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PMID:18305 | Determination of the Tamm and Horsfall glycoprotein in human urine. | A method for the routine determination of the Tamm and Horsfall glycoprotein in human urine is presented. The method is based on quantitative electroimmunoassay. It was demonstrated that the solubility of the Tamm and Horsfall glycoprotein was dependent on urinary ionic strength and pH. To minimize these variables all urines were buffered to pH 5.5 and then preincubated at 37 degrees C in 0.3% sodium dodecyl sulphate. This procedure solubilized and maintained the Tamm and Horsfall glycoprotein in solution. Excess sodium dodecyl sulphate was removed by gel filtration before the quantities of the Tamm and Horsfall glycoprotein were determined by electroimmunoassay. The method is simple, reproducible, has high precision and compares favourably with earlier published methods for the determinations of the Tamm and Horsfall glycoprotein. | Determination of the Tamm and Horsfall glycoprotein in human urine. A method for the routine determination of the Tamm and Horsfall glycoprotein in human urine is presented. The method is based on quantitative electroimmunoassay. It was demonstrated that the solubility of the Tamm and Horsfall glycoprotein was dependent on urinary ionic strength and pH. To minimize these variables all urines were buffered to pH 5.5 and then preincubated at 37 degrees C in 0.3% sodium dodecyl sulphate. This procedure solubilized and maintained the Tamm and Horsfall glycoprotein in solution. Excess sodium dodecyl sulphate was removed by gel filtration before the quantities of the Tamm and Horsfall glycoprotein were determined by electroimmunoassay. The method is simple, reproducible, has high precision and compares favourably with earlier published methods for the determinations of the Tamm and Horsfall glycoprotein. | [
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PMID:18306 | Heterogeneity of erythrocyte pyruvate kinase deficiency and related metabolic disorders in patients with hematological diseases. | In several patients suffering from congenital non-spherocytic hemolytic anemia or from malignant hemotological disorder associated with erythrocyte pyruvate kinase (PK) deficiency, a metabolic study has been carried out involving the following biochemical determinations: assay of red cell enzyme activities; estimation of glucose consumption; measurement of the rate of glycolytic intermediates; and, in some cases, enzyme purification and characterization of the PK variant. Metabolic equilibrium most probably does not depend on kinetic characteristics of PK molecules. Furthermore, the data obtained allow separation of cases with congenital non-spherocytic hemolytic anemia (hereditary defect) and acquired PK deficiencies. | Heterogeneity of erythrocyte pyruvate kinase deficiency and related metabolic disorders in patients with hematological diseases. In several patients suffering from congenital non-spherocytic hemolytic anemia or from malignant hemotological disorder associated with erythrocyte pyruvate kinase (PK) deficiency, a metabolic study has been carried out involving the following biochemical determinations: assay of red cell enzyme activities; estimation of glucose consumption; measurement of the rate of glycolytic intermediates; and, in some cases, enzyme purification and characterization of the PK variant. Metabolic equilibrium most probably does not depend on kinetic characteristics of PK molecules. Furthermore, the data obtained allow separation of cases with congenital non-spherocytic hemolytic anemia (hereditary defect) and acquired PK deficiencies. | [
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PMID:18312 | Clonidine and the vasodilating beta blocker antihypertensive drug interaction. | Because propranolol is contraindicated in some patients and since clonidine can decrease heart rate and renin release, clonidine was substituted for propranolol in 14 severely hypertensive minoxidil-treated outpatients. Clonidine induced weight loss which, since plasma concentrations were not suppressed, was not due to inhibition of release of antidiuretic hormone or renin. These endocrine interrelations were confirmed by later administration of clonidine to 4 of the subjects under controlled circumstances in our General Clinical Research Center. When substituted for propranolol, clonidine controlled blood pressure and heart rate in 8 of the 9 outpatients whose blood pressure had been previously well controlled. Clonidine and propranolol had additive antihypertensive effects in the other 5 patients. Thus, clonidine can substitute for propranolol or when added to the propranolol-vasodilator combination supply an additional blood pressure-lowering effects. This substitution or addition results in an increase in side effects. In addition, clonidine has a diuretic action under these circumstances by an unknown mechanism. | Clonidine and the vasodilating beta blocker antihypertensive drug interaction. Because propranolol is contraindicated in some patients and since clonidine can decrease heart rate and renin release, clonidine was substituted for propranolol in 14 severely hypertensive minoxidil-treated outpatients. Clonidine induced weight loss which, since plasma concentrations were not suppressed, was not due to inhibition of release of antidiuretic hormone or renin. These endocrine interrelations were confirmed by later administration of clonidine to 4 of the subjects under controlled circumstances in our General Clinical Research Center. When substituted for propranolol, clonidine controlled blood pressure and heart rate in 8 of the 9 outpatients whose blood pressure had been previously well controlled. Clonidine and propranolol had additive antihypertensive effects in the other 5 patients. Thus, clonidine can substitute for propranolol or when added to the propranolol-vasodilator combination supply an additional blood pressure-lowering effects. This substitution or addition results in an increase in side effects. In addition, clonidine has a diuretic action under these circumstances by an unknown mechanism. | [
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PMID:18310 | Plasmalemmal calcium in cardiac excitation-contraction coupling. | 1. Mammalian heart muscle is extremely sensitive to the external calcium concentration. It reacts to alterations of the external calcium concentration with an immediate adaptation of contractile force. 2. In mammalian heart there is a network of large transverse tubules throughout the cell. These structures are regularly arranged at the level of the sarcomeric Z- and I-lines and increase the cell surface by a factor of ten. 3. Experimental evidence favours the assumption that the plasmalemma could be the site of a loosely bound superficial Ca fraction which becomes ionized upon depolarization and is again bound upon repolarization of the cardiac cell membrane. 4. A mechanism is discussed which bases the excitation-contraction coupling process on a physicochemical interaction of calcium with membrane phospholipids. The degree of interaction is thought to be governed by the transmembrane electric field, the induced dipole moment of membrane constituents, and proton activity within the membrane. | Plasmalemmal calcium in cardiac excitation-contraction coupling. 1. Mammalian heart muscle is extremely sensitive to the external calcium concentration. It reacts to alterations of the external calcium concentration with an immediate adaptation of contractile force. 2. In mammalian heart there is a network of large transverse tubules throughout the cell. These structures are regularly arranged at the level of the sarcomeric Z- and I-lines and increase the cell surface by a factor of ten. 3. Experimental evidence favours the assumption that the plasmalemma could be the site of a loosely bound superficial Ca fraction which becomes ionized upon depolarization and is again bound upon repolarization of the cardiac cell membrane. 4. A mechanism is discussed which bases the excitation-contraction coupling process on a physicochemical interaction of calcium with membrane phospholipids. The degree of interaction is thought to be governed by the transmembrane electric field, the induced dipole moment of membrane constituents, and proton activity within the membrane. | [
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PMID:18313 | Metabolism of flurazepam by the small intestine. | The metabolism of flurazepam-5-14C has been studied in man following catheterization of the portal and hepatic veins. Flurazepam was administered through a tube into the stomach in one patient and into the duodenum in two patients. Thin-layer chromatographs of portal vein blood showed that there was a rapid and early appearance of metabolites of flurazepam consistent with the metabolism of the flurazepam by the intestinal mucosa and at times when the concentrations in the hepatic vein and peripheral blood were very much lower than those in the portal vein. The major metabolites identified in portal vein blood were the mono- and didesetyl metabolies of flurazepam. Considerable hepatic uptake of flurazepam and its metabolites occurred, as evidenced by the lower concentrations of the parent compound and metabolites in the hepatic vein. Thus, "first-pass" metabolism of flurazepam following oral administration occurs in the small bowel mucosa of man as well as in the liver. | Metabolism of flurazepam by the small intestine. The metabolism of flurazepam-5-14C has been studied in man following catheterization of the portal and hepatic veins. Flurazepam was administered through a tube into the stomach in one patient and into the duodenum in two patients. Thin-layer chromatographs of portal vein blood showed that there was a rapid and early appearance of metabolites of flurazepam consistent with the metabolism of the flurazepam by the intestinal mucosa and at times when the concentrations in the hepatic vein and peripheral blood were very much lower than those in the portal vein. The major metabolites identified in portal vein blood were the mono- and didesetyl metabolies of flurazepam. Considerable hepatic uptake of flurazepam and its metabolites occurred, as evidenced by the lower concentrations of the parent compound and metabolites in the hepatic vein. Thus, "first-pass" metabolism of flurazepam following oral administration occurs in the small bowel mucosa of man as well as in the liver. | [
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PMID:18311 | A study in mice of bromo and chloro acylurea analogues of the sedative-hypnotic bromureides. | 1. A series of 1-(2-chloroacyl)ureas, related to the sedative-hypnotic drugs bromvaletone and carbromal, was synthesized and tested in mice to determine central depressant and acute toxic effects. Four 1-(2-bromoacyl)ureas and two 3-halo compounds were included for comparison. 2. Large variations in potency were seen between the compounds. Much of this can be ascribed to differences in lipophilicity. Among homologous 1-(2-chloroacyl)ureas, those with 6 acyl carbons had maximal potency. Among groups of structural isomers, the most potent were 2-halo, 3-alkyl substituted compounds. 3. The most potent compounds were also those with the largest ratios of hypnotic to lethal activity. 4. The variation in the onset and duration of action of these compounds enables a choice to be made for a compound with a particular set of characteristics. | A study in mice of bromo and chloro acylurea analogues of the sedative-hypnotic bromureides. 1. A series of 1-(2-chloroacyl)ureas, related to the sedative-hypnotic drugs bromvaletone and carbromal, was synthesized and tested in mice to determine central depressant and acute toxic effects. Four 1-(2-bromoacyl)ureas and two 3-halo compounds were included for comparison. 2. Large variations in potency were seen between the compounds. Much of this can be ascribed to differences in lipophilicity. Among homologous 1-(2-chloroacyl)ureas, those with 6 acyl carbons had maximal potency. Among groups of structural isomers, the most potent were 2-halo, 3-alkyl substituted compounds. 3. The most potent compounds were also those with the largest ratios of hypnotic to lethal activity. 4. The variation in the onset and duration of action of these compounds enables a choice to be made for a compound with a particular set of characteristics. | [
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|
PMID:18316 | The effect of chronic frusemide administration on intracellular potassium, sodium and pH of cardiac and skeletal muscle. | 1. Chronic administration of frusemide in large doses of 4 mg day-1 kg-1 for 3 weeks caused a significant reduction of cell water in rabbit cardiac and skeletal muscle. Intracellular Na+ concentration, intracellular pH and extracellular space was unchanged in both tissues. Intracellular K+ concentration increased slightly in both cardiac and skeletal muscle. 2. It is concluded that frusemide does not reduce intracellular K+ concentration in cardiac or skeletal muscle of normal animals receiving a normal oral potassium intake. | The effect of chronic frusemide administration on intracellular potassium, sodium and pH of cardiac and skeletal muscle. 1. Chronic administration of frusemide in large doses of 4 mg day-1 kg-1 for 3 weeks caused a significant reduction of cell water in rabbit cardiac and skeletal muscle. Intracellular Na+ concentration, intracellular pH and extracellular space was unchanged in both tissues. Intracellular K+ concentration increased slightly in both cardiac and skeletal muscle. 2. It is concluded that frusemide does not reduce intracellular K+ concentration in cardiac or skeletal muscle of normal animals receiving a normal oral potassium intake. | [
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|
PMID:18319 | Essential facets of radiological diagnosis of extremity trauma. | Trauma to the extremities is sometimes dismissed as relatively unimportant in certain radiologic circles. In many instances, radiologic responsibility is denied and relegated to the orthopedist. Although the radiologic manifestations of much trauma to the extremities is clear cut and does not require sophisticated techniques or interpretation, there are many diagnoses which require innovative techniques and a knowledge of mechanisms of injury as well as the subtle oseous and soft tissue manifestations which may be confronted radiologically. The above factors will be stressed along with the need for the fundamental knowledge of radiologic anatomy which is required in order to appreciate some of the more subtle changes of extremity trauma. | Essential facets of radiological diagnosis of extremity trauma. Trauma to the extremities is sometimes dismissed as relatively unimportant in certain radiologic circles. In many instances, radiologic responsibility is denied and relegated to the orthopedist. Although the radiologic manifestations of much trauma to the extremities is clear cut and does not require sophisticated techniques or interpretation, there are many diagnoses which require innovative techniques and a knowledge of mechanisms of injury as well as the subtle oseous and soft tissue manifestations which may be confronted radiologically. The above factors will be stressed along with the need for the fundamental knowledge of radiologic anatomy which is required in order to appreciate some of the more subtle changes of extremity trauma. | [
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PMID:18320 | Some factors affecting testosterone, dihydrotestosterone, 5 alpha-androstan-3 alpha,17 beta-diol and 5 alpha-androstan-3 beta,17 beta-diol secretion by invitro perfused rabbit testes. | Intra-arterial infusion of testosterone-3H gave rise to tritiated dihydrotestosterone, 5 alpha-androstan-3 alpha,17 beta-diol and 5 alpha-androstan-3beta,17 beta-diol in spermatic venous effluent of the perfused rabbit testis-epididymis. Mass spectrometric measurements confirmed that these four androgens were present in spermatic venous effluent of the perfused rabbit testis-epididymis. Gas liquid chromatographic measurement showed that testosterone, dihydrotestosterone, 5 alpha-androstan-3 alpha,17 beta-diol and 5 alpha-androstan-3 beta,17 beta-diol were secreted in similar amounts by the in vitro perfused and in situ rabbit testis-epididymis results obtained by perfusing the testis minus the epididymis suggested that the bulk of these androgens originate from the catabolism of testosterone within the testis rather than the epididymis. Suprisingly, germinal epithelium destruction by heat failed to alter the testosterone, dihydrotestosterone and 5 alpha-androstan-3 alpha,17 beta-diol secretion by the in vitro perfused rabbit testis. In contrast, the secretion of 5 alpha-androstan-3 beta,17 beta-diol was significantly (P less than 0.05) reduced in the same cryptorchid compared to control testes. | Some factors affecting testosterone, dihydrotestosterone, 5 alpha-androstan-3 alpha,17 beta-diol and 5 alpha-androstan-3 beta,17 beta-diol secretion by invitro perfused rabbit testes. Intra-arterial infusion of testosterone-3H gave rise to tritiated dihydrotestosterone, 5 alpha-androstan-3 alpha,17 beta-diol and 5 alpha-androstan-3beta,17 beta-diol in spermatic venous effluent of the perfused rabbit testis-epididymis. Mass spectrometric measurements confirmed that these four androgens were present in spermatic venous effluent of the perfused rabbit testis-epididymis. Gas liquid chromatographic measurement showed that testosterone, dihydrotestosterone, 5 alpha-androstan-3 alpha,17 beta-diol and 5 alpha-androstan-3 beta,17 beta-diol were secreted in similar amounts by the in vitro perfused and in situ rabbit testis-epididymis results obtained by perfusing the testis minus the epididymis suggested that the bulk of these androgens originate from the catabolism of testosterone within the testis rather than the epididymis. Suprisingly, germinal epithelium destruction by heat failed to alter the testosterone, dihydrotestosterone and 5 alpha-androstan-3 alpha,17 beta-diol secretion by the in vitro perfused rabbit testis. In contrast, the secretion of 5 alpha-androstan-3 beta,17 beta-diol was significantly (P less than 0.05) reduced in the same cryptorchid compared to control testes. | [
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PMID:18322 | Testicular androgen binding protein (ABP) - a parameter of Sertoli cell secretory function. | Using ABP as an index of Sertoli cell secretory function, several important features of the Sertoli cell have emerged: 1. The stimulation of ABP production by FSH clearly points to the Sertoli cell as a target cell for FSH (3,4,9-16,21,24). 2. The dramatic effects of androgens on ABP production both in immature and mature rats also suggest that the Sertoli cell is a target cell for androgen (3,12,14,16,25). 3. The striking reduction in ABP production in the cryptorchid testis raises the question whether impairment of Sertoli cell function is the primary reason for the loss of germ cells that occurs in this condition (20). 4. Drugs like nitrofurazone or ethionine, or X-irradiation only slightly affect the secretory function of the Sertoli cells (ABP production), indicating that these treatments most probably have direct effects on the germ cells as well. Thus, measurement of ABP production rate is a very important tool in order to evaluate how hormones, drugs and physical injuries might affect the secretory function of the Sertoli cell. This test system might be of great use in order to study the physiology and hormonal regulation of the Sertoli cells. It might also be valuable in pharmacological and toxicological studies. | Testicular androgen binding protein (ABP) - a parameter of Sertoli cell secretory function. Using ABP as an index of Sertoli cell secretory function, several important features of the Sertoli cell have emerged: 1. The stimulation of ABP production by FSH clearly points to the Sertoli cell as a target cell for FSH (3,4,9-16,21,24). 2. The dramatic effects of androgens on ABP production both in immature and mature rats also suggest that the Sertoli cell is a target cell for androgen (3,12,14,16,25). 3. The striking reduction in ABP production in the cryptorchid testis raises the question whether impairment of Sertoli cell function is the primary reason for the loss of germ cells that occurs in this condition (20). 4. Drugs like nitrofurazone or ethionine, or X-irradiation only slightly affect the secretory function of the Sertoli cells (ABP production), indicating that these treatments most probably have direct effects on the germ cells as well. Thus, measurement of ABP production rate is a very important tool in order to evaluate how hormones, drugs and physical injuries might affect the secretory function of the Sertoli cell. This test system might be of great use in order to study the physiology and hormonal regulation of the Sertoli cells. It might also be valuable in pharmacological and toxicological studies. | [
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|
PMID:18324 | Regional assignment of seven genes on chromosome 1 of man by use of man-Chinese hamster somatic cell hybrids. I. Results obtained after hybridization of human cells carrying reciprocal translocations involving chromosome 1. | Regional localization studies of genes coding for human PGD, PPH1, PGM1, UGPP, GuK1, Pep-C, and FH, which have been assigned to chromosome 1, were performed with man-Chinese hamster somatic cell hybrids, Informative hybrids that retained fragments of the human chromosome 1 were produced by fusion of hamster cells with human cells carrying reciprocal translocations involving chromosome 1. Analysis of the hybrids that retained one of the translocation chromosomes or de novo rearrangements involving the human 1 revealed the following gene positions: PGD and PPH1 in 1pter leads to 1p32, PGM1 in 1p32 leads to 1p22, UGPP and GuK1 in 1q21 leads to 1q42, FH in 1qter leads to 1q42, and Pep-C probably in 1q42. | Regional assignment of seven genes on chromosome 1 of man by use of man-Chinese hamster somatic cell hybrids. I. Results obtained after hybridization of human cells carrying reciprocal translocations involving chromosome 1. Regional localization studies of genes coding for human PGD, PPH1, PGM1, UGPP, GuK1, Pep-C, and FH, which have been assigned to chromosome 1, were performed with man-Chinese hamster somatic cell hybrids, Informative hybrids that retained fragments of the human chromosome 1 were produced by fusion of hamster cells with human cells carrying reciprocal translocations involving chromosome 1. Analysis of the hybrids that retained one of the translocation chromosomes or de novo rearrangements involving the human 1 revealed the following gene positions: PGD and PPH1 in 1pter leads to 1p32, PGM1 in 1p32 leads to 1p22, UGPP and GuK1 in 1q21 leads to 1q42, FH in 1qter leads to 1q42, and Pep-C probably in 1q42. | [
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PMID:18325 | Brain monoamines act through the prostaglandin release to influence the body temperature. | In the present study, the thermal responses induced by intraventicular administration of pyrogen prostaglandin E1, the brain monoamines norepinephrine and serotonin, and the antipyretic sodium acetylsalicylate (aspirin) were measured in conscious rabbits to assess the possible involvement of these substances in fever production. The body temperatures, metabolic rate, respiratory evaporative heat loss and vasomotor activity in response to the administration of these drugs were measured. The results showed that sodium acetylsalicylate, an inhibitor of prostaglandin synthetase, antagonizes the norepinephrine induced fever but not the prostaglandin fever. The data also showed that the serotonin induced hypothermia was reversed by prostaglandin administration. Thus, the fact strongly suggest that the prostaglandin E1 serves as a fever-prducing mediator in the central nervous system. Also, the norepinephrine fever and serotonin hpyothermia, respectively, may be associated with an increase and a decrease in prostaglandin synthesis in the brain. | Brain monoamines act through the prostaglandin release to influence the body temperature. In the present study, the thermal responses induced by intraventicular administration of pyrogen prostaglandin E1, the brain monoamines norepinephrine and serotonin, and the antipyretic sodium acetylsalicylate (aspirin) were measured in conscious rabbits to assess the possible involvement of these substances in fever production. The body temperatures, metabolic rate, respiratory evaporative heat loss and vasomotor activity in response to the administration of these drugs were measured. The results showed that sodium acetylsalicylate, an inhibitor of prostaglandin synthetase, antagonizes the norepinephrine induced fever but not the prostaglandin fever. The data also showed that the serotonin induced hypothermia was reversed by prostaglandin administration. Thus, the fact strongly suggest that the prostaglandin E1 serves as a fever-prducing mediator in the central nervous system. Also, the norepinephrine fever and serotonin hpyothermia, respectively, may be associated with an increase and a decrease in prostaglandin synthesis in the brain. | [
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|
