing impairment. Microscopically, the involved cartilages showed degenerative and slight inflammatory changes and were eventually replaced by fibrous tissue. Histochemical studies, utilizing staining with Alcian blue at controlled electrolyte concentrations (Scott technique) and at controlled pH:s, with or without digestion with bacterial chondroitinase ABC; and staining with the PAS-method, with or without diastase digestion, revealed a complete or relative loss of glucosaminoglycans and glycogen. A biosynthetic defect is considered unlikely to be the primary pathogenetic mechanism of relapsing polychondritis. Histological and histochemical examination of biopsies from involved cartilages contribute to a definite diagnosis.
|
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] |
PMID:21509
|
A dental splint for use during peroral endoscopy.
|
Damage to teeth is a well known complication of peroral endoscopy. A case is described in which potentially fatal complications of such damage occurred. This spurred on the development of a dental splint as an aid in the prevention of such damage. This splint is described, and evidence for the beneficial results of its use is presented.
|
A dental splint for use during peroral endoscopy. Damage to teeth is a well known complication of peroral endoscopy. A case is described in which potentially fatal complications of such damage occurred. This spurred on the development of a dental splint as an aid in the prevention of such damage. This splint is described, and evidence for the beneficial results of its use is presented.
|
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] |
PMID:21513
|
Differetial staining of glycosaminoglycans, utilizing bacterial chondroitinase and chondrosulphatase.
|
A histochemical method for the differentiation of glucosaminoglycans, utilizing bacterial chondroitinase ABC and chondro-4- and -6 sulphatases, and staining with Alcian blue, is presented. The method is applied on human tissues with known glucosaminoglycan content (ganglion cyst, umbilical cord, foetal cartilage, adult cartilage) and the results are compared with the results obtained by staining with Alcian blue at controlled pH levels, with or without prior digestion with bovine testicular hyaloronidase, and the Scott method, utilizing Alcian blue at varying concentrations of MgCl2. It is concluded that chondroitinase ABC digest chondroitin-4 and -6 sulphate and to some extent also hyaluronic acid and dermatan sulphate, but not heparin and keratosulphate.
|
Differetial staining of glycosaminoglycans, utilizing bacterial chondroitinase and chondrosulphatase. A histochemical method for the differentiation of glucosaminoglycans, utilizing bacterial chondroitinase ABC and chondro-4- and -6 sulphatases, and staining with Alcian blue, is presented. The method is applied on human tissues with known glucosaminoglycan content (ganglion cyst, umbilical cord, foetal cartilage, adult cartilage) and the results are compared with the results obtained by staining with Alcian blue at controlled pH levels, with or without prior digestion with bovine testicular hyaloronidase, and the Scott method, utilizing Alcian blue at varying concentrations of MgCl2. It is concluded that chondroitinase ABC digest chondroitin-4 and -6 sulphate and to some extent also hyaluronic acid and dermatan sulphate, but not heparin and keratosulphate.
|
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] |
PMID:21517
|
Cerebral blood flow and oxygen consumption in rat, measured with microspheres or xenon.
|
The cerebral blood flow and, in some rats, the cerebral rate of oxygen consumption were measured in three groups of male rats. Fractionation of radioisotope-labeled microspheres was used to measure regional cerebral blood flow in four parts of the rat brain. The arterial and cerebral venous concentrations of radioactive xenon during desaturation were used to measure the blood flow and oxygen consumption of cortex when venous blood was collected from the superior sagittal sinus, or of whole brain when the transverse sinus was sampled. The regional cerebral flow measured with microspheres had a large standard error reflecting the technical difficulty of this method. The cerebral blood flow measured with xenon was higher when venous blood was sampled from the superior sagittal sinus than when sampled from the transverse sinus, but cerebral oxygen consumption rates were similar. The difference reflects the greater trauma involved in the superior sagittal approach and possible extracerebral contamination present in the transverse sinus approach.
|
Cerebral blood flow and oxygen consumption in rat, measured with microspheres or xenon. The cerebral blood flow and, in some rats, the cerebral rate of oxygen consumption were measured in three groups of male rats. Fractionation of radioisotope-labeled microspheres was used to measure regional cerebral blood flow in four parts of the rat brain. The arterial and cerebral venous concentrations of radioactive xenon during desaturation were used to measure the blood flow and oxygen consumption of cortex when venous blood was collected from the superior sagittal sinus, or of whole brain when the transverse sinus was sampled. The regional cerebral flow measured with microspheres had a large standard error reflecting the technical difficulty of this method. The cerebral blood flow measured with xenon was higher when venous blood was sampled from the superior sagittal sinus than when sampled from the transverse sinus, but cerebral oxygen consumption rates were similar. The difference reflects the greater trauma involved in the superior sagittal approach and possible extracerebral contamination present in the transverse sinus approach.
|
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] |
PMID:21518
|
Serum inorganic phosphate in middle-aged men. I. Inverse relation to body weight.
|
Serum inorganic phosphate was determined in 752 men born in 1926. An inverse correlation was found between serum inorganic phosphate levels and body weight. Other parameters of possible relevance to this finding, such as serum calcium, serum albumin, serum lambda-glutamyltransferase, and the occurrence of upper gastrointestinal disorders were analysed, and no correlations to the phosphate level were detected.
|
Serum inorganic phosphate in middle-aged men. I. Inverse relation to body weight. Serum inorganic phosphate was determined in 752 men born in 1926. An inverse correlation was found between serum inorganic phosphate levels and body weight. Other parameters of possible relevance to this finding, such as serum calcium, serum albumin, serum lambda-glutamyltransferase, and the occurrence of upper gastrointestinal disorders were analysed, and no correlations to the phosphate level were detected.
|
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] |
PMID:21534
|
Introduction of artificial crosslinks into proteins.
|
In this chapter, we present a brief overview of the current status of protein crosslinking technology. An attempt is made to compare the natural crosslinks and natural crosslinking agents to the artificial ones, and a brief section is devoted to the potential use of enzymes (transglutaminase and peroxidase) as crosslinking agents in vitro. Homobifunctional (x-R-x) and heterobifunctional (x-R-y) reagents are considered in terms of the kinds of functional groups and R-groups that have been used in protein crosslinking, and some examples of reagents and applications from the recent literature are tabulated.
|
Introduction of artificial crosslinks into proteins. In this chapter, we present a brief overview of the current status of protein crosslinking technology. An attempt is made to compare the natural crosslinks and natural crosslinking agents to the artificial ones, and a brief section is devoted to the potential use of enzymes (transglutaminase and peroxidase) as crosslinking agents in vitro. Homobifunctional (x-R-x) and heterobifunctional (x-R-y) reagents are considered in terms of the kinds of functional groups and R-groups that have been used in protein crosslinking, and some examples of reagents and applications from the recent literature are tabulated.
|
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] |
PMID:21535
|
Comparison of hydrophobic and strongly hydrophilic cleavable crosslinking reagents in intermolecular bond formation in aggregates of proteins or protein-RNA.
|
Most of the bifunctional reagents in protein chemistry possess a strongly hydrophobic backbone, derived from aliphatic or aromatic hydrocarbons. Even bifunctionals of more than 30 A in length of this sort form intramolecular bridges preferentially. In recent years, the intermolecular crosslinking of physiological protein aggregates has gained in importance. As shown in the crosslinking of hemoglobin with two sets of hydrophobic and strongly hydrophilic reagents, derived from azo dyes and tartaric acid, respectively, in this case it is not primarily the length of the bifunctional, but the hydrophilic structure that will enhance intermolecular crosslinking. Artificial dimers of native structure may be obtained. For the crosslinking of RNA to protein, we have synthesized a new reagent, 3-(2-bromo-3-oxobutane-1-sulphonyl)-propionic acid p-nitrophenyl ester. In a two step reaction, it is attached to adenine and cytosine moieties at pH 6 first, and to lysine side chains at pH 7,5. The reagent has been applied to the poly-A sequence of globin messenger RNA nucleoprotein.
|
Comparison of hydrophobic and strongly hydrophilic cleavable crosslinking reagents in intermolecular bond formation in aggregates of proteins or protein-RNA. Most of the bifunctional reagents in protein chemistry possess a strongly hydrophobic backbone, derived from aliphatic or aromatic hydrocarbons. Even bifunctionals of more than 30 A in length of this sort form intramolecular bridges preferentially. In recent years, the intermolecular crosslinking of physiological protein aggregates has gained in importance. As shown in the crosslinking of hemoglobin with two sets of hydrophobic and strongly hydrophilic reagents, derived from azo dyes and tartaric acid, respectively, in this case it is not primarily the length of the bifunctional, but the hydrophilic structure that will enhance intermolecular crosslinking. Artificial dimers of native structure may be obtained. For the crosslinking of RNA to protein, we have synthesized a new reagent, 3-(2-bromo-3-oxobutane-1-sulphonyl)-propionic acid p-nitrophenyl ester. In a two step reaction, it is attached to adenine and cytosine moieties at pH 6 first, and to lysine side chains at pH 7,5. The reagent has been applied to the poly-A sequence of globin messenger RNA nucleoprotein.
|
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] |
PMID:21536
|
Comparison of wool reactions with selected mono and bifunctional reagents.
|
The molecular structure of wool is discussed in relation to chemical reactivity and the role of disulfide crosslinks. Ideal characteristics of an effective medium (e.g. dimethylformamide) for modifying wool include the ability to penetrate and swell wool without interfering with reagents used. The extent of reaction of wool or reduced wool is compared for mono-and bifunctional activated vinyl compounds, isocyanates, acid chlorides, acid anhydrides, sulfonyl chlorides, and alkyl halides. The degree of crosslinking is assessed by solubility, supercontraction, and tensile tests. Optical and electron scanning microscopy can give evidence of external polymer deposition in contrast to internal chemical modification. Effects of crosslinking by bifunctional reagents are related to changes in mechanical, chemical, and biological (moth-resisting) properties of the modified wool.
|
Comparison of wool reactions with selected mono and bifunctional reagents. The molecular structure of wool is discussed in relation to chemical reactivity and the role of disulfide crosslinks. Ideal characteristics of an effective medium (e.g. dimethylformamide) for modifying wool include the ability to penetrate and swell wool without interfering with reagents used. The extent of reaction of wool or reduced wool is compared for mono-and bifunctional activated vinyl compounds, isocyanates, acid chlorides, acid anhydrides, sulfonyl chlorides, and alkyl halides. The degree of crosslinking is assessed by solubility, supercontraction, and tensile tests. Optical and electron scanning microscopy can give evidence of external polymer deposition in contrast to internal chemical modification. Effects of crosslinking by bifunctional reagents are related to changes in mechanical, chemical, and biological (moth-resisting) properties of the modified wool.
|
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] |
PMID:21537
|
Kinetic studies of immobilized alpha-chymotrypsin in apolar solvents.
|
The mechanism of alpha-chymotrypsin action has been probed by extending studies of native chymotrypsin to immobilized chymotrypsin, where the organic content of the solution can be raised to much higher levels and thus one can explicitly look at the role of water. When one does this, one finds that water only appears in the deacylation reaction. The premise that one can go from native chymotrypsin (souble) to immobilized chymotrypsin (insoluble) has been tested by several criteria. It has been found in many instances that the two are identical: in absolute rate, in pKa. They are, however, not identical to one another in binding, due to differences in diffusion, which is to be expected. Thus, mechanistically immobilized and native chymotrypsin are identical to one another and the use of immobilized chymotrypsin can be used to specify the mechanism even more: it must proceed through two tetrahedral intermediates and two acyl-enzyme intermediates.
|
Kinetic studies of immobilized alpha-chymotrypsin in apolar solvents. The mechanism of alpha-chymotrypsin action has been probed by extending studies of native chymotrypsin to immobilized chymotrypsin, where the organic content of the solution can be raised to much higher levels and thus one can explicitly look at the role of water. When one does this, one finds that water only appears in the deacylation reaction. The premise that one can go from native chymotrypsin (souble) to immobilized chymotrypsin (insoluble) has been tested by several criteria. It has been found in many instances that the two are identical: in absolute rate, in pKa. They are, however, not identical to one another in binding, due to differences in diffusion, which is to be expected. Thus, mechanistically immobilized and native chymotrypsin are identical to one another and the use of immobilized chymotrypsin can be used to specify the mechanism even more: it must proceed through two tetrahedral intermediates and two acyl-enzyme intermediates.
|
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] |
PMID:21538
|
Factors affecting cyanoborohydride reduction of aromatic Schiff's bases in proteins.
|
Reductive alkylation of bovine serum albumin (BSA) and wool by a aromatic aldehydes and sodium cyanoborohydride has been investigated. The aldehydes used were chosen to allow convenient quantitative measurement of binding by ultraviolet spectroscopy. Alkylation of BSA occurred primarily at two highly reactive sites. Variation of time, PH, reactant concentration, and addition of urea had little effect on the extent of alkylation of BSA. However, more extensive alkylation was achieved in buffered aqueous dimethyl sulfoxide. The unusual reactivity of two epsilon-amino groups on the BSA molecule is attributed to closely placed lysine residues in the primary sequence rather than to favorable placement of unrelated, distant reactive centers. Similarly, only a few of the potentially available epsilon-amino groups of wool were observed to react.
|
Factors affecting cyanoborohydride reduction of aromatic Schiff's bases in proteins. Reductive alkylation of bovine serum albumin (BSA) and wool by a aromatic aldehydes and sodium cyanoborohydride has been investigated. The aldehydes used were chosen to allow convenient quantitative measurement of binding by ultraviolet spectroscopy. Alkylation of BSA occurred primarily at two highly reactive sites. Variation of time, PH, reactant concentration, and addition of urea had little effect on the extent of alkylation of BSA. However, more extensive alkylation was achieved in buffered aqueous dimethyl sulfoxide. The unusual reactivity of two epsilon-amino groups on the BSA molecule is attributed to closely placed lysine residues in the primary sequence rather than to favorable placement of unrelated, distant reactive centers. Similarly, only a few of the potentially available epsilon-amino groups of wool were observed to react.
|
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] |
PMID:21540
|
Electron microscopy of an oligomeric protein stabilized by polyfunctional cross-linking.
|
Oligomeric proteins can be intramolecularly cross-linked with polylysine in a reaction in which a water soluble carbodiimide mediates an amide linkage between the protein carboxyl groups and the epsilon-amino groups of polylysine. Studies carried out with a cytochrome p-450 indicate that a small number of molecules in a population which has been cross-linked in this way retain important features of their tertiary and quaternary structure when negatively stained and examined in the electron microscope. Use of the method in determining the subunit geometry of oligomeric proteins is discussed.
|
Electron microscopy of an oligomeric protein stabilized by polyfunctional cross-linking. Oligomeric proteins can be intramolecularly cross-linked with polylysine in a reaction in which a water soluble carbodiimide mediates an amide linkage between the protein carboxyl groups and the epsilon-amino groups of polylysine. Studies carried out with a cytochrome p-450 indicate that a small number of molecules in a population which has been cross-linked in this way retain important features of their tertiary and quaternary structure when negatively stained and examined in the electron microscope. Use of the method in determining the subunit geometry of oligomeric proteins is discussed.
|
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] |
PMID:21546
|
Cell transfer studies with the DK/or inbred chicken lines.
|
The histoincompatibility in the DK-OR-1 (B13B13) and DK/OR-3 (B19B19) inbred White Leghorn chicken lines was assessed with the Simonsen's spleen assay, GVHR mortality counts and antibody production by transferred cells in embryonic in vivo hosts. The two lines appear to be sufficiently histocompatible for use in cell transfer experiments. The occurrence of antigen responsive cells (ARC) and precursor cells in bone marrow as assessed with the in vivo culture and baby chick immune enhancement assays. Bone marrow of unimmunized donors possessed very little anti-MRBC immune responsiveness. Bone marrow cells from MRBC-primed donors, however, showed low anti-MRBC activity in the in vivo culture which could be elevated with a second antigen stimulation. Chicks grafted as embryos with bone marrow from immunized donors responded to MRBC immunization in an enhanced manner as compared to the control untreated chicks which responded poorly if at all to the antigen. Immunization appears (a) to increase the pool of potential ARC in the bone marrow of the donor and (b) to augment the responsiveness of bone marrow cells transferred into embryonic hosts.
|
Cell transfer studies with the DK/or inbred chicken lines. The histoincompatibility in the DK-OR-1 (B13B13) and DK/OR-3 (B19B19) inbred White Leghorn chicken lines was assessed with the Simonsen's spleen assay, GVHR mortality counts and antibody production by transferred cells in embryonic in vivo hosts. The two lines appear to be sufficiently histocompatible for use in cell transfer experiments. The occurrence of antigen responsive cells (ARC) and precursor cells in bone marrow as assessed with the in vivo culture and baby chick immune enhancement assays. Bone marrow of unimmunized donors possessed very little anti-MRBC immune responsiveness. Bone marrow cells from MRBC-primed donors, however, showed low anti-MRBC activity in the in vivo culture which could be elevated with a second antigen stimulation. Chicks grafted as embryos with bone marrow from immunized donors responded to MRBC immunization in an enhanced manner as compared to the control untreated chicks which responded poorly if at all to the antigen. Immunization appears (a) to increase the pool of potential ARC in the bone marrow of the donor and (b) to augment the responsiveness of bone marrow cells transferred into embryonic hosts.
|
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] |
PMID:21547
|
Role of the major histocompatibility complex in resistance to Marek's disease: restriction of the growth of JMV-MD tumor cells in genetically resistant birds.
|
B21 is associated with resistance to Marek's disease (MD). Forty populations of chickens from all over the world were examined for the presence of the B21 allele. B21 was found in twelve of these populations and it's presence was confirmed by GVH testing in all ten populations which were tested. The populations in which B21 was detected represent the extreme production types of the species and include the progenitor of the species, the Red Jungle Fowl. Our studies suggest that B21 may have strong survival value for the species. An allogeneic transplantable lymphoma of MD, the JMV tumor cell line, grows more slowly in MD resistant (B21/B21) chicks than in MD susceptible (B2/B2) chicks. This is the first direct evidence that genetic resistance to MD may involve an active (immunological?) restriction of tumor cell growth. JMV cells were further characterized as a transplant of B1 carrying lymphoblastoid cells, an allele which may be associated with susceptibility to MD.
|
Role of the major histocompatibility complex in resistance to Marek's disease: restriction of the growth of JMV-MD tumor cells in genetically resistant birds. B21 is associated with resistance to Marek's disease (MD). Forty populations of chickens from all over the world were examined for the presence of the B21 allele. B21 was found in twelve of these populations and it's presence was confirmed by GVH testing in all ten populations which were tested. The populations in which B21 was detected represent the extreme production types of the species and include the progenitor of the species, the Red Jungle Fowl. Our studies suggest that B21 may have strong survival value for the species. An allogeneic transplantable lymphoma of MD, the JMV tumor cell line, grows more slowly in MD resistant (B21/B21) chicks than in MD susceptible (B2/B2) chicks. This is the first direct evidence that genetic resistance to MD may involve an active (immunological?) restriction of tumor cell growth. JMV cells were further characterized as a transplant of B1 carrying lymphoblastoid cells, an allele which may be associated with susceptibility to MD.
|
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] |
PMID:21552
|
[Development cooperation in the development of a basic medical service].
|
Team work in building up a basic medical service. - This article reports on a project of Swiss aid to the developing country of the Peruvian Andes, whose organization is completely integrated into the national Peruvian ministry of health. After having stated the social and medical problems the article explains, how a team of Swiss doctors tries to reach to remote parts of a developing country by means of a medicine adapted to the economical situation of the poor farmer population. This is achieved by training local health auxiliaries and teaching them preventive medicine. On the other hand the curative medicine has to take second place and is concentrated mainly on a large scale tuberculosis programme. Ideas, experiences, problems and first evaluations are set out and discussed.
|
[Development cooperation in the development of a basic medical service]. Team work in building up a basic medical service. - This article reports on a project of Swiss aid to the developing country of the Peruvian Andes, whose organization is completely integrated into the national Peruvian ministry of health. After having stated the social and medical problems the article explains, how a team of Swiss doctors tries to reach to remote parts of a developing country by means of a medicine adapted to the economical situation of the poor farmer population. This is achieved by training local health auxiliaries and teaching them preventive medicine. On the other hand the curative medicine has to take second place and is concentrated mainly on a large scale tuberculosis programme. Ideas, experiences, problems and first evaluations are set out and discussed.
|
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] |
PMID:21553
|
[Rural health center in Bali].
|
The rural Health Centre of Mengwi, on the island of Bali, was built and equipped for US $ 15,000 by Project Concern, a non-sectarian, medical relief organization. It has been run so successfully by them since it opened in 1974, that not only has the Centre gained the trust of the local community into which it is now fully integrated, but it is also regarded by the Indonesian Government as a pilot project. Amongst the factors leading to this success, the author underlines the following points: a suitable location for the Centre, easily accessible by the villagers; a local staff recruited from the villages of Mengwi District, well trained and sympathetic to the population; constant retraining of the staff leading to better medical care; the availability of a doctor and nurse around the clock; an adequate supply of drugs. Evaluation of the operation is best done by the acceptance of special programmes such as the Under-Five Clinic, the Family Planning Scheme, Ante-Natal Clinic and vaccinations. The mere success of the general clinic would not mean that the Centre had been integrated into the community and become a factor in rural development. The success of these special clinics, as well as the out-reach programmes, demonstrates that through adequate delivery of medical care, the Health Centre of Mengwi can now fulfil its role of being an important factor in the development of a rural community in the tropics.
|
[Rural health center in Bali]. The rural Health Centre of Mengwi, on the island of Bali, was built and equipped for US $ 15,000 by Project Concern, a non-sectarian, medical relief organization. It has been run so successfully by them since it opened in 1974, that not only has the Centre gained the trust of the local community into which it is now fully integrated, but it is also regarded by the Indonesian Government as a pilot project. Amongst the factors leading to this success, the author underlines the following points: a suitable location for the Centre, easily accessible by the villagers; a local staff recruited from the villages of Mengwi District, well trained and sympathetic to the population; constant retraining of the staff leading to better medical care; the availability of a doctor and nurse around the clock; an adequate supply of drugs. Evaluation of the operation is best done by the acceptance of special programmes such as the Under-Five Clinic, the Family Planning Scheme, Ante-Natal Clinic and vaccinations. The mere success of the general clinic would not mean that the Centre had been integrated into the community and become a factor in rural development. The success of these special clinics, as well as the out-reach programmes, demonstrates that through adequate delivery of medical care, the Health Centre of Mengwi can now fulfil its role of being an important factor in the development of a rural community in the tropics.
|
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] |
PMID:21554
|
Development aid through information. Trying a new concept in Columbia.
|
The origin, execution, and preliminary results of a development project in Cali (Colombia) are presented. This project, which involves a special concept, aims at breaking new ground in the field of development aid. The aim of the Vivamos Mejor foundation, established in Colombia, is to provide practical help for poor families. The foundation is carrying out two model projects at the moment, which provide urgently required help and which at the same time investigate the feasibility and effectiveness of the projects in a scientific manner. Under the first project, teams of native inhabitants provide information and instruct the population thoroughly on questions of nutrition, hygiene, and birth control. Under the second project, children who are suffering from disease and malnutrition are temporarily rehabilitated in crèches, while at the same time their mothers learn how to provide the child and its brothers and sisters with a more healthy home. On one hand the Swiss projects in Cali aim at helping families in the poor areas there, while on the other, they are being scientifically analyzed as model projects in the hope that they can provide examples for the large cities in other developing countries.
|
Development aid through information. Trying a new concept in Columbia. The origin, execution, and preliminary results of a development project in Cali (Colombia) are presented. This project, which involves a special concept, aims at breaking new ground in the field of development aid. The aim of the Vivamos Mejor foundation, established in Colombia, is to provide practical help for poor families. The foundation is carrying out two model projects at the moment, which provide urgently required help and which at the same time investigate the feasibility and effectiveness of the projects in a scientific manner. Under the first project, teams of native inhabitants provide information and instruct the population thoroughly on questions of nutrition, hygiene, and birth control. Under the second project, children who are suffering from disease and malnutrition are temporarily rehabilitated in crèches, while at the same time their mothers learn how to provide the child and its brothers and sisters with a more healthy home. On one hand the Swiss projects in Cali aim at helping families in the poor areas there, while on the other, they are being scientifically analyzed as model projects in the hope that they can provide examples for the large cities in other developing countries.
|
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] |
PMID:21555
|
[Water resources and health problems in developing countries].
|
The rapidly increasing use of water for agricultural and hydroelectric development in developing countries is causing serious health problems. These can be divided into two categories; the diseases whose transmission depends on water; and the changes in the life style of the population. We review briefly the principal health problems and analyze the principles to be followed in solving them, and the reasons of the success or failure of health programs. Among the topics we consider, are the development versus health, the collaboration of health service personnel with the rest of the personnel participating in water development schemes, the necessity of ensuring a multidisciplinary approach to the solutions of these health problems, the necessity of integration of the public health component within the administrative and budgetary levels and the coordination of public health measures with the development of project from the planning phase through the completion and maintenance phases, the purely medical aspects of the health problems involved appearing as being of secondary importance. Common sense, and the use of simple, cheap methods adapted to the local conditions, and applied focaly, provide better solutions than the sophisticated, more general and expensive approaches. The time and budget accorded for surveys and research, usually excessive, should be limited. It is necessary to adhere to a disciplined use of water. Ambitions and utopian dreams and strategies of disease eradication should, in general, give way to more limited, but accessible objectives, whose attainment would lead to a decrease of the clinical and socioeconomic gravity of the health problems to a "reasonable" level.
|
[Water resources and health problems in developing countries]. The rapidly increasing use of water for agricultural and hydroelectric development in developing countries is causing serious health problems. These can be divided into two categories; the diseases whose transmission depends on water; and the changes in the life style of the population. We review briefly the principal health problems and analyze the principles to be followed in solving them, and the reasons of the success or failure of health programs. Among the topics we consider, are the development versus health, the collaboration of health service personnel with the rest of the personnel participating in water development schemes, the necessity of ensuring a multidisciplinary approach to the solutions of these health problems, the necessity of integration of the public health component within the administrative and budgetary levels and the coordination of public health measures with the development of project from the planning phase through the completion and maintenance phases, the purely medical aspects of the health problems involved appearing as being of secondary importance. Common sense, and the use of simple, cheap methods adapted to the local conditions, and applied focaly, provide better solutions than the sophisticated, more general and expensive approaches. The time and budget accorded for surveys and research, usually excessive, should be limited. It is necessary to adhere to a disciplined use of water. Ambitions and utopian dreams and strategies of disease eradication should, in general, give way to more limited, but accessible objectives, whose attainment would lead to a decrease of the clinical and socioeconomic gravity of the health problems to a "reasonable" level.
|
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] |
PMID:21556
|
[Community health aide. Critical estimation of his task within the nutrition program].
|
Community Health Aides (CHA), locally recruited and trained, visit households and identify malnourished children by means of weighing them in monthly intervals, recording the results on "Gomez" weight-for-age charts. The CHA acts as the people's nearest adviser. In order to become a useful if not the most important member of the health team, some common mistakes and distorted views should be corrected early in her career. The weight-for-age chart is an invaluable tool to record the child's state of health. It is the trend in weight gain that is relevant and not an isolated weight point on the graph. Maternal, perinatal and neonatal histories should be taken as they help to classify the low weight child. 3/4 of the children in the Young Child Nutrition Programme (YCNP) are underweight but also underheight for age. The designation of malnutrition grade I/II/III is misleading. Either one speaks of "undernutrition" if one considers weight-for-age only or one takes other anthropometric measurements such as the height or length in order to classify Protein-Energy-Malnutrition. A physical examination and clinical records are essential in the evaluation of malnutrition - one should not rely on the graph only. By measuring the height of children, one may well be surprised to discover that many children in St. James are on the obese side. Obesity is another form of malnutrition prevalent in the wealthy societies of western industrialized countries. It is paradox that we should increase the number of obese people in a world which is threatened by shortage of food energies and proteins.
|
[Community health aide. Critical estimation of his task within the nutrition program]. Community Health Aides (CHA), locally recruited and trained, visit households and identify malnourished children by means of weighing them in monthly intervals, recording the results on "Gomez" weight-for-age charts. The CHA acts as the people's nearest adviser. In order to become a useful if not the most important member of the health team, some common mistakes and distorted views should be corrected early in her career. The weight-for-age chart is an invaluable tool to record the child's state of health. It is the trend in weight gain that is relevant and not an isolated weight point on the graph. Maternal, perinatal and neonatal histories should be taken as they help to classify the low weight child. 3/4 of the children in the Young Child Nutrition Programme (YCNP) are underweight but also underheight for age. The designation of malnutrition grade I/II/III is misleading. Either one speaks of "undernutrition" if one considers weight-for-age only or one takes other anthropometric measurements such as the height or length in order to classify Protein-Energy-Malnutrition. A physical examination and clinical records are essential in the evaluation of malnutrition - one should not rely on the graph only. By measuring the height of children, one may well be surprised to discover that many children in St. James are on the obese side. Obesity is another form of malnutrition prevalent in the wealthy societies of western industrialized countries. It is paradox that we should increase the number of obese people in a world which is threatened by shortage of food energies and proteins.
|
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] |
PMID:21557
|
Smallpox eradication in Rajasthan, India.
|
In 1962, when the smallpox eradication programme was launched in India, there were 3909 reported cases in Rajasthan. Mass vaccination was introduced but had little impact on the incidence of the disease; in 1970, 4074 cases were reported and in 1971, the reported incidence (4821 cases) was the highest in India. Changes in the vaccination technique, the use of a new and potent vaccine and the introduction of a new strategy based on surveillance and containment of outbreaks, rather than mass vaccination, brought good results. By 1972, only 1970 cases were reported and endemic smallpox was eliminated from Rajasthan before the launching of the intensified all-India compaign in July, 1973. In 1974, epidemics were occurring in the neighbouring states of Uttar Pradesh and Madhya Pradesh as well as in Bihar and West Bengal. But in that year, there were only 10 new smallpox outbreaks in Rajasthan, all originating from imported cases. The organization and progress of the eradication campaign and the history of smallpox eradication in Rajasthan are described in this communication.
|
Smallpox eradication in Rajasthan, India. In 1962, when the smallpox eradication programme was launched in India, there were 3909 reported cases in Rajasthan. Mass vaccination was introduced but had little impact on the incidence of the disease; in 1970, 4074 cases were reported and in 1971, the reported incidence (4821 cases) was the highest in India. Changes in the vaccination technique, the use of a new and potent vaccine and the introduction of a new strategy based on surveillance and containment of outbreaks, rather than mass vaccination, brought good results. By 1972, only 1970 cases were reported and endemic smallpox was eliminated from Rajasthan before the launching of the intensified all-India compaign in July, 1973. In 1974, epidemics were occurring in the neighbouring states of Uttar Pradesh and Madhya Pradesh as well as in Bihar and West Bengal. But in that year, there were only 10 new smallpox outbreaks in Rajasthan, all originating from imported cases. The organization and progress of the eradication campaign and the history of smallpox eradication in Rajasthan are described in this communication.
|
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] |
PMID:21558
|
Malaria in the Republic of the Philippines. A review.
|
Malaria in the Republic of the Philippines is caused principally by P. falciparum and P. vivax, with the former as predominant species. P. malariae is occasionally reported, while P. ovale is very rare and has been reported only in the island of Palawan. Malaria is widespread in distribution with prevalence varying from one area to the other. In 1970, the malaria morbidity rate was reported to be 77.6 per 100,000 while the mortality rate was 1.8 per 100,000. Case detection activities revealed that, in 1973, the slide parasite rate was 7.2%, the annual parasite index was 6.1% and the annual blood examination rate was 8.4%. The principal vector of malaria in the Philippines is An. minimus flavirostris which breeds in clear, fresh-water streams in foothills and mountain slopes. An. mangyanus and An. maculatus appear to play a secondary role. The vectorial capacity of the former appears to be confined only where conditions are primitive, while the latter is associated with malaria transmission in high altitudes. In the absence of fresh-water streams, the salt-water breeder mosquito, An. litoralis, assumes the vectorial role. The epidemiology of malaria in the Philippines is discussed. Emergence of strains of P. falciparum with diminished sensitivity to the commonly used antimalarial drugs is reported in Palawan and Rizal provinces. The development of malaria control activities in the Philippines are presented. As of 1972, Cagayan Valley, Palawan, Mindoro, Sulu and circumscribed areas in Mindanao are still considered hard-core malarious areas with on-going persistent transmission.
|
Malaria in the Republic of the Philippines. A review. Malaria in the Republic of the Philippines is caused principally by P. falciparum and P. vivax, with the former as predominant species. P. malariae is occasionally reported, while P. ovale is very rare and has been reported only in the island of Palawan. Malaria is widespread in distribution with prevalence varying from one area to the other. In 1970, the malaria morbidity rate was reported to be 77.6 per 100,000 while the mortality rate was 1.8 per 100,000. Case detection activities revealed that, in 1973, the slide parasite rate was 7.2%, the annual parasite index was 6.1% and the annual blood examination rate was 8.4%. The principal vector of malaria in the Philippines is An. minimus flavirostris which breeds in clear, fresh-water streams in foothills and mountain slopes. An. mangyanus and An. maculatus appear to play a secondary role. The vectorial capacity of the former appears to be confined only where conditions are primitive, while the latter is associated with malaria transmission in high altitudes. In the absence of fresh-water streams, the salt-water breeder mosquito, An. litoralis, assumes the vectorial role. The epidemiology of malaria in the Philippines is discussed. Emergence of strains of P. falciparum with diminished sensitivity to the commonly used antimalarial drugs is reported in Palawan and Rizal provinces. The development of malaria control activities in the Philippines are presented. As of 1972, Cagayan Valley, Palawan, Mindoro, Sulu and circumscribed areas in Mindanao are still considered hard-core malarious areas with on-going persistent transmission.
|
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] |
PMID:21559
|
Microfilarial polyarthritis in a massive Loa loa infestation. A case report.
|
A Cameroonian affected with massive Loa loa infection developed febrile arthritis with involvement of both knees and the left ankle. Although the patient was first seen by us after one month of treatment with Indomethacin, at this time the joints were still inflamed and microfilariae of Loa loa were found in the synovial fluid. No other etiological mechanism was identified. Following the articular puncture and treatment with Ketoprofen, the arthritis subsisted within a week. This is the first case to be studied in which arthritis during loasis has been explicitly documented by the presence of intra-articular microfilariae.
|
Microfilarial polyarthritis in a massive Loa loa infestation. A case report. A Cameroonian affected with massive Loa loa infection developed febrile arthritis with involvement of both knees and the left ankle. Although the patient was first seen by us after one month of treatment with Indomethacin, at this time the joints were still inflamed and microfilariae of Loa loa were found in the synovial fluid. No other etiological mechanism was identified. Following the articular puncture and treatment with Ketoprofen, the arthritis subsisted within a week. This is the first case to be studied in which arthritis during loasis has been explicitly documented by the presence of intra-articular microfilariae.