PMID:18328 | [Recurrent periarteritis nodosa with predominantly cerebral signs in children (author's transl)]. | Periarteritis nodosa occurs also in children and has the same wide spectrum of signs and symptoms as in adults. In a 4 1/2 year-old girl the disease at first was characterised by cerebral, encephalitis-like symptoms, arterial hypertension, intestinal wall necrosis and, finally, pericardial tamponade as a result of rupture of an arteritic coronary artery aneurysm. An earlier attack had involved liver and skeletal muscle, with necrosis in the liver. Peculiar morphological features within the "classical" course of periarteritis nodosa are endarteritic changes of single retroperitoneal and coronary arteries. | [Recurrent periarteritis nodosa with predominantly cerebral signs in children (author's transl)]. Periarteritis nodosa occurs also in children and has the same wide spectrum of signs and symptoms as in adults. In a 4 1/2 year-old girl the disease at first was characterised by cerebral, encephalitis-like symptoms, arterial hypertension, intestinal wall necrosis and, finally, pericardial tamponade as a result of rupture of an arteritic coronary artery aneurysm. An earlier attack had involved liver and skeletal muscle, with necrosis in the liver. Peculiar morphological features within the "classical" course of periarteritis nodosa are endarteritic changes of single retroperitoneal and coronary arteries. | [
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|
PMID:18329 | Drug-induced cardiovascular diseases. | A wide variety of drugs may be associated with serious cardiovascular toxicity. Toxicity due to drugs primarily used for treating cardiovascular toxicity. Toxicity due to drugs primarily used for treating cardiac disorders is the most extensively documented, especially the arrhythmias due to digitalis glycosides. Various arrhythmias are also caused by toxic levels of many antiarrhythmic agents including quinidine, procainamide and phenytotin. Myocardial depression and heart failure are serious side-effects of beta-adrenoceptor blocking agents and myocardial ischaemia due to sympathominetic amines may result from both direct and indirect mechanisms. The many toxic reactions in the cardiovascular system due to non-cardiac drugs are less widely known and for the most part less clearly understood. Many remain controversial at the current time; for example, the diathesis toward thromboembolism in women taking oral contraceptives. Potential cardiac toxicity due to drugs used in the rapidly expanding sphere of anti-neoplastic chemotherapy is exemplified by the cardiomyopathy-like toxicities of doxorubicin and daunorubicin. Many of the psychotherapeutic drugs including phenothiazine antipsychotics and tricyclic antidepressants have arrhythmogenic potential. | Drug-induced cardiovascular diseases. A wide variety of drugs may be associated with serious cardiovascular toxicity. Toxicity due to drugs primarily used for treating cardiovascular toxicity. Toxicity due to drugs primarily used for treating cardiac disorders is the most extensively documented, especially the arrhythmias due to digitalis glycosides. Various arrhythmias are also caused by toxic levels of many antiarrhythmic agents including quinidine, procainamide and phenytotin. Myocardial depression and heart failure are serious side-effects of beta-adrenoceptor blocking agents and myocardial ischaemia due to sympathominetic amines may result from both direct and indirect mechanisms. The many toxic reactions in the cardiovascular system due to non-cardiac drugs are less widely known and for the most part less clearly understood. Many remain controversial at the current time; for example, the diathesis toward thromboembolism in women taking oral contraceptives. Potential cardiac toxicity due to drugs used in the rapidly expanding sphere of anti-neoplastic chemotherapy is exemplified by the cardiomyopathy-like toxicities of doxorubicin and daunorubicin. Many of the psychotherapeutic drugs including phenothiazine antipsychotics and tricyclic antidepressants have arrhythmogenic potential. | [
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|
PMID:18331 | [Telemetry of the tissue Ph value under the effect of local anesthetics. Preliminary report]. | After discussing the problems involved in in vivo pH measurements, pH telemetry is presented as a new method. Long-term in vivo measurements of the pH are intended to be performed by means of a miniature transmitter implanted into the subcutaneous tissue of guinea pigs. | [Telemetry of the tissue Ph value under the effect of local anesthetics. Preliminary report]. After discussing the problems involved in in vivo pH measurements, pH telemetry is presented as a new method. Long-term in vivo measurements of the pH are intended to be performed by means of a miniature transmitter implanted into the subcutaneous tissue of guinea pigs. | [
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|
PMID:18332 | [The salivary fluoride level after administration of fluorides and fluoride complex salts]. | Compounds of fluorine and Si or Fe are found in nature and are sometimes formed from F ions in the stomach. The chemistry of the release of resorbable F ions from such compounds in the intestinal tract is described. To demonstrate fluorine availability at the enamel surface daily profiles of fluorine concentration in the saliva of seven subjects after a single administration of a NaF and Na3FeF6 tablet are presented and their effect on the remineralization of the dental enamel described. | [The salivary fluoride level after administration of fluorides and fluoride complex salts]. Compounds of fluorine and Si or Fe are found in nature and are sometimes formed from F ions in the stomach. The chemistry of the release of resorbable F ions from such compounds in the intestinal tract is described. To demonstrate fluorine availability at the enamel surface daily profiles of fluorine concentration in the saliva of seven subjects after a single administration of a NaF and Na3FeF6 tablet are presented and their effect on the remineralization of the dental enamel described. | [
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|
PMID:18334 | [Change in the mental contents in the paradoxical stage and in stage 2 under the effects of a benzodiazepine, flunitrazepam]. | In some insomniacs under flunitrazepam treatment is noted on one hand a global increase of dream memories, and on the other hand an increase of dreams with unpleasant or anxious contents. The present investigation is intended first to test the hypothesis of a possible increase of mental contents in sleep outside the paradoxical phases, to account for the increase of dream memories without concomitant increase of paradoxical stage. Second it intends to investigate if the increase of unpleasant dreams may be found in the normal subject in laboratory. | [Change in the mental contents in the paradoxical stage and in stage 2 under the effects of a benzodiazepine, flunitrazepam]. In some insomniacs under flunitrazepam treatment is noted on one hand a global increase of dream memories, and on the other hand an increase of dreams with unpleasant or anxious contents. The present investigation is intended first to test the hypothesis of a possible increase of mental contents in sleep outside the paradoxical phases, to account for the increase of dream memories without concomitant increase of paradoxical stage. Second it intends to investigate if the increase of unpleasant dreams may be found in the normal subject in laboratory. | [
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|
PMID:18338 | Influence of temperature on the determination of enzyme activities in human serum. gamma-Glutamyl transferase. | Optimum reaction conditions for determination of gamma-glutamyl transferase were studied at 25, 30 and 37 degrees C using a kinetic test and gamma-glutamyl-3-carboxy-4-nitranilide as substrate. There was no dependence on temperature of half saturation constants of gamma-glutamyl-3-carboxy-4-nitranilide. The corresponding constants for glycylglycine were influenced by temperature and the pH. The optimum showed a dependence upon temperature. In Arrhenius' plot, a deviation from straight line can be observed only above 35 degrees C. The influence of temperature on the determination of enzyme activities in human serum are discussed. | Influence of temperature on the determination of enzyme activities in human serum. gamma-Glutamyl transferase. Optimum reaction conditions for determination of gamma-glutamyl transferase were studied at 25, 30 and 37 degrees C using a kinetic test and gamma-glutamyl-3-carboxy-4-nitranilide as substrate. There was no dependence on temperature of half saturation constants of gamma-glutamyl-3-carboxy-4-nitranilide. The corresponding constants for glycylglycine were influenced by temperature and the pH. The optimum showed a dependence upon temperature. In Arrhenius' plot, a deviation from straight line can be observed only above 35 degrees C. The influence of temperature on the determination of enzyme activities in human serum are discussed. | [
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|
PMID:18339 | Guanosine triphosphate: 5-hydroxylysine phosphotransferase in rat kidney cortex. | An enzyme which catalyzes the transfer of the gamma-phosphate from GTP onto 5-hydroxylysine was partially purified from rat kidney cortex by means of acid precipitation and DEAE-Sephadex A-50 column chromatography. The enzyme activity was assayed by measuring the transfer of [32P] from gamma-[32P]-GTP to materials not adsorbed by charcoal. This partially purified enzyme showed essentially no GTP phosphohydrolase activity and an optimal pH of 8.0. An apparent Km of about 23.8 mumol/1 was obtained with respect to 5-hydroxylysine. Mg2+ was required for the activity of this enzyme. Ethanolamine, L-lysine, L-ornithine and choline inhibited the enzyme but L-threonine, L-serine and hydroxy-L-proline did not. None of these compounds severed as substrate for this enzyme. | Guanosine triphosphate: 5-hydroxylysine phosphotransferase in rat kidney cortex. An enzyme which catalyzes the transfer of the gamma-phosphate from GTP onto 5-hydroxylysine was partially purified from rat kidney cortex by means of acid precipitation and DEAE-Sephadex A-50 column chromatography. The enzyme activity was assayed by measuring the transfer of [32P] from gamma-[32P]-GTP to materials not adsorbed by charcoal. This partially purified enzyme showed essentially no GTP phosphohydrolase activity and an optimal pH of 8.0. An apparent Km of about 23.8 mumol/1 was obtained with respect to 5-hydroxylysine. Mg2+ was required for the activity of this enzyme. Ethanolamine, L-lysine, L-ornithine and choline inhibited the enzyme but L-threonine, L-serine and hydroxy-L-proline did not. None of these compounds severed as substrate for this enzyme. | [
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|
PMID:18340 | Isatin enzyme interactions. V. Activation of rat liver acid phosphatase. | Influence of Isatin on rat tissue acid phosphatase has been studied. It has an organ-specific effect, the liver enzyme is activated, the brain enzyme is inhibited while those of kidney and intestine are not affected. Isatin activation of the liver enzyme is of mixed type and pH dependent. Rat liver enzyme appears to require intact amino and sulfhydryl groups for activity. Isatin seems to combine with the enzyme through the sulfhydryl group of the latter. | Isatin enzyme interactions. V. Activation of rat liver acid phosphatase. Influence of Isatin on rat tissue acid phosphatase has been studied. It has an organ-specific effect, the liver enzyme is activated, the brain enzyme is inhibited while those of kidney and intestine are not affected. Isatin activation of the liver enzyme is of mixed type and pH dependent. Rat liver enzyme appears to require intact amino and sulfhydryl groups for activity. Isatin seems to combine with the enzyme through the sulfhydryl group of the latter. | [
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|
PMID:18341 | Effect of cofactor depletion on liver tyrosine aminotransferase expression. | Hepatic tyrosine aminotransferase (L-tyrosine: 2-oxoglutarate aminotransferase, EC 2.6.1.5) was partially purified from pyridoxine depleted and control rats and subsequently resolved by electrophoresis on polyacrylamide gels. Enzyme activity was detected histochemically in situ on the gel. Six enzymatically active forms were detected. Cofactor depletion effected further resolution of the enzyme into seven active forms as revealed by the bifurcation of the major active peak. | Effect of cofactor depletion on liver tyrosine aminotransferase expression. Hepatic tyrosine aminotransferase (L-tyrosine: 2-oxoglutarate aminotransferase, EC 2.6.1.5) was partially purified from pyridoxine depleted and control rats and subsequently resolved by electrophoresis on polyacrylamide gels. Enzyme activity was detected histochemically in situ on the gel. Six enzymatically active forms were detected. Cofactor depletion effected further resolution of the enzyme into seven active forms as revealed by the bifurcation of the major active peak. | [
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|
PMID:18342 | Interactions of citrate synthases from osmoconforming and osmoregulating animals with salt: possible signs of molecular eco-adaptation? | This study considers differential sensitivity of citrate synthase (citrate oxaloacetatelyase [CoA acetylating]) EC 4.1.3.7. from an osmoconforming animal (sea anemone) and an osmoregulating animal (the pig) to salt. Attention is drawn to the fact that the osmoconforming sea anemone is in essence a sessile creature while the pig is readily mobile and able to change its ionic environment at will. It had been shown earlier that citrate synthase from another osmoconformer (oyster) is also not sensitive to ionic strength while citrate synthase from osmoregulating white shrimp is sensitive to increasing levels of salt. However, these enzymes are characteristically regulated by ATP and alpha-ketoglutarate. Both forms of citrate synthase are denatured by 6 M guanidine hydrochloride and are aided by salt levels in their refolding but the rate and extent of refolding of the osmoconformer citrate synthase are greater than those of the osmoregulator citrate synthase. Catalytic activity of both forms of citrate synthase is inhibited by incubation in distilled water; osmoconformer citrate synthase was inhibited completely in 7 h while osmoregulator citrate synthase was inhibited only 60% in this time and 80% after 22 h in distilled water. The eco-adaptive and evolutionary implications of these findings are discussed. | Interactions of citrate synthases from osmoconforming and osmoregulating animals with salt: possible signs of molecular eco-adaptation? This study considers differential sensitivity of citrate synthase (citrate oxaloacetatelyase [CoA acetylating]) EC 4.1.3.7. from an osmoconforming animal (sea anemone) and an osmoregulating animal (the pig) to salt. Attention is drawn to the fact that the osmoconforming sea anemone is in essence a sessile creature while the pig is readily mobile and able to change its ionic environment at will. It had been shown earlier that citrate synthase from another osmoconformer (oyster) is also not sensitive to ionic strength while citrate synthase from osmoregulating white shrimp is sensitive to increasing levels of salt. However, these enzymes are characteristically regulated by ATP and alpha-ketoglutarate. Both forms of citrate synthase are denatured by 6 M guanidine hydrochloride and are aided by salt levels in their refolding but the rate and extent of refolding of the osmoconformer citrate synthase are greater than those of the osmoregulator citrate synthase. Catalytic activity of both forms of citrate synthase is inhibited by incubation in distilled water; osmoconformer citrate synthase was inhibited completely in 7 h while osmoregulator citrate synthase was inhibited only 60% in this time and 80% after 22 h in distilled water. The eco-adaptive and evolutionary implications of these findings are discussed. | [
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|
PMID:18343 | A deoxyribonuclease from Chlamydomonas reinhardii. 1. Purification and properties. | A deoxyribonuclease has been purified more than 2000-fold from the green algae, Chlamydomonas reinhardii. The enzyme is most active on denatured DNA. Optimum activity is at pH 8.5, in 80 mM Tris-HCl buffer and 2 mM CaCl2. Other divalent cations can replace Ca2+ with varying lower efficiency. EDTA and inorganic phosphate are strongly inhibitory, while ATP and high concentrations of 2-mercaptoethanol are slightly inhibitory. The molecular weight is approximately 35 000, the Stokes radius is 2.7 nm, and the sedimentation coefficient 2.8 S. It is a single polypeptide chain, and the frictional ratio of 1.27 suggests it is only slightly asymetrical. The isoelectric point is 9.5. This enzyme has been termed exonuclease 1. | A deoxyribonuclease from Chlamydomonas reinhardii. 1. Purification and properties. A deoxyribonuclease has been purified more than 2000-fold from the green algae, Chlamydomonas reinhardii. The enzyme is most active on denatured DNA. Optimum activity is at pH 8.5, in 80 mM Tris-HCl buffer and 2 mM CaCl2. Other divalent cations can replace Ca2+ with varying lower efficiency. EDTA and inorganic phosphate are strongly inhibitory, while ATP and high concentrations of 2-mercaptoethanol are slightly inhibitory. The molecular weight is approximately 35 000, the Stokes radius is 2.7 nm, and the sedimentation coefficient 2.8 S. It is a single polypeptide chain, and the frictional ratio of 1.27 suggests it is only slightly asymetrical. The isoelectric point is 9.5. This enzyme has been termed exonuclease 1. | [
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|
PMID:18344 | Dihydroxyacetone reductase from Mucor javanicus. 1. Isolation and properties. | An NADPH-dependent oxidoreductase has been extracted from the mycelium of the fungus Mucor Javanicus (Wehmer) and enriched 1000-fold with respect to the protein contained in the crude extract after centrifugation at 2600 X g. The molecular weight of the enzyme was estimated by gel filtration to be about 100 000; electrophoresis under dissociating conditions indicates four subunits of molecular weight about 28 000. Data on stability and activity of the enzyme as a function of pH and temperature are reported. From a kinetic study and product analysis of the reduction of the two enantiomeric trans-1-decalones and also from a kinetic study of the oxidation of the two diastereomeric pairs of trans-1-decalols it follows that the enzymes is an e-Si oxidoreductase (according to the nomenclature proposed by Dutler et al., Eur. J. Biochem. 22 [1971]203-212 and Prelog and Helmchen, Helv. Chim. Acta, 55 [1972] 2581-2598). This classification is amply confirmed by the kinetic behaviour of a large number of alicyclic substrates. Using (4-2HSi-labelled coenzyme to reduce (9S)-trans-1,4-decalindione, it was shown that the enzyme is HSi (= HS = HB)-stereospecific with respect to the coenzyme. It is demonstrated that the oxidoreductase from Mucor javanicus can be used for the preparation of optically pure chiral alcohols and ketones. In the following paper evidence is presented that the natural substrate of the enzyme is dihydroxyacetone. | Dihydroxyacetone reductase from Mucor javanicus. 1. Isolation and properties. An NADPH-dependent oxidoreductase has been extracted from the mycelium of the fungus Mucor Javanicus (Wehmer) and enriched 1000-fold with respect to the protein contained in the crude extract after centrifugation at 2600 X g. The molecular weight of the enzyme was estimated by gel filtration to be about 100 000; electrophoresis under dissociating conditions indicates four subunits of molecular weight about 28 000. Data on stability and activity of the enzyme as a function of pH and temperature are reported. From a kinetic study and product analysis of the reduction of the two enantiomeric trans-1-decalones and also from a kinetic study of the oxidation of the two diastereomeric pairs of trans-1-decalols it follows that the enzymes is an e-Si oxidoreductase (according to the nomenclature proposed by Dutler et al., Eur. J. Biochem. 22 [1971]203-212 and Prelog and Helmchen, Helv. Chim. Acta, 55 [1972] 2581-2598). This classification is amply confirmed by the kinetic behaviour of a large number of alicyclic substrates. Using (4-2HSi-labelled coenzyme to reduce (9S)-trans-1,4-decalindione, it was shown that the enzyme is HSi (= HS = HB)-stereospecific with respect to the coenzyme. It is demonstrated that the oxidoreductase from Mucor javanicus can be used for the preparation of optically pure chiral alcohols and ketones. In the following paper evidence is presented that the natural substrate of the enzyme is dihydroxyacetone. | [
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|
PMID:18348 | On the structure of flavin-oxygen intermediates involved in enzymatic reactions. | During the catalytic reactions of flavoprotein hydroxylases and bacterial luciferase, flavin peroxides are formed as intermediates [see Massey, V. and Hemmerich, P. (1976) in The Enzymes, 3rd edn (P. Boyer, ed.) pp. 421--505, Academic Press, New York]. These intermediates have been postulated to be C(4a) derivatives of the flavin coenzyme. To test this hypothesis, modified flavin coenzymes carrying an oxygen substituent at position C(4a) of the isoalloxazine ring were synthesized. They are tightly bound by the apoenzymes of D-amino acid oxidase, p-hydroxybenzoate hydroxylase and lactate oxidase; the resulting complexes show spectral properties closely similar to those of the transient oxygen adducts of the hydroxylases. The optical spectra of the lumiflavin model compounds were found to be highly dependent on the solvent environment and nature of the subsituents. Under appropriate conditions they simulate satisfactorily the spectra of the transient enzymatic oxygen adducts. The results support the proposal that the primary oxygen adducts formed with these flavoproteins on reaction of the reduced enzymes with oxygen are flavin C(4a) peroxides. | On the structure of flavin-oxygen intermediates involved in enzymatic reactions. During the catalytic reactions of flavoprotein hydroxylases and bacterial luciferase, flavin peroxides are formed as intermediates [see Massey, V. and Hemmerich, P. (1976) in The Enzymes, 3rd edn (P. Boyer, ed.) pp. 421--505, Academic Press, New York]. These intermediates have been postulated to be C(4a) derivatives of the flavin coenzyme. To test this hypothesis, modified flavin coenzymes carrying an oxygen substituent at position C(4a) of the isoalloxazine ring were synthesized. They are tightly bound by the apoenzymes of D-amino acid oxidase, p-hydroxybenzoate hydroxylase and lactate oxidase; the resulting complexes show spectral properties closely similar to those of the transient oxygen adducts of the hydroxylases. The optical spectra of the lumiflavin model compounds were found to be highly dependent on the solvent environment and nature of the subsituents. Under appropriate conditions they simulate satisfactorily the spectra of the transient enzymatic oxygen adducts. The results support the proposal that the primary oxygen adducts formed with these flavoproteins on reaction of the reduced enzymes with oxygen are flavin C(4a) peroxides. | [
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|
PMID:18351 | A phosphorus-magnetic-resonance study of the interaction of Mg2+ with adenyl-5'-yl imidodiphosphate. Binding sites of Mg2+ ion on the phosphate chain. | The interaction of Mg2+ ions with adenyl-5'-yl imidodiphosphate, AMP-P(NH)P, has been studied at basic and acidic pH values by phosphorus magnetic resonance spectroscopy in aqueous solution. The results suggest that Mg2+ binds simultaneously to one (or both) of the two free oxygen atoms of the beta-phosphate moiety and to the nitrogen atom of the phosphate chain (P alpha-O-P beta-N-P gamma). The interaction arises from 1: 1 complexing of Mg2+ to AMP-P(NH)P. The mode of the Mg2+ binding on the phosphate chain remains the same at both basic and acidic pH values. As in the case of ATP and ADP, the association of Mg2+ reduces the pK by about 1.5 units. On the other hand phosphorus titration curves showed that when the phosphate chain does not possess the regular periodicity (O-P alpha-O-P beta-X-P gamma-O,X not equal to O) as in the case of ATP, protonation of the terminal phosphate group may induce a 31P chemical shift variation less important for this group than for the preceding one. | A phosphorus-magnetic-resonance study of the interaction of Mg2+ with adenyl-5'-yl imidodiphosphate. Binding sites of Mg2+ ion on the phosphate chain. The interaction of Mg2+ ions with adenyl-5'-yl imidodiphosphate, AMP-P(NH)P, has been studied at basic and acidic pH values by phosphorus magnetic resonance spectroscopy in aqueous solution. The results suggest that Mg2+ binds simultaneously to one (or both) of the two free oxygen atoms of the beta-phosphate moiety and to the nitrogen atom of the phosphate chain (P alpha-O-P beta-N-P gamma). The interaction arises from 1: 1 complexing of Mg2+ to AMP-P(NH)P. The mode of the Mg2+ binding on the phosphate chain remains the same at both basic and acidic pH values. As in the case of ATP and ADP, the association of Mg2+ reduces the pK by about 1.5 units. On the other hand phosphorus titration curves showed that when the phosphate chain does not possess the regular periodicity (O-P alpha-O-P beta-X-P gamma-O,X not equal to O) as in the case of ATP, protonation of the terminal phosphate group may induce a 31P chemical shift variation less important for this group than for the preceding one. | [