|
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] |
PMID:21561
|
Ambulatory management in alcoholism.
|
In the acute phase, management begins with ruling out conditions that would require hospitalization. Moderate withdrawal symptoms in susceptible patients require substitution medication which can be monitored on an outpatient basis. In the subacute phase, after withdrawal, frequent visits, family education and judicious use of drugs are required. Depressive reactions are common. There are no reliable indexes for predicting the success of a return to controlled drinking.
|
Ambulatory management in alcoholism. In the acute phase, management begins with ruling out conditions that would require hospitalization. Moderate withdrawal symptoms in susceptible patients require substitution medication which can be monitored on an outpatient basis. In the subacute phase, after withdrawal, frequent visits, family education and judicious use of drugs are required. Depressive reactions are common. There are no reliable indexes for predicting the success of a return to controlled drinking.
|
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] |
PMID:21563
|
Lower-esophageal sphincter function does not determine resting upper-esophageal sphincter pressure.
|
Records of 269 esophageal motility studies were reviewed to determine the relationship between lower-esophageal sphincter (LES) function and upper-esophageal sphincter (UES) pressure. Average and greatest UES pressures were similar in patients with LES pressures less than 10 mm Hg or greater than 20 mm Hg, and in patients with and without gastroesophageal reflux as determined by an intraesophageal pH electrode test. Although teliologically appealing, the belief that patients with weak lower-esophageal sphincters and gastroesophageal reflux have stronger upper-esophageal sphincters to guard against pharyngeal reflux and aspiration cannot be confirmed by current manometric techniques.
|
Lower-esophageal sphincter function does not determine resting upper-esophageal sphincter pressure. Records of 269 esophageal motility studies were reviewed to determine the relationship between lower-esophageal sphincter (LES) function and upper-esophageal sphincter (UES) pressure. Average and greatest UES pressures were similar in patients with LES pressures less than 10 mm Hg or greater than 20 mm Hg, and in patients with and without gastroesophageal reflux as determined by an intraesophageal pH electrode test. Although teliologically appealing, the belief that patients with weak lower-esophageal sphincters and gastroesophageal reflux have stronger upper-esophageal sphincters to guard against pharyngeal reflux and aspiration cannot be confirmed by current manometric techniques.
|
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] |
PMID:21564
|
Effect of multiple-stress procedures on monkey gastroduodenal mucosa, serum gastrin, and hydrogen ion kinetics.
|
By arranging a series of psychological contingencies (unpredictability, uncontrollability, conflict), coupled with delivery of a physical stimulus (electric shock), we produced gastroduodenal mucosal lesions in 7 of 8 rhesus monkeys. The most severe conflict paradigm most consistently produced lesions across subjects. Of the 30 lesions observed by endoscopy, 80% occurred near the anatomic junction of gastric body and antrum, in the antrum, or in the duodenum. Lesions varied in severity from discolorations of the mucosa to disruptions of mucosal integrity. Lesions in the stomach generally disappeared in several days despite the continuation of stress; some duodenal lesions were equally evanescent, but in 2 monkeys, lesions lasted over a week. Hydrogen ion kinetics were measured in 2 monkeys that developed gastric lesions and 2 that developed duodenal lesions. The rate at which hydrogen ion entered the duodenum was uniformly suppressed for all 4 monkeys during their first session of shock avoidance; during their last session, the gastric subgroup continued to show suppression while the duodenal subgroup returned towards control levels. Serum gastrin levels were unchanged by the multiple-stress procedures. Our finding of consistently producible, stress-induced gastroduodenal pathology in anatomic areas similar to those involved in man suggests that the subhuman primate is suitable for further efforts to produce an animal model of psychosomatic ulcer disease.
|
Effect of multiple-stress procedures on monkey gastroduodenal mucosa, serum gastrin, and hydrogen ion kinetics. By arranging a series of psychological contingencies (unpredictability, uncontrollability, conflict), coupled with delivery of a physical stimulus (electric shock), we produced gastroduodenal mucosal lesions in 7 of 8 rhesus monkeys. The most severe conflict paradigm most consistently produced lesions across subjects. Of the 30 lesions observed by endoscopy, 80% occurred near the anatomic junction of gastric body and antrum, in the antrum, or in the duodenum. Lesions varied in severity from discolorations of the mucosa to disruptions of mucosal integrity. Lesions in the stomach generally disappeared in several days despite the continuation of stress; some duodenal lesions were equally evanescent, but in 2 monkeys, lesions lasted over a week. Hydrogen ion kinetics were measured in 2 monkeys that developed gastric lesions and 2 that developed duodenal lesions. The rate at which hydrogen ion entered the duodenum was uniformly suppressed for all 4 monkeys during their first session of shock avoidance; during their last session, the gastric subgroup continued to show suppression while the duodenal subgroup returned towards control levels. Serum gastrin levels were unchanged by the multiple-stress procedures. Our finding of consistently producible, stress-induced gastroduodenal pathology in anatomic areas similar to those involved in man suggests that the subhuman primate is suitable for further efforts to produce an animal model of psychosomatic ulcer disease.
|
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] |
PMID:21565
|
The natural history of neoplasia. Newer insights into an old problem.
|
The stages of initiation and promotion in the natural history of epidermal carcinogenesis have been known for many years. Recently, experimental systems other than skin have been shown to exhibit similar, if not completely analogous, stages in the natural history of neoplasia. In particular, the demonstration by Peraino and his associates that phenobarbital may enhance the production of hepatomas by a relatively subcarcinogenic dose of acetylaminofluorene was one of the first demonstrations of stages occurring in an extraepidermal neoplasm. Studies reported in this paper have demonstrated that administration of phenobarbital (0.05% in the diet) for 6 months following a single dose of diethylnitrosamine (5 to 10 mg/kg) given within 24 hours after partial hepatectomy resulted in a marked increase in the number of enzyme-altered foci in the liver as well as in the production of hepatocellular carcinomas. This was compared to animals receiving only a single dose of diethylnitrosamine following partial hepatectomy with no further treatment, in which only a relatively small number of foci were evident in the absence of phenobarbital feeding. Using three different enzyme markers, a distinct degree of phenotypic heterogeneity of the enzyme-altered foci in liver was demonstrated. These studies have shown that liver carcinogensis can be readily divided into two stages: a) initiation by a single dose of diethylnitrosamine following partial hepatectomy and b) promotion by the continuous feeding of phenobarbital. Furthermore, the immediate progeny of the initiated cells, the enzyme-altered focus, may be recognized by suitable microscopic means prior to the formation of gross lesions as required in the skin system. These initiated cell populations exhibit a degree of biochemical heterogeneity which reflects that seen in fully developed hepatic neoplasms, suggesting that promotion and progression in this system does not significantly alter the basic biochemical characteristics of the initiated cell.
|
The natural history of neoplasia. Newer insights into an old problem. The stages of initiation and promotion in the natural history of epidermal carcinogenesis have been known for many years. Recently, experimental systems other than skin have been shown to exhibit similar, if not completely analogous, stages in the natural history of neoplasia. In particular, the demonstration by Peraino and his associates that phenobarbital may enhance the production of hepatomas by a relatively subcarcinogenic dose of acetylaminofluorene was one of the first demonstrations of stages occurring in an extraepidermal neoplasm. Studies reported in this paper have demonstrated that administration of phenobarbital (0.05% in the diet) for 6 months following a single dose of diethylnitrosamine (5 to 10 mg/kg) given within 24 hours after partial hepatectomy resulted in a marked increase in the number of enzyme-altered foci in the liver as well as in the production of hepatocellular carcinomas. This was compared to animals receiving only a single dose of diethylnitrosamine following partial hepatectomy with no further treatment, in which only a relatively small number of foci were evident in the absence of phenobarbital feeding. Using three different enzyme markers, a distinct degree of phenotypic heterogeneity of the enzyme-altered foci in liver was demonstrated. These studies have shown that liver carcinogensis can be readily divided into two stages: a) initiation by a single dose of diethylnitrosamine following partial hepatectomy and b) promotion by the continuous feeding of phenobarbital. Furthermore, the immediate progeny of the initiated cells, the enzyme-altered focus, may be recognized by suitable microscopic means prior to the formation of gross lesions as required in the skin system. These initiated cell populations exhibit a degree of biochemical heterogeneity which reflects that seen in fully developed hepatic neoplasms, suggesting that promotion and progression in this system does not significantly alter the basic biochemical characteristics of the initiated cell.
|
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] |
PMID:21566
|
Kinetic differences between uterine and renal renins in the dog.
|
It has previously been reported that in the dog the Michaelis constant (Km) of renal renin with respect to homologous plasma substrate is the same as that for the angiotensin-generating enzyme extractable from the myometrium during pregnancy. We have reexamined the kinetics with a different methodology. Enzyme preparations from dog kidneys, the uterus of a nonnephrectomized pregnant bitch, and the uterus of a nephrectomized pregnant bitch were used. The substrate, prepared by diluting plasma from a nephrectomized, hysterectomized pregnant bitch, had a maximum endogenous substrate concentration of 1,750 ng/ml angiotensin I equivalents. Lineweaver-Burk kinetic analysis was used to calculate the Michaelis constants at pH 6.2. The Km of the renal enzyme was 720 ng/ml, of the enzyme prepared from the nonnephrectomized uterus 1,570 ng/ml, and of the nephrectomized dog uterus 1,500 ng/ml. The uterine enzyme was not activated by transient acid treatment, in contrast to renins reported from other sources. The data support the concept of biochemical heterogeneity of the two enzymes in the dog.
|
Kinetic differences between uterine and renal renins in the dog. It has previously been reported that in the dog the Michaelis constant (Km) of renal renin with respect to homologous plasma substrate is the same as that for the angiotensin-generating enzyme extractable from the myometrium during pregnancy. We have reexamined the kinetics with a different methodology. Enzyme preparations from dog kidneys, the uterus of a nonnephrectomized pregnant bitch, and the uterus of a nephrectomized pregnant bitch were used. The substrate, prepared by diluting plasma from a nephrectomized, hysterectomized pregnant bitch, had a maximum endogenous substrate concentration of 1,750 ng/ml angiotensin I equivalents. Lineweaver-Burk kinetic analysis was used to calculate the Michaelis constants at pH 6.2. The Km of the renal enzyme was 720 ng/ml, of the enzyme prepared from the nonnephrectomized uterus 1,570 ng/ml, and of the nephrectomized dog uterus 1,500 ng/ml. The uterine enzyme was not activated by transient acid treatment, in contrast to renins reported from other sources. The data support the concept of biochemical heterogeneity of the two enzymes in the dog.
|
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] |
PMID:21567
|
pH-temperature dependence of organic acid transport in rat kidney slices.
|
Uptake of p-aminohippurate (PAH) by rat kidney slices was optimal at a pH of around 7.3 when incubation temperature was 37 degrees C. At 25 degrees C, however, optimal pH was increased to about 7.5. When the uptake was evaluated as a function of the OH/H ratio of the medium, it was found that optimal uptake is at a constant OH/H. Data showing the same characteristics of OH/H dependence were also observed for phenol red. Passive uptake of PAH was pH independent. Efflux rates for PAH also showed the pH optimal which was identical to that of uptake. At both 25 and 37 degrees C the Km appeared to be independent of pH while the maximum rate of active influx. An optimal OH/H ratio determines maximal velocity and this ratio is fixed and independent of temperature.
|
pH-temperature dependence of organic acid transport in rat kidney slices. Uptake of p-aminohippurate (PAH) by rat kidney slices was optimal at a pH of around 7.3 when incubation temperature was 37 degrees C. At 25 degrees C, however, optimal pH was increased to about 7.5. When the uptake was evaluated as a function of the OH/H ratio of the medium, it was found that optimal uptake is at a constant OH/H. Data showing the same characteristics of OH/H dependence were also observed for phenol red. Passive uptake of PAH was pH independent. Efflux rates for PAH also showed the pH optimal which was identical to that of uptake. At both 25 and 37 degrees C the Km appeared to be independent of pH while the maximum rate of active influx. An optimal OH/H ratio determines maximal velocity and this ratio is fixed and independent of temperature.
|
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] |
PMID:21568
|
Ionic composition of arterial and mixed venous plasma in the unanesthetized dog.
|
The use of permanent catheters in the aorta and pulmonary artery permitted the establishment of normal values for hemoglobin concentration in blood and -or pH, PCO2, osmolality, and protein and electrolyte concentrations in the plasma of arterial and venous samples from unanesthetized, undisturbed dogs, and the comparison of the ionic composition of simultaneously taken arterial and venous samples. Arterial samples yielded the following mean values: CHb, 143 g liter-1; pHP, 7.427; PCO2, 32.5 mmHg; CPosmol, 295 mmol kg-1; CPpr, 73.1 g liter-1, CPNa+, 148.0; CPK+, 3.9; CPCa2+, 2.38; CPMg2+, 0.85; CPCl-, 116.0; CPHCO3-, 21.1; CPlact-, 1.4; CPphosph, 1.21; net cation equivalency, 16.4; and anion gap, 1.03 mmol liter-1 in eight male mongrel dogs with seven or eight samplings from each dog. The anion gap in arterial and venous plasma was small, indicating that the contribution of sulfate and organic acids to the ionic composition of dog plasma is quantitatively unimportant. In simultaneously taken arterial and venous samples the following significant arteriovenous differences were found: HP, +0.038; PCO2, -5.6 mmHg; CPosmol, -1.8 mmol kg-1; protein, -0.8 g liter-1; CPNa+, -1.0; CPK+, -0.1; CPCl-, +1.3; and CPHCO3-, -1.7 mmol liter-1. These differences are explained on the basis of the changes that occur in blood upon the addition of CO2 and the ensuing chloride and water shifts.
|
Ionic composition of arterial and mixed venous plasma in the unanesthetized dog. The use of permanent catheters in the aorta and pulmonary artery permitted the establishment of normal values for hemoglobin concentration in blood and -or pH, PCO2, osmolality, and protein and electrolyte concentrations in the plasma of arterial and venous samples from unanesthetized, undisturbed dogs, and the comparison of the ionic composition of simultaneously taken arterial and venous samples. Arterial samples yielded the following mean values: CHb, 143 g liter-1; pHP, 7.427; PCO2, 32.5 mmHg; CPosmol, 295 mmol kg-1; CPpr, 73.1 g liter-1, CPNa+, 148.0; CPK+, 3.9; CPCa2+, 2.38; CPMg2+, 0.85; CPCl-, 116.0; CPHCO3-, 21.1; CPlact-, 1.4; CPphosph, 1.21; net cation equivalency, 16.4; and anion gap, 1.03 mmol liter-1 in eight male mongrel dogs with seven or eight samplings from each dog. The anion gap in arterial and venous plasma was small, indicating that the contribution of sulfate and organic acids to the ionic composition of dog plasma is quantitatively unimportant. In simultaneously taken arterial and venous samples the following significant arteriovenous differences were found: HP, +0.038; PCO2, -5.6 mmHg; CPosmol, -1.8 mmol kg-1; protein, -0.8 g liter-1; CPNa+, -1.0; CPK+, -0.1; CPCl-, +1.3; and CPHCO3-, -1.7 mmol liter-1. These differences are explained on the basis of the changes that occur in blood upon the addition of CO2 and the ensuing chloride and water shifts.
|
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] |
PMID:21569
|
Mechanism of active chloride transport by urinary bladder of the Colombian toad.
|
In order to investigate the mechanism of active chloride transport, experiments were performed to verify the previous report that removal of potassium from the solutions bathing the urinary bladders of Colombian toads induced a reversal of short-circuit current (RSCC) and active chloride transport. The present experiments confirmed these findings. However, there was no correlation between the magnitude of the RSCC and the chloride transport. Furthermore, removal of chloride from the mucosal bathing solutions did not markedly affect the RSCC, indicating that the chloride transport was electrically silent. Removal of bicarbonate from the bathing solutions eliminated the RSCC, and acetazolamide inhibited both the RSCC and net chloride transport. These findings suggest a relationship between chloride and hydrogen ion transport. Inhibition of sodium transport by removal of sodium or addition of ouabain or amiloride has previously been shown to produce an RSCC. In the present experiments these same maneuvers caused an RSCC and induced net chloride transport. It is concluded that active chloride transport by Colombian toad bladder is stimulated by inhibition of sodium transport. The inhibition of active chloride transport by acetazolamide suggests a possible role for carbonic anhydrase in this process.
|
Mechanism of active chloride transport by urinary bladder of the Colombian toad. In order to investigate the mechanism of active chloride transport, experiments were performed to verify the previous report that removal of potassium from the solutions bathing the urinary bladders of Colombian toads induced a reversal of short-circuit current (RSCC) and active chloride transport. The present experiments confirmed these findings. However, there was no correlation between the magnitude of the RSCC and the chloride transport. Furthermore, removal of chloride from the mucosal bathing solutions did not markedly affect the RSCC, indicating that the chloride transport was electrically silent. Removal of bicarbonate from the bathing solutions eliminated the RSCC, and acetazolamide inhibited both the RSCC and net chloride transport. These findings suggest a relationship between chloride and hydrogen ion transport. Inhibition of sodium transport by removal of sodium or addition of ouabain or amiloride has previously been shown to produce an RSCC. In the present experiments these same maneuvers caused an RSCC and induced net chloride transport. It is concluded that active chloride transport by Colombian toad bladder is stimulated by inhibition of sodium transport. The inhibition of active chloride transport by acetazolamide suggests a possible role for carbonic anhydrase in this process.