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|
PMID:18352 | Nickel cytochrome c. Effect of protein moiety on the metal ion coordination. | Nickel cytochrome c has been synthesized by the reaction of metal-free porphyrin cytochrome c with Ni(II) ions in 0.6 Mglycylglycine and 4 M KSCN. Electronic spectra and susceptibility measurement showed the nickel to be in a high-spin octahedral configuration exemplifying the strong influence of the protein moiety as a macrocyclic ligand on the coordination chemistry of the metal ion. Nickel cytochrome c has the same electrophoretic mobility, helicity and pK values of conformational transitions as the native enzyme. At high pH, the partially denatured nickel cytochrome c becomes dimeric. Nitric oxide reacts with nickel cytochrome c to form the nitrosyl derivative with (formula: see text). Reaction of NO with nickel protoporphyrin IX dimethyl ester in toluene, pyridine, or methylthioethanol produced no stable nitrosyl products, clearly demonstrating the effect of protein on metal ion ligation. | Nickel cytochrome c. Effect of protein moiety on the metal ion coordination. Nickel cytochrome c has been synthesized by the reaction of metal-free porphyrin cytochrome c with Ni(II) ions in 0.6 Mglycylglycine and 4 M KSCN. Electronic spectra and susceptibility measurement showed the nickel to be in a high-spin octahedral configuration exemplifying the strong influence of the protein moiety as a macrocyclic ligand on the coordination chemistry of the metal ion. Nickel cytochrome c has the same electrophoretic mobility, helicity and pK values of conformational transitions as the native enzyme. At high pH, the partially denatured nickel cytochrome c becomes dimeric. Nitric oxide reacts with nickel cytochrome c to form the nitrosyl derivative with (formula: see text). Reaction of NO with nickel protoporphyrin IX dimethyl ester in toluene, pyridine, or methylthioethanol produced no stable nitrosyl products, clearly demonstrating the effect of protein on metal ion ligation. | [
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|
PMID:18353 | Anorchi. | Anorchia was diagnosed in 3 patients after a hormonal study entailing the basic determination of FSH, LH and testosterone by a radioimmunoassay technique and by later stimulation with HCG. 2,000 IU are administered during 3 consecutive days with a new testosterone determination 24 and 72 h after the last administration. If the result is negative, the test is repeated but the HCG dose is doubled. A new negative response confirms the diagnosis of anorchia. If the results are positive a spermatic phlebography allows localization of the testicles or the remains of Wolffian origin in order to simplify surgery. | Anorchi. Anorchia was diagnosed in 3 patients after a hormonal study entailing the basic determination of FSH, LH and testosterone by a radioimmunoassay technique and by later stimulation with HCG. 2,000 IU are administered during 3 consecutive days with a new testosterone determination 24 and 72 h after the last administration. If the result is negative, the test is repeated but the HCG dose is doubled. A new negative response confirms the diagnosis of anorchia. If the results are positive a spermatic phlebography allows localization of the testicles or the remains of Wolffian origin in order to simplify surgery. | [
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|
PMID:18354 | Regional changes in the activities of aminergic biosynthetic enzymes in the brains of hypertensive rats. | The activities of monoamine biosynthetic enzymes were measured in brain regions of several hypertensive rat models at various ages. The types of hypertensive rats were the spontaneously hypertensive rat (SHR) and a stroke-prone substrain of the SHR as well as DOCA-salt and renal hypertensive rats. The genetically hypertensive rats had significantly elevated blood pressures as compared to the Wistar-Kyoto control rat after 5 weeks of age. During the early development of hypertension in the SHR, the activities of tyrosine hydroxylase in the hypothalamus and corpus striatum and of dopamine-beta-hydroxylase in the hypothalamus and pons-medulla were significantly higher than in the control rats. Tryptophan-hydroxylase was also elevated in the hypothalamus in SHR. From 3 to 8 weeks of age there appeared to be a significant correlation between hypothalamic dopamine-beta-hydroxylase activity and blood pressure in the hypertensive rats. In contrast, the activities of tyrosine hydroxylase and dopamine-beta-hydroxylase were slightly decreased in the DOCA-salt and renal hypertensive rats. It is suggested that noradrenergic or adrenergic neurons in the hypothalamus may participate in the initiation of elevated blood pressure in the genetic, but not in the DOCA-salt or renal hypertensive rats. | Regional changes in the activities of aminergic biosynthetic enzymes in the brains of hypertensive rats. The activities of monoamine biosynthetic enzymes were measured in brain regions of several hypertensive rat models at various ages. The types of hypertensive rats were the spontaneously hypertensive rat (SHR) and a stroke-prone substrain of the SHR as well as DOCA-salt and renal hypertensive rats. The genetically hypertensive rats had significantly elevated blood pressures as compared to the Wistar-Kyoto control rat after 5 weeks of age. During the early development of hypertension in the SHR, the activities of tyrosine hydroxylase in the hypothalamus and corpus striatum and of dopamine-beta-hydroxylase in the hypothalamus and pons-medulla were significantly higher than in the control rats. Tryptophan-hydroxylase was also elevated in the hypothalamus in SHR. From 3 to 8 weeks of age there appeared to be a significant correlation between hypothalamic dopamine-beta-hydroxylase activity and blood pressure in the hypertensive rats. In contrast, the activities of tyrosine hydroxylase and dopamine-beta-hydroxylase were slightly decreased in the DOCA-salt and renal hypertensive rats. It is suggested that noradrenergic or adrenergic neurons in the hypothalamus may participate in the initiation of elevated blood pressure in the genetic, but not in the DOCA-salt or renal hypertensive rats. | [
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|
PMID:18355 | Some cardiovascular effects of ST-91 and clonidine. | St-91, 2(2,6-diethylphenylamino)-2-imidazoline, is a clonidine derivative which does not penetrate the blood-brain barrier. In spontaneously hypertensive (SH) rats is acutely increased arterial pressure and reduced heart rate while at 8 to 12 h after oral administration, it slightly lowered arterial pressure. In contrast, clonidine had acute antihypertensive activity at all doses used. By intracerebroventricular administration to SH rats, both drugs (St-91 and clonidine) reduced arterial pressure and heart rate; in this respect, clonidine was more potent then St-91. Cardiac acceleration induced by low frequency electrical stimulation of right cardiac sympathetic nerves in anesthetized and vagotomized dogs was reduced by St-91 at the same doses by clonidine. Phenoxybenzamine, phentolamine and desipramine antagonized the inhibitory effects of St-91 on electrically induced cardiac acceleration. It was concluded that St-91, like clonidine, stimulates inhibitory alpha-adrenergic receptors at the sympathetic nerve endings but, unlike clonidine, is substantially devoid of acute antihypertensive activity. This suggests that stimulation of peripheral presynaptic inhibitory alpha-adrenergic receptors is not likely to represent the sole mechanism of antihypertensive action of clonidine. | Some cardiovascular effects of ST-91 and clonidine. St-91, 2(2,6-diethylphenylamino)-2-imidazoline, is a clonidine derivative which does not penetrate the blood-brain barrier. In spontaneously hypertensive (SH) rats is acutely increased arterial pressure and reduced heart rate while at 8 to 12 h after oral administration, it slightly lowered arterial pressure. In contrast, clonidine had acute antihypertensive activity at all doses used. By intracerebroventricular administration to SH rats, both drugs (St-91 and clonidine) reduced arterial pressure and heart rate; in this respect, clonidine was more potent then St-91. Cardiac acceleration induced by low frequency electrical stimulation of right cardiac sympathetic nerves in anesthetized and vagotomized dogs was reduced by St-91 at the same doses by clonidine. Phenoxybenzamine, phentolamine and desipramine antagonized the inhibitory effects of St-91 on electrically induced cardiac acceleration. It was concluded that St-91, like clonidine, stimulates inhibitory alpha-adrenergic receptors at the sympathetic nerve endings but, unlike clonidine, is substantially devoid of acute antihypertensive activity. This suggests that stimulation of peripheral presynaptic inhibitory alpha-adrenergic receptors is not likely to represent the sole mechanism of antihypertensive action of clonidine. | [
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|
PMID:18356 | Specific alpha-adrenoceptor blocking effect of droperidol on isolated smooth muscles. | The present study was conducted so as to obtain more insight into the controversies concerning the alpha-adrenoceptor blocking properties of droperidol, a short-acting neuroleptic agent used in neuroleptanalgesia. The effect of droperidol on the vasoconstriction induced by norepinephrine, sympathetic nerve stimulation, histamine and potassium ions was studied on isolated, perfused ear arteries; its effect on norepinephrine-induced contraction was studied on isolated aorta, spleen and vas deferens. In addition, the onset and duration of action of droperidol was studied. Low doses of droperidol inhibit the vasoconstriction induced by norepinephrine and sympathetic nerve stimulation in the ear artery of the rabbit (3.3 X 10(-9) M and 1.3 X 10(-8) M respectively). At similar low doses, droperidol inhibits norepinephrine-induced contractions in the other tissues studied and has a potency comparable to that of phentolamine; its action is rapid in onset and of short duration. High doses of droperidol (10(-6) M) also inhibit the vasoconstriction of the ear artery induced by histamine and by potassium ions. These findings indicate that a low doses, droperidol has specific and competitive alpha-adrenoceptor blocking effects. | Specific alpha-adrenoceptor blocking effect of droperidol on isolated smooth muscles. The present study was conducted so as to obtain more insight into the controversies concerning the alpha-adrenoceptor blocking properties of droperidol, a short-acting neuroleptic agent used in neuroleptanalgesia. The effect of droperidol on the vasoconstriction induced by norepinephrine, sympathetic nerve stimulation, histamine and potassium ions was studied on isolated, perfused ear arteries; its effect on norepinephrine-induced contraction was studied on isolated aorta, spleen and vas deferens. In addition, the onset and duration of action of droperidol was studied. Low doses of droperidol inhibit the vasoconstriction induced by norepinephrine and sympathetic nerve stimulation in the ear artery of the rabbit (3.3 X 10(-9) M and 1.3 X 10(-8) M respectively). At similar low doses, droperidol inhibits norepinephrine-induced contractions in the other tissues studied and has a potency comparable to that of phentolamine; its action is rapid in onset and of short duration. High doses of droperidol (10(-6) M) also inhibit the vasoconstriction of the ear artery induced by histamine and by potassium ions. These findings indicate that a low doses, droperidol has specific and competitive alpha-adrenoceptor blocking effects. | [
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|
PMID:18357 | Withdrawal syndrome upon cessation of chronic clonidine treatment in rats. | Clinical reports indicate that cessation of treatment with the antihypertensive agent clonidine is associated with a withdrawal syndrome which may include a hypertensive overshoot of critical proportions. We have attempted to produce an animal model of this syndrome in rats. Rats were treated with clonidine in the drinking water (5 microgram/ml; total dose 300-500 microgram/kg/day) which produced a significant (approx. 20%) decrease in heart rate and blood pressure. Within 24 h of cessation of treatment a significantly greater (approximately 100 beats/min) heart rate was seen in treated animals than in control animals when measurements were made in conscious animals. No hypertensive overshoot was observed. Cessation of treatment was associated with an increase in sympatho-adrenal tone as shown by a trans-synaptic induction of adrenal tyrosine hydroxylase activity. Adrenal denervation prevented the rise in adrenal tyrosine hydroxylase seen after cessation of treatment. Administration of clonidine to pregnant rats (10th day until term) did not alter the development of adrenal tyrosine hydroxylase in the offspring. The data indicate that a withdrawal syndrome is produced upon cessation of chronic clonidine treatment. | Withdrawal syndrome upon cessation of chronic clonidine treatment in rats. Clinical reports indicate that cessation of treatment with the antihypertensive agent clonidine is associated with a withdrawal syndrome which may include a hypertensive overshoot of critical proportions. We have attempted to produce an animal model of this syndrome in rats. Rats were treated with clonidine in the drinking water (5 microgram/ml; total dose 300-500 microgram/kg/day) which produced a significant (approx. 20%) decrease in heart rate and blood pressure. Within 24 h of cessation of treatment a significantly greater (approximately 100 beats/min) heart rate was seen in treated animals than in control animals when measurements were made in conscious animals. No hypertensive overshoot was observed. Cessation of treatment was associated with an increase in sympatho-adrenal tone as shown by a trans-synaptic induction of adrenal tyrosine hydroxylase activity. Adrenal denervation prevented the rise in adrenal tyrosine hydroxylase seen after cessation of treatment. Administration of clonidine to pregnant rats (10th day until term) did not alter the development of adrenal tyrosine hydroxylase in the offspring. The data indicate that a withdrawal syndrome is produced upon cessation of chronic clonidine treatment. | [
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|
PMID:18366 | [Effect of various analeptics on the outcome of acute microwave lesion in mice]. | The survival of albino mice irradiated by microwaves till the terminal state (wave length of 12.5 cm, intensity-62 +/- 5 microvat, for 14-16 minutes), given directly after irradiation diethylamide of nicotinic acid (cordiamine) in a dose of 50 mg/kg intraperitoneally and strychnine nitrate in a dose of 1 mg/kg, subcutaneously, i. e. nearly 1.5 times as much as received by controls, was studied. The application of caffeine sodium benzoate, camphor, metrasol, lobeline hydrochloride and cytisine, employed in different doses, proved to be little effective. | [Effect of various analeptics on the outcome of acute microwave lesion in mice]. The survival of albino mice irradiated by microwaves till the terminal state (wave length of 12.5 cm, intensity-62 +/- 5 microvat, for 14-16 minutes), given directly after irradiation diethylamide of nicotinic acid (cordiamine) in a dose of 50 mg/kg intraperitoneally and strychnine nitrate in a dose of 1 mg/kg, subcutaneously, i. e. nearly 1.5 times as much as received by controls, was studied. The application of caffeine sodium benzoate, camphor, metrasol, lobeline hydrochloride and cytisine, employed in different doses, proved to be little effective. | [
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|
PMID:18370 | [Effect of corticotropin on functional and metabolic processes in muscles]. | Changes of excitability, contractility, temperature, activity of oxydation-reduction enzymes, pyridine nucleotides, and of free fatty acids were studied in m. m. gastrocnemii of rabbits and rats during activity, after administration of 1 unit per 100 g of corticotropin (ACTH). Within 30 min after administration, the ACTH did not alter the excitability of a neural-muscular system but increased its efficiency by means of stimulation of the free fatty acids usage. Besides, the ACTH elicits no regular changes of the oxydation--reduction enzymes activity during a short-lasting muscular activity. Its regulating effect on the metabolic processes in muscles is realised at the level of anaerobic processes. | [Effect of corticotropin on functional and metabolic processes in muscles]. Changes of excitability, contractility, temperature, activity of oxydation-reduction enzymes, pyridine nucleotides, and of free fatty acids were studied in m. m. gastrocnemii of rabbits and rats during activity, after administration of 1 unit per 100 g of corticotropin (ACTH). Within 30 min after administration, the ACTH did not alter the excitability of a neural-muscular system but increased its efficiency by means of stimulation of the free fatty acids usage. Besides, the ACTH elicits no regular changes of the oxydation--reduction enzymes activity during a short-lasting muscular activity. Its regulating effect on the metabolic processes in muscles is realised at the level of anaerobic processes. | [
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|
PMID:18367 | [Relationship between the pain-relieving action of narcotic analgesics and their effect on respiration]. | Experiments with rabbits (30) and albino rats (110) demonstrated that morphine (1 mg/kg), promedol (trimeperidine) (2 mg/kg) and phentanyl (0.02 mg/kg), while raising by 21-24 per cent of the algesia threshold, produced an analgesic effect differeing in its duration (morphine-130 min, trimeperidine-70 min, phentanyl-17 min). This is attended by changes in respiration (greater on introduction of phentanyl, and lesser, following administration of morphine) and by shifts in the functional state of the tissues (greater on introduction of trimeperidine and lesser after aministration of morphine). | [Relationship between the pain-relieving action of narcotic analgesics and their effect on respiration]. Experiments with rabbits (30) and albino rats (110) demonstrated that morphine (1 mg/kg), promedol (trimeperidine) (2 mg/kg) and phentanyl (0.02 mg/kg), while raising by 21-24 per cent of the algesia threshold, produced an analgesic effect differeing in its duration (morphine-130 min, trimeperidine-70 min, phentanyl-17 min). This is attended by changes in respiration (greater on introduction of phentanyl, and lesser, following administration of morphine) and by shifts in the functional state of the tissues (greater on introduction of trimeperidine and lesser after aministration of morphine). | [
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|
PMID:18371 | Nucleolar and functional characterization of lymphocytes following cyclophosphamide treatment. | Profound lymphocyte depletion occurs in thymus, lymph nodes and spleens of normal and thymectomized mice early after a single high sublethal dose of cyclophosphamide. Among individual lymphocytes characterized by the different types of nucleolus, the "non-activable" lymphocytes with micronucleoli are affected most markedly by CY, whereas the proportion of the "active" lymphocytes is increased (up to 70% in the spleens). During the period of regeneration lymphocytes with micronucleoli increase rapidly, the increment thereof is very high on day 14 in the spleens of all mice, and still higher in the blood and lymph nodes of thymectomized animals. During the period of lymphocytopenia, in spite of the increased proportion of the "active" lymphocytes, the ability of residual cells of elicit GVH reactions and to incorporate 14C-uridine decreases. In contrast to changes in the early period, the recovery of the "active" lymphocytes and GVH reactivity, which is still incomplete on day 14, depends on the presence of an intact thymus. | Nucleolar and functional characterization of lymphocytes following cyclophosphamide treatment. Profound lymphocyte depletion occurs in thymus, lymph nodes and spleens of normal and thymectomized mice early after a single high sublethal dose of cyclophosphamide. Among individual lymphocytes characterized by the different types of nucleolus, the "non-activable" lymphocytes with micronucleoli are affected most markedly by CY, whereas the proportion of the "active" lymphocytes is increased (up to 70% in the spleens). During the period of regeneration lymphocytes with micronucleoli increase rapidly, the increment thereof is very high on day 14 in the spleens of all mice, and still higher in the blood and lymph nodes of thymectomized animals. During the period of lymphocytopenia, in spite of the increased proportion of the "active" lymphocytes, the ability of residual cells of elicit GVH reactions and to incorporate 14C-uridine decreases. In contrast to changes in the early period, the recovery of the "active" lymphocytes and GVH reactivity, which is still incomplete on day 14, depends on the presence of an intact thymus. | [
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|
PMID:18374 | Evidence for improved cardiac performance after beta-blockade in patients with coronary artery disease. | The study was undertaken to investigate the acute haemodynamic effects of bunitrolol (0-2-hydroxy-3-(tert.butylamino)-propoxy)-bity. Right and left heart catheterization was performed in eleven patients with documented coronary artery disease. After bunitrolol (10 mg i.v.), there was a statistically significant decrease in left ventricular and aortic systolic pressures left ventricular end-diastolic pressure, aortic diastolic and mean pressures, pressure-rate product and compliance index (delta P/delta V). Left ventricular dp/dt, left ventricular dp/dt over isovolumic pressure, systemic resistance and heart rate tended to decrease, stroke volume and left ventricular stroke work index tended to increase, without statistical significance. Cardiac index showed individual variations, the mean values for the group being unchanged. Correlation of left ventricular end-diastolic pressure and left ventricular stroke work index showed a shift toward improved ventricular function curve in most cases, deterioration in no instance. Supine exercise was performed in ten patients. Angina occurred in nine patients; in five only before and in four before and after beta-blockade. Post-drug exercise heart rate, pressure-rate product and left ventricular end-diastolic pressure were significantly lower, the latter also in the four patients who still presented exercise angina. It is concluded that certain beta-blockers can improve cardiac performance at rest and during exercise in patients with coronary artery disease. This is explainable on the basis of a more favourable balance between oxygen supply and demand, together with a less marked negative inotropic effect due to the partial agonist activity of the agent used in the study. | Evidence for improved cardiac performance after beta-blockade in patients with coronary artery disease. The study was undertaken to investigate the acute haemodynamic effects of bunitrolol (0-2-hydroxy-3-(tert.butylamino)-propoxy)-bity. Right and left heart catheterization was performed in eleven patients with documented coronary artery disease. After bunitrolol (10 mg i.v.), there was a statistically significant decrease in left ventricular and aortic systolic pressures left ventricular end-diastolic pressure, aortic diastolic and mean pressures, pressure-rate product and compliance index (delta P/delta V). Left ventricular dp/dt, left ventricular dp/dt over isovolumic pressure, systemic resistance and heart rate tended to decrease, stroke volume and left ventricular stroke work index tended to increase, without statistical significance. Cardiac index showed individual variations, the mean values for the group being unchanged. Correlation of left ventricular end-diastolic pressure and left ventricular stroke work index showed a shift toward improved ventricular function curve in most cases, deterioration in no instance. Supine exercise was performed in ten patients. Angina occurred in nine patients; in five only before and in four before and after beta-blockade. Post-drug exercise heart rate, pressure-rate product and left ventricular end-diastolic pressure were significantly lower, the latter also in the four patients who still presented exercise angina. It is concluded that certain beta-blockers can improve cardiac performance at rest and during exercise in patients with coronary artery disease. This is explainable on the basis of a more favourable balance between oxygen supply and demand, together with a less marked negative inotropic effect due to the partial agonist activity of the agent used in the study. | [
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|
PMID:18375 | The use of flurazepam (dalmane) as a substitute for barbiturates and methaqualone/diphenhydramine (mandrax) in general practice. | A twelve-week study involving fifty-three patients is described as taking place in a practice with a higher than average geriatric population. The purpose of the study was to substitute flurazepam for habitually used barbiturates or methaqualone/diphenhydramine. Of the original fifty-three patients admitted to the study, fifty-one completed; the two drop-outs resulting from concomitant physical illness. Eighty-four per cent of patients were successfully changed to flurazepam. Of those who did not accept flurazepam, eight per cent accepted nitrazepam, while six per cent of patients were motivated to stop all hypnotics. During the three month period of the study none of the well-known disadvantages of the barbiturates and methaqualone/diphenhydramine were seen with flurazepam. The author found flurazepam to be a very efficient hypnotic of relatively low toxicity which could be easily substituted fro barbiturates and methaqualone/diphenhydramine in the treatment of long-term insomnia. | The use of flurazepam (dalmane) as a substitute for barbiturates and methaqualone/diphenhydramine (mandrax) in general practice. A twelve-week study involving fifty-three patients is described as taking place in a practice with a higher than average geriatric population. The purpose of the study was to substitute flurazepam for habitually used barbiturates or methaqualone/diphenhydramine. Of the original fifty-three patients admitted to the study, fifty-one completed; the two drop-outs resulting from concomitant physical illness. Eighty-four per cent of patients were successfully changed to flurazepam. Of those who did not accept flurazepam, eight per cent accepted nitrazepam, while six per cent of patients were motivated to stop all hypnotics. During the three month period of the study none of the well-known disadvantages of the barbiturates and methaqualone/diphenhydramine were seen with flurazepam. The author found flurazepam to be a very efficient hypnotic of relatively low toxicity which could be easily substituted fro barbiturates and methaqualone/diphenhydramine in the treatment of long-term insomnia. | [
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|
PMID:18376 | A trial of benorylate tablets in the symptomatic relief of osteoarthritis. | A two-week assessment of Benoral tablets was carried out in general practice in 171 patients with degenerative joint disease to see how symptomatic response related to selected presenting features of the disease. A short history, less severe initial state and multiple joint involvement were each associated with a better response. Overall 84-4% of patients reported Benoral tablets to have helped relieve their symptoms and a high proportion preferred Benoral to their previous anti-arthritic medication. | A trial of benorylate tablets in the symptomatic relief of osteoarthritis. A two-week assessment of Benoral tablets was carried out in general practice in 171 patients with degenerative joint disease to see how symptomatic response related to selected presenting features of the disease. A short history, less severe initial state and multiple joint involvement were each associated with a better response. Overall 84-4% of patients reported Benoral tablets to have helped relieve their symptoms and a high proportion preferred Benoral to their previous anti-arthritic medication. | [
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|
PMID:18377 | Temperature dependence of adenylate cyclase activity from rat white adipocytes. | The effects of temperature on adenylate cyclase (AC) activity from rat white adipocytes were studied. Arrhenius plots of the data were found to be biphasic for basal AC activity, with a break near 27 degrees C. Noradrenaline and corticotropin induced a shift in the break with a rise in energy of activation (Ea) on both sides of the break. Aabove break point only, Ea increased with respect to hormone does as a hyperbolic function. The maximum value was in the range 17-21 Kcal x mol-1. Temperature was shown to have only a slight effect on the binding capacity of corticotropin to its receptor sites. The possibility of the existence of multiple thermodynamic states for the enzyme is envisaged. Basal AC activity is thermodynamically different from the hormone-stimulated enzyme. Hormones induce changes in the basal conformation of the enzyme, and this is reflected in modifications of Arrhenius plots. The maximal state of activation reached with high doses of hormones could be interpreted as a 'desensitization' of enzyme and/or enzyme systems to membrane lipid interactions. | Temperature dependence of adenylate cyclase activity from rat white adipocytes. The effects of temperature on adenylate cyclase (AC) activity from rat white adipocytes were studied. Arrhenius plots of the data were found to be biphasic for basal AC activity, with a break near 27 degrees C. Noradrenaline and corticotropin induced a shift in the break with a rise in energy of activation (Ea) on both sides of the break. Aabove break point only, Ea increased with respect to hormone does as a hyperbolic function. The maximum value was in the range 17-21 Kcal x mol-1. Temperature was shown to have only a slight effect on the binding capacity of corticotropin to its receptor sites. The possibility of the existence of multiple thermodynamic states for the enzyme is envisaged. Basal AC activity is thermodynamically different from the hormone-stimulated enzyme. Hormones induce changes in the basal conformation of the enzyme, and this is reflected in modifications of Arrhenius plots. The maximal state of activation reached with high doses of hormones could be interpreted as a 'desensitization' of enzyme and/or enzyme systems to membrane lipid interactions. | [
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|
PMID:18380 | Amino acid replacements resulting from suppression and missense reversion of a chain-terminator mutation in Neurospora. | The Neurospora crassa super-suppressor mutation, ssu-1, suppresses the auxotrophic phenotype of the mutant am(17) by inserting tyrosine at residue 313 of NADP-specific glutamate dehydrogenase, a position occupied in the wild type by glutamate. Two classes of am(17) revertants due to further mutation within the am gene have, respectively, tyrosine and leucine at residue 313. These replacements are consistent with a chain-terminating codon in am(17) of either the amber (UAG) or the ochre type (UAA), but are inconsistent with UGA. The Leu313 and Tyr313 variants of the enzyme have effective activity but are grossly different from the wild type in Michaelis constants (especially for ammonium) and heat stabilities at two different pH values. They show smaller but significant differences in these respects from each other. | Amino acid replacements resulting from suppression and missense reversion of a chain-terminator mutation in Neurospora. The Neurospora crassa super-suppressor mutation, ssu-1, suppresses the auxotrophic phenotype of the mutant am(17) by inserting tyrosine at residue 313 of NADP-specific glutamate dehydrogenase, a position occupied in the wild type by glutamate. Two classes of am(17) revertants due to further mutation within the am gene have, respectively, tyrosine and leucine at residue 313. These replacements are consistent with a chain-terminating codon in am(17) of either the amber (UAG) or the ochre type (UAA), but are inconsistent with UGA. The Leu313 and Tyr313 variants of the enzyme have effective activity but are grossly different from the wild type in Michaelis constants (especially for ammonium) and heat stabilities at two different pH values. They show smaller but significant differences in these respects from each other. | [
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|
PMID:18381 | Qualitative changes in fibrinogen following exposure to agents used for preparation of fibrin monomers. | The solubility, clottability, thrombin clotting time and agarose gel chromatography pattern of human fibrinogen were studied after exposure to solvents commonly used to prepare fibrin monomers (0.0167 M acetic acid with 2.5 mM EDTA; 1 M NaBr, pH 5.2; 3.3 M urea, pH 7.4; 5 M urea, pH 7.4). When exposed to acetic acid at room temperature, fibrinogen precipitated almost immediately and quantitatively. Subsequent dialysis for 72 h against 0.3 M NaCl, pH 7.4, caused resolution of fibrinogen to a varying degree, the amount depending on the time of exposure. The redissolved fibrinogen showed reduced clottability, markedly shortened thrombin clotting time and a chromatographic profile that indicated large amounts of aggregates. Fibrinogen exposed to 1 M NaBr, pH 5.2, at room temperature for 1 h showed a slightly shortened thrombin clotting time and a broadened chromatographic profile. Exposure for 24 h to the same agent resulted in reduced solubility and clottability, a prolonged thrombin clotting time and progressive broadening of the chromatographic profile. Similar findings were obtained with fibrinogen exposed to 5 M urea, pH 7.4. Exposure to 3.3 M urea at pH 7.4 for 24 h, room temperature, led only to a moderate increase in solubility. | Qualitative changes in fibrinogen following exposure to agents used for preparation of fibrin monomers. The solubility, clottability, thrombin clotting time and agarose gel chromatography pattern of human fibrinogen were studied after exposure to solvents commonly used to prepare fibrin monomers (0.0167 M acetic acid with 2.5 mM EDTA; 1 M NaBr, pH 5.2; 3.3 M urea, pH 7.4; 5 M urea, pH 7.4). When exposed to acetic acid at room temperature, fibrinogen precipitated almost immediately and quantitatively. Subsequent dialysis for 72 h against 0.3 M NaCl, pH 7.4, caused resolution of fibrinogen to a varying degree, the amount depending on the time of exposure. The redissolved fibrinogen showed reduced clottability, markedly shortened thrombin clotting time and a chromatographic profile that indicated large amounts of aggregates. Fibrinogen exposed to 1 M NaBr, pH 5.2, at room temperature for 1 h showed a slightly shortened thrombin clotting time and a broadened chromatographic profile. Exposure for 24 h to the same agent resulted in reduced solubility and clottability, a prolonged thrombin clotting time and progressive broadening of the chromatographic profile. Similar findings were obtained with fibrinogen exposed to 5 M urea, pH 7.4. Exposure to 3.3 M urea at pH 7.4 for 24 h, room temperature, led only to a moderate increase in solubility. | [
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|
PMID:18382 | Ditazole and platelets. II. Effect of ditazole on in vivo platelet aggregation and bleeding time in rats. | Ditazole (4,5-diphenyl-2-bis-(2-hydroxyethyl)-aminoxazol) is a new drug shown to inhibit prostaglandin release from rat platelets. The present study indicates that ditazole, at doses similar to those inhibiting prostaglandin formation in rat, inhibits in vivo collagen--but not ADP-induced rat platelet aggregation. In addition, ditazole does not prolong the bleeding time in rats, but even tends to shorten it. This effect could be due to the inhibition of prostaglandin formation at the side of the vascular injury produced to induce bleeding. | Ditazole and platelets. II. Effect of ditazole on in vivo platelet aggregation and bleeding time in rats. Ditazole (4,5-diphenyl-2-bis-(2-hydroxyethyl)-aminoxazol) is a new drug shown to inhibit prostaglandin release from rat platelets. The present study indicates that ditazole, at doses similar to those inhibiting prostaglandin formation in rat, inhibits in vivo collagen--but not ADP-induced rat platelet aggregation. In addition, ditazole does not prolong the bleeding time in rats, but even tends to shorten it. This effect could be due to the inhibition of prostaglandin formation at the side of the vascular injury produced to induce bleeding. | [
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|
PMID:18383 | Reduction of nitro-BT by some components of the chloroform-methanol extract of the grey matter of the rat brain. | An attempt was made to extract the substance present in the nerve fibres of the cerebral cortex and basal ganglia which reduced Nitro-BT in alkaline medium pH 9-5 [4,5]. The investigations were performed on 40 white Wistar rats of both sexes and ca. 200 g of body weight. The brains taken for studies were fixed in Baker's formalin for 1,5 hr, thereafter grinded and homogenized in chloroform-methanol solution supplemented up to 100 ml of final volume after homogenisation. Thus prepared solution was left for 24 hr at 4 degrees C temperature and than centrifuged 2000 g/min. The extract was evaporated and the sediment further investigated. The following fraction were received: 1. fraction soluble in aceton, 2. fraction soluble in ethylalcohol, 3. fraction soluble in ether, 4. fraction soluble in water and 5. the sediment. Each fraction was incubated with a standard medium containing Nitro-BT. After incubation the amount of reduced Nitro-BT in each of the incubation medium was spectrophotometrically measured. The most intense reduction was found in the incubation medium with the water fraction. The spectrophotometrically determined reduction of Nitro-BT of the water fraction compared with that of the gangliosides solution were similar. The addition into the incubation media biogenic amines and N,N-diethyl-p-phenylenediamine enhances the reduction of Nitro-BT significantly. According to the results obtained an interference of gangliosides and of the investigated amines in the reduction of Nitro-BT is suggested. | Reduction of nitro-BT by some components of the chloroform-methanol extract of the grey matter of the rat brain. An attempt was made to extract the substance present in the nerve fibres of the cerebral cortex and basal ganglia which reduced Nitro-BT in alkaline medium pH 9-5 [4,5]. The investigations were performed on 40 white Wistar rats of both sexes and ca. 200 g of body weight. The brains taken for studies were fixed in Baker's formalin for 1,5 hr, thereafter grinded and homogenized in chloroform-methanol solution supplemented up to 100 ml of final volume after homogenisation. Thus prepared solution was left for 24 hr at 4 degrees C temperature and than centrifuged 2000 g/min. The extract was evaporated and the sediment further investigated. The following fraction were received: 1. fraction soluble in aceton, 2. fraction soluble in ethylalcohol, 3. fraction soluble in ether, 4. fraction soluble in water and 5. the sediment. Each fraction was incubated with a standard medium containing Nitro-BT. After incubation the amount of reduced Nitro-BT in each of the incubation medium was spectrophotometrically measured. The most intense reduction was found in the incubation medium with the water fraction. The spectrophotometrically determined reduction of Nitro-BT of the water fraction compared with that of the gangliosides solution were similar. The addition into the incubation media biogenic amines and N,N-diethyl-p-phenylenediamine enhances the reduction of Nitro-BT significantly. According to the results obtained an interference of gangliosides and of the investigated amines in the reduction of Nitro-BT is suggested. | [
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|
PMID:18384 | [Simple on-line digital determination of the cardiac action potential and its application]. | A system of digital measuring of the cardiac action potential was constructed in order to measure quickly and accurately the resting potential or maximum diastolic potential, overshoot potential, amplitude, time for 50% repolarization and time for 90% repolarization. This system consists of a dual-beam oscilloscope, a digital voltage meter, a digital interval meter and a simple circuit constructed by operational amplifiers. The measurement of the action potential was achieved by tracing only the resting potential or the maximum diastolic potential and the crest of the action potential on the oscilloscope using another beam. The time for measurement is usually 10-15 sec and errors in measurement were considered to be negligible. This system is considered to be useful for digital measurement of the action potential of various cardiac tissues. This system was applied for observation of effects of bufetolol, an adrenergic beta-receptor blocking drug on the action potential of the dog Purkinje fibres. Bufetolol (10(-5) M) caused a slight depolarization of the maximum diastolic potential, decreased overshoot potential, amplitude and maximum rate of rise, and shortened the time for 50% repolarization of the action potential. Bufetolol (10(-4) M) additionally prolonged the time for 90% repolarization. | [Simple on-line digital determination of the cardiac action potential and its application]. A system of digital measuring of the cardiac action potential was constructed in order to measure quickly and accurately the resting potential or maximum diastolic potential, overshoot potential, amplitude, time for 50% repolarization and time for 90% repolarization. This system consists of a dual-beam oscilloscope, a digital voltage meter, a digital interval meter and a simple circuit constructed by operational amplifiers. The measurement of the action potential was achieved by tracing only the resting potential or the maximum diastolic potential and the crest of the action potential on the oscilloscope using another beam. The time for measurement is usually 10-15 sec and errors in measurement were considered to be negligible. This system is considered to be useful for digital measurement of the action potential of various cardiac tissues. This system was applied for observation of effects of bufetolol, an adrenergic beta-receptor blocking drug on the action potential of the dog Purkinje fibres. Bufetolol (10(-5) M) caused a slight depolarization of the maximum diastolic potential, decreased overshoot potential, amplitude and maximum rate of rise, and shortened the time for 50% repolarization of the action potential. Bufetolol (10(-4) M) additionally prolonged the time for 90% repolarization. | [
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|
PMID:18385 | Phosphate uptake and involvement of binding protein in Tween-80 supplemented culture of Aspergillus fumigatus. | Tween-80 supplementation in submerged culture of Aspergillus fumigatus resulted in an increase of phosphate uptake. The uptake system was characterized as saturable, energy-dependent and operating against the concentration gradient. Control and Tween 80 cultures showed similar Km values for phosphate uptake (50 micrometer). Cold osmotic shock treatment of the cultures was found to cause considerable reduction in the ability to take up phosphorus with concomitant release of the binding protein into the shock fluid. Binding protein preparation from Tween-80 supplemented cells showed more activity than that from control cells. | Phosphate uptake and involvement of binding protein in Tween-80 supplemented culture of Aspergillus fumigatus. Tween-80 supplementation in submerged culture of Aspergillus fumigatus resulted in an increase of phosphate uptake. The uptake system was characterized as saturable, energy-dependent and operating against the concentration gradient. Control and Tween 80 cultures showed similar Km values for phosphate uptake (50 micrometer). Cold osmotic shock treatment of the cultures was found to cause considerable reduction in the ability to take up phosphorus with concomitant release of the binding protein into the shock fluid. Binding protein preparation from Tween-80 supplemented cells showed more activity than that from control cells. | [
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|
PMID:18386 | Variation in age at puberty in monkeys. | 5 female and 3 male patas monkeys and 6 female and 3 male talapoin monkeys matured in a captive breeding colony. Age at puberty is given, and some variation discussed. The talapoin, a very small monkey, becomes adult at 4 1/2 years for females, 1 or 2 years later for males. The patas, a rather large monkey, becomes adult at 2 1/2 years, for females, and 1 or 2 years later for males. Both these ages for puberty differ from data for the rhesus monkey which has been accepted as generalizable to all Old World monkeys. Possible causes of differences between species in average age at puberty are discussed, including nutrition, environmental inconstancy, and relative size of infant and mother. It is suggested that age at first conception, a biologically more relevant index than menarche, should be considered as a potentially important adaptive variable when describing primate species. | Variation in age at puberty in monkeys. 5 female and 3 male patas monkeys and 6 female and 3 male talapoin monkeys matured in a captive breeding colony. Age at puberty is given, and some variation discussed. The talapoin, a very small monkey, becomes adult at 4 1/2 years for females, 1 or 2 years later for males. The patas, a rather large monkey, becomes adult at 2 1/2 years, for females, and 1 or 2 years later for males. Both these ages for puberty differ from data for the rhesus monkey which has been accepted as generalizable to all Old World monkeys. Possible causes of differences between species in average age at puberty are discussed, including nutrition, environmental inconstancy, and relative size of infant and mother. It is suggested that age at first conception, a biologically more relevant index than menarche, should be considered as a potentially important adaptive variable when describing primate species. | [
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|
PMID:18388 | [Acupuncture analgesia]. | The possibility of abolishing pain during operations by needling acupuncture points was detected in China 20 years ago. During the last years the Western World showed great interest in this method, which was tested in a great number of surgical operations. Acupuncture was successful, especially when it was introduced by a short conventional anesthesia. Of special importance seems the possible reduction of anesthetic agents. Though the mode of action of acupuncture cannot yet be explained completely, there exist three different hypotheses: Hypnosis and suggestion, neurophysiological and humoral mechanisms. An actual review on experiments concerning these theories is given. | [Acupuncture analgesia]. The possibility of abolishing pain during operations by needling acupuncture points was detected in China 20 years ago. During the last years the Western World showed great interest in this method, which was tested in a great number of surgical operations. Acupuncture was successful, especially when it was introduced by a short conventional anesthesia. Of special importance seems the possible reduction of anesthetic agents. Though the mode of action of acupuncture cannot yet be explained completely, there exist three different hypotheses: Hypnosis and suggestion, neurophysiological and humoral mechanisms. An actual review on experiments concerning these theories is given. | [
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|
PMID:18389 | [Treatment of concommitant depression in neurologic diseases with dibenzepin infusions]. | A definition of concommitant depression is given. The results of infusorial therapy of Dibenzepin in 53 patients with neurological diseases and concommitant depression are reported. In 86% of all cases good or very good results could be obtained. The preparation is excellently suited for this therapy. The necessity of double therapy, namely the therapy of the basic disease as well as the therapy of the depressive syndrome is stressed. | [Treatment of concommitant depression in neurologic diseases with dibenzepin infusions]. A definition of concommitant depression is given. The results of infusorial therapy of Dibenzepin in 53 patients with neurological diseases and concommitant depression are reported. In 86% of all cases good or very good results could be obtained. The preparation is excellently suited for this therapy. The necessity of double therapy, namely the therapy of the basic disease as well as the therapy of the depressive syndrome is stressed. | [
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PMID:18390 | [Blood picture in lactate acidosis. Part 2: acid-base equilibrium and lactate]. | A differentiation between lactate emia (lactic acid emia) and lactate acidosis (lactic acidosis) is made. The normal value for blood lactate concentration is 1-2 mmol/1. The term lactate emia is used for lactate values between 2-6 mmol/1. The limiting value for the diagnosis of lactate acidosis should be more than 7-8 mmol/1 for the blood lactate concentration. Furthermore the different buffer mechanisms are evaluated in respect to their influence on the pH of the blood and to lactate metabolism. Especially the mechanism of respiratory compensation for metabolic acidosis is discussed. It is stated that for the diagnosis of lactate acidosis the blood-pH and the bicarbonate concentration should be measured. | [Blood picture in lactate acidosis. Part 2: acid-base equilibrium and lactate]. A differentiation between lactate emia (lactic acid emia) and lactate acidosis (lactic acidosis) is made. The normal value for blood lactate concentration is 1-2 mmol/1. The term lactate emia is used for lactate values between 2-6 mmol/1. The limiting value for the diagnosis of lactate acidosis should be more than 7-8 mmol/1 for the blood lactate concentration. Furthermore the different buffer mechanisms are evaluated in respect to their influence on the pH of the blood and to lactate metabolism. Especially the mechanism of respiratory compensation for metabolic acidosis is discussed. It is stated that for the diagnosis of lactate acidosis the blood-pH and the bicarbonate concentration should be measured. | [