|
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] |
PMID:21570
|
The brain as a dream state generator: an activation-synthesis hypothesis of the dream process.
|
Recent research in the neurobiology of dreaming sleep provides new evidence for possible structural and functional substrates of formal aspects of the dream process. The data suggest that dreaming sleep is physiologically determined and shaped by a brain stem neuronal mechanism that can be modeled physiologically and mathematically. Formal features of the generator processes with strong implications for dream theory include periodicity and automaticity of forebrain activation, suggesting a preprogrammed neural basis for dream mentation in sleep; intense and sporadic activation of brain stem sensorimotor circuits including reticular, oculomotor, and vestibular neurons, possibly determining spatiotemporal aspects of dream imagery; and shifts in transmitter ratios, possibly accounting for dream amnesia. The authors suggest that the automatically activated forebrain synthesizes the dream by comparing information generated in specific brain stem circuits with information stored in memory.
|
The brain as a dream state generator: an activation-synthesis hypothesis of the dream process. Recent research in the neurobiology of dreaming sleep provides new evidence for possible structural and functional substrates of formal aspects of the dream process. The data suggest that dreaming sleep is physiologically determined and shaped by a brain stem neuronal mechanism that can be modeled physiologically and mathematically. Formal features of the generator processes with strong implications for dream theory include periodicity and automaticity of forebrain activation, suggesting a preprogrammed neural basis for dream mentation in sleep; intense and sporadic activation of brain stem sensorimotor circuits including reticular, oculomotor, and vestibular neurons, possibly determining spatiotemporal aspects of dream imagery; and shifts in transmitter ratios, possibly accounting for dream amnesia. The authors suggest that the automatically activated forebrain synthesizes the dream by comparing information generated in specific brain stem circuits with information stored in memory.
|
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] |
PMID:21571
|
An evaluation of a unique new antipsychotic agent, sulpiride: effects on serum prolactin and growth hormone levels.
|
Sulpiride is a new and chemically different neuroleptic. In a study of 16 severely ill schizophrenic patients, this compound displayed definite evidence of antipsychotic activity while producing few adverse reactions. The relative absence of extrapyramidal side effects may indicate that sulpride has a low potential for producing tardive dyskinesia. Two patients who did not develop significant increases in prolactin levels did show a definite therapeutic response to sulpiride. Thus it appears that central dopaminergic blockade in the hypothalamic area is not a prerequisite for antipsychotic activity.
|
An evaluation of a unique new antipsychotic agent, sulpiride: effects on serum prolactin and growth hormone levels. Sulpiride is a new and chemically different neuroleptic. In a study of 16 severely ill schizophrenic patients, this compound displayed definite evidence of antipsychotic activity while producing few adverse reactions. The relative absence of extrapyramidal side effects may indicate that sulpride has a low potential for producing tardive dyskinesia. Two patients who did not develop significant increases in prolactin levels did show a definite therapeutic response to sulpiride. Thus it appears that central dopaminergic blockade in the hypothalamic area is not a prerequisite for antipsychotic activity.
|
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] |
PMID:21572
|
[Long-term study on the influence of running exercise, food restriction, running exercise + food restriction and of parenterally applicated testis cells on age-parameters of the rat (author's transl)].
|
Preliminary results of a long-term study on 1100 male Sprague-Dawley rats are presented, showing the influence of running exercise, of restricted diet, of both of these together and of the s.c. application of lypholized testis cells. Up to now animals aged 9, 15 and 24 mths were investigated. The test-animals were exposed to the experimental conditions from their 6th month of life. The running exercise was carried out on a treadmill (30 m at a speed of 25 m/min, horizontal) 5 days a week. A food restriction of about 20% was achieved by 2 fasting-days a week. The lyophilized testis cells were injected for the first time at an age of 9 mths and afterwards in 6 mths intervals. Between the injection and the assessment of the age-parameters there was an interval of 6 mths. S o fare the following parameters of the comprehensive test-program have been evaluated: 1) running performance in m (treadmill, 20 m/min, ascent 15 degrees), 2) motor activity (Animex Activity Meter), 3) chemical contraction-relaxation of tail-tendon-fibers, 4) total lipids and total cholesterol in the plasma. The results obtained so far show that a mild regular training, moderate food restriction and the s.c. application of testis-cells are able to cause a significant shift in the dirction of a younger biological age in at least some of the age parameters. The action of the testis-cells seems to affect most of the age parameters but shows the tendency to be more distinct at a higher age. More concrete statements about the influence on the biological age or the vitality will only be possible after the multivariate analysis of the results.
|
[Long-term study on the influence of running exercise, food restriction, running exercise + food restriction and of parenterally applicated testis cells on age-parameters of the rat (author's transl)]. Preliminary results of a long-term study on 1100 male Sprague-Dawley rats are presented, showing the influence of running exercise, of restricted diet, of both of these together and of the s.c. application of lypholized testis cells. Up to now animals aged 9, 15 and 24 mths were investigated. The test-animals were exposed to the experimental conditions from their 6th month of life. The running exercise was carried out on a treadmill (30 m at a speed of 25 m/min, horizontal) 5 days a week. A food restriction of about 20% was achieved by 2 fasting-days a week. The lyophilized testis cells were injected for the first time at an age of 9 mths and afterwards in 6 mths intervals. Between the injection and the assessment of the age-parameters there was an interval of 6 mths. S o fare the following parameters of the comprehensive test-program have been evaluated: 1) running performance in m (treadmill, 20 m/min, ascent 15 degrees), 2) motor activity (Animex Activity Meter), 3) chemical contraction-relaxation of tail-tendon-fibers, 4) total lipids and total cholesterol in the plasma. The results obtained so far show that a mild regular training, moderate food restriction and the s.c. application of testis-cells are able to cause a significant shift in the dirction of a younger biological age in at least some of the age parameters. The action of the testis-cells seems to affect most of the age parameters but shows the tendency to be more distinct at a higher age. More concrete statements about the influence on the biological age or the vitality will only be possible after the multivariate analysis of the results.
|
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] |
PMID:21573
|
Effect of aminoglutethimide on calcitonin sensitivity.
|
Studying the influence of aminoglutethimide treatment on calcitonin sensitivity, serum calcium levels were measured in normal rats, aminoglutethimide treated rats, adrenalectomized and castrated rats, following i.v. injection of porcine calcitonin. The aminoglutethimide administration caused a more distinct calcitonin resistance than surgical sex hormone deprivation. It was found that castration in most of experiments failed to enhance calcitonin resistance in aminoglutethimide treated animals. The aminoglutethimide administration may influence the steroid synthesis of adrenals, accessory adrenals and testes too. Results obtained with castrated, aminoglutethimide plus dehydroepiandrosterone treated and castrated, aminoglutethimide plus epiandrosterone treated rats affirm the earlier observations, i.e. in state of sex hormone deficiency the exogeneous androgens have an enhancing or restoring effect on calcitonin sensitivity.
|
Effect of aminoglutethimide on calcitonin sensitivity. Studying the influence of aminoglutethimide treatment on calcitonin sensitivity, serum calcium levels were measured in normal rats, aminoglutethimide treated rats, adrenalectomized and castrated rats, following i.v. injection of porcine calcitonin. The aminoglutethimide administration caused a more distinct calcitonin resistance than surgical sex hormone deprivation. It was found that castration in most of experiments failed to enhance calcitonin resistance in aminoglutethimide treated animals. The aminoglutethimide administration may influence the steroid synthesis of adrenals, accessory adrenals and testes too. Results obtained with castrated, aminoglutethimide plus dehydroepiandrosterone treated and castrated, aminoglutethimide plus epiandrosterone treated rats affirm the earlier observations, i.e. in state of sex hormone deficiency the exogeneous androgens have an enhancing or restoring effect on calcitonin sensitivity.
|
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] |
PMID:21574
|
[General directions for the care of the dying patient by the Swiss Academy of Medical Sciences (author's transl)].
|
The medical ethical and legal principles for the care of the dying are given, different for the patient with judgement and for the inconscieous patient. Contrary to the patient in danger of death (Moriturus) where the doctor has the duty to save the live, in the case of the dying (Moribundus), where the disease is irreversible and the prognosis irremediable, passive euthanasy is permitted. Here the human assistance from man to man is essential. In a separate paper of the directions answer is given to special medical questions in the care of the dying.
|
[General directions for the care of the dying patient by the Swiss Academy of Medical Sciences (author's transl)]. The medical ethical and legal principles for the care of the dying are given, different for the patient with judgement and for the inconscieous patient. Contrary to the patient in danger of death (Moriturus) where the doctor has the duty to save the live, in the case of the dying (Moribundus), where the disease is irreversible and the prognosis irremediable, passive euthanasy is permitted. Here the human assistance from man to man is essential. In a separate paper of the directions answer is given to special medical questions in the care of the dying.
|
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] |
PMID:21575
|
[Somatical performance and environment in the elderly (author's transl)].
|
The arrangement with the envirement requires somatical performance of men. We developed a mathematical model by recording data of performance. This model shows a decreasing capacity of performance correlated with age. The real meassured data are vastly different to theoretical conception. In difference to the assumption equal increasing load in elder and younger people leads to equal acceleration of pulse rate we are able to demonstrate decreasing physical strength is clutched to shortage of weight. The loss of performance in elderly men requires a restriction either of social contacts or of supply and consumption. For preserve of physical strength there is required to treat malnutrition and especially to promote vigour, substance an strength of muscles.
|
[Somatical performance and environment in the elderly (author's transl)]. The arrangement with the envirement requires somatical performance of men. We developed a mathematical model by recording data of performance. This model shows a decreasing capacity of performance correlated with age. The real meassured data are vastly different to theoretical conception. In difference to the assumption equal increasing load in elder and younger people leads to equal acceleration of pulse rate we are able to demonstrate decreasing physical strength is clutched to shortage of weight. The loss of performance in elderly men requires a restriction either of social contacts or of supply and consumption. For preserve of physical strength there is required to treat malnutrition and especially to promote vigour, substance an strength of muscles.
|
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] |
PMID:21576
|
Thermal contraction-relaxation and dissolution of rat tail tendon collagen in different ages.
|
Here thermal contraction-relaxation and dissolution of collagen from rat tail tendon fibers were studied simultaneously. It is concluded that both processes change their character in the same manner with increasing age and that the relaxation phase probably is a result of the dissolution of collagen from the fibers.
|
Thermal contraction-relaxation and dissolution of rat tail tendon collagen in different ages. Here thermal contraction-relaxation and dissolution of collagen from rat tail tendon fibers were studied simultaneously. It is concluded that both processes change their character in the same manner with increasing age and that the relaxation phase probably is a result of the dissolution of collagen from the fibers.
|
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] |
PMID:21577
|
[Acute confusional states in the elderly (author's transl)].
|
It is described that the acute confusional state in elderly people can be regarded as the combination of a catastrophe-reaction, superimposed on the primary symptoms of an underlying physical disorder. The catastrophe-reaction, the secondary symptoms, are described as the manifestation of the failure of the hypothalamo-hypophyso-adrenocortical axis, comparable with a relative glucocorticosteroid deficiency. The therapy of an acute confusional state should aim at the treatment of the underlying physical disorder and at the treatment of the catastrophereaction by injecting intramuscularly for two days high doses of methyl-prednisolon. It is demonstrated that frequently in elderly people a subacute confusion can be developed, due to a catastrophe-reaction, superimposed on an underlying masked depression. A combined treatment, giving for two days high doses of methyl-prednisolon in order to treat the catastrophe-reaction and giving low doses of tricyclic anti-depressants to treat the depression, seems to result in amazing improvement.
|
[Acute confusional states in the elderly (author's transl)]. It is described that the acute confusional state in elderly people can be regarded as the combination of a catastrophe-reaction, superimposed on the primary symptoms of an underlying physical disorder. The catastrophe-reaction, the secondary symptoms, are described as the manifestation of the failure of the hypothalamo-hypophyso-adrenocortical axis, comparable with a relative glucocorticosteroid deficiency. The therapy of an acute confusional state should aim at the treatment of the underlying physical disorder and at the treatment of the catastrophereaction by injecting intramuscularly for two days high doses of methyl-prednisolon. It is demonstrated that frequently in elderly people a subacute confusion can be developed, due to a catastrophe-reaction, superimposed on an underlying masked depression. A combined treatment, giving for two days high doses of methyl-prednisolon in order to treat the catastrophe-reaction and giving low doses of tricyclic anti-depressants to treat the depression, seems to result in amazing improvement.
|
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] |
PMID:21596
|
Estimation of plasma lorazepam by gas-liquid chromatography and a benzene extraction.
|
Plasma lorazepam can be easily and reliably estimated using a simple single stage benzene extraction and gas-lizuid chromatography with flunitrazepam as the internal standard. Reproducible results within the clinical range of plasma levels were obtained. This method permits the extraction and chromatographing of 40 samples in approximately 7 1/2 hours.
|
Estimation of plasma lorazepam by gas-liquid chromatography and a benzene extraction. Plasma lorazepam can be easily and reliably estimated using a simple single stage benzene extraction and gas-lizuid chromatography with flunitrazepam as the internal standard. Reproducible results within the clinical range of plasma levels were obtained. This method permits the extraction and chromatographing of 40 samples in approximately 7 1/2 hours.
|
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] |
PMID:21603
|
Determinants of lung bacterial clearance in mice after acute hypoxia.
|
Net lung bacterial clearance in normal mice is determined by the balance of in vivo bacterial multiplication on the one hand, and the defense mechanisms of mucociliary clearance and phagocytosis and killing by the oxygen-dependent alveolar macrophage on the other. The bactericidal function of the macrophage is the major component of the defense mechanism. The effect of acute hypoxia on the defense mechanism was studied in mice exposed to aerosols of Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, and Streptococcus pneumoniae. Physical clearance was not impaired by acute hypoxia, and bacterial replication was not stimulated by the low oxygen atmosphere. Clearance of Staphylococcus aureus, Klebsiella pneumoniae, and Escherichia coli was impaired during acute hypoxia due to decreased phagocytosis or killing by the alveolar macrophage. The important human pathogen Streptococcus pneumoniae was cleared normally in the presence of acute hypoxia. This observation suggests that an oxygen-independent clearance mechanism is important in lung defense against the pneumococcus. This may be a separate mechanism within the alveolar macrophage or a system as yet unidentified.
|
Determinants of lung bacterial clearance in mice after acute hypoxia. Net lung bacterial clearance in normal mice is determined by the balance of in vivo bacterial multiplication on the one hand, and the defense mechanisms of mucociliary clearance and phagocytosis and killing by the oxygen-dependent alveolar macrophage on the other. The bactericidal function of the macrophage is the major component of the defense mechanism. The effect of acute hypoxia on the defense mechanism was studied in mice exposed to aerosols of Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, and Streptococcus pneumoniae. Physical clearance was not impaired by acute hypoxia, and bacterial replication was not stimulated by the low oxygen atmosphere. Clearance of Staphylococcus aureus, Klebsiella pneumoniae, and Escherichia coli was impaired during acute hypoxia due to decreased phagocytosis or killing by the alveolar macrophage. The important human pathogen Streptococcus pneumoniae was cleared normally in the presence of acute hypoxia. This observation suggests that an oxygen-independent clearance mechanism is important in lung defense against the pneumococcus. This may be a separate mechanism within the alveolar macrophage or a system as yet unidentified.
|
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] |
PMID:21604
|
Detection of penumococcal antigens in the sputum in pneumococcal pneumonia.
|
Forty-seven patients with bacterial pneumonia were grouped by use of clinical criteria according to the relative certainty of a diagnosis of pneumococcal pneumonia. Sputums were tested for pneumococcal antigens by counterimmunoelectrophoresis with polyvalent pneumococcal antiserum. Antigens were detected in the sputum of 29 of 39 patients with evidence of pneumococcal pneumonia, and there was good correlation between the detection of antigens and the degree of certainty of the clinical diagnosis. Antigens persisted briefly in the sputum during therapy with antimicrobial drugs and could be detected during the first 48 hours of therapy in most cases of pneumococcal pneumonia. Pneumococci were isolated from the sputum in only 18 of 39 cases of pneumococcal pneumonia, and sputum cultures did not correlate as well as counterimmunoelectrophoresis with clinical diagnoses. In studies of 27 patients with chronic bronchitis without pneumonia, pneumococci were isolated from sputum in 10 cases, whereas counterimmunoelectrophoresis was positive in 5 cases. Counterimmunoelectrophoresis provides a simple and rapid method for detecting pneumococcal antigens in sputum, and it appears to be more reliable than sputum cultures in establishing a presumptive diagnosis in pneumococcal pneumonia.
|
Detection of penumococcal antigens in the sputum in pneumococcal pneumonia. Forty-seven patients with bacterial pneumonia were grouped by use of clinical criteria according to the relative certainty of a diagnosis of pneumococcal pneumonia. Sputums were tested for pneumococcal antigens by counterimmunoelectrophoresis with polyvalent pneumococcal antiserum. Antigens were detected in the sputum of 29 of 39 patients with evidence of pneumococcal pneumonia, and there was good correlation between the detection of antigens and the degree of certainty of the clinical diagnosis. Antigens persisted briefly in the sputum during therapy with antimicrobial drugs and could be detected during the first 48 hours of therapy in most cases of pneumococcal pneumonia. Pneumococci were isolated from the sputum in only 18 of 39 cases of pneumococcal pneumonia, and sputum cultures did not correlate as well as counterimmunoelectrophoresis with clinical diagnoses. In studies of 27 patients with chronic bronchitis without pneumonia, pneumococci were isolated from sputum in 10 cases, whereas counterimmunoelectrophoresis was positive in 5 cases. Counterimmunoelectrophoresis provides a simple and rapid method for detecting pneumococcal antigens in sputum, and it appears to be more reliable than sputum cultures in establishing a presumptive diagnosis in pneumococcal pneumonia.