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|
PMID:18391 | [The disease entity of lactate acidosis. 3. Lactate as a metabolic product]. | High blood lactate concentrations can be achieved by means of intravenous bicarbonate infusion. Metabolic production of lactic acid in this case is a compensation mechanism for the alcalosis induced by bicarbonate. This metabolic condition is called lactate alcalosis. The meaning and the diagnostic value of the lactate/pyruvate quotient and of excess lactate are discussed. A metabolic increase of the lactate/pyruvate quotient (normal values being 10-20) can be attained during the intravenous application of polyalcohols (like xylitol or sorbitol) or of ethanol. In these cases blood lactate concentration remains approximately normal. The alterations are due to the metabolism of the alcohols predominantly in the cytoplasmic compartment of the hepatocytes. The anion-gap is caused by the fact that the anions are considered only in part. However, the diagnostic value of the anion-gap is only minimum. An increase in the anion-gap with a simultaneous decrease in blood-pH is not significant for a lactate acidosis. | [The disease entity of lactate acidosis. 3. Lactate as a metabolic product]. High blood lactate concentrations can be achieved by means of intravenous bicarbonate infusion. Metabolic production of lactic acid in this case is a compensation mechanism for the alcalosis induced by bicarbonate. This metabolic condition is called lactate alcalosis. The meaning and the diagnostic value of the lactate/pyruvate quotient and of excess lactate are discussed. A metabolic increase of the lactate/pyruvate quotient (normal values being 10-20) can be attained during the intravenous application of polyalcohols (like xylitol or sorbitol) or of ethanol. In these cases blood lactate concentration remains approximately normal. The alterations are due to the metabolism of the alcohols predominantly in the cytoplasmic compartment of the hepatocytes. The anion-gap is caused by the fact that the anions are considered only in part. However, the diagnostic value of the anion-gap is only minimum. An increase in the anion-gap with a simultaneous decrease in blood-pH is not significant for a lactate acidosis. | [
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|
PMID:18392 | Disparity between insulin and proinsulin on stimulation of hepatic tyrosine transaminase. | The ability of insulin and proinsulin to stimulate tyrosine transaminase in dexamethasone-treated cultured rat liver cells was compared. Insulin increased this enzyme whereas proinsulin was seemingly without effect. Since proinsulin was not degraded by these cells, failure of stimulation of hepatic tyrosine transaminase could not be due to rapid destruction of the prohormone. This is the first demonstration of an action of insulin that cannot be duplicated by proinsulin. | Disparity between insulin and proinsulin on stimulation of hepatic tyrosine transaminase. The ability of insulin and proinsulin to stimulate tyrosine transaminase in dexamethasone-treated cultured rat liver cells was compared. Insulin increased this enzyme whereas proinsulin was seemingly without effect. Since proinsulin was not degraded by these cells, failure of stimulation of hepatic tyrosine transaminase could not be due to rapid destruction of the prohormone. This is the first demonstration of an action of insulin that cannot be duplicated by proinsulin. | [
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|
PMID:18393 | Iodotyrosine deiodination in the normal and acutely TSH-stimulated thyroid. | The deiodination of L-MIT-125I was measured in rat thyroid homogenates and slices before and after acute TSH stimulation. Slices and homogenates were incubated with identical concentrations of tissue and substrate in the presence and absence of NADPH. 1 USP unit TSH added in vitro to thyroid slices failed to stimulate deiodination; a single in vivo ip injection of 3 USP units TSH was also unable to raise deiodinating activity. In contrast to TSH, NADPH added to homogenates and slices enhanced deiodination significantly. However, several arguments, including a review of the literature, strongly militate against the hypothesis of an increased intracellular concentration of the coenzyme NADPH being the prerequisite to enhanced deiodination. The results suggest that deiodinase activity in acutely stimulated thyroids is not limited by the intracellular concentration of the enzyme itself nor by the availability of co-enzyme. Therefore, the increased iodide release induced by acute TSH stimulation is a mere consequence of the enhanced thyroglobulin proteolysis and does not require higher enzyme concentration. It will be shown subsequently that a different conclusion must be drawn in experiments with chronic TSH stimulation. | Iodotyrosine deiodination in the normal and acutely TSH-stimulated thyroid. The deiodination of L-MIT-125I was measured in rat thyroid homogenates and slices before and after acute TSH stimulation. Slices and homogenates were incubated with identical concentrations of tissue and substrate in the presence and absence of NADPH. 1 USP unit TSH added in vitro to thyroid slices failed to stimulate deiodination; a single in vivo ip injection of 3 USP units TSH was also unable to raise deiodinating activity. In contrast to TSH, NADPH added to homogenates and slices enhanced deiodination significantly. However, several arguments, including a review of the literature, strongly militate against the hypothesis of an increased intracellular concentration of the coenzyme NADPH being the prerequisite to enhanced deiodination. The results suggest that deiodinase activity in acutely stimulated thyroids is not limited by the intracellular concentration of the enzyme itself nor by the availability of co-enzyme. Therefore, the increased iodide release induced by acute TSH stimulation is a mere consequence of the enhanced thyroglobulin proteolysis and does not require higher enzyme concentration. It will be shown subsequently that a different conclusion must be drawn in experiments with chronic TSH stimulation. | [
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|
PMID:18395 | Specific, water-soluble polypeptides in identified neurons of Aplysia californica. | Application of an ethylene glycol lysis technique to extract water-soluble, low molecular weight polypeptides in Aplysia neurons, was used in conjunction with microgradient gel electrophoresis and micro-isoelectric focusing, to identify unique polypeptides in specific, identified neurons. The polypeptides found in neurons R15, R3-13, R14, and the bag cells were particularly abundant, consistent with the previously suggested neurosecretory role for these cells. Water extraction of the strongly basic polypeptides (pI 10.7) in R3-13 and R14 required an acidic lysis medium. | Specific, water-soluble polypeptides in identified neurons of Aplysia californica. Application of an ethylene glycol lysis technique to extract water-soluble, low molecular weight polypeptides in Aplysia neurons, was used in conjunction with microgradient gel electrophoresis and micro-isoelectric focusing, to identify unique polypeptides in specific, identified neurons. The polypeptides found in neurons R15, R3-13, R14, and the bag cells were particularly abundant, consistent with the previously suggested neurosecretory role for these cells. Water extraction of the strongly basic polypeptides (pI 10.7) in R3-13 and R14 required an acidic lysis medium. | [
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|
PMID:18396 | Distribution of beta-N-acetylglucosaminidase, hyaluronoglucosaminidase and acrosin in buffalo and goat spermatozoa. | The distribution of beta-N-acetylglucosaminidase, hyaluronoglucosaminidase and acrosin in buffalo and goat sperm acrosomes was studied. The three hydrolases were found to occur in soluble and bound forms. In the bound form, they were associated with the denuded sperm and were maximally solubilized at pH 3.0. The possible role of beta-N-acetylglucosaminidase in fertilization is discussed. | Distribution of beta-N-acetylglucosaminidase, hyaluronoglucosaminidase and acrosin in buffalo and goat spermatozoa. The distribution of beta-N-acetylglucosaminidase, hyaluronoglucosaminidase and acrosin in buffalo and goat sperm acrosomes was studied. The three hydrolases were found to occur in soluble and bound forms. In the bound form, they were associated with the denuded sperm and were maximally solubilized at pH 3.0. The possible role of beta-N-acetylglucosaminidase in fertilization is discussed. | [
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|
PMID:18397 | [Purification and properties of the tetrahydropteroylglutamate methyltransferase from green beans (Phaseolus vulgaris) (author's transl)]. | The tetrahydropteroylglutamate methyltransferase from green beans (Phaseolus vulgaris) has been purified 80-fold by ion exchange chromatography and gel filtration. Optimal methyl transfer is found at pH 6.5 and 39 degrees C. Even at 0 degrees C, however, a considerable catalytic rate is observed. The Michaelis-Menten constants for homocysteine and 5-methyltetrahydropteroylglutamate are 0.43mM and 2.4 mM, respectively. Magnesium ions enhance the activity. Even purified preparations appear to contain traces of magnesium ions firmly bound, since a residual activity is found without addition of magnesium salts. Though the reaction requires anaerobiosis, an excess of reducing agents is inhibitory. The molecular weight of the transferase, determined by gel filtration, is 40 000 +/- 6%. | [Purification and properties of the tetrahydropteroylglutamate methyltransferase from green beans (Phaseolus vulgaris) (author's transl)]. The tetrahydropteroylglutamate methyltransferase from green beans (Phaseolus vulgaris) has been purified 80-fold by ion exchange chromatography and gel filtration. Optimal methyl transfer is found at pH 6.5 and 39 degrees C. Even at 0 degrees C, however, a considerable catalytic rate is observed. The Michaelis-Menten constants for homocysteine and 5-methyltetrahydropteroylglutamate are 0.43mM and 2.4 mM, respectively. Magnesium ions enhance the activity. Even purified preparations appear to contain traces of magnesium ions firmly bound, since a residual activity is found without addition of magnesium salts. Though the reaction requires anaerobiosis, an excess of reducing agents is inhibitory. The molecular weight of the transferase, determined by gel filtration, is 40 000 +/- 6%. | [
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|
PMID:18400 | Induction of placental alkaline phosphatase in choriocarcinoma cells by 5-bromo-2'-deoxyuridine. | Growth of choriocarcinoma cells in the presence of 5-bromo-2'-deoxyuridine (BrdUrd) results in a 30- to 40-fold increase in alkaline phosphatase activity. The effects of BrdUrd is specific for phosphatase with an alkaline pH optimum. The induction by BrdUrd is probably not due to the production of an altered enzyme, since the induced enzyme resembles the basal enzyme in thermal denaturation and kinetic properties. Enzyme induction can be prevented by thymidine but not by deoxycytidine or deoxyuridine. The induction of alkaline phosphatase appears to require incorporation of the BrdUrd into cellular DNA. The presence of BrdUrd in the growth medium is not necessary for alkaline phosphatase induction in proliferating cells containing BrdUrd-substituted genomes. However, enzyme induction and maintenance of the induced levels of alkaline phosphatase in nonproliferating cells containing BrdUrd-substituted DNA requires the presence of the analogues in the medium. The induction of alkaline phosphatase by BrdUrd in probably an indirect process. | Induction of placental alkaline phosphatase in choriocarcinoma cells by 5-bromo-2'-deoxyuridine. Growth of choriocarcinoma cells in the presence of 5-bromo-2'-deoxyuridine (BrdUrd) results in a 30- to 40-fold increase in alkaline phosphatase activity. The effects of BrdUrd is specific for phosphatase with an alkaline pH optimum. The induction by BrdUrd is probably not due to the production of an altered enzyme, since the induced enzyme resembles the basal enzyme in thermal denaturation and kinetic properties. Enzyme induction can be prevented by thymidine but not by deoxycytidine or deoxyuridine. The induction of alkaline phosphatase appears to require incorporation of the BrdUrd into cellular DNA. The presence of BrdUrd in the growth medium is not necessary for alkaline phosphatase induction in proliferating cells containing BrdUrd-substituted genomes. However, enzyme induction and maintenance of the induced levels of alkaline phosphatase in nonproliferating cells containing BrdUrd-substituted DNA requires the presence of the analogues in the medium. The induction of alkaline phosphatase by BrdUrd in probably an indirect process. | [
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|
PMID:18403 | Effects of trypan blue treatment on the immune responses of mice. | It has been reported that trypan blue treatment decreases the nonspecific resistance of mice to transplanted tumors and inhibits the in vitro cytotoxic activity of activated macrophages. We wished to determine whether this effect of trypan blue could be due to a selective inhibition of certain macrophage functions or whether it reflected a broader form of immunosuppression. We therefore tested the effects of trypan blue on a variety of immunological responses. Treatment of mice with trypan blue delayed their rejection of skin allografts and transplants of a highly antigenic syngeneic ultraviolet light-induced tumor. Trypan blue treatment of either donor or recipient decreased the local graft-versus-host reaction. Filtration of lymph node cells from trypan blue-treated donors on a nylon wool column before use in the graft-versus-host assay abrogated the depressive effect of trypan blue. A transient reduction in the blastogenic response of spleen cells to concanavalin A and lipopolysaccharide mitogens was observed after a single injection of trypan blue, but the response of lymph node cells was unaffected. The depressed response of splenic lymphocytes was not entirely reversed by removal of adherent cells. The primary and secondary hemagglutinin responses to sheep erythrocytes were unaffected in trypan blue-treated mice, and the proportion and phagocytic activity of thioglycolate-induced peritoneal macrophages were also unaltered. We conclude that treatment of mice with trypan blue selectively inhibits certain macrophage functions but, at high doses, it can also inhibit some lymphocyte activities. | Effects of trypan blue treatment on the immune responses of mice. It has been reported that trypan blue treatment decreases the nonspecific resistance of mice to transplanted tumors and inhibits the in vitro cytotoxic activity of activated macrophages. We wished to determine whether this effect of trypan blue could be due to a selective inhibition of certain macrophage functions or whether it reflected a broader form of immunosuppression. We therefore tested the effects of trypan blue on a variety of immunological responses. Treatment of mice with trypan blue delayed their rejection of skin allografts and transplants of a highly antigenic syngeneic ultraviolet light-induced tumor. Trypan blue treatment of either donor or recipient decreased the local graft-versus-host reaction. Filtration of lymph node cells from trypan blue-treated donors on a nylon wool column before use in the graft-versus-host assay abrogated the depressive effect of trypan blue. A transient reduction in the blastogenic response of spleen cells to concanavalin A and lipopolysaccharide mitogens was observed after a single injection of trypan blue, but the response of lymph node cells was unaffected. The depressed response of splenic lymphocytes was not entirely reversed by removal of adherent cells. The primary and secondary hemagglutinin responses to sheep erythrocytes were unaffected in trypan blue-treated mice, and the proportion and phagocytic activity of thioglycolate-induced peritoneal macrophages were also unaltered. We conclude that treatment of mice with trypan blue selectively inhibits certain macrophage functions but, at high doses, it can also inhibit some lymphocyte activities. | [
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|
PMID:18404 | Lung bacterial clearance in murine pneumococcal pneumonia. | We studied the bactericidal capacity of the rat lung during the development of pneumococcal pneumonia. Pneumonia was produced in a lower lobe by the intrabronchial instillation of 10(4)Streptococcus pneumoniae cells in buffer. Lung bacterial counts progressively increased, reaching 10(7) per lung within 48 h, and the increase was associated with localized atelectasis and consolidation. Bacterial multiplication was inhibited with tetracycline at various intervals after infection, and the subsequent clearance of pneumococci was determined. Viable pneumococci were rapidly killed by lung defenses if bacterial multiplication was inhibited within 12 h of the onset of infection. No change occurred in the bacterial populationif tetracycline was delayed until 24 h after infection, indicating that pneumococcal killing by lung defenses had ceased. This effect could be reproduced with the addition of pneumococcal capsular polysaccharide to the inoculum, which produced a dose-related inhibition of pneumococcal clearance. The clearance of S. epidermidis was not impaired in the presence of pneumococcal pneumonia or by administration of exogenous capsular polysaccharide. These data indicate that pneumococcal pneumonia causes a marked impairment in lung antipneumococcal defenses within 24 h of the onset of infection. This acquired defect in antibacterial defenses may be due to the accumulation of pneumococcal capsular material in the lungs of infected animals. | Lung bacterial clearance in murine pneumococcal pneumonia. We studied the bactericidal capacity of the rat lung during the development of pneumococcal pneumonia. Pneumonia was produced in a lower lobe by the intrabronchial instillation of 10(4)Streptococcus pneumoniae cells in buffer. Lung bacterial counts progressively increased, reaching 10(7) per lung within 48 h, and the increase was associated with localized atelectasis and consolidation. Bacterial multiplication was inhibited with tetracycline at various intervals after infection, and the subsequent clearance of pneumococci was determined. Viable pneumococci were rapidly killed by lung defenses if bacterial multiplication was inhibited within 12 h of the onset of infection. No change occurred in the bacterial populationif tetracycline was delayed until 24 h after infection, indicating that pneumococcal killing by lung defenses had ceased. This effect could be reproduced with the addition of pneumococcal capsular polysaccharide to the inoculum, which produced a dose-related inhibition of pneumococcal clearance. The clearance of S. epidermidis was not impaired in the presence of pneumococcal pneumonia or by administration of exogenous capsular polysaccharide. These data indicate that pneumococcal pneumonia causes a marked impairment in lung antipneumococcal defenses within 24 h of the onset of infection. This acquired defect in antibacterial defenses may be due to the accumulation of pneumococcal capsular material in the lungs of infected animals. | [
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|
PMID:18405 | Mild alkaline hydrolysis of lipopolysaccharide endotoxin enhances its mitogencity for murine B cells. | Mild alkaline hydrolysis was found to enhance the mitogenicity of lipopolysaccharide endotoxin for murine B lymphocytes. Alkaline treated lipopolysaccharide also retained its property as a polyclonal activator. Whereas this treatment reduced the lethality of endotoxin for mice, its toxicity for lymphocytes cultured in the absence of fetal calf serum was increased. Lipid analysis indicated that there were no significant changes in the fatty acids of lipid A, but particle size was significantly reduced and the material was more homogeneous and soluble than untreated lipopolysaccharide. The relationship of these effect on the structure of lipopolysaccharide endotoxin to the mechanism of B-lymphocyte activation is discussed. | Mild alkaline hydrolysis of lipopolysaccharide endotoxin enhances its mitogencity for murine B cells. Mild alkaline hydrolysis was found to enhance the mitogenicity of lipopolysaccharide endotoxin for murine B lymphocytes. Alkaline treated lipopolysaccharide also retained its property as a polyclonal activator. Whereas this treatment reduced the lethality of endotoxin for mice, its toxicity for lymphocytes cultured in the absence of fetal calf serum was increased. Lipid analysis indicated that there were no significant changes in the fatty acids of lipid A, but particle size was significantly reduced and the material was more homogeneous and soluble than untreated lipopolysaccharide. The relationship of these effect on the structure of lipopolysaccharide endotoxin to the mechanism of B-lymphocyte activation is discussed. | [
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|
PMID:18402 | Effect of some antihistaminic drugs on the oestrous cycyle of rats. | The effect of some antihistaminic drugs namely antazoline hydrochloride, diphenhy, dramine hydrochloride and mepyramine maleate has been investigated on the oestrous cycle in albino rats. On daily intraperitoneal administration, for 6 days, all the three drugs (10 mg/kg) significantly (P less than 0.001) prolonged the duration of oestrous cycle. The duration of subsequent cycle returned to near normal. | Effect of some antihistaminic drugs on the oestrous cycyle of rats. The effect of some antihistaminic drugs namely antazoline hydrochloride, diphenhy, dramine hydrochloride and mepyramine maleate has been investigated on the oestrous cycle in albino rats. On daily intraperitoneal administration, for 6 days, all the three drugs (10 mg/kg) significantly (P less than 0.001) prolonged the duration of oestrous cycle. The duration of subsequent cycle returned to near normal. | [
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|
PMID:18406 | [Assay of aminoglycosides in serum using the urease method (author's transl)]. | The urease method developed by Noone et al. for rapid bioassay of aminoglycoside antibiotics in serum is described in detail. The accuracy of the method was improved by using a BM 1-09 electrode assembly (Metrohm, CH-9100 Herisau, Switzerland) in conjunction with a digital pH-Meter (Metrohm E 500). The mean difference between spiked serum samples and measured concentrations was +0.37 +/-0.78 microgram/ml. The coefficient of determination between the urease and agar diffusion method was 0.94. Disadvantages of the method are a low sensitivity at concentrations below 1.5 microgram/ml and a large serum sample (1.5 ml). Advantages are its simplicity and rapidity. | [Assay of aminoglycosides in serum using the urease method (author's transl)]. The urease method developed by Noone et al. for rapid bioassay of aminoglycoside antibiotics in serum is described in detail. The accuracy of the method was improved by using a BM 1-09 electrode assembly (Metrohm, CH-9100 Herisau, Switzerland) in conjunction with a digital pH-Meter (Metrohm E 500). The mean difference between spiked serum samples and measured concentrations was +0.37 +/-0.78 microgram/ml. The coefficient of determination between the urease and agar diffusion method was 0.94. Disadvantages of the method are a low sensitivity at concentrations below 1.5 microgram/ml and a large serum sample (1.5 ml). Advantages are its simplicity and rapidity. | [
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|
PMID:18407 | Circulating prolactin levels. I. Normal females. | A specific, homologous radioimmunoassay was used to quantitate circulating levels of prolactin in females in various stages of life. They were grouped in periods of newborns to peripubertal females, reproductive, gravid and puerperal, and postmenopausal females. The results reveal dynamic changes of prolactin throughout various stages of life and support the mounting evidence that prolactin's role in humans might mimic that of animals. The role of prolactin in the ovarian cycle is unclear and will have to be ascertained at the molecular level. | Circulating prolactin levels. I. Normal females. A specific, homologous radioimmunoassay was used to quantitate circulating levels of prolactin in females in various stages of life. They were grouped in periods of newborns to peripubertal females, reproductive, gravid and puerperal, and postmenopausal females. The results reveal dynamic changes of prolactin throughout various stages of life and support the mounting evidence that prolactin's role in humans might mimic that of animals. The role of prolactin in the ovarian cycle is unclear and will have to be ascertained at the molecular level. | [