|
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] |
PMID:21606
|
Regional quality control survey of blood-gas analysis.
|
We undertook an external quality control survey of blood-gas analysis in 16 laboratories at 13 hospitals. All samples were prepared in the laboratories under investigation by equilibration of blood or serum with gas mixtures of known composition. pH of serum was measured with no significant bias but with an SD of random error 0.026 pH units, which was almost twice the SD of the reference range (0.015). An acceptable random error (half SD of reference range) was not obtained in a longitudinal internal quality control suvey although there were acceptable results for buffer pH in both field and internal surveys. Blood PO2 was measured with no significant bias but with SD of random error 1.38 kPa which reduced to 0.72 kPa by excluding one egregious result. The latter value was just over half of the SD of the reference range (1.2 kPa). PCO2 of blood was also measured without significant bias but with a much smaller SD of random error of 0.28 kPa (by excluding one egregious result), which was again just over half the SD of the reference range (0.51 kPa). Measurements of blood PO2 and PCO2 seem generally acceptable in relation to their respective reference ranges but measurements of pH were unsatisfactory in both internal and external trials.
|
Regional quality control survey of blood-gas analysis. We undertook an external quality control survey of blood-gas analysis in 16 laboratories at 13 hospitals. All samples were prepared in the laboratories under investigation by equilibration of blood or serum with gas mixtures of known composition. pH of serum was measured with no significant bias but with an SD of random error 0.026 pH units, which was almost twice the SD of the reference range (0.015). An acceptable random error (half SD of reference range) was not obtained in a longitudinal internal quality control suvey although there were acceptable results for buffer pH in both field and internal surveys. Blood PO2 was measured with no significant bias but with SD of random error 1.38 kPa which reduced to 0.72 kPa by excluding one egregious result. The latter value was just over half of the SD of the reference range (1.2 kPa). PCO2 of blood was also measured without significant bias but with a much smaller SD of random error of 0.28 kPa (by excluding one egregious result), which was again just over half the SD of the reference range (0.51 kPa). Measurements of blood PO2 and PCO2 seem generally acceptable in relation to their respective reference ranges but measurements of pH were unsatisfactory in both internal and external trials.
|
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] |
PMID:21607
|
A long-term evaluation of the CentrifiChem centrifugal microanalyser during routine use in the clinical biochemistry laboratory.
|
The CentrifiChem centrifugal microanalyser (model 300) has been used routinely in this laboratory for two years. Its reliability for performing total bilirubin, aspartate aminotransferase alkaline phosphatase and gamma-glutamyl transferase analyses during this time is reported, using methods modified in a preliminary study. Accuracy of the analyses has been assessed by comparing the results with those from other analytical systems and by using commercial control sera. Determinations have been made for within-batch, between-day (20 days), and long term (100 weeks) precision. Other aspects evaluated were the range over which methods were linear and the cost of operation.
|
A long-term evaluation of the CentrifiChem centrifugal microanalyser during routine use in the clinical biochemistry laboratory. The CentrifiChem centrifugal microanalyser (model 300) has been used routinely in this laboratory for two years. Its reliability for performing total bilirubin, aspartate aminotransferase alkaline phosphatase and gamma-glutamyl transferase analyses during this time is reported, using methods modified in a preliminary study. Accuracy of the analyses has been assessed by comparing the results with those from other analytical systems and by using commercial control sera. Determinations have been made for within-batch, between-day (20 days), and long term (100 weeks) precision. Other aspects evaluated were the range over which methods were linear and the cost of operation.
|
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] |
PMID:21608
|
The genetic and biochemical properties of the D-amino acid oxidases in human tissues.
|
1. Two distinct forms of oxidases catalysing the oxidative deamidation of D-alpha-amino acids have been identified in human tissues: D-amino acid oxidase (DAMOX) and D-aspartate oxidase (DASOX). 2. The enzymes differ in their electrophoretic properties, tissue distribution, binding with flavine adenine denucleotide, heat stability, molecular size and possibly in subunit structure. 3. Neither enzyme exhibits genetic polymorphism in European populations, but a rare electrophoretic variant phenotype (DASOX 2-1) was identified which suggests that the DASOX locus is autosomal and independent of the DAMOX locus.
|
The genetic and biochemical properties of the D-amino acid oxidases in human tissues. 1. Two distinct forms of oxidases catalysing the oxidative deamidation of D-alpha-amino acids have been identified in human tissues: D-amino acid oxidase (DAMOX) and D-aspartate oxidase (DASOX). 2. The enzymes differ in their electrophoretic properties, tissue distribution, binding with flavine adenine denucleotide, heat stability, molecular size and possibly in subunit structure. 3. Neither enzyme exhibits genetic polymorphism in European populations, but a rare electrophoretic variant phenotype (DASOX 2-1) was identified which suggests that the DASOX locus is autosomal and independent of the DAMOX locus.
|
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] |
PMID:21605
|
[Anhidrotic activity of anticholinergics in mice. Influence of mode of administration].
|
Anhidrotic concentrations 50 of four anticholinergics were determined in mice using an original skin palmar conductance method. Intra-peritoneal, intra-palmar and topical routes were compared. Relative anhidrotic activities according to the mode of administration of the different anticholinergics showed definite differences, e.g. hydroxyzine, which was the less active per i.p. route, was the most active per topical route. The method is likely to be a useful test for selecting the most active anticholinergic and its most efficient route of administration for the treatment of hyperhidrosis.
|
[Anhidrotic activity of anticholinergics in mice. Influence of mode of administration]. Anhidrotic concentrations 50 of four anticholinergics were determined in mice using an original skin palmar conductance method. Intra-peritoneal, intra-palmar and topical routes were compared. Relative anhidrotic activities according to the mode of administration of the different anticholinergics showed definite differences, e.g. hydroxyzine, which was the less active per i.p. route, was the most active per topical route. The method is likely to be a useful test for selecting the most active anticholinergic and its most efficient route of administration for the treatment of hyperhidrosis.
|
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-0.021693624556064606,
-0.018608616665005684,
-0.021027833223342896,
0.0029802669305354357,
-0.019829686731100082,
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0.01963939517736435,
0.0516178235411644,
0.03094874881207943,
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0.036568429321050644,
0.1308089941740036
] |
PMID:21610
|
Hepatotoxicity of erythromycin estolate during pregnancy.
|
Women in the second half of pregnancy, who were infected with genital mycoplasmas and who gave written informed consent, were randomly assigned to receive capsules of identical appearance containing erythromycin estolate, clindamycin hydrochloride, or a placebo for 6 weeks. Levels of serum glutamic oxalacetic transaminase (SGOT) were determined before and during treatment by a fluorometric method. All pretreatment levels of SGOT were normal (<41 units). Participants who received erythromycin estolate had significantly more abnormally elevated levels of SGOT (16/161, 9.9%) than did those who received clindamycin (4/168, 2.4%, P < 0.01) or those who received placebo (3/165, 1.8%, P < 0.01). Elevated levels of SGOT ranged from 44 to 130 U. Serum bilirubin levels were normal. Gamma-glutamyl transpeptidase activity was abnormal in six of six participants who had abnormal levels of SGOT while receiving erythromycin estolate. There were few associated symptoms, and all levels of SGOT returned to normal after cessation of treatment. The treatment of pregnant women with erythromycin estolate may be inadvisable.
|
Hepatotoxicity of erythromycin estolate during pregnancy. Women in the second half of pregnancy, who were infected with genital mycoplasmas and who gave written informed consent, were randomly assigned to receive capsules of identical appearance containing erythromycin estolate, clindamycin hydrochloride, or a placebo for 6 weeks. Levels of serum glutamic oxalacetic transaminase (SGOT) were determined before and during treatment by a fluorometric method. All pretreatment levels of SGOT were normal (<41 units). Participants who received erythromycin estolate had significantly more abnormally elevated levels of SGOT (16/161, 9.9%) than did those who received clindamycin (4/168, 2.4%, P < 0.01) or those who received placebo (3/165, 1.8%, P < 0.01). Elevated levels of SGOT ranged from 44 to 130 U. Serum bilirubin levels were normal. Gamma-glutamyl transpeptidase activity was abnormal in six of six participants who had abnormal levels of SGOT while receiving erythromycin estolate. There were few associated symptoms, and all levels of SGOT returned to normal after cessation of treatment. The treatment of pregnant women with erythromycin estolate may be inadvisable.
|
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] |
PMID:21611
|
Mutation of an inosine-producing strain of Bacillus subtilis to DL-methionine sulfoxide resistance for guanosine production.
|
An inosine-producing strain of Bacillus subtilis was mutated to resistance against the antagonist of glutamine, DL-methionine sulfoxide. Among the mutants derived, guanosine producers were observed frequently. The best strain, 14119, produced 9.6 g of guanosine per liter at a weight yield of 12% from consumed sugar. Inosine production decreased concomitantly. When resistance was increased further by exposure to higher doses of DL-methionine sulfoxide, another strain, AG169, was obtained that did not excrete inosine but produced increased amounts of xanthosine. In these strains, the specific activity of 5'-nucleotidase was lower and that of inosine 5'-monophosphate (IMP) dehydrogenase was higher than the parent strain. It is speculated that the metabolic flow from IMP to xanthosine 5'-monophosphate proceeds more smoothly than that from IMP to inosine and yields more xanthosine and guanosine.
|
Mutation of an inosine-producing strain of Bacillus subtilis to DL-methionine sulfoxide resistance for guanosine production. An inosine-producing strain of Bacillus subtilis was mutated to resistance against the antagonist of glutamine, DL-methionine sulfoxide. Among the mutants derived, guanosine producers were observed frequently. The best strain, 14119, produced 9.6 g of guanosine per liter at a weight yield of 12% from consumed sugar. Inosine production decreased concomitantly. When resistance was increased further by exposure to higher doses of DL-methionine sulfoxide, another strain, AG169, was obtained that did not excrete inosine but produced increased amounts of xanthosine. In these strains, the specific activity of 5'-nucleotidase was lower and that of inosine 5'-monophosphate (IMP) dehydrogenase was higher than the parent strain. It is speculated that the metabolic flow from IMP to xanthosine 5'-monophosphate proceeds more smoothly than that from IMP to inosine and yields more xanthosine and guanosine.
|
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] |
PMID:21612
|
Production, purification, and characterization of alpha-amylase from Thermomonospora curvata.
|
Thermomonospora curvata produces an extracellular alpha-amylase. Maximal amylase production by cultures in a starch-mineral salts medium occurred at pH 7.5 and 53 degrees C. The crude enzyme was unstable to heating (65 degrees C) at pH 4 to 6, and was activated when heated at pH 8. The enzyme was purified 66-fold with a 9% yield and appeared homogeneous on discontinuous gel electrophoresis. The pH and temperature optima for activity of the purified enzyme were 5.5 to 6.0 and 65 degrees C. The molecular weight was calculated to be 62,000. The Km for starch was 0.39 mg/ml. The amylolytic pattern consisted of a mixture of maltotetraose and maltopentaose.
|
Production, purification, and characterization of alpha-amylase from Thermomonospora curvata. Thermomonospora curvata produces an extracellular alpha-amylase. Maximal amylase production by cultures in a starch-mineral salts medium occurred at pH 7.5 and 53 degrees C. The crude enzyme was unstable to heating (65 degrees C) at pH 4 to 6, and was activated when heated at pH 8. The enzyme was purified 66-fold with a 9% yield and appeared homogeneous on discontinuous gel electrophoresis. The pH and temperature optima for activity of the purified enzyme were 5.5 to 6.0 and 65 degrees C. The molecular weight was calculated to be 62,000. The Km for starch was 0.39 mg/ml. The amylolytic pattern consisted of a mixture of maltotetraose and maltopentaose.
|
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0.015892697498202324,
-0.08206737041473389,
-0.033316440880298615,
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-0.022898970171809196,
-0.011894059367477894,
0.10134866088628769,
-0.042530033737421036
] |
PMID:21613
|
Bacterial lipopolysaccharides as inducers of disease resistance in tobacco.
|
The cell wall component of Pseudomonas solanacearum that induces disease resistance in tobacco was highly heat stable at neutral or alkaline pH but highly labile at acid pH. Activity was unaffected by nucleases and proteases but destroyed by a mixture of beta-glycosidases. Washing of bacterial cell walls released a lipopolysaccharide (LPS) fraction with high inducer activity. Purified LPS, extracted by a variety of procedures from whole cells, isolated cell walls, and culture filtrates of both smooth and rough forms of P. solanacearum, induced disease resistance in tobacco at concentrations as low as 50 microgram/ml. The LPS from the non-plant pathogens Escherichia coli B, E. coli K, and Serratia marcescens was also active. Cell wall protein, free phospholipid, and nucleic acids were not necessary for activity. Moreover, since LPS from rough forms was active, the O-specific polysaccharide of the LPS was not required for activity. Hydrolysis of the remaining core-lipid A linkage or deacylation of lipid A destroyed inducer activity. When injected into tobacco leaves, purified LPS attached to tobacco mesophyll cell walls and induced ultrastructural changes in the host cell similar to those induced by attachment of whole heat-killed bacteria.
|
Bacterial lipopolysaccharides as inducers of disease resistance in tobacco. The cell wall component of Pseudomonas solanacearum that induces disease resistance in tobacco was highly heat stable at neutral or alkaline pH but highly labile at acid pH. Activity was unaffected by nucleases and proteases but destroyed by a mixture of beta-glycosidases. Washing of bacterial cell walls released a lipopolysaccharide (LPS) fraction with high inducer activity. Purified LPS, extracted by a variety of procedures from whole cells, isolated cell walls, and culture filtrates of both smooth and rough forms of P. solanacearum, induced disease resistance in tobacco at concentrations as low as 50 microgram/ml. The LPS from the non-plant pathogens Escherichia coli B, E. coli K, and Serratia marcescens was also active. Cell wall protein, free phospholipid, and nucleic acids were not necessary for activity. Moreover, since LPS from rough forms was active, the O-specific polysaccharide of the LPS was not required for activity. Hydrolysis of the remaining core-lipid A linkage or deacylation of lipid A destroyed inducer activity. When injected into tobacco leaves, purified LPS attached to tobacco mesophyll cell walls and induced ultrastructural changes in the host cell similar to those induced by attachment of whole heat-killed bacteria.
|
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] |
PMID:21634
|
Effect of blood transfusion in low birthweight infants.
|
143 fresh blood transfusions were given to 32 low birthweight babies, 28 of whom had hyaline membrane disease. The arterial or central venous pressure was raised by a blood transfusion if before transfusion the mean arterial pressure was less than 35 mmHg or if the diastolic central venous pressure was less than -- 0-5 mmHg. There was no effect of blood transfusion on pH. It therefore appears either that metabolic acidosis in hyaline membrane disease is not caused by poor peripheral perfusion or that blood transfusion does not increase peripheral blood flow in this condition. The safety of the procedure is assessed.
|
Effect of blood transfusion in low birthweight infants. 143 fresh blood transfusions were given to 32 low birthweight babies, 28 of whom had hyaline membrane disease. The arterial or central venous pressure was raised by a blood transfusion if before transfusion the mean arterial pressure was less than 35 mmHg or if the diastolic central venous pressure was less than -- 0-5 mmHg. There was no effect of blood transfusion on pH. It therefore appears either that metabolic acidosis in hyaline membrane disease is not caused by poor peripheral perfusion or that blood transfusion does not increase peripheral blood flow in this condition. The safety of the procedure is assessed.
|
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] |
PMID:21637
|
Effect of glucocorticoids on the hexose monophosphate pathway in human rheumatoid synovial lining cells in vitro and in vivo.
|
Human rheumatoid synovial lining cells have up to four times the capacity to oxidize glucose 6-phosphate, the first step of the hexose monophosphate pathway, as do the nonrheumatoid cells. The reducing equivalents produced by this system have many significant metabolic effects. Exposure of these cells by 10(-5) M prednisolone in vitro, or to 6 mg/day in vivo, causes some depression of this activity in the rheumatoid synovial lining cells; less than this dose of steroid, or the administration of nonsteroidal drugs in vivo, has little or no effect. The depression of activity produced by 6 mg/day does not bring this activity down to the value found in nonrheumatoid synoviocytes.
|
Effect of glucocorticoids on the hexose monophosphate pathway in human rheumatoid synovial lining cells in vitro and in vivo. Human rheumatoid synovial lining cells have up to four times the capacity to oxidize glucose 6-phosphate, the first step of the hexose monophosphate pathway, as do the nonrheumatoid cells. The reducing equivalents produced by this system have many significant metabolic effects. Exposure of these cells by 10(-5) M prednisolone in vitro, or to 6 mg/day in vivo, causes some depression of this activity in the rheumatoid synovial lining cells; less than this dose of steroid, or the administration of nonsteroidal drugs in vivo, has little or no effect. The depression of activity produced by 6 mg/day does not bring this activity down to the value found in nonrheumatoid synoviocytes.
|
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] |
PMID:21638
|
Release of antral and duodenal gastrin in response to an intestinal meal.
|
In dogs prepared with isolated, innervated antral pouches, intraduodenal perfusion with liver extract at pH 7, with the atrum buffered at pH 7, resulted in a significant release of gastrin selectively from the antrum and from the duodenum. Acidification of the meal to pH 1 abolished both antral and duodenal gastrin release, whereas acidification of the antrum abolished only the antral gastrin response. After antrectomy, liver extract at pH 7 caused a diminished but significant release of duodenal gastrin. These studies provide evidence that an intestinal meal may release (in addition to a specific intestinal phase hormone) gastrin from the intestine, and from antrum, by means of a pH-sensitive mechanism which may involve a humoral agent (enterobombesin?) from the small bowel.
|
Release of antral and duodenal gastrin in response to an intestinal meal. In dogs prepared with isolated, innervated antral pouches, intraduodenal perfusion with liver extract at pH 7, with the atrum buffered at pH 7, resulted in a significant release of gastrin selectively from the antrum and from the duodenum. Acidification of the meal to pH 1 abolished both antral and duodenal gastrin release, whereas acidification of the antrum abolished only the antral gastrin response. After antrectomy, liver extract at pH 7 caused a diminished but significant release of duodenal gastrin. These studies provide evidence that an intestinal meal may release (in addition to a specific intestinal phase hormone) gastrin from the intestine, and from antrum, by means of a pH-sensitive mechanism which may involve a humoral agent (enterobombesin?) from the small bowel.
|
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] |
PMID:21639
|
Hydroxylation of 5,5-diphenylhydantoin (phenytoin) by dog liver microsomes.
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Chang and Glazko in 1972 had reported failure to demonstrate any production from 5-5-diphenylhydantoin (phenytoin, DPH) by dog liver microsomes of either 5-(m-hydroxyphenyl-5-phenylhydantoin (m-HPPH) or 5-(P-hydroxyphenyl-5-phenylhydantoin (p-HPPH), metabolites of DPH produced by the dog in vivo. We have incubated DPH with 9,000 X g supermatants of dog liver homogenates and with suspensions of separated microsomes with added NADPH generating system, Mg(2+), and nicotinamide and have demonstrated the production of both m-HPPH and p-HPPH. Both metabolities were detected by thin-layer chromatography of extracts of the incubation mixtures. Detection and roughly quantitative measurement of m-HPPH were also accomplished with gas chromatography.
|
Hydroxylation of 5,5-diphenylhydantoin (phenytoin) by dog liver microsomes. Chang and Glazko in 1972 had reported failure to demonstrate any production from 5-5-diphenylhydantoin (phenytoin, DPH) by dog liver microsomes of either 5-(m-hydroxyphenyl-5-phenylhydantoin (m-HPPH) or 5-(P-hydroxyphenyl-5-phenylhydantoin (p-HPPH), metabolites of DPH produced by the dog in vivo. We have incubated DPH with 9,000 X g supermatants of dog liver homogenates and with suspensions of separated microsomes with added NADPH generating system, Mg(2+), and nicotinamide and have demonstrated the production of both m-HPPH and p-HPPH. Both metabolities were detected by thin-layer chromatography of extracts of the incubation mixtures. Detection and roughly quantitative measurement of m-HPPH were also accomplished with gas chromatography.
|
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] |
PMID:21641
|
[pH and lactic acid values in the cerebrospinal fluid as a diagnostic aid for differentiating viral and purulent meningoencephalitis].
|
Concentrations of lactate and pH were determined in the CSF of 20 normal children, 36 with viral meningitis, 16 with purulent meningitis and six with tuberculous meningitis. No differences were found between normal cases and those with viral meningitis. In patients with purulent meningitis concentration of lactate was above 30 mg/100 ml and pH below 7.3. Both determinations allowed differentiation between viral meningitis and purulent meningitis after 48 hr of treatment with antibiotics in 80% of the cases.
|
[pH and lactic acid values in the cerebrospinal fluid as a diagnostic aid for differentiating viral and purulent meningoencephalitis]. Concentrations of lactate and pH were determined in the CSF of 20 normal children, 36 with viral meningitis, 16 with purulent meningitis and six with tuberculous meningitis. No differences were found between normal cases and those with viral meningitis. In patients with purulent meningitis concentration of lactate was above 30 mg/100 ml and pH below 7.3. Both determinations allowed differentiation between viral meningitis and purulent meningitis after 48 hr of treatment with antibiotics in 80% of the cases.
|
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] |
PMID:21642
|
[Acid-base status in the blood of domestic pigs during exhausting locomotor stress of small and medium intensity].
|
Twenty-two fattening pigs were stressed until their rectal temperatures had reached 42 degrees C. The means of stress included walking on moving conveyor belt at ambient temperatures between 22 degrees C and 27 degrees C and relative humidities between 69 and 98%, belt speeds of 0.7 m/s, 1.3 m/s or 0.7 m/s, and stimulation by means of a forcing rod. Rectal temperature, respiratory frequency, lactic acid level in blood plasma, pH, as well as pCO2 and base excess in the blood were measured prior to, during, and after stressing. Gasping was strongly pronounced due to severe alkalisis, but it was somewhat mitigated on running on the belt with exposure to additional stress on account of lactic acid accumulation in the plasma.
|
[Acid-base status in the blood of domestic pigs during exhausting locomotor stress of small and medium intensity]. Twenty-two fattening pigs were stressed until their rectal temperatures had reached 42 degrees C. The means of stress included walking on moving conveyor belt at ambient temperatures between 22 degrees C and 27 degrees C and relative humidities between 69 and 98%, belt speeds of 0.7 m/s, 1.3 m/s or 0.7 m/s, and stimulation by means of a forcing rod. Rectal temperature, respiratory frequency, lactic acid level in blood plasma, pH, as well as pCO2 and base excess in the blood were measured prior to, during, and after stressing. Gasping was strongly pronounced due to severe alkalisis, but it was somewhat mitigated on running on the belt with exposure to additional stress on account of lactic acid accumulation in the plasma.
|
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] |
PMID:21644
|
Monoamine metabolite levels in cerebrospinal fluid of psychotic women treated with melperone or thiothixene.
|
Psychotic women with schizophrenic symptoms were treated with melperone 100 mg X 3 (n = 29) or thiothixene 10 mg X 3 (N = 34) USING A DOUBLE-BLIND PROCEDURE. Before and during treatment, levels of HVA, MOPEG, and 5-HIAA, the major metabolites of DA, NE, and 5-HT, were determined in lumbar cerebrospinal fluid by a mass fragmentographic technique. Both treatments resulted in an elevation of the HVA levels after 2 weeks, thiothixene having a more marked effect. The effect of thiothixene but not of melperone persisted after 4 weeks. Thiothixene did not influence the MOPEG level, but melperone reduced it after 4 weeks of treatment. The 5-HIAA levels were not significantly altered by the drugs. The HVA/MOPEG and the HVA/5-HIAA ratios were highly significantly elevated by both drugs after 2 as well as 4 weeks. Thiothixene induced a significantly greater change of these ratios than melperone. The results supply evidence that thiothixene accelerates central dopamine metabolism in man, presumably by blocking DA receptors. Melperone appears to act similarly, but has an effect which is weaker and/or of shorter duration. During long-term treatment with melperone the receptors develop tolerance to it. The acceleration in DA metabolism declines and the effect of melperone switches instead to central NA metabolism. The results indicate that both drugs cause long-term changes in the activity ratios of central monoamine systems. It is suggested that such changes in several systems rather than single biochemical events may be related to the antipsychotic effects of neuroleptic drugs. This study also demonstrated the versatility of using monoamine metabolite analysis of the CSF as a tool for the quantification of biochemical effects of neuroleptic drugs on the human CNS.
|
Monoamine metabolite levels in cerebrospinal fluid of psychotic women treated with melperone or thiothixene. Psychotic women with schizophrenic symptoms were treated with melperone 100 mg X 3 (n = 29) or thiothixene 10 mg X 3 (N = 34) USING A DOUBLE-BLIND PROCEDURE. Before and during treatment, levels of HVA, MOPEG, and 5-HIAA, the major metabolites of DA, NE, and 5-HT, were determined in lumbar cerebrospinal fluid by a mass fragmentographic technique. Both treatments resulted in an elevation of the HVA levels after 2 weeks, thiothixene having a more marked effect. The effect of thiothixene but not of melperone persisted after 4 weeks. Thiothixene did not influence the MOPEG level, but melperone reduced it after 4 weeks of treatment. The 5-HIAA levels were not significantly altered by the drugs. The HVA/MOPEG and the HVA/5-HIAA ratios were highly significantly elevated by both drugs after 2 as well as 4 weeks. Thiothixene induced a significantly greater change of these ratios than melperone. The results supply evidence that thiothixene accelerates central dopamine metabolism in man, presumably by blocking DA receptors. Melperone appears to act similarly, but has an effect which is weaker and/or of shorter duration. During long-term treatment with melperone the receptors develop tolerance to it. The acceleration in DA metabolism declines and the effect of melperone switches instead to central NA metabolism. The results indicate that both drugs cause long-term changes in the activity ratios of central monoamine systems. It is suggested that such changes in several systems rather than single biochemical events may be related to the antipsychotic effects of neuroleptic drugs. This study also demonstrated the versatility of using monoamine metabolite analysis of the CSF as a tool for the quantification of biochemical effects of neuroleptic drugs on the human CNS.
|
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] |
PMID:21645
|
Adult cryptorchids: Sexual development and activity.
|
Plasma testosterone in 72 unilateral and 83 bilateral cryptorchids, 11-45 years of age, revealed that, from 13 years on , values of male hormone were lower than in the control group of 69 normal boys and 85 fertile and potent men. Analysis of heterosexual development by means of the HTDM questionnaire in 49 unilateral and 57 bilateral cryptorchids, aged 21-40 years, who were examined for sterility, revealed only slight social developmental retardation compared to 184 normospermic men from sterile marriages. On the other hand, examination of sexual activity by means of the SAM questionnaire in the same three groups revealed that in cryptorchids the first ejaculation is slightly later and that the frequency of nocturnal emissions in pubescence is lower. In adulthood the patients with cryptorchidism exhibit fewer signs of high sexual activity suca as repeated coitus on the same day or a high frequency of sexual intercourse. Also, there is an earlier appearance of prolonged periods of sexual abstinence.
|
Adult cryptorchids: Sexual development and activity. Plasma testosterone in 72 unilateral and 83 bilateral cryptorchids, 11-45 years of age, revealed that, from 13 years on , values of male hormone were lower than in the control group of 69 normal boys and 85 fertile and potent men. Analysis of heterosexual development by means of the HTDM questionnaire in 49 unilateral and 57 bilateral cryptorchids, aged 21-40 years, who were examined for sterility, revealed only slight social developmental retardation compared to 184 normospermic men from sterile marriages. On the other hand, examination of sexual activity by means of the SAM questionnaire in the same three groups revealed that in cryptorchids the first ejaculation is slightly later and that the frequency of nocturnal emissions in pubescence is lower. In adulthood the patients with cryptorchidism exhibit fewer signs of high sexual activity suca as repeated coitus on the same day or a high frequency of sexual intercourse. Also, there is an earlier appearance of prolonged periods of sexual abstinence.
|
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] |
PMID:21646
|
[Ultrastructure of B-cell tumors of the pancreas (insulinomas - insular carcinoid tumors)].
|
The subcellular organization of 6 B-cell tumours of the human pancreas was studied. The tumour parenchyma consisted of processed light, dark, and moderately electron dense cells containing insulin granules. The presence of villi in the cytoplasm of tumour cells and intranuclear fibrills was demonstrated. The participation of tumour cells in formation of amyloid-like thin-fiber substance filling the intercellular and perivascular spaces was established. The similarity between endothelial cells of tumour vessels and parenchymatous cells was shown. A hypothesis on neuroectodermal origin of the endocrine tissue of the pancreas is supported.
|
[Ultrastructure of B-cell tumors of the pancreas (insulinomas - insular carcinoid tumors)]. The subcellular organization of 6 B-cell tumours of the human pancreas was studied. The tumour parenchyma consisted of processed light, dark, and moderately electron dense cells containing insulin granules. The presence of villi in the cytoplasm of tumour cells and intranuclear fibrills was demonstrated. The participation of tumour cells in formation of amyloid-like thin-fiber substance filling the intercellular and perivascular spaces was established. The similarity between endothelial cells of tumour vessels and parenchymatous cells was shown. A hypothesis on neuroectodermal origin of the endocrine tissue of the pancreas is supported.
|
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] |
PMID:21648
|
Polyarteritis nodosa and deafness. A human temporal bone study.
|
Temporal bone changes were described in a 66 year old woman with polyarteritis nodosa who became deaf 7 months before death. Polyarteritis nodosa of the left internal auditory artery was demonstrated with fibrosis and bone formation involving the cochlea and vestibular system. Endolymphatic hydrops of the basal turn of the cochlea was also present, as well as a chronic perforation of the free wall of the saccule.
|
Polyarteritis nodosa and deafness. A human temporal bone study. Temporal bone changes were described in a 66 year old woman with polyarteritis nodosa who became deaf 7 months before death. Polyarteritis nodosa of the left internal auditory artery was demonstrated with fibrosis and bone formation involving the cochlea and vestibular system. Endolymphatic hydrops of the basal turn of the cochlea was also present, as well as a chronic perforation of the free wall of the saccule.
|
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] |
PMID:21652
|
Biochemical and genetic factors in the heat inactivation of murine beta-glucuronidase.
|
The enzymes coded for by two alleles at the glucuronidase structural locus (Gus) were compared in their response to pH, buffering anion, buffer molarity, ionic strength, and temperature. The heat-labile Gush gene product responded in a qualitatively similar but quantitatively reduced manner compared to the relatively heat-stable Gusb gene product. In all buffers tested, the enzyme was most heat stable at pH 5.0. Ranking of the various buffer anions tested, according to increasing heat stabilization, was water less than acetate less than or equal to phosphate less than citrate. Varying the molarity of the buffers from 0.01 to 0.6 M at pH 5.0 revealed further differences among the buffers. Increasing ionic strength exerted a destabilizing force on the protein. The half-life of the enzyme decreased by as much as a hundredfold between 71 and 75 C. The Gush/Gush genotype also results in decreased activity levels in all tissues, reportedly because of decreased synthesis. The heat inactivation curves of Gusb/Gush heterozygotes were incompatible with any theoretical curve based on the assumption that the Gusb and Gush chromosomes in the heterozygote behave in a manner similar to that seen in the homozygotes.
|
Biochemical and genetic factors in the heat inactivation of murine beta-glucuronidase. The enzymes coded for by two alleles at the glucuronidase structural locus (Gus) were compared in their response to pH, buffering anion, buffer molarity, ionic strength, and temperature. The heat-labile Gush gene product responded in a qualitatively similar but quantitatively reduced manner compared to the relatively heat-stable Gusb gene product. In all buffers tested, the enzyme was most heat stable at pH 5.0. Ranking of the various buffer anions tested, according to increasing heat stabilization, was water less than acetate less than or equal to phosphate less than citrate. Varying the molarity of the buffers from 0.01 to 0.6 M at pH 5.0 revealed further differences among the buffers. Increasing ionic strength exerted a destabilizing force on the protein. The half-life of the enzyme decreased by as much as a hundredfold between 71 and 75 C. The Gush/Gush genotype also results in decreased activity levels in all tissues, reportedly because of decreased synthesis. The heat inactivation curves of Gusb/Gush heterozygotes were incompatible with any theoretical curve based on the assumption that the Gusb and Gush chromosomes in the heterozygote behave in a manner similar to that seen in the homozygotes.
|
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] |
PMID:21653
|
Studies on the female sterile mutant rudimentary of Drosophila melanogaster. II. An analysis of aspartate transcarbamylase and dihydroorotase activities in wild-type and rudimentary strains.
|
The activities of the enzymes aspartate transcarbamylase (ATCase) and dihydroorotase (DHOase) were determined in adult females from a wild-type strain and from eight different alleles of the X-linked mutation rudimentary (r) of Drosophila melanogaster. The alleles chosen span the genetic map of the r locus. The characteristics of the DHOase-catalyzed reaction which converts carbamyl aspartate to dehydroorotate are briefly described. Of all of the r strains tested, only one, r9, has wild-type levels of aspartate transcarbamylase and dihydroorotase activities. The other seven show either intermediate or very low levels of activity for both enzymes. The lowered ATCase and DHOase activities observed in mutants which do not map in the region of the structural gene for these enzymes are interpreted in light of recent evidence that ATCase and DHOase are part of a three-enzyme complex.
|
Studies on the female sterile mutant rudimentary of Drosophila melanogaster. II. An analysis of aspartate transcarbamylase and dihydroorotase activities in wild-type and rudimentary strains. The activities of the enzymes aspartate transcarbamylase (ATCase) and dihydroorotase (DHOase) were determined in adult females from a wild-type strain and from eight different alleles of the X-linked mutation rudimentary (r) of Drosophila melanogaster. The alleles chosen span the genetic map of the r locus. The characteristics of the DHOase-catalyzed reaction which converts carbamyl aspartate to dehydroorotate are briefly described. Of all of the r strains tested, only one, r9, has wild-type levels of aspartate transcarbamylase and dihydroorotase activities. The other seven show either intermediate or very low levels of activity for both enzymes. The lowered ATCase and DHOase activities observed in mutants which do not map in the region of the structural gene for these enzymes are interpreted in light of recent evidence that ATCase and DHOase are part of a three-enzyme complex.
|
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] |
PMID:21654
|
Properties of soluble somatostatin-binding protein.