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|
PMID:18408 | Follicular development during late human pregnancy. | In contrast to the accepted view of ovarian quiescence during pregnancy, the ovaries of preparturient women are covered with a dense population of small superficial follicles. In this study we have measured the follicular size and status of the oocytes and cumulus oophorus in 298 follicles from the ovaries of 30 women. The samples were taken at cesarean section and were examined with Nomarski differential interference microscopy. No oocyte was recovered from 50% of the follicles; 79% of the recovered oocytes and 78% of their cumuli were degenerative. Degeneration was correlated with appearance of phagocytes in the follicular fluid. These findings suggest that the endocrine status of gestation does not prevent early follicular development, but induces premature atresia. | Follicular development during late human pregnancy. In contrast to the accepted view of ovarian quiescence during pregnancy, the ovaries of preparturient women are covered with a dense population of small superficial follicles. In this study we have measured the follicular size and status of the oocytes and cumulus oophorus in 298 follicles from the ovaries of 30 women. The samples were taken at cesarean section and were examined with Nomarski differential interference microscopy. No oocyte was recovered from 50% of the follicles; 79% of the recovered oocytes and 78% of their cumuli were degenerative. Degeneration was correlated with appearance of phagocytes in the follicular fluid. These findings suggest that the endocrine status of gestation does not prevent early follicular development, but induces premature atresia. | [
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|
PMID:18409 | Successful separation of X and Y and spermatozoa in human and bull semen. | Using particle free semen extender and applying a low temperature thermal convection laminar flow to bring the sperm cells in vertical orientation and allowing them to act through the center of buoyancy, a bimodal distribution was achieved. Laser scanner for observing sedimentation pattern and F-body technique for identification of male sperm were used. Specimens from five human donors and ten bulls were separated, checked for purity, and frozen in liquid nitrogen. The lighter fractions contained 59.4% male (P less than 0.01), the heavier fractions 63.5% female (P less than 0.01), and the nonprocessed semen 48.0% male cells. Over 10,000 cells were counted to arrive at this statistically significant result. Observed sex ratio closely matched the predicted purity figures. Lighter and heavier fractions were further purified using forced convection galvanic method. Seventy to eighty percent purity in both fractions was confirmed by F-body and small cell galvanic migration tests with human as well as with bull semen. | Successful separation of X and Y and spermatozoa in human and bull semen. Using particle free semen extender and applying a low temperature thermal convection laminar flow to bring the sperm cells in vertical orientation and allowing them to act through the center of buoyancy, a bimodal distribution was achieved. Laser scanner for observing sedimentation pattern and F-body technique for identification of male sperm were used. Specimens from five human donors and ten bulls were separated, checked for purity, and frozen in liquid nitrogen. The lighter fractions contained 59.4% male (P less than 0.01), the heavier fractions 63.5% female (P less than 0.01), and the nonprocessed semen 48.0% male cells. Over 10,000 cells were counted to arrive at this statistically significant result. Observed sex ratio closely matched the predicted purity figures. Lighter and heavier fractions were further purified using forced convection galvanic method. Seventy to eighty percent purity in both fractions was confirmed by F-body and small cell galvanic migration tests with human as well as with bull semen. | [
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|
PMID:18410 | Autoradiographic studies of rabbit ova after fertilization with thymidine-H3 and -C14 labelled spermatozoa originating from different bucks. | Rabbit spermatozoa were labelled in vivo injecting male rabbits with thymidine-H3 and thymidine-C14. Ova fertilized with H3-labelled spermatozoa were found to contain more silver grains when they had cleaved several times than before cleavage. Fertilized ova originating from rabbits that had been mated with both an H3- and C14-thymidine labelled buck carried both labels. An explanation for this phenomenon may be that additional spermatozoa in the perivitelline space of fertilized ova pass labelled DNA-bases to the cleaving blastomeres. | Autoradiographic studies of rabbit ova after fertilization with thymidine-H3 and -C14 labelled spermatozoa originating from different bucks. Rabbit spermatozoa were labelled in vivo injecting male rabbits with thymidine-H3 and thymidine-C14. Ova fertilized with H3-labelled spermatozoa were found to contain more silver grains when they had cleaved several times than before cleavage. Fertilized ova originating from rabbits that had been mated with both an H3- and C14-thymidine labelled buck carried both labels. An explanation for this phenomenon may be that additional spermatozoa in the perivitelline space of fertilized ova pass labelled DNA-bases to the cleaving blastomeres. | [
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|
PMID:18411 | Absence of a prenidatory effect of luteinizing hormone releasing hormone (LHRH) in hamsters. | Luteinizing hormone releasing hormone at high doses will terminate gestation in rats during early and midpregancy (ED50 approximately equal to 100 microgram/day) and rabbits during early pregnancy. Early pregnancy in hamsters, in contradistinction, seems refractory to this effect. Administration of LHRH up to massive doses (10 mg/day) over the first 3 or 7 days of pregnancy failed to affect the pregnancies in meaningful fashion. Further, a single injection (100 mg) on day 5 had no effect on pregnancy; this system has been employed for the assay of prostaglandins because hamsters are remarkably sensitive to PG's (PGF2alpha, ED50 approximately equal to 17 microgram, PGE2, ED50 approximately equal to 210 microgram). The absence of response of hamsters to LHRH cannot be interpreted at present. | Absence of a prenidatory effect of luteinizing hormone releasing hormone (LHRH) in hamsters. Luteinizing hormone releasing hormone at high doses will terminate gestation in rats during early and midpregancy (ED50 approximately equal to 100 microgram/day) and rabbits during early pregnancy. Early pregnancy in hamsters, in contradistinction, seems refractory to this effect. Administration of LHRH up to massive doses (10 mg/day) over the first 3 or 7 days of pregnancy failed to affect the pregnancies in meaningful fashion. Further, a single injection (100 mg) on day 5 had no effect on pregnancy; this system has been employed for the assay of prostaglandins because hamsters are remarkably sensitive to PG's (PGF2alpha, ED50 approximately equal to 17 microgram, PGE2, ED50 approximately equal to 210 microgram). The absence of response of hamsters to LHRH cannot be interpreted at present. | [
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|
PMID:18412 | The role of prostaglandins on the increase in motility of the rat-uterine horn containing a silk suture. | The presence of a silk suture in one uterine horn of the rats leads to an increase in spontaneous motility when compared with the contralateral control horn. Indomethacin, an inhibitor of prostaglandin synthetase inhibits the motility suggesting a role of prostaglandins in this process. | The role of prostaglandins on the increase in motility of the rat-uterine horn containing a silk suture. The presence of a silk suture in one uterine horn of the rats leads to an increase in spontaneous motility when compared with the contralateral control horn. Indomethacin, an inhibitor of prostaglandin synthetase inhibits the motility suggesting a role of prostaglandins in this process. | [
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|
PMID:18413 | Paper chromatographic estimation of fructose and myo-inositol in human seminal fluid: a method for evaluating seminal vesicle and prostatic function. | A semiquantitative paper chromatographic method for the separation of myo-inositol and fructose in human seminal fluid was described. The intensity of the chromatographic zones characteristic of myo-inositol, which is a marker for human prostatic secretion and for fructose, which is useful in evaluating seminal vesicle function, were estimated after staining with an alkaline silver nitrate reagent. Characteristic patterns were found for patients with prostatic or vesicular dysfunction, and for patients with agenesis or blockage of the vas deferens. Paper chromatographic analysis of split ejaculate samples allowed evaluation of the sequence of ejaculation of the prostate and seminal vesicles. | Paper chromatographic estimation of fructose and myo-inositol in human seminal fluid: a method for evaluating seminal vesicle and prostatic function. A semiquantitative paper chromatographic method for the separation of myo-inositol and fructose in human seminal fluid was described. The intensity of the chromatographic zones characteristic of myo-inositol, which is a marker for human prostatic secretion and for fructose, which is useful in evaluating seminal vesicle function, were estimated after staining with an alkaline silver nitrate reagent. Characteristic patterns were found for patients with prostatic or vesicular dysfunction, and for patients with agenesis or blockage of the vas deferens. Paper chromatographic analysis of split ejaculate samples allowed evaluation of the sequence of ejaculation of the prostate and seminal vesicles. | [
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|
PMID:18414 | Gonadal function following vasectomy in the rat. | Adult rats were studied at four, eight, and 12 months following vasectomy and sham-operation. The weights of the seminal vesicles, ventral prostate, pituitary, and kidneys were not significantly affected by vasectomy. Testicular endocrine function in vasectotomized rat was transiently stimulated as witnessed by elevation in testicular venous testosterone and androstenedione after four months. There then occurred signs of decline in gametogenic function and atrophy of the testis after 12 months whereas hormonogenesis appeared to remain at normal levels. There was no alteration in the morphology of the epididymis at any of the time intervals of study after vasectomy. | Gonadal function following vasectomy in the rat. Adult rats were studied at four, eight, and 12 months following vasectomy and sham-operation. The weights of the seminal vesicles, ventral prostate, pituitary, and kidneys were not significantly affected by vasectomy. Testicular endocrine function in vasectotomized rat was transiently stimulated as witnessed by elevation in testicular venous testosterone and androstenedione after four months. There then occurred signs of decline in gametogenic function and atrophy of the testis after 12 months whereas hormonogenesis appeared to remain at normal levels. There was no alteration in the morphology of the epididymis at any of the time intervals of study after vasectomy. | [
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|
PMID:18415 | Testosterone production and metabolism in laboratory-maintained male rhesus monkeys. | Plasma production rates (PR), metabolic clearance rates (MCR) and plasma levels of testosterone were determined in 10 male, laboratory-maintained rhesus monkeys on two occasions out-of-season (April and August) and once in-season (October). Plasma testosterone levels in October were higher than those in April and August. The plasma PR was unchanged in August, but markedly increased in October, as compared to April. The MCR showed a parallel increase betueen August and October. Thus changes in testosterone production and metabolism can be observed between in- and out-of-season male rhesus monkeys, maintained under strict laboratory conditions. These changes must be taken into consideration when the male rhesus minkey is used as an experimental model for human reproductive endocrinology. | Testosterone production and metabolism in laboratory-maintained male rhesus monkeys. Plasma production rates (PR), metabolic clearance rates (MCR) and plasma levels of testosterone were determined in 10 male, laboratory-maintained rhesus monkeys on two occasions out-of-season (April and August) and once in-season (October). Plasma testosterone levels in October were higher than those in April and August. The plasma PR was unchanged in August, but markedly increased in October, as compared to April. The MCR showed a parallel increase betueen August and October. Thus changes in testosterone production and metabolism can be observed between in- and out-of-season male rhesus monkeys, maintained under strict laboratory conditions. These changes must be taken into consideration when the male rhesus minkey is used as an experimental model for human reproductive endocrinology. | [
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|
PMID:18416 | Pituitary and ovarian response to acute stimulation with LH-RH in normal and anovulatory women. | The LH FSH estradiol and progesterone responses to acute stimulation with LH-RH were studied in 12 normal women with ovulatory cycles (4 in the initial follicular phase, 4 in the mid-follicular phase and 4 in the late follicular phase) and in two castrated women, two under hormonal contraception, two with ovarian amenorrhea, twelve with central amenorrhea of no detectable origin (6 with normal and 6 with low basal gonadotrophins), eleven anovulatory patients with pseudomenstruation, two with anorexia nervosa, and two with pituitary amenorrhea. Each woman received a rapid i.v. injection of 100 microgram synthetic LH-RH at 9:00 a.m. Serum levels of LH, FSH, estradiol and progesterone were determined by radioimmunoassay in samples collected before and 60, 120, 240 and 480 minutes after injection. The findings were : 1) A significant rise in estradiol and progesterone levels, in addition to LH and FSH elevation, in normal women; 2) A lack of ovarian steroid response in the castrated women and in ovarian amenorrheas, which suggests that the source of steroid response to stimulation is not extragonadal; 3) Significant differences in the responses of the four hormones to LH-RH in the women with central amenorrhea in comparison with the normal group with great variability of results; the steroid response in the presence of a positive LH response might correlate with the severity and/or prognosis of the disorder, a point deserving further study; 4) In anovulatory women with pseudomenstruation, LH responses for the most part normal, and particularly, progesterone responses. | Pituitary and ovarian response to acute stimulation with LH-RH in normal and anovulatory women. The LH FSH estradiol and progesterone responses to acute stimulation with LH-RH were studied in 12 normal women with ovulatory cycles (4 in the initial follicular phase, 4 in the mid-follicular phase and 4 in the late follicular phase) and in two castrated women, two under hormonal contraception, two with ovarian amenorrhea, twelve with central amenorrhea of no detectable origin (6 with normal and 6 with low basal gonadotrophins), eleven anovulatory patients with pseudomenstruation, two with anorexia nervosa, and two with pituitary amenorrhea. Each woman received a rapid i.v. injection of 100 microgram synthetic LH-RH at 9:00 a.m. Serum levels of LH, FSH, estradiol and progesterone were determined by radioimmunoassay in samples collected before and 60, 120, 240 and 480 minutes after injection. The findings were : 1) A significant rise in estradiol and progesterone levels, in addition to LH and FSH elevation, in normal women; 2) A lack of ovarian steroid response in the castrated women and in ovarian amenorrheas, which suggests that the source of steroid response to stimulation is not extragonadal; 3) Significant differences in the responses of the four hormones to LH-RH in the women with central amenorrhea in comparison with the normal group with great variability of results; the steroid response in the presence of a positive LH response might correlate with the severity and/or prognosis of the disorder, a point deserving further study; 4) In anovulatory women with pseudomenstruation, LH responses for the most part normal, and particularly, progesterone responses. | [
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|
PMID:18417 | Seminal fructose and acid phosphatase in vasectomised men. | Seminal fructose and acid phosphatase levels have been determined in 30 vasectomised and 30 nonvasectomised healthy adults, both the groups being closely matched for age (32 to 45 years). The fructose and acid phosphatase values in the vasectomised men, 3 to 6 months after the operation, were 316 +/- 36 mg% and 2,098 +/- 112 K.A. units/ml, respectively, while in the control group they were 251 +/- 28 mg% and 1,932 +/- 92 K.A. units/ml. The differences are statistically significant (P less than 0.001) and the possible implications of the post-vasectomy rise in these androgen dependent seminal constituents are discussed. | Seminal fructose and acid phosphatase in vasectomised men. Seminal fructose and acid phosphatase levels have been determined in 30 vasectomised and 30 nonvasectomised healthy adults, both the groups being closely matched for age (32 to 45 years). The fructose and acid phosphatase values in the vasectomised men, 3 to 6 months after the operation, were 316 +/- 36 mg% and 2,098 +/- 112 K.A. units/ml, respectively, while in the control group they were 251 +/- 28 mg% and 1,932 +/- 92 K.A. units/ml. The differences are statistically significant (P less than 0.001) and the possible implications of the post-vasectomy rise in these androgen dependent seminal constituents are discussed. | [
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|
PMID:18418 | The 'in vivo' effects of oxytocin and vasopressin on spontaneous contractility of the rat epididymis. | The spontaneous contractility of the epididymis in the rat was recorded in vivo and the effects of the neurohypophyseal hormones were studied. Oxytocin (50 muU and 500 muU/100 g body weight) produced a progressive increase in tonus together with an increase in amplitude and frequency of the contractions. Vasopressin (100 muU and 1000 muU/100 g body weight) showed similar effects. No differences were apparent at the doses studied. | The 'in vivo' effects of oxytocin and vasopressin on spontaneous contractility of the rat epididymis. The spontaneous contractility of the epididymis in the rat was recorded in vivo and the effects of the neurohypophyseal hormones were studied. Oxytocin (50 muU and 500 muU/100 g body weight) produced a progressive increase in tonus together with an increase in amplitude and frequency of the contractions. Vasopressin (100 muU and 1000 muU/100 g body weight) showed similar effects. No differences were apparent at the doses studied. | [
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|
PMID:18420 | Interpretation of thermal perturbation spectra of proteins. | The thermal perturbation difference spectrum of reduced lysozyme has a long wave length extremum at 304 nm at pH 6.15 and a very small extremum at 306 nm at pH 1.5. These results differ from those of Leach & Smith (1972), which showed an extremum at 293 nm, the same as for model tryptophyl compounds. Our result may arise from a conformational difference between the two sample temperatures. The interpretation of thermal perturbation spectra of proteins is discussed. Contributions from thermally induced concentration differences, buried chromophores, and chromophores in crevices are considered in the interpretation of the thermal perturbation spectrum of bovine serum albumin. It is suggested that chromophores in pauci-aqueous crevices may appear buried toward thermal perturbation spectroscopy but accessible toward solvent perturbation and chemical reagents. | Interpretation of thermal perturbation spectra of proteins. The thermal perturbation difference spectrum of reduced lysozyme has a long wave length extremum at 304 nm at pH 6.15 and a very small extremum at 306 nm at pH 1.5. These results differ from those of Leach & Smith (1972), which showed an extremum at 293 nm, the same as for model tryptophyl compounds. Our result may arise from a conformational difference between the two sample temperatures. The interpretation of thermal perturbation spectra of proteins is discussed. Contributions from thermally induced concentration differences, buried chromophores, and chromophores in crevices are considered in the interpretation of the thermal perturbation spectrum of bovine serum albumin. It is suggested that chromophores in pauci-aqueous crevices may appear buried toward thermal perturbation spectroscopy but accessible toward solvent perturbation and chemical reagents. | [
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|
PMID:18421 | Stabilisation of enzyme structures by inhibitors. A nuclear magnetic resonance study of the effect of phosphate on the acid unfolding of ribonuclease A. | The acid denaturation of bovine pancreatic ribonuclease A in the presence of 0.2M sodium dihydrogen phosphate has been studied by n.m.r. spectroscopy. Phenylalanine, tyrosine and methionine resonances serve as monitors of the unfolding process. It is shown that the inhibitor shifts the equilibrium towards the native structure at acid pH. Exchange broadening of the C-2 resonances of the active site histidines, 12 and 119, occurs in the presence of phosphate, suggesting an equilibrium between native and unfolded structures. Stabilisation of the partially unfolded protein is observed at pH 1.5, as evidenced by the lack of the histidine resonance due to random coil protein. A scheme of the equilibria relating the various states of the protein is proposed. | Stabilisation of enzyme structures by inhibitors. A nuclear magnetic resonance study of the effect of phosphate on the acid unfolding of ribonuclease A. The acid denaturation of bovine pancreatic ribonuclease A in the presence of 0.2M sodium dihydrogen phosphate has been studied by n.m.r. spectroscopy. Phenylalanine, tyrosine and methionine resonances serve as monitors of the unfolding process. It is shown that the inhibitor shifts the equilibrium towards the native structure at acid pH. Exchange broadening of the C-2 resonances of the active site histidines, 12 and 119, occurs in the presence of phosphate, suggesting an equilibrium between native and unfolded structures. Stabilisation of the partially unfolded protein is observed at pH 1.5, as evidenced by the lack of the histidine resonance due to random coil protein. A scheme of the equilibria relating the various states of the protein is proposed. | [
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|
PMID:18422 | Levels of neurotransmitters in brain of vitamin B12 deficient rats. | The levels of norepinephrine, dopamine and 5-hydroxytryptamine in brain homogenates of vitamin B12-deficient rats have been investigated. The norepinephrine levels were significantly decreased in the deficient animals compared to controls. The two major catabolic pathways of norepinephrine e.g. monoamine oxidase and catechol-O-methyl transferase did not show significant variations. Both acetyl cholinesterase and butiryl-cholinesterase markedly decreased in the plasma of the vitamin B12-deficient rats. | Levels of neurotransmitters in brain of vitamin B12 deficient rats. The levels of norepinephrine, dopamine and 5-hydroxytryptamine in brain homogenates of vitamin B12-deficient rats have been investigated. The norepinephrine levels were significantly decreased in the deficient animals compared to controls. The two major catabolic pathways of norepinephrine e.g. monoamine oxidase and catechol-O-methyl transferase did not show significant variations. Both acetyl cholinesterase and butiryl-cholinesterase markedly decreased in the plasma of the vitamin B12-deficient rats. | [
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|