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A soluble somatostatin-binding protein was detected in the cytosol fractions of various rat, human and bovine tissues. Maximum binding occurred at pH8.0-8.5 and was Ca(2+)-dependent. The specific binding of somatostatin per 10mug of cytosol protein from 12 rat tissues ranged between 36 and 15%, and 3% for peripheral blood cells. There was also substantial binding in cytosol from human anterior pituitary and liver, and bovine anterior pituitary. The specific binding in rat and human plasma in the presence of EDTA was only 1%. Gel filtration suggested a molecular weight of approx. 80000 for the somatostatin-binding protein from several sources. Exposure of the binding protein to trypsin eliminates somatostatin-binding activity but ribonuclease and deoxyribonuclease have no effect. The binding protein is thermolabile, ethanol-precipitable, and not completely specific for somatostatin. Bound (125)I-labelled [Tyr(1)]somatostatin is not easily displaced by excess of unlabelled somatostatin. The effects of dithiothreitol and mercaptoethanol on the binding of (125)I-labelled [Tyr(1)]somatostatin to the binding protein suggests that binding involves two sequential steps, first loose binding, then disulphide linkage. Since semipurified somatostatin-binding protein causes a dose-related inhibition of the binding of (125)I-labelled [Tyr(1)]somatostatin in radioimmunoassays for somatostatin, estimates of somatostatin content of tissue extracts by radioimmunoassay in some cases may be spuriously high. It is not yet clear whether the binding protein is a true cytosol protein or an easily solubilized membrane protein.
|
Properties of soluble somatostatin-binding protein. A soluble somatostatin-binding protein was detected in the cytosol fractions of various rat, human and bovine tissues. Maximum binding occurred at pH8.0-8.5 and was Ca(2+)-dependent. The specific binding of somatostatin per 10mug of cytosol protein from 12 rat tissues ranged between 36 and 15%, and 3% for peripheral blood cells. There was also substantial binding in cytosol from human anterior pituitary and liver, and bovine anterior pituitary. The specific binding in rat and human plasma in the presence of EDTA was only 1%. Gel filtration suggested a molecular weight of approx. 80000 for the somatostatin-binding protein from several sources. Exposure of the binding protein to trypsin eliminates somatostatin-binding activity but ribonuclease and deoxyribonuclease have no effect. The binding protein is thermolabile, ethanol-precipitable, and not completely specific for somatostatin. Bound (125)I-labelled [Tyr(1)]somatostatin is not easily displaced by excess of unlabelled somatostatin. The effects of dithiothreitol and mercaptoethanol on the binding of (125)I-labelled [Tyr(1)]somatostatin to the binding protein suggests that binding involves two sequential steps, first loose binding, then disulphide linkage. Since semipurified somatostatin-binding protein causes a dose-related inhibition of the binding of (125)I-labelled [Tyr(1)]somatostatin in radioimmunoassays for somatostatin, estimates of somatostatin content of tissue extracts by radioimmunoassay in some cases may be spuriously high. It is not yet clear whether the binding protein is a true cytosol protein or an easily solubilized membrane protein.
|
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] |
PMID:21655
|
Microbial metabolism of aromatic nitriles. Enzymology of C-N cleavage by Nocardia sp. (Rhodochrous group) N.C.I.B. 11216.
|
1. An organism utilizing benzonitrile as sole carbon and nitrogen source was isolated by the enrichment-culture technique and identified as a Nocardia sp. of the rhodochrous group. 2. Respiration studies indicate that nitrile degradation proceeds through benzoic acid and catechol. 3. Cell-free extracts of benzonitrile-grown cells contain an enzyme that catalyses the conversion of benzonitrile directly into benzoic acid without intermediate formation of benzamide. 4. This nitrilase enzyme was purified by DEAE-cellulose chromatography and gel filtration on Sephadex G-100 in the presence and absence of substrate. The purity of the enzyme was confirmed by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and isoelectric focusing on polyacrylamide gel. 5. The enzyme shows a time-dependent substrate-activation process in which the substrate catalyses the association of inactive subunits of mol.wt. 45000 to form the polymeric 12-unit active enzyme of mol.wt. 560000. The time required for complete association is highly dependent on the concentration of the enzyme, temperature and pH. 6. The associated enzyme has a pH optimum of 8.0 and K(m) with benzonitrile as substrate of 4mm. The activation energy of the reaction as deduced from the Arrhenius plot is 51.8kJ/mol. 7. Enzyme activity is inhibited by thiol-specific reagents and several metal ions. 8. Studies with different substrates indicate that the nitrilase is specific for nitrile groups directly attached to the benzene ring. Various substituents in the ring are compatible with activity, though ortho-substitution, except by fluorine, renders the nitrile invulnerable to attack. 9. The environmental implications of these findings and the possible significance of the enzyme in the regulation of metabolism are discussed.
|
Microbial metabolism of aromatic nitriles. Enzymology of C-N cleavage by Nocardia sp. (Rhodochrous group) N.C.I.B. 11216. 1. An organism utilizing benzonitrile as sole carbon and nitrogen source was isolated by the enrichment-culture technique and identified as a Nocardia sp. of the rhodochrous group. 2. Respiration studies indicate that nitrile degradation proceeds through benzoic acid and catechol. 3. Cell-free extracts of benzonitrile-grown cells contain an enzyme that catalyses the conversion of benzonitrile directly into benzoic acid without intermediate formation of benzamide. 4. This nitrilase enzyme was purified by DEAE-cellulose chromatography and gel filtration on Sephadex G-100 in the presence and absence of substrate. The purity of the enzyme was confirmed by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and isoelectric focusing on polyacrylamide gel. 5. The enzyme shows a time-dependent substrate-activation process in which the substrate catalyses the association of inactive subunits of mol.wt. 45000 to form the polymeric 12-unit active enzyme of mol.wt. 560000. The time required for complete association is highly dependent on the concentration of the enzyme, temperature and pH. 6. The associated enzyme has a pH optimum of 8.0 and K(m) with benzonitrile as substrate of 4mm. The activation energy of the reaction as deduced from the Arrhenius plot is 51.8kJ/mol. 7. Enzyme activity is inhibited by thiol-specific reagents and several metal ions. 8. Studies with different substrates indicate that the nitrilase is specific for nitrile groups directly attached to the benzene ring. Various substituents in the ring are compatible with activity, though ortho-substitution, except by fluorine, renders the nitrile invulnerable to attack. 9. The environmental implications of these findings and the possible significance of the enzyme in the regulation of metabolism are discussed.
|
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] |
PMID:21656
|
Kinetic properties of a magnesium ion- and calcium ion-stimulated adenosine triphosphatase from the outer-membrane fraction of rat spleen mitochondria.
|
1. Isolated outer membranes from rat spleen mitochondria can be stored in liquid N(2) for several weeks without significant loss of ATPase (adenosine triphosphatase) activity. 2. The ATPase reaction has a broad pH optimum centering on neutral pH, with little significant activity above pH9.0 or below pH5.5. 3. A sigmoidal response of the ATPase activity to temperature is observed between 0 and 55 degrees C, with complete inactivation at 60 degrees C. The Arrhenius plot shows that the activation energy above the transition temperature (22 degrees C) (E(a)=144kJ/mol) is one-third of that calculated for below the transition temperature (E'(a)=408kJ/mol). 4. The outer-membrane ATPase (K(m) for MgATP=50mum) is inactive unless Mg(2+) is added, whereas the inner-membrane ATPase (K(m) for ATP=11mum) is active without added Mg(2+) unless the mitochondria have been depleted of all endogenous Mg(2+) (by using ionophore A23187). 5. The substrate for the outer-membrane ATPase is a bivalent metal ion-nucleoside triphosphate complex in which Mg(2+) (K(m)=50mum) can be replaced effectively by Ca(2+) (K(m)=6.7mum) or Mn(2+), and ATP by ITP. Cu(2+), Co(2+), Sr(2+), Ba(2+), Ni(2+), Cd(2+) and Zn(2+) support very little ATP hydrolysis. 6. Univalent metal ions (Na(+), K(+), Rb(+), Cs(+) and NH(4) (+), but not Li(+)) stimulate the MgATPase activity (<10%) at low concentrations (50mm), but, except for K(+), are slightly inhibitory (20-30%) at higher concentrations (500mm). 7. The Mg(2+)-stimulated ATPase activity is significantly inhibited by Cu(2+) (K(i)=90mum), Ni(2+) (K(i)=510mum), Zn(2+) (K(i)=680mum) and Co(2+) (K(i)=1020mum), but not by Mg(2+), Ca(2+), Ba(2+) or Sr(2+). 8. The outer-membrane ATPase is insensitive to the inhibitors oligomycin, NN'-dicyclohexylcarbodiimide, NaN(3), ouabain and thiol-specific reagents. A significant inhibition is observed at high concentrations of AgNO(3) (0.5mm) and NaF (10mm). 9. The activity towards MgATP is competitively inhibited by the product MgADP (K(i)=0.7mm) but not by the second product P(i) or by 5'-AMP.
|
Kinetic properties of a magnesium ion- and calcium ion-stimulated adenosine triphosphatase from the outer-membrane fraction of rat spleen mitochondria. 1. Isolated outer membranes from rat spleen mitochondria can be stored in liquid N(2) for several weeks without significant loss of ATPase (adenosine triphosphatase) activity. 2. The ATPase reaction has a broad pH optimum centering on neutral pH, with little significant activity above pH9.0 or below pH5.5. 3. A sigmoidal response of the ATPase activity to temperature is observed between 0 and 55 degrees C, with complete inactivation at 60 degrees C. The Arrhenius plot shows that the activation energy above the transition temperature (22 degrees C) (E(a)=144kJ/mol) is one-third of that calculated for below the transition temperature (E'(a)=408kJ/mol). 4. The outer-membrane ATPase (K(m) for MgATP=50mum) is inactive unless Mg(2+) is added, whereas the inner-membrane ATPase (K(m) for ATP=11mum) is active without added Mg(2+) unless the mitochondria have been depleted of all endogenous Mg(2+) (by using ionophore A23187). 5. The substrate for the outer-membrane ATPase is a bivalent metal ion-nucleoside triphosphate complex in which Mg(2+) (K(m)=50mum) can be replaced effectively by Ca(2+) (K(m)=6.7mum) or Mn(2+), and ATP by ITP. Cu(2+), Co(2+), Sr(2+), Ba(2+), Ni(2+), Cd(2+) and Zn(2+) support very little ATP hydrolysis. 6. Univalent metal ions (Na(+), K(+), Rb(+), Cs(+) and NH(4) (+), but not Li(+)) stimulate the MgATPase activity (<10%) at low concentrations (50mm), but, except for K(+), are slightly inhibitory (20-30%) at higher concentrations (500mm). 7. The Mg(2+)-stimulated ATPase activity is significantly inhibited by Cu(2+) (K(i)=90mum), Ni(2+) (K(i)=510mum), Zn(2+) (K(i)=680mum) and Co(2+) (K(i)=1020mum), but not by Mg(2+), Ca(2+), Ba(2+) or Sr(2+). 8. The outer-membrane ATPase is insensitive to the inhibitors oligomycin, NN'-dicyclohexylcarbodiimide, NaN(3), ouabain and thiol-specific reagents. A significant inhibition is observed at high concentrations of AgNO(3) (0.5mm) and NaF (10mm). 9. The activity towards MgATP is competitively inhibited by the product MgADP (K(i)=0.7mm) but not by the second product P(i) or by 5'-AMP.
|
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] |
PMID:21657
|
Evidence for two-step binding of reduced nicotinamide-adenine dinucleotide to aldehyde dehydrogenase.
|
The displacement of NADH from cytoplasmic aldehyde dehydrogenase (EC 1.2.1.3) from sheep liver was studied by using NAD+, 1,10-phenanthroline, ADP-ribose, deamino-NAD+ and pyridine-3-aldehyde-adenine dinucleotide as displacing agents, by following the decrease in fluorescence as a function of time. The data obtained could be fitted by assuming two first-order processes were occurring, a faster process with an apparent rate constant of 0.85 +/- 0.20 s-1 and a relative amplitude of 60 +/- 10% and a slower process with an apparent rate constant of 0.20 +/- 0.05 s-1 and a relative amplitude of 40 +/- 10% (except for pyridine-3-aldehyde-adenine dinucleotide, where the apparent rate constant for the slow process was 0.05 s-1). The displacement rates did not change significantly when the pH was varied from 6.0 to 9.0. Kinetic data are also reported for the dependence of the rate of binding of NADH to the enzyme on the total concentration of NADH. Detailed arguments are presented based on the isolation and purification procedures, the equilibrium coenzyme-binding studies and the kinetic data, which lead to the following model for the release of NADH from the enzyme: (formula: see article). The parameters that best fit the data are: k + 1 = 0.2 s-1; k - 1 = 0.05 s-1; k + 2 = 0.8 s-1 and k - 2 = 5 X 10(5)litre-mol-1-s-1. The slow phase of the NADH release is similar to the steady-state turnover number for substrates such as acetaldehyde and propionaldehyde and appears to contribute significantly to the limitation of the steady-state rate.
|
Evidence for two-step binding of reduced nicotinamide-adenine dinucleotide to aldehyde dehydrogenase. The displacement of NADH from cytoplasmic aldehyde dehydrogenase (EC 1.2.1.3) from sheep liver was studied by using NAD+, 1,10-phenanthroline, ADP-ribose, deamino-NAD+ and pyridine-3-aldehyde-adenine dinucleotide as displacing agents, by following the decrease in fluorescence as a function of time. The data obtained could be fitted by assuming two first-order processes were occurring, a faster process with an apparent rate constant of 0.85 +/- 0.20 s-1 and a relative amplitude of 60 +/- 10% and a slower process with an apparent rate constant of 0.20 +/- 0.05 s-1 and a relative amplitude of 40 +/- 10% (except for pyridine-3-aldehyde-adenine dinucleotide, where the apparent rate constant for the slow process was 0.05 s-1). The displacement rates did not change significantly when the pH was varied from 6.0 to 9.0. Kinetic data are also reported for the dependence of the rate of binding of NADH to the enzyme on the total concentration of NADH. Detailed arguments are presented based on the isolation and purification procedures, the equilibrium coenzyme-binding studies and the kinetic data, which lead to the following model for the release of NADH from the enzyme: (formula: see article). The parameters that best fit the data are: k + 1 = 0.2 s-1; k - 1 = 0.05 s-1; k + 2 = 0.8 s-1 and k - 2 = 5 X 10(5)litre-mol-1-s-1. The slow phase of the NADH release is similar to the steady-state turnover number for substrates such as acetaldehyde and propionaldehyde and appears to contribute significantly to the limitation of the steady-state rate.
|
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] |
PMID:21658
|
Purification and some properties of 1-aspartamido-beta-N-acetylglucosamine amidohydrolase from human liver.
|
Human liver 1-aspartamido-beta-N-acetylglucosamine amidohydrolase (aspartylglucosylaminase, EC 3.5.1.26) was purified 17 500-fold to apparent homogeneity as judged from polyacrylamide-gel disc electrophoresis. A pH optimum of 7.7-9.0 was found. The Km value was pH- and temperature-dependent. At 37 degrees C and pH 7.7, Km was 0.16 mM and it increased to 0.29 at pH 6.0 and 0.23 at pH 9.0. At 25 degrees C and pH 7.7, a Km value of 0.99 mM was obtained. When the substrate concentration was varied, apparent Michaelis-Menten kinetics were obtained. p-Hydroxymercuribenzoate, glutathione or cysteine had no effect on the enzyme activity; 5 mM-N-acetylcysteine inhibited about 47% of the total enzyme activity. Apart from Cu2+, other bivalent ions were virtually ineffective at 1 mM. The kinetic study differentiates this enzyme from aspartylglucosylaminase from other sources.
|
Purification and some properties of 1-aspartamido-beta-N-acetylglucosamine amidohydrolase from human liver. Human liver 1-aspartamido-beta-N-acetylglucosamine amidohydrolase (aspartylglucosylaminase, EC 3.5.1.26) was purified 17 500-fold to apparent homogeneity as judged from polyacrylamide-gel disc electrophoresis. A pH optimum of 7.7-9.0 was found. The Km value was pH- and temperature-dependent. At 37 degrees C and pH 7.7, Km was 0.16 mM and it increased to 0.29 at pH 6.0 and 0.23 at pH 9.0. At 25 degrees C and pH 7.7, a Km value of 0.99 mM was obtained. When the substrate concentration was varied, apparent Michaelis-Menten kinetics were obtained. p-Hydroxymercuribenzoate, glutathione or cysteine had no effect on the enzyme activity; 5 mM-N-acetylcysteine inhibited about 47% of the total enzyme activity. Apart from Cu2+, other bivalent ions were virtually ineffective at 1 mM. The kinetic study differentiates this enzyme from aspartylglucosylaminase from other sources.
|
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] |
PMID:21659
|
Effect of substrates and effectors on the reversible inactivation of pig spleen phosphofructokinase by adenosine triphosphate.
|
1. To investigate the mechanism of the reversible inactivation of pig spleen phosphofructokinase by ATP, the effect of order of addition of reactants (substrates, effectors and enzyme solution) was studied by preincubating the enzyme before assay with various combinations of its substrates and effectors. 2. Preincubation of the enzyme with MgATP or ATP at pH7.0 before addition of fructose 6-phosphate caused a rapid and much greater inhibition of activity than that observed when the reaction (carried out at identical substrate concentrations) was initiated with enzyme. 3. The rapid inhibition caused by preincubation with ATP, together with the sigmoidal response to fructose 6-phosphate and activation by AMP, were all blocked by prior photo-oxidation of the enzyme with Methylene Blue, which selectively destroys the inhibitory binding site for ATP [Ahlfors & Mansour (1969) J. Biol. Chem.244, 1247-1251]. 4. Fructose 6-phosphate, but not Mg(2+), protected phosphofructokinase from inhibition during preincubation with ATP in a manner that was sigmoidally dependent on the fructose 6-phosphate concentration. 5. Mg(2+), by protecting the enzyme from the inhibitory effect of preincubation at low pH (7.0) and by preventing its activation during preincubation with fructose 6-phosphate, demonstrated both a weak activating effect in the absence of the other substrates and a stronger inhibitory effect in the presence of fructose 6-phosphate. 6. Positive effectors (K(+), NH(4) (+), AMP and aspartate) protected the enzyme from inhibition during preincubation with MgATP in proportion to their potency as activators, but citrate potentiated the ATP inhibition. P(i) significantly slowed the inactivation process without itself acting as a positive effector. 7. The non-linear dependence of the initial rate of the unmodified enzyme on protein concentration (associated with increased positive homotropic co-operativity to fructose 6-phosphate) was intensified by preincubation with ATP and abolished by photo-oxidation. 8. The results are interpreted in terms of an association-dissociation model which postulates that protonation, at low pH, of a photo-oxidation-sensitive inhibitory site for ATP allows more rapid dissociation of an active tetramer to an inactive dimeric species.
|
Effect of substrates and effectors on the reversible inactivation of pig spleen phosphofructokinase by adenosine triphosphate. 1. To investigate the mechanism of the reversible inactivation of pig spleen phosphofructokinase by ATP, the effect of order of addition of reactants (substrates, effectors and enzyme solution) was studied by preincubating the enzyme before assay with various combinations of its substrates and effectors. 2. Preincubation of the enzyme with MgATP or ATP at pH7.0 before addition of fructose 6-phosphate caused a rapid and much greater inhibition of activity than that observed when the reaction (carried out at identical substrate concentrations) was initiated with enzyme. 3. The rapid inhibition caused by preincubation with ATP, together with the sigmoidal response to fructose 6-phosphate and activation by AMP, were all blocked by prior photo-oxidation of the enzyme with Methylene Blue, which selectively destroys the inhibitory binding site for ATP [Ahlfors & Mansour (1969) J. Biol. Chem.244, 1247-1251]. 4. Fructose 6-phosphate, but not Mg(2+), protected phosphofructokinase from inhibition during preincubation with ATP in a manner that was sigmoidally dependent on the fructose 6-phosphate concentration. 5. Mg(2+), by protecting the enzyme from the inhibitory effect of preincubation at low pH (7.0) and by preventing its activation during preincubation with fructose 6-phosphate, demonstrated both a weak activating effect in the absence of the other substrates and a stronger inhibitory effect in the presence of fructose 6-phosphate. 6. Positive effectors (K(+), NH(4) (+), AMP and aspartate) protected the enzyme from inhibition during preincubation with MgATP in proportion to their potency as activators, but citrate potentiated the ATP inhibition. P(i) significantly slowed the inactivation process without itself acting as a positive effector. 7. The non-linear dependence of the initial rate of the unmodified enzyme on protein concentration (associated with increased positive homotropic co-operativity to fructose 6-phosphate) was intensified by preincubation with ATP and abolished by photo-oxidation. 8. The results are interpreted in terms of an association-dissociation model which postulates that protonation, at low pH, of a photo-oxidation-sensitive inhibitory site for ATP allows more rapid dissociation of an active tetramer to an inactive dimeric species.
|
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] |
PMID:21667
|
[Synthesis and physico-chemical properties of the antihypertensive agent urapidil (author's transl)].
|
6-(3-[4-(o-Methoxyphenyl)-1-piperazinyl]-propylamino)-1,3-dimethyluracil (Urapidil, Ebrantil) is a new antihypertensive agent whose synthesis and physico-chemical properties are described. The size of the permeation constant KD of the passive transport through collodion-lecithin membranes and the value of the partition coefficient P in the neutral area indicate a good absorption and uniform distribution of the substance in the organism.
|
[Synthesis and physico-chemical properties of the antihypertensive agent urapidil (author's transl)]. 6-(3-[4-(o-Methoxyphenyl)-1-piperazinyl]-propylamino)-1,3-dimethyluracil (Urapidil, Ebrantil) is a new antihypertensive agent whose synthesis and physico-chemical properties are described. The size of the permeation constant KD of the passive transport through collodion-lecithin membranes and the value of the partition coefficient P in the neutral area indicate a good absorption and uniform distribution of the substance in the organism.
|
[
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] |
PMID:21668
|
Effect of dihydroergotoxine on activities of beta-receptor stimulators.
|
The effect of dihydroergotoxine (DET), an alpha-receptor blocker, on the bronchodilating effect of various beta-receptor stimulators was examined using the isolated guinea-pig tracheal smooth muscle. Relaxation of isolated guinea-pig tracheal smooth muscle with isoproterenol hydrochloride, salbutamol hemisulfate and clorprenaline hydrochloride, was markedly increased by continous infusion of DET. This result may suggest that DET, an alpha-receptor blocker, can be used together with bronchodilators such as beta-receptor stimulator for the treatment of various chronic obstructive pulmonary diseases.
|
Effect of dihydroergotoxine on activities of beta-receptor stimulators. The effect of dihydroergotoxine (DET), an alpha-receptor blocker, on the bronchodilating effect of various beta-receptor stimulators was examined using the isolated guinea-pig tracheal smooth muscle. Relaxation of isolated guinea-pig tracheal smooth muscle with isoproterenol hydrochloride, salbutamol hemisulfate and clorprenaline hydrochloride, was markedly increased by continous infusion of DET. This result may suggest that DET, an alpha-receptor blocker, can be used together with bronchodilators such as beta-receptor stimulator for the treatment of various chronic obstructive pulmonary diseases.
|
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] |
PMID:21669
|
A study of antidepressant activity of some indole alkylamines.
|
The antidepressant characteristics of three indole alkylamines were investigated and compared with phenelzine and imipramine by utilising specific pharmacological tools like reserpine, amphetamine, tryptamine and tetrabenazine for determining their possible mechanism of action. Amongst the three indole compounds investigated, indole-3-(2-aminopropyl)-acetate (U-14 164E), indole-3(2-aminobutyl)-d-acetate (u-17 312E) and beta-phenethylhydrazine (phenelzine) produced complete antagonism to reserpine induced sedation, hypothermia as well as facilitation of convulsive seizures. Some of these features suggest that MAO inhibition might be a common mechanism of action of these indoles. The potentiation of CNS effects of tryptamine by these compounds is an outstanding feature of MAO inhibitors, while imipramine is ineffective. Qualitative differences between these indoles and imipramine are evident in the tetrabenazine test. The potentiation of amphetamine induced motor excitation and pentobarbitone narcosis has been explained.
|
A study of antidepressant activity of some indole alkylamines. The antidepressant characteristics of three indole alkylamines were investigated and compared with phenelzine and imipramine by utilising specific pharmacological tools like reserpine, amphetamine, tryptamine and tetrabenazine for determining their possible mechanism of action. Amongst the three indole compounds investigated, indole-3-(2-aminopropyl)-acetate (U-14 164E), indole-3(2-aminobutyl)-d-acetate (u-17 312E) and beta-phenethylhydrazine (phenelzine) produced complete antagonism to reserpine induced sedation, hypothermia as well as facilitation of convulsive seizures. Some of these features suggest that MAO inhibition might be a common mechanism of action of these indoles. The potentiation of CNS effects of tryptamine by these compounds is an outstanding feature of MAO inhibitors, while imipramine is ineffective. Qualitative differences between these indoles and imipramine are evident in the tetrabenazine test. The potentiation of amphetamine induced motor excitation and pentobarbitone narcosis has been explained.
|
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] |
PMID:21671
|
Bioavailability of quinidine in slow-release form. A comparison between two preparations containing quinidine bisulphate as the active constituent.
|
Two different slow-release preparations of quinidine bisulphate (A and B) have been tested. The in vitro dissolution rate of preparation B was substantially lower in intestinal than in gastric juice, whereas the release rate of quinidine from preparation A was virtually unaffected by the pH of the dissolution medium. After a single dose of two tablets of each of the preparations to 6 healthy volunteers, corresponding to 386 mg (B) and 320 mg of quinidine base (A), the maximum plasma concentration was attained after about 4.5 h. The peak concentration was 5.2 +/- 0.5 mumol/l for preparation A and 4.1 +/- 0.4 mumol/l for B. A similar difference was found in the area under the plasma concentration curve (AUC), which was 68 +/- 10 mumol-h/l and 54 +/- 5 mumol-h/l, respectively. Taking into consideration that preparation B contained 20.6% more active drug per tablet these values indicate that the extent of bioavailability is about 50% higher for tablet A than for tablet B.
|
Bioavailability of quinidine in slow-release form. A comparison between two preparations containing quinidine bisulphate as the active constituent. Two different slow-release preparations of quinidine bisulphate (A and B) have been tested. The in vitro dissolution rate of preparation B was substantially lower in intestinal than in gastric juice, whereas the release rate of quinidine from preparation A was virtually unaffected by the pH of the dissolution medium. After a single dose of two tablets of each of the preparations to 6 healthy volunteers, corresponding to 386 mg (B) and 320 mg of quinidine base (A), the maximum plasma concentration was attained after about 4.5 h. The peak concentration was 5.2 +/- 0.5 mumol/l for preparation A and 4.1 +/- 0.4 mumol/l for B. A similar difference was found in the area under the plasma concentration curve (AUC), which was 68 +/- 10 mumol-h/l and 54 +/- 5 mumol-h/l, respectively. Taking into consideration that preparation B contained 20.6% more active drug per tablet these values indicate that the extent of bioavailability is about 50% higher for tablet A than for tablet B.
|
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] |
PMID:21677
|
Flunitrazepam compared with althesin as an induction agent in balanced anaesthesia.
|
The effects of flunitrazepam and Althesin as anaesthetic induction agents were studied in a double-blind trial in 97 patients undergoing abdominal or gynaecological surgery. There was no difference between the two preparations in respect of the quality of induction. A marked difference in the quality of maintenance of anaesthesia were seen, in favour of flunitrazepam. Recovery was more rapid from Althesin anaesthesia. A low incidence of nausea was observed in both groups.
|
Flunitrazepam compared with althesin as an induction agent in balanced anaesthesia. The effects of flunitrazepam and Althesin as anaesthetic induction agents were studied in a double-blind trial in 97 patients undergoing abdominal or gynaecological surgery. There was no difference between the two preparations in respect of the quality of induction. A marked difference in the quality of maintenance of anaesthesia were seen, in favour of flunitrazepam. Recovery was more rapid from Althesin anaesthesia. A low incidence of nausea was observed in both groups.
|
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] |
PMID:21678
|
Studies of drugs given before anaesthesia XXVI: lorazepam.
|
Lorazepam has been studied as preanaesthetic medication given by mouth, i.m. and i.v. Sediation and side-effects and the incidence of anterograde amnesia in patients having a standard operation under methohexitone-nitrous oxide-oxygen anaesthesia were assessed. In a preliminary study of three i.m. (2-, 4- and 8-mg) and six oral (1-,2-,2.5-,4-,5- and 8-mg) doses, the optimum dose was found to be 4 mg for patients with an average weight of 60 kg. This dose was studied in detail when given by all three routes and compared with the commercially available 2.5- and 5-mg tablets. Even when given i.v., there was a delay of 30-40 min in the onset of maximum sedative effect and drowsiness persisted for at least 4 h. Although the onset of action by i.m. injection was slightly faster than when the drug was given by mouth this advantage was more than offset by the high frequencies of pain at the site of injection and restlessness which persisted for 20-40 min. Oral lorazepam in doses of 2.5-5.0 mg was a reliable, effective sedative which could be recommended for routine preanaesthetic medication, provided rapid recovery was not essential. Its soporific effect was accompanied by an appreciable incidence of anterograde amnesia.
|
Studies of drugs given before anaesthesia XXVI: lorazepam. Lorazepam has been studied as preanaesthetic medication given by mouth, i.m. and i.v. Sediation and side-effects and the incidence of anterograde amnesia in patients having a standard operation under methohexitone-nitrous oxide-oxygen anaesthesia were assessed. In a preliminary study of three i.m. (2-, 4- and 8-mg) and six oral (1-,2-,2.5-,4-,5- and 8-mg) doses, the optimum dose was found to be 4 mg for patients with an average weight of 60 kg. This dose was studied in detail when given by all three routes and compared with the commercially available 2.5- and 5-mg tablets. Even when given i.v., there was a delay of 30-40 min in the onset of maximum sedative effect and drowsiness persisted for at least 4 h. Although the onset of action by i.m. injection was slightly faster than when the drug was given by mouth this advantage was more than offset by the high frequencies of pain at the site of injection and restlessness which persisted for 20-40 min. Oral lorazepam in doses of 2.5-5.0 mg was a reliable, effective sedative which could be recommended for routine preanaesthetic medication, provided rapid recovery was not essential. Its soporific effect was accompanied by an appreciable incidence of anterograde amnesia.
|
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] |
PMID:21679
|
Substrate and substrate analogue binding properties of Renilla luciferase.
|
Luciferase from the anthozoan coelenterate Renilla reniformis catalyzes the oxidative decarboxylation of luciferin consuming 1 mol of O2 per mol of luciferin oxidized and producing 1 mol of CO2, 1 mol of oxyluciferin, and light (lambdaB, 480 nm) with a 5.5% quantum yield. In this work we have examined the binding characteristics of luciferin, luciferin analogues, and competitive inhibitors of the luciferin-luciferase reaction. The results show that luciferin binding and orientation in the single luciferin binding site of luciferase are highly specific for and dependent upon the three group substituents of the luciferin molecule while the imidazolone-pyrazine nucleus of luciferin is not directly involved in binding. Anaerobic luciferin binding promotes a rapid concentration-dependent aggregation of luciferase which results in irreversible inactivation of the enzyme. This aggregation phenomenon is not observed upon binding of oxyluciferin, luciferyl sulfate, or luciferin analogues in which the substituent at the 2 position of the imidazolone-pyrazine ring has been substantially altered.
|
Substrate and substrate analogue binding properties of Renilla luciferase. Luciferase from the anthozoan coelenterate Renilla reniformis catalyzes the oxidative decarboxylation of luciferin consuming 1 mol of O2 per mol of luciferin oxidized and producing 1 mol of CO2, 1 mol of oxyluciferin, and light (lambdaB, 480 nm) with a 5.5% quantum yield. In this work we have examined the binding characteristics of luciferin, luciferin analogues, and competitive inhibitors of the luciferin-luciferase reaction. The results show that luciferin binding and orientation in the single luciferin binding site of luciferase are highly specific for and dependent upon the three group substituents of the luciferin molecule while the imidazolone-pyrazine nucleus of luciferin is not directly involved in binding. Anaerobic luciferin binding promotes a rapid concentration-dependent aggregation of luciferase which results in irreversible inactivation of the enzyme. This aggregation phenomenon is not observed upon binding of oxyluciferin, luciferyl sulfate, or luciferin analogues in which the substituent at the 2 position of the imidazolone-pyrazine ring has been substantially altered.
|
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] |
PMID:21680
|
Pyridoxamine-pyruvate transaminase. 1. Determination of the active site stoichiometry and the pH dependence of the dissociation constant for 5'-deoxypyridoxal.
|
Spectrophotometric titration of pyridoxamine-pyruvate transaminase (EC 2.6.1.30) with pyridoxal at pH 7.15 gives four equivalent binding sites per tetramer. The pH dependence of the equilibrium constant for the association of 5'-deoxypyridoxal with the active site lysine residue was determined spectrophotometrically. These dissociation constants increase with increasing pH over the range pH 7.5-9 and are correlated with the values obtained from fast reactions kinetics (Gilmer, P. J., and Kirsch, J. F. (1977), Biochemistry 16 (following paper in this issue)). In addition to this specific reaction at an active site lysine residue, a second slower reaction at non-active site residues is observable at pH values greater than 8. The pH dependencies of the association and dissociation rate constants for this slow reaction were studied over the pH range 8 to 9 after blocking the active site by NaBH4 reduction of the pyridoxal adduct. The enzyme is stabilized and markedly activated by potassium ion.
|
Pyridoxamine-pyruvate transaminase. 1. Determination of the active site stoichiometry and the pH dependence of the dissociation constant for 5'-deoxypyridoxal. Spectrophotometric titration of pyridoxamine-pyruvate transaminase (EC 2.6.1.30) with pyridoxal at pH 7.15 gives four equivalent binding sites per tetramer. The pH dependence of the equilibrium constant for the association of 5'-deoxypyridoxal with the active site lysine residue was determined spectrophotometrically. These dissociation constants increase with increasing pH over the range pH 7.5-9 and are correlated with the values obtained from fast reactions kinetics (Gilmer, P. J., and Kirsch, J. F. (1977), Biochemistry 16 (following paper in this issue)). In addition to this specific reaction at an active site lysine residue, a second slower reaction at non-active site residues is observable at pH values greater than 8. The pH dependencies of the association and dissociation rate constants for this slow reaction were studied over the pH range 8 to 9 after blocking the active site by NaBH4 reduction of the pyridoxal adduct. The enzyme is stabilized and markedly activated by potassium ion.
|
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