PMID:18423 | The buccal absorption of ascorbic acid and its passage through lipoid membrane. | Ascorbic acid concentration was measured in centrifuged and uncentrifuged saliva from normal young adult men and women. The salivary ascorbic acid content of uncentrifuged saliva was significantly higher in women on account of the ascorbic acid contained in the cellular sediment. Ascorbic acid absorption into the buccal mucosa was measured from solutions of pH range 3.4-9.0 held in the mouth for periods of 1-9 minutes. For a constant mouth contact time, increase in pH of the loading solutions resulted in reduction of buccal absorption, and diminishing transfer into the buccal epithelium. Percentage absorption was greater in males than females throughout the pH range. At pH 5, 80% of the loading solution was absorbed after 5 minutes. Diffusion of ascorbic acid through the lipoid membrane was more rapid in males. Female buccal cells had significantly higher ascorbic acid concentrations than male cells. Absorption of laevo and dextro ascorbic acid in human beings can be considered as partitioning into, or passage through, a lipoid phase into the buccal cells. The rate of passage of ascorbic acid into the buccal lining cells under controlled conditions of pH and buccal contact time is pH and concentration dependent in human beings. | The buccal absorption of ascorbic acid and its passage through lipoid membrane. Ascorbic acid concentration was measured in centrifuged and uncentrifuged saliva from normal young adult men and women. The salivary ascorbic acid content of uncentrifuged saliva was significantly higher in women on account of the ascorbic acid contained in the cellular sediment. Ascorbic acid absorption into the buccal mucosa was measured from solutions of pH range 3.4-9.0 held in the mouth for periods of 1-9 minutes. For a constant mouth contact time, increase in pH of the loading solutions resulted in reduction of buccal absorption, and diminishing transfer into the buccal epithelium. Percentage absorption was greater in males than females throughout the pH range. At pH 5, 80% of the loading solution was absorbed after 5 minutes. Diffusion of ascorbic acid through the lipoid membrane was more rapid in males. Female buccal cells had significantly higher ascorbic acid concentrations than male cells. Absorption of laevo and dextro ascorbic acid in human beings can be considered as partitioning into, or passage through, a lipoid phase into the buccal cells. The rate of passage of ascorbic acid into the buccal lining cells under controlled conditions of pH and buccal contact time is pH and concentration dependent in human beings. | [
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PMID:18425 | Physicochemical characteristics, morphology and morphogenesis of virions of the causative agent of Crimean hemorrhagic fever. | Similar physicochemical characteristics were found with strains of Crimean hemorrhagic fever (CHF) and Congo viruses. The particle size, sedimentation coefficient, buoyant density, weight of the particles as well as morphology and morphogenesis of these viruses were similar to those of other members of the Bunyaviridae family. Data are presented on reproduction of CHF virus in cell cultures and on the inner structure of its virion. | Physicochemical characteristics, morphology and morphogenesis of virions of the causative agent of Crimean hemorrhagic fever. Similar physicochemical characteristics were found with strains of Crimean hemorrhagic fever (CHF) and Congo viruses. The particle size, sedimentation coefficient, buoyant density, weight of the particles as well as morphology and morphogenesis of these viruses were similar to those of other members of the Bunyaviridae family. Data are presented on reproduction of CHF virus in cell cultures and on the inner structure of its virion. | [
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|
PMID:18428 | Studies of binding C3-substitute rifamycins to human and bovine serum albumin. | The interactions of a series of C3-substituted rifamycins with human and bovine serum albumins were studied in order to find possible correlations between the degree of binding and the structural features of the various molecules. The results obtained indicate some of the physicochemical properties and, therefore, of the structural requirements which appear to determine or influence the bonding mechanisms of this series of rifamycins. Two types of interaction were found to exist, ionic and hydrophobic types. The findings suggest that the inhibition by protein of the antibacterial activities of these antibiotics depends on the type of bonding mechanism rather than the degree of binding. | Studies of binding C3-substitute rifamycins to human and bovine serum albumin. The interactions of a series of C3-substituted rifamycins with human and bovine serum albumins were studied in order to find possible correlations between the degree of binding and the structural features of the various molecules. The results obtained indicate some of the physicochemical properties and, therefore, of the structural requirements which appear to determine or influence the bonding mechanisms of this series of rifamycins. Two types of interaction were found to exist, ionic and hydrophobic types. The findings suggest that the inhibition by protein of the antibacterial activities of these antibiotics depends on the type of bonding mechanism rather than the degree of binding. | [
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|
PMID:18429 | On RNA-polymerases of leukemia L 1210 origin and an enzymatic method to screen antitumor antibiotics. | Four DNA-dependent RNA-polymerases were separated from the cell homogenate of moust leukemia L1210 cell by DEAE-cellulose column chromatography and tentatively designated as Peaks I, II, III and IV in the elution order. Peak II was inactivated by the addition of alpha-amanitin and effects of antibiotics and enzymes on the RNA-polymerase activity using Peaks, I, II and a mixture of Peaks I and II were examined. The RNA-polymerases were used to screen for enzyme inhibitors produced by microbes. This enzymatic method was successfully proved to select antitumor antibiotics. | On RNA-polymerases of leukemia L 1210 origin and an enzymatic method to screen antitumor antibiotics. Four DNA-dependent RNA-polymerases were separated from the cell homogenate of moust leukemia L1210 cell by DEAE-cellulose column chromatography and tentatively designated as Peaks I, II, III and IV in the elution order. Peak II was inactivated by the addition of alpha-amanitin and effects of antibiotics and enzymes on the RNA-polymerase activity using Peaks, I, II and a mixture of Peaks I and II were examined. The RNA-polymerases were used to screen for enzyme inhibitors produced by microbes. This enzymatic method was successfully proved to select antitumor antibiotics. | [
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|
PMID:18426 | Pig plasma benzylamine oxidase: some considerations on the mechanism of the reaction. | The kinetics of benzaldehyde formation during the oxidation of benzylamine by pig plasma benzylamine oxidase have been studied at different pH values. It has been shown that the first step of the reaction catalyzed by this enzyme is the formation of a Schiff base between the amino group of the substrate and the aldehyde group of the enzyme. Present kinetic studies are consistent with this mechanism and demonstrate the existence of two steps which are alternatively rate-limiting one below pH 6.0, the other above this pH value. The rate-limiting step above pH 6.0 requires the participation of OH-. A mechanism of reaction has been proposed in which the release of products follows the sequence: hydrogen peroxide, aldehyde, ammonia. | Pig plasma benzylamine oxidase: some considerations on the mechanism of the reaction. The kinetics of benzaldehyde formation during the oxidation of benzylamine by pig plasma benzylamine oxidase have been studied at different pH values. It has been shown that the first step of the reaction catalyzed by this enzyme is the formation of a Schiff base between the amino group of the substrate and the aldehyde group of the enzyme. Present kinetic studies are consistent with this mechanism and demonstrate the existence of two steps which are alternatively rate-limiting one below pH 6.0, the other above this pH value. The rate-limiting step above pH 6.0 requires the participation of OH-. A mechanism of reaction has been proposed in which the release of products follows the sequence: hydrogen peroxide, aldehyde, ammonia. | [
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|
PMID:18436 | Oxygen consumption and ventilation of dogs during passive and active exercise. | A comparison was made between the influence of passive and active exercise on the energy metabolism (EM) and respiratory parameters (RP) of dogs: respiratory minute volume and ventilatory equivalent for O2. Passive exercise was performed in 24 dogs, anesthetized with pentobarbital sodium and morphine or ketamine hydrochloride and pentobarbital sodium, by moving the four limbs at constant frequencies of 25, 45, or 80 movements/min before and after curarization. Active exercise was induced in 35 animals by electric stimulation of the quadriceps and triceps muscles or of the distal ends of femoral and radial nerves at the same frequencies previously indicated. Values obtained for EM and RP during passive exercise were not significantly different from those obtained during resting conditions; active exercise caused a significant increase which was directly proportional to the frequencies of limb movement. Our results indicate that passive muscle movement induces no alteration in O2 consumption and the possible relfexes generated in these experimental conditions were not apparently important on ventilatory control. In active movements, over the range of the frequencies studied, there is a proportional increase in O2 consumption followed by a corresponding increase in ventilation. | Oxygen consumption and ventilation of dogs during passive and active exercise. A comparison was made between the influence of passive and active exercise on the energy metabolism (EM) and respiratory parameters (RP) of dogs: respiratory minute volume and ventilatory equivalent for O2. Passive exercise was performed in 24 dogs, anesthetized with pentobarbital sodium and morphine or ketamine hydrochloride and pentobarbital sodium, by moving the four limbs at constant frequencies of 25, 45, or 80 movements/min before and after curarization. Active exercise was induced in 35 animals by electric stimulation of the quadriceps and triceps muscles or of the distal ends of femoral and radial nerves at the same frequencies previously indicated. Values obtained for EM and RP during passive exercise were not significantly different from those obtained during resting conditions; active exercise caused a significant increase which was directly proportional to the frequencies of limb movement. Our results indicate that passive muscle movement induces no alteration in O2 consumption and the possible relfexes generated in these experimental conditions were not apparently important on ventilatory control. In active movements, over the range of the frequencies studied, there is a proportional increase in O2 consumption followed by a corresponding increase in ventilation. | [
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|
PMID:18437 | Rate of pH changes in blood plasma in vitro and in vivo. | The rate of pH decrease in dog blood after addition of CO2 (dissolved in saline) was measured in vitro with a combination pH electrode and fast-response amplifier. The pH change was found to have an apparent half time of 6.0 +/- 0.5 s at 38 degrees C, similar to that predicted by previous mathematical simulations on the basis of uncatalyzed CO2 hydration kinetics and the buffering characteristics of plasma. Measurements were also made in vivo in dogs by withdrawing blood rapidly from a carotid artery through a temperature-controlled chamber containing a pH electrode. When the flow was stopped, we measured the pH change which occurs in blood (after it leaves the lungs) as the slow dehydration of bicarbonate continues. The measured half time was 7.3 +/- 0.6 s, again agreeing well with predictions. Essentially the same half time was found when the direction of approach to chemical equilibrium was reversed by adding CO2 to the inspired air. The rate of the reactions could be increased markedly by hemolyzing the blood to release carbonic anhydrase into the plasma. | Rate of pH changes in blood plasma in vitro and in vivo. The rate of pH decrease in dog blood after addition of CO2 (dissolved in saline) was measured in vitro with a combination pH electrode and fast-response amplifier. The pH change was found to have an apparent half time of 6.0 +/- 0.5 s at 38 degrees C, similar to that predicted by previous mathematical simulations on the basis of uncatalyzed CO2 hydration kinetics and the buffering characteristics of plasma. Measurements were also made in vivo in dogs by withdrawing blood rapidly from a carotid artery through a temperature-controlled chamber containing a pH electrode. When the flow was stopped, we measured the pH change which occurs in blood (after it leaves the lungs) as the slow dehydration of bicarbonate continues. The measured half time was 7.3 +/- 0.6 s, again agreeing well with predictions. Essentially the same half time was found when the direction of approach to chemical equilibrium was reversed by adding CO2 to the inspired air. The rate of the reactions could be increased markedly by hemolyzing the blood to release carbonic anhydrase into the plasma. | [
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PMID:18438 | Urease of Klebsiella aerogenes: control of its synthesis by glutamine synthetase. | Urease was purified 24-fold from extracts of Klebsiella aerogenes. The enzyme has a molecular weight of 230,000 as determined by gel filtration, is highly substrate specific, and has a Km for urea of 0.7 mM. A mutant strain lacking urease was isolated; it failed to grow with urea as the sole source of nitrogen but did grow on media containing other nitrogen sources such as ammonia, histidine, or arginine. Urease was present at a high level when the cells were starved for nitrogen; its synthesis was repressed when the external ammonia concentration was high. Formation of urease did not require induction by urea and was not subject to catabolite repression. Its synthesis was controlled by glutamine synthetase. Mutants lacking glutamine synthetase failed to produce urease, and mutants forming glutamine synthetase at a high constitutive level also formed urease constitutively. Thus, the formation of urease is regulated like that of other enzymes of K. aerogenes capable of supplying the cell with ammonia or glutamate. | Urease of Klebsiella aerogenes: control of its synthesis by glutamine synthetase. Urease was purified 24-fold from extracts of Klebsiella aerogenes. The enzyme has a molecular weight of 230,000 as determined by gel filtration, is highly substrate specific, and has a Km for urea of 0.7 mM. A mutant strain lacking urease was isolated; it failed to grow with urea as the sole source of nitrogen but did grow on media containing other nitrogen sources such as ammonia, histidine, or arginine. Urease was present at a high level when the cells were starved for nitrogen; its synthesis was repressed when the external ammonia concentration was high. Formation of urease did not require induction by urea and was not subject to catabolite repression. Its synthesis was controlled by glutamine synthetase. Mutants lacking glutamine synthetase failed to produce urease, and mutants forming glutamine synthetase at a high constitutive level also formed urease constitutively. Thus, the formation of urease is regulated like that of other enzymes of K. aerogenes capable of supplying the cell with ammonia or glutamate. | [
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|
PMID:18439 | Fermentative metabolism of pyruvate by Rhodospirillum rubrum after anaerobic growth in darkness. | Rhodospirillum rubrum grew anaerobically in darkness and fermented sodium pyruvate by a pyruvate formate-lyase reaction. During 30 min of anaerobic dark or light incubation with sodium pyrivate, crude extracts from fermentatively grown cells produced about 6 micronmol of acetylphosphate and formate per mg of protein in reactions performed at pH 8.3. Cell extracts also catalyzed the exchange of sodium [14C]formate into sodium pyruvate at an apparent pH optimum of 7.3 to 7.5, but only about 2.5 micronmol of acetylphosphate was produced at this lower pH value. R. rubrum may also form pyruvate:ferredoxin oxidoreductase activity, as evidenced by low bicarbonate exchange activity. However, its participation in pyruvate metabolism in anaerobic dark-grown cells was not understood. During anaerobic, dark growth with pyruvate, formate was an intermediate in H2 and CO2 gas evolution. In contrast with H2 production by a light-dependent H2-nitrogenase system in photosynthetically grown cells, H2 formation in fermenting R. rubrum occurred through a carbon monoxide-sensitive formic hydrogenlyase reaction not influenced by light. | Fermentative metabolism of pyruvate by Rhodospirillum rubrum after anaerobic growth in darkness. Rhodospirillum rubrum grew anaerobically in darkness and fermented sodium pyruvate by a pyruvate formate-lyase reaction. During 30 min of anaerobic dark or light incubation with sodium pyrivate, crude extracts from fermentatively grown cells produced about 6 micronmol of acetylphosphate and formate per mg of protein in reactions performed at pH 8.3. Cell extracts also catalyzed the exchange of sodium [14C]formate into sodium pyruvate at an apparent pH optimum of 7.3 to 7.5, but only about 2.5 micronmol of acetylphosphate was produced at this lower pH value. R. rubrum may also form pyruvate:ferredoxin oxidoreductase activity, as evidenced by low bicarbonate exchange activity. However, its participation in pyruvate metabolism in anaerobic dark-grown cells was not understood. During anaerobic, dark growth with pyruvate, formate was an intermediate in H2 and CO2 gas evolution. In contrast with H2 production by a light-dependent H2-nitrogenase system in photosynthetically grown cells, H2 formation in fermenting R. rubrum occurred through a carbon monoxide-sensitive formic hydrogenlyase reaction not influenced by light. | [
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|
PMID:18440 | Characterization of an endonuclease associated with the drug resistance plasmid pKM101. | An endonuclease was detected in strains of Salmonella typhimurium containing the drug resistance plasmid pKM101. The enzyme was not detectable in strains lacking this plasmid, but it was present in strains containing mutants of pKM101 that were no longer able to enhance host cell mutagenesis. The endonuclease had a molecular weight of roughly 75,000 and, at pH 7.0, was equally active on single-stranded and duplex deoxyribonucleic acid (DNA). The reaction with single-stranded DNA was optimal at pH 5.5, whereas with duplex DNA the optimum was pH 6.8. The enzyme required a divalent cation for activity, and it had no detectable exonuclease activity with single-stranded or duplex DNA. The endonuclease extensively degraded DNA with no apparent base specificity, forming 5'-phosphomonoester termini. Although characterization of the endonuclease has not revealed its function, the enzyme does not appear to be a restriction endonuclease. | Characterization of an endonuclease associated with the drug resistance plasmid pKM101. An endonuclease was detected in strains of Salmonella typhimurium containing the drug resistance plasmid pKM101. The enzyme was not detectable in strains lacking this plasmid, but it was present in strains containing mutants of pKM101 that were no longer able to enhance host cell mutagenesis. The endonuclease had a molecular weight of roughly 75,000 and, at pH 7.0, was equally active on single-stranded and duplex deoxyribonucleic acid (DNA). The reaction with single-stranded DNA was optimal at pH 5.5, whereas with duplex DNA the optimum was pH 6.8. The enzyme required a divalent cation for activity, and it had no detectable exonuclease activity with single-stranded or duplex DNA. The endonuclease extensively degraded DNA with no apparent base specificity, forming 5'-phosphomonoester termini. Although characterization of the endonuclease has not revealed its function, the enzyme does not appear to be a restriction endonuclease. | [
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|
PMID:18441 | Enzymatic reduction of mercurous ions in Escherichia coli bearing R factor. | Mercurous ion (Hg(+))-dependent reduced nicotinamide adenine dinucleotide phosphate oxidation was demonstrated in an extract from cells of Escherichia coli W2252 that bear R factor. | Enzymatic reduction of mercurous ions in Escherichia coli bearing R factor. Mercurous ion (Hg(+))-dependent reduced nicotinamide adenine dinucleotide phosphate oxidation was demonstrated in an extract from cells of Escherichia coli W2252 that bear R factor. | [
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|
PMID:18446 | Base specificity of polyamine binding to synthetic polynucleotides. | The binding of polyamines and magnesium to synthetic polynucleotides has been studied by gel filtration on a Sephadex G-50 column. Among the single-stranded polynucleotides examined [poly(A), poly(C), and poly(U)], polyamines were found to bind to poly(C) and poly(U) preferentially, while the binding of Mg2+ was greatest with poly(A). Spermine bound to poly(U) was displaced completely by NH4+ but incompletely by Mg2+, while Mg2+ bound to poly(A) was displaced completely be spermine but incompletely by NH4+. The optimal pH for the binding of spermine to poly(U) was found to be about 7.9, while Mg2+ could bind to poly(A) over a broad pH range (7.1--8.7). | Base specificity of polyamine binding to synthetic polynucleotides. The binding of polyamines and magnesium to synthetic polynucleotides has been studied by gel filtration on a Sephadex G-50 column. Among the single-stranded polynucleotides examined [poly(A), poly(C), and poly(U)], polyamines were found to bind to poly(C) and poly(U) preferentially, while the binding of Mg2+ was greatest with poly(A). Spermine bound to poly(U) was displaced completely by NH4+ but incompletely by Mg2+, while Mg2+ bound to poly(A) was displaced completely be spermine but incompletely by NH4+. The optimal pH for the binding of spermine to poly(U) was found to be about 7.9, while Mg2+ could bind to poly(A) over a broad pH range (7.1--8.7). | [
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|
PMID:18447 | Morphology of lipid micelles containing lysolecithin. | Mixtures of lysolecithin with various phospholipids were studied by electron microscopy using negative staining. Mixtures of dipalmitoyllecithin and lysolecithin produced disc-shaped structures which were stacked in aggregates with a 6.0--6.4 nm repeat. The disc were 10--50 nm in diameter. The disc-shaped structures were best observed in equimolar mixtures of dipalmitoyllecithin and lysolecithin. When dipalmitoyllecithin was replaced by dimyristoyllecithin, the structures were rather different from those observed in the system containing dipalmitoyllecithin; a cylindrical micellar phase was predominant. Equimolar mixtures of egg lecithin and lysolecithin formed the more usual smectic, concentric lamellae (liposomes) and elongated rod-like micelles which might be bimolecular fragments of spherules. The radius of the rod-like micelles was about 6 nm. Structures of rod-like micelles were observed more frequently in the samples after incubation at room temperature and then further incubation at 0 degrees C. Equimolar mixtures of didecanoyllecithin and lysolecithin produced large amounts of elongated rod-like micelles. Beef brain sphingoymyelin showed disc-shaped structures when mixed with lysolecithin. Incorporation of cholesterol into the mixtures of dipalmitoyllecithin and lysolecithin changed the morphological structure; the size of the disc became larger and eventually liposomes were formed with an increase of cholesterol content. The structures observed in mixtures of dipalmitoyllecithin or sphingomyelin and lysolecithin closely resembled those observed in complexes of apolipoprotein and lipid. | Morphology of lipid micelles containing lysolecithin. Mixtures of lysolecithin with various phospholipids were studied by electron microscopy using negative staining. Mixtures of dipalmitoyllecithin and lysolecithin produced disc-shaped structures which were stacked in aggregates with a 6.0--6.4 nm repeat. The disc were 10--50 nm in diameter. The disc-shaped structures were best observed in equimolar mixtures of dipalmitoyllecithin and lysolecithin. When dipalmitoyllecithin was replaced by dimyristoyllecithin, the structures were rather different from those observed in the system containing dipalmitoyllecithin; a cylindrical micellar phase was predominant. Equimolar mixtures of egg lecithin and lysolecithin formed the more usual smectic, concentric lamellae (liposomes) and elongated rod-like micelles which might be bimolecular fragments of spherules. The radius of the rod-like micelles was about 6 nm. Structures of rod-like micelles were observed more frequently in the samples after incubation at room temperature and then further incubation at 0 degrees C. Equimolar mixtures of didecanoyllecithin and lysolecithin produced large amounts of elongated rod-like micelles. Beef brain sphingoymyelin showed disc-shaped structures when mixed with lysolecithin. Incorporation of cholesterol into the mixtures of dipalmitoyllecithin and lysolecithin changed the morphological structure; the size of the disc became larger and eventually liposomes were formed with an increase of cholesterol content. The structures observed in mixtures of dipalmitoyllecithin or sphingomyelin and lysolecithin closely resembled those observed in complexes of apolipoprotein and lipid. | [
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|
PMID:18448 | Affinity labeling of adenine nucleotide-related enzymes with reactive adenine nucleotide analogs. I. Affinity labeling of glyceraldehyde 3-phosphate dehydrogenase and myokinase with a reactive AMP analog. | Rabbit muscle glyceraldehyde 3-phosphate dehydrogenase (GPD) and myokinase (MK) were rapidly inactivated by a reactive AMP analog, N6-(p-bromoacetaminobenzyl)-AMP, under mild conditions. Complete inactivation was observed when 4 and 0.3 mol of the reagent with respect to enzyme were reacted with GPD and MK, respectively. The inactivation of both enzymes were favored at higher pH and the enzymes were protected by addition of adenine nucleotide substrate. Modified GPD or MK had no affinity for AMP-Sepharose, in contrast to the native enzymes. From these results, the inactivation of GPD and MK by the reactive AMP analog can be regarded as an affinity labeling. The posibility that the present AMP analog may be used as a general affinity labeling reagent for various adenine nucleotide-related enzymes is discussed based on the results obtained. | Affinity labeling of adenine nucleotide-related enzymes with reactive adenine nucleotide analogs. I. Affinity labeling of glyceraldehyde 3-phosphate dehydrogenase and myokinase with a reactive AMP analog. Rabbit muscle glyceraldehyde 3-phosphate dehydrogenase (GPD) and myokinase (MK) were rapidly inactivated by a reactive AMP analog, N6-(p-bromoacetaminobenzyl)-AMP, under mild conditions. Complete inactivation was observed when 4 and 0.3 mol of the reagent with respect to enzyme were reacted with GPD and MK, respectively. The inactivation of both enzymes were favored at higher pH and the enzymes were protected by addition of adenine nucleotide substrate. Modified GPD or MK had no affinity for AMP-Sepharose, in contrast to the native enzymes. From these results, the inactivation of GPD and MK by the reactive AMP analog can be regarded as an affinity labeling. The posibility that the present AMP analog may be used as a general affinity labeling reagent for various adenine nucleotide-related enzymes is discussed based on the results obtained. | [
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|
PMID:18442 | Survey for alpha-(1 leads to 3)-glucanase activity among yeasts. | Activity of alpha-(1 leads to 3)-glucanase was found in species of Cryptococcus, Rhodotorula, and Endomyces. Observations on the expression and stability of this enzyme in Rhodotorula minuta var. texensis was presented. | Survey for alpha-(1 leads to 3)-glucanase activity among yeasts. Activity of alpha-(1 leads to 3)-glucanase was found in species of Cryptococcus, Rhodotorula, and Endomyces. Observations on the expression and stability of this enzyme in Rhodotorula minuta var. texensis was presented. | [
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|
PMID:18449 | Stereochemical studies of hydrogen incorporation from nucleotides with fatty acid synthetase from Brevibacterium ammoniagenes. | The biosynthesis of fatty acids from malonyl-CoA and acetyl-CoA was investigated with an enzyme preparation which was purified 100-fold from Brevibacterium ammoniagenes. Fatty acids synthesized in the presence of D2O and stereospecifically deuterated NADPH and NADH were isolated and analyzed by mass chromatography to examine the localization of deuterium in the molecule. The following results were obtained: 1) HB hydrogen of NADPH was used for beta-ketoacyl reductase. 2) HB hydrogen of NADH was used for enoyl reductase. 3) Hydrogen atoms from water were found on the even-numbered methylene carbon atoms (2-hydrogen atoms per carbon atom) and some were also found on the odd-numbered methylene carbon atoms. 4) Hydrogen atoms from NADPH was found on the odd-numbered methylene carbon atoms (1 hydrogen per carbon). 5) Hydrogen atoms from NADH was also found on the odd-numbered methylene carbon atoms, but the number of incorporated hydrogen atoms was less than expected. The exchange of HB hydrogen of NADH with water catalyzed by enoyl reductase was suspected. 6) The exchange of methylene hydrogen atoms of malonyl-CoA with protons of water was suggested by 13C NMR analysis. | Stereochemical studies of hydrogen incorporation from nucleotides with fatty acid synthetase from Brevibacterium ammoniagenes. The biosynthesis of fatty acids from malonyl-CoA and acetyl-CoA was investigated with an enzyme preparation which was purified 100-fold from Brevibacterium ammoniagenes. Fatty acids synthesized in the presence of D2O and stereospecifically deuterated NADPH and NADH were isolated and analyzed by mass chromatography to examine the localization of deuterium in the molecule. The following results were obtained: 1) HB hydrogen of NADPH was used for beta-ketoacyl reductase. 2) HB hydrogen of NADH was used for enoyl reductase. 3) Hydrogen atoms from water were found on the even-numbered methylene carbon atoms (2-hydrogen atoms per carbon atom) and some were also found on the odd-numbered methylene carbon atoms. 4) Hydrogen atoms from NADPH was found on the odd-numbered methylene carbon atoms (1 hydrogen per carbon). 5) Hydrogen atoms from NADH was also found on the odd-numbered methylene carbon atoms, but the number of incorporated hydrogen atoms was less than expected. The exchange of HB hydrogen of NADH with water catalyzed by enoyl reductase was suspected. 6) The exchange of methylene hydrogen atoms of malonyl-CoA with protons of water was suggested by 13C NMR analysis. | [
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|
PMID:18450 | Chemical modifications of ribonuclease U1. | In order to obtain information on the nature of the amino acid residues involved in the activity of ribonuclease U1 [EC 3.1.4.8], various chemical modifications of the enzyme were carried out. RNase U1 was inactivated by reaction with iodoacetate at pH 5.5 with concomitant incorporation of 1 carboxymethyl group per molecule of the enzyme. The residue specifically modified by iodoacetate was identified as one of the glutamic acid residues, as in the case of RNase T1. The enzyme was also inactivated extensively by reaction with iodoacetamide at pH 8.0 with the loss of about one residue each of histidine and lysine. When RNase U1 was treated with a large excess of phenylglyoxal, the enzymatic activity and binding ability toward 3'-GMP were lost, with simultaneous modification of about 1 residue of arginine. The reaction of citraconic anhydride with RNase U1 led to the loss of enzymatic activity and modification of about 1 residue of lysine. The inactivated enzyme, however, retained binding ability toward 3'-GMP. These results indicate that there are marked similarities in the active sites of RNases T1 and U1. | Chemical modifications of ribonuclease U1. In order to obtain information on the nature of the amino acid residues involved in the activity of ribonuclease U1 [EC 3.1.4.8], various chemical modifications of the enzyme were carried out. RNase U1 was inactivated by reaction with iodoacetate at pH 5.5 with concomitant incorporation of 1 carboxymethyl group per molecule of the enzyme. The residue specifically modified by iodoacetate was identified as one of the glutamic acid residues, as in the case of RNase T1. The enzyme was also inactivated extensively by reaction with iodoacetamide at pH 8.0 with the loss of about one residue each of histidine and lysine. When RNase U1 was treated with a large excess of phenylglyoxal, the enzymatic activity and binding ability toward 3'-GMP were lost, with simultaneous modification of about 1 residue of arginine. The reaction of citraconic anhydride with RNase U1 led to the loss of enzymatic activity and modification of about 1 residue of lysine. The inactivated enzyme, however, retained binding ability toward 3'-GMP. These results indicate that there are marked similarities in the active sites of RNases T1 and U1. | [
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|
PMID:18451 | Purification and some properties of cyclohexylamine oxidase from a Pseudomonas sp. | Cyclohexylamine oxidase was purified 90-fold from cell-free extracts of Pseudomonas sp. capable of assimilating sodium cyclamate. The purified enzyme was homogeneous in disc electrophoresis, and the molecular weight was found to be approximately 80,000 by gel filtration. The enzyme catalyzed the following reaction: cyclohexylamine+O2+H2O leads to cyclohexanone+NH3+H2O2. The enzyme thus can be classified as an amine oxidase; it utilized oxygen as the ultimate electron acceptor. The pH optimum of the reaction was 6.8 and the apparent Km value for cyclohexylamine was 2.5 X 10(-4) M. The enzyme was highly specific for the deamination of alicyclic primary amines such as cyclohexylamine, but was found to be inactive toward ordinary amines used as substrates for amine oxidases. The enzyme solution was yellow in color and showed a typical flavoprotein spectrum; the addition of cyclohexylamine under anaerobic conditions caused reduction of the flavin in the native enzyme. The flavin of the prosthetic group was identified as FAD by thin layer chromatography. The participation of sulfhydryl groups in the enzymic action was also suggested by the observation that the enzyme activity was inhibited in the presence of PCMB and could be recovered by the addition of glutathione. | Purification and some properties of cyclohexylamine oxidase from a Pseudomonas sp. Cyclohexylamine oxidase was purified 90-fold from cell-free extracts of Pseudomonas sp. capable of assimilating sodium cyclamate. The purified enzyme was homogeneous in disc electrophoresis, and the molecular weight was found to be approximately 80,000 by gel filtration. The enzyme catalyzed the following reaction: cyclohexylamine+O2+H2O leads to cyclohexanone+NH3+H2O2. The enzyme thus can be classified as an amine oxidase; it utilized oxygen as the ultimate electron acceptor. The pH optimum of the reaction was 6.8 and the apparent Km value for cyclohexylamine was 2.5 X 10(-4) M. The enzyme was highly specific for the deamination of alicyclic primary amines such as cyclohexylamine, but was found to be inactive toward ordinary amines used as substrates for amine oxidases. The enzyme solution was yellow in color and showed a typical flavoprotein spectrum; the addition of cyclohexylamine under anaerobic conditions caused reduction of the flavin in the native enzyme. The flavin of the prosthetic group was identified as FAD by thin layer chromatography. The participation of sulfhydryl groups in the enzymic action was also suggested by the observation that the enzyme activity was inhibited in the presence of PCMB and could be recovered by the addition of glutathione. | [
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|
PMID:18455 | Purification and properties of phenylalanine ammonia-lyase in cut-injured sweet potato. | L-Phenylalanine ammonia-lyase (PAL) activity was developed in response to cut injury in sweet potato root tissue. The enzyme was purified from tissue incubated for 1 day after slicing by ammonium sulfate fractionation, column chromatographies on L-phenylalanyl Sepharose 4B, phosphocellulose. Sephadex G-200 and Sepharose 6B and preparative polyacrylamide gel electrophoresis. The molecular weight and sedimentation coefficient were estimated to be 285,000 to 320,000 and 11.6 to 11.9 S, respectively. Electrophoresis on sodium dodecyl sulfate-polyacrylamide gel yielded a single stained protein band which corresponded to a subunit weight of 80,000. Thus, the enzyme seems to be composed of four subunits of the same size. Neither L-tyrosine nor D-phenylalanine served as a substrate. Two Km values for the PAL were observed above and below 30 micrometers at various temperatures and were lower than those for PALs of other plants. The slope of the Arrhenius plot had a discontinuity at 17 degrees C. The values of activation energy were calculated to be 15,000 cal and 19,000 cal above and below 17 degrees C, respectively. Similar discontinuities were also observed in the effect of temperature on the Km values and the Hill coefficients. Negative cooperativity was observed at 10 degrees C (n = 0.83), but was not marked above 20 degrees C (n = 0.94). | Purification and properties of phenylalanine ammonia-lyase in cut-injured sweet potato. L-Phenylalanine ammonia-lyase (PAL) activity was developed in response to cut injury in sweet potato root tissue. The enzyme was purified from tissue incubated for 1 day after slicing by ammonium sulfate fractionation, column chromatographies on L-phenylalanyl Sepharose 4B, phosphocellulose. Sephadex G-200 and Sepharose 6B and preparative polyacrylamide gel electrophoresis. The molecular weight and sedimentation coefficient were estimated to be 285,000 to 320,000 and 11.6 to 11.9 S, respectively. Electrophoresis on sodium dodecyl sulfate-polyacrylamide gel yielded a single stained protein band which corresponded to a subunit weight of 80,000. Thus, the enzyme seems to be composed of four subunits of the same size. Neither L-tyrosine nor D-phenylalanine served as a substrate. Two Km values for the PAL were observed above and below 30 micrometers at various temperatures and were lower than those for PALs of other plants. The slope of the Arrhenius plot had a discontinuity at 17 degrees C. The values of activation energy were calculated to be 15,000 cal and 19,000 cal above and below 17 degrees C, respectively. Similar discontinuities were also observed in the effect of temperature on the Km values and the Hill coefficients. Negative cooperativity was observed at 10 degrees C (n = 0.83), but was not marked above 20 degrees C (n = 0.94). | [
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PMID:18457 | The behavior of 9-aminoacridine as an indicator of transmembrane pH difference in liposomes of natural bacterial phospholipids. | The behavior of 9-aminoacridine as an indicator of pH differences artificially set across a membrane has been reexamined in liposomes prepared from bacterial phospholipids extracted from chromatophores of Rhodopseudomonas capsulata grown photoheterotrophically. The dye behaves as an ideal indicator for pH differences lower than about three units; at higher pH's the expected linear dependence of Q/(100-Q) vs. pH is no longer strictly observed. Similarly a linear dependence upon the volume of the liposomes added has been verified. The amine ceases to respond to pH changes when the pH of the external medium exceeds the value of 10, corresponsing to the pKa of 9-aminoacridine. The apparent volume of the inner phase of liposomes, as calculated from fluorescence quenching, but not the slope of dependence of fluorescence on pH, appears to be affected by several factors, including the ionic composition, the osmolarity of the external medium, and the microscopic structure of the liposomes. Millimolar concentrations of earth-alkaline cations diminish the apparent internal volume of liposomes, in agreement with the complexing effect of these ions on phospholipid bilayers. The osmotic response of the apparent inner volume has also been verified; this parameter decreases linearly with the reciprocal of the external osmolarity, as expected from the van't Hoff relation; an osmolarity exceeding 0.3 M is, however, necessary in order to observe this effect. | The behavior of 9-aminoacridine as an indicator of transmembrane pH difference in liposomes of natural bacterial phospholipids. The behavior of 9-aminoacridine as an indicator of pH differences artificially set across a membrane has been reexamined in liposomes prepared from bacterial phospholipids extracted from chromatophores of Rhodopseudomonas capsulata grown photoheterotrophically. The dye behaves as an ideal indicator for pH differences lower than about three units; at higher pH's the expected linear dependence of Q/(100-Q) vs. pH is no longer strictly observed. Similarly a linear dependence upon the volume of the liposomes added has been verified. The amine ceases to respond to pH changes when the pH of the external medium exceeds the value of 10, corresponsing to the pKa of 9-aminoacridine. The apparent volume of the inner phase of liposomes, as calculated from fluorescence quenching, but not the slope of dependence of fluorescence on pH, appears to be affected by several factors, including the ionic composition, the osmolarity of the external medium, and the microscopic structure of the liposomes. Millimolar concentrations of earth-alkaline cations diminish the apparent internal volume of liposomes, in agreement with the complexing effect of these ions on phospholipid bilayers. The osmotic response of the apparent inner volume has also been verified; this parameter decreases linearly with the reciprocal of the external osmolarity, as expected from the van't Hoff relation; an osmolarity exceeding 0.3 M is, however, necessary in order to observe this effect. | [
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|
PMID:18462 | CDP-diglyceride:inositol transferase from rat liver. Purification and properties. | CDP-diglyceride:inositol transferase, which catalyzes the final step of the de novo synthesis of phosphatidylinositol, was solubilized by sodium cholate from microsomes prepared from rat liver and purified by ammonium sulfate fractionation, sucrose density gradient centrifugation, and DEAE-cellulose column chromatography. Addition of phospholipid during the purification and the assay procedures prevented irreversible loss of the enzyme activity to some extent. The resulting preparation was nearly homogeneous as judged by polyacrylamide gel electrophoresis. The recovery of the purified enzyme from the microsomal fraction was 3 to 3.3% with respect to activity and 0.12% with respect to amount of protein. The molecular weight of the enzyme was estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be 60,000. The purified enzyme required exogenous phospholipds for its activity. Various phospholipid classes activated the enzyme rather nonspecifically. The Km for myo-inositol was 2.5 X 10(-3) M and that for CDP-diglyceride was 1.7 X 10(-4) M. The pH optimum was 8.6. The enzyme required Mm2+ or Mg2+ for activity. The optimal concentration of Mn2+ for activation was 0.5 mM, while the activity in the presence of Mg2+ increased up to 20 mM. The enzyme was inhibited by thiol-reactive reagents. There was a competition for inositol by inosose-2 but not by scyllitol. | CDP-diglyceride:inositol transferase from rat liver. Purification and properties. CDP-diglyceride:inositol transferase, which catalyzes the final step of the de novo synthesis of phosphatidylinositol, was solubilized by sodium cholate from microsomes prepared from rat liver and purified by ammonium sulfate fractionation, sucrose density gradient centrifugation, and DEAE-cellulose column chromatography. Addition of phospholipid during the purification and the assay procedures prevented irreversible loss of the enzyme activity to some extent. The resulting preparation was nearly homogeneous as judged by polyacrylamide gel electrophoresis. The recovery of the purified enzyme from the microsomal fraction was 3 to 3.3% with respect to activity and 0.12% with respect to amount of protein. The molecular weight of the enzyme was estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be 60,000. The purified enzyme required exogenous phospholipds for its activity. Various phospholipid classes activated the enzyme rather nonspecifically. The Km for myo-inositol was 2.5 X 10(-3) M and that for CDP-diglyceride was 1.7 X 10(-4) M. The pH optimum was 8.6. The enzyme required Mm2+ or Mg2+ for activity. The optimal concentration of Mn2+ for activation was 0.5 mM, while the activity in the presence of Mg2+ increased up to 20 mM. The enzyme was inhibited by thiol-reactive reagents. There was a competition for inositol by inosose-2 but not by scyllitol. | [
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|
PMID:18463 | Characterization of the glutamine site of Escherichia coli guanosine 5'-monophosphate synthetase. | Alkylation of guanosine 5'-monophosphate (GMP) synthetase with the glutamine analogs L-2-amino-4-oxo-5-chloropentanoic acid (chloroketon) and 6-diazo-5-oxonorleucine (DON) inactivated glutamine- and NH3-dependent GMP synthetase. Inactivation exhibited second order kinetics. Complete inactivation was accompanied by covalent attachment of 0.4 to 0.5 equivalent of chloroketon/subunit. Alkylation of GMP synthetase with iodacetamide selectively inactivated glutamine-dependent activity. The NH3-dependent activity was relatively unaffected. Approximately 1 equivalent of carboxamidomethyl group was incorporated per subunit. Carboxymethylcysteine was the only modified amino acid hydrolysis. Prior treatment with chloroketone decreased the capacity for alkylation by iodacetamide, suggesting that both reagents alkylate the same residue. GMP synthetase exhibits glutaminase activity when ATP is replaced by adenosine plus PPi. Iodoacetamide inactivates glutaminase concomitant with glutamine-dependent GMP synthetase. Analysis of pH versus velocity and Km data indicates that the amide of glutamine remains enzyme bound and does not mix with exogenous NH3 in the synthesis of GMP. | Characterization of the glutamine site of Escherichia coli guanosine 5'-monophosphate synthetase. Alkylation of guanosine 5'-monophosphate (GMP) synthetase with the glutamine analogs L-2-amino-4-oxo-5-chloropentanoic acid (chloroketon) and 6-diazo-5-oxonorleucine (DON) inactivated glutamine- and NH3-dependent GMP synthetase. Inactivation exhibited second order kinetics. Complete inactivation was accompanied by covalent attachment of 0.4 to 0.5 equivalent of chloroketon/subunit. Alkylation of GMP synthetase with iodacetamide selectively inactivated glutamine-dependent activity. The NH3-dependent activity was relatively unaffected. Approximately 1 equivalent of carboxamidomethyl group was incorporated per subunit. Carboxymethylcysteine was the only modified amino acid hydrolysis. Prior treatment with chloroketone decreased the capacity for alkylation by iodacetamide, suggesting that both reagents alkylate the same residue. GMP synthetase exhibits glutaminase activity when ATP is replaced by adenosine plus PPi. Iodoacetamide inactivates glutaminase concomitant with glutamine-dependent GMP synthetase. Analysis of pH versus velocity and Km data indicates that the amide of glutamine remains enzyme bound and does not mix with exogenous NH3 in the synthesis of GMP. | [
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PMID:18464 | Nature of the lectin-induced activation of plasma membrane Mg2+ATPase. | The Mg2+ATPase activity of liver plasma membranes decreases markedly with increasing temperature above 30 degrees. This negative temperature dependency is counteracted by the binding of wheat germ agglutinin, concanavalin A, or Ricinus communis agglutinin (at concentrations greater than or equal 0.5 mg/ml) to membranes prior to assay of the enzyme. With one of these lectins bound, the enzyme has a single energy of activation between 20 degrees and 45 degrees. The binding of dimeric succinyl concanavalin A, soybean agglutinin, fucose-binding lectin from Lotus tetragonolobus, or the leucoagglutinin from Phaseolus vulgaris does not alter the temperature dependency of the enzyme. The latter two lectins, however, do prevent the concanavalin A-induced activation of the enzyme at 37 degrees. At saturating substrate concentrations, the enzyme is not inhibited by any of the lectins tested over a wide range of concentrations. Cytochalasin B and colchicine separately or in combination have little influence on the lectin-induced enhancement of enzyme activity. Chlorpromazine and vinblastine sulfate each partially prevent the activation and in combination do so completely. Treatment of the membranes with the detergent Lubrol-PX or phospholipase A prevents activation of the enzyme by concanavalin A. The results are consistent with a restriction by the lectin of an environment which is normally too disordered for maximal enzyme activity above 30 degrees. | Nature of the lectin-induced activation of plasma membrane Mg2+ATPase. The Mg2+ATPase activity of liver plasma membranes decreases markedly with increasing temperature above 30 degrees. This negative temperature dependency is counteracted by the binding of wheat germ agglutinin, concanavalin A, or Ricinus communis agglutinin (at concentrations greater than or equal 0.5 mg/ml) to membranes prior to assay of the enzyme. With one of these lectins bound, the enzyme has a single energy of activation between 20 degrees and 45 degrees. The binding of dimeric succinyl concanavalin A, soybean agglutinin, fucose-binding lectin from Lotus tetragonolobus, or the leucoagglutinin from Phaseolus vulgaris does not alter the temperature dependency of the enzyme. The latter two lectins, however, do prevent the concanavalin A-induced activation of the enzyme at 37 degrees. At saturating substrate concentrations, the enzyme is not inhibited by any of the lectins tested over a wide range of concentrations. Cytochalasin B and colchicine separately or in combination have little influence on the lectin-induced enhancement of enzyme activity. Chlorpromazine and vinblastine sulfate each partially prevent the activation and in combination do so completely. Treatment of the membranes with the detergent Lubrol-PX or phospholipase A prevents activation of the enzyme by concanavalin A. The results are consistent with a restriction by the lectin of an environment which is normally too disordered for maximal enzyme activity above 30 degrees. | [
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|
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