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PMID:23892 | Histopathology of veins after intravenous lorazepam and RO 21-3981. | A previously established rat model has been utilized to demonstrate that an acute inflammatory response occurs after high intravenous doses of lorazepam. This occurs only with high concentrations of drug equivalent to 20 times the normal clinical dosage in man. In contract, water soluble RO 21-3981 produces no vascular pathology in any dosage evaluated. It appears that propylene glycol may play a role in the pathogenesis of the intravascular injury observed. | Histopathology of veins after intravenous lorazepam and RO 21-3981. A previously established rat model has been utilized to demonstrate that an acute inflammatory response occurs after high intravenous doses of lorazepam. This occurs only with high concentrations of drug equivalent to 20 times the normal clinical dosage in man. In contract, water soluble RO 21-3981 produces no vascular pathology in any dosage evaluated. It appears that propylene glycol may play a role in the pathogenesis of the intravascular injury observed. | [
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PMID:23894 | Detection and prevalence of pneumococci with increased resistance to penicillin. | Susceptibility to penicillin was determined for 6000 strains of pneumococci isolated during 1974--76 from patients in Alberta and the adjacent region of the Northwest Territories. Strains were considered to be relatively resistant if the minimum inhibitory concentration (MIC) of penicillin was 0.16 microgram (0.26 U)/mL or more, which is eight or more times greater than the MIC for fully susceptible strains. Resistance was detected in 143 strains (2.4%) isolated from 122 patients and belonging to four capsular types. The MIC of the most resistant strains was 0.32 microgram (0.53 U/mL. Penicillin-resistant strains were highly resistant to oxacillin, the MIC being at least 30 times greater than that for penicillin-susceptible strains. Pneumococci resistant to penicillin may readily be detected by the narrowness or absence of a zone of inhibition around a 1-microgram oxacillin disc in susceptibility tests on blood agar. The degree of resistance reported here is relative and does not necessarily preclude successful treatment with full therapeutic doses of penicillin G, but penicillin preparations that give low blood concentrations may not be suitable for treating infections caused by these strains. | Detection and prevalence of pneumococci with increased resistance to penicillin. Susceptibility to penicillin was determined for 6000 strains of pneumococci isolated during 1974--76 from patients in Alberta and the adjacent region of the Northwest Territories. Strains were considered to be relatively resistant if the minimum inhibitory concentration (MIC) of penicillin was 0.16 microgram (0.26 U)/mL or more, which is eight or more times greater than the MIC for fully susceptible strains. Resistance was detected in 143 strains (2.4%) isolated from 122 patients and belonging to four capsular types. The MIC of the most resistant strains was 0.32 microgram (0.53 U/mL. Penicillin-resistant strains were highly resistant to oxacillin, the MIC being at least 30 times greater than that for penicillin-susceptible strains. Pneumococci resistant to penicillin may readily be detected by the narrowness or absence of a zone of inhibition around a 1-microgram oxacillin disc in susceptibility tests on blood agar. The degree of resistance reported here is relative and does not necessarily preclude successful treatment with full therapeutic doses of penicillin G, but penicillin preparations that give low blood concentrations may not be suitable for treating infections caused by these strains. | [
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PMID:23897 | Gamma-glutamyltransferase in putative premalignant liver cell populations during hepatocarcinogenesis. | The activity of gamma-glutamyltransferase, as measured quantitatively and by histochemical staining, was studied in different cell populations during the induction of liver cancer with 2-acetylaminofluorene (2-AAF) or diethylnitrosamine and compared with findings in fetal and in intact and regenerating adult liver. The enzyme activity is 20-fold higher in 12-week nodules than in control livers and 30-fold higher in 20-week nodules than in controls. A similar 30-fold increase in activity relative to control is present in hepatomas, induced by either 2-AAF or diethylnitrosamine, and in fetal hepatocytes. The enzyme shows increases in activity in foci of very early putative preneoplastic hepatocytes induced by a single dose of diethylnitrosamine and selected by low doses of 2-AAF plus partial hepatectomy. By 7 days, the foci show a 4-fold increase in enzyme activity, and by 3 weeks they are 40-fold higher than in the control liver. Histochemically, the foci are strongly positive for gamma-glutamyltransferase, especially in the bile canaliculi. By 21 days, the ductular (oval) cells induced by 2-AAF have disappeared. When stained for the enzyme activity, the foci stand out clearly against the negative background of the liver, allowing easy quantitation. It appears that gamma-glutamyltransferase is a useful marker for preneoplastic hepatocytes. | Gamma-glutamyltransferase in putative premalignant liver cell populations during hepatocarcinogenesis. The activity of gamma-glutamyltransferase, as measured quantitatively and by histochemical staining, was studied in different cell populations during the induction of liver cancer with 2-acetylaminofluorene (2-AAF) or diethylnitrosamine and compared with findings in fetal and in intact and regenerating adult liver. The enzyme activity is 20-fold higher in 12-week nodules than in control livers and 30-fold higher in 20-week nodules than in controls. A similar 30-fold increase in activity relative to control is present in hepatomas, induced by either 2-AAF or diethylnitrosamine, and in fetal hepatocytes. The enzyme shows increases in activity in foci of very early putative preneoplastic hepatocytes induced by a single dose of diethylnitrosamine and selected by low doses of 2-AAF plus partial hepatectomy. By 7 days, the foci show a 4-fold increase in enzyme activity, and by 3 weeks they are 40-fold higher than in the control liver. Histochemically, the foci are strongly positive for gamma-glutamyltransferase, especially in the bile canaliculi. By 21 days, the ductular (oval) cells induced by 2-AAF have disappeared. When stained for the enzyme activity, the foci stand out clearly against the negative background of the liver, allowing easy quantitation. It appears that gamma-glutamyltransferase is a useful marker for preneoplastic hepatocytes. | [
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PMID:23898 | Psychotropic drugs as potential antitumor agents: a selective screening study. | Compounds with known psychotropic properties were tested for activity in murine ip L1210 leukemia and B 16 melanoma in a protocol designed to obtain leads for new antitumor agents which might also possess central nervous system (CNS) antitumor properties. Barbiturates and hallucinogenic compounds were the only compound types deliberately excluded. Representatives from most of the other known CNS agent classes were included among the 297 psychotropic drugs evaluated. Sixteen of these agents were reproducibly active against the L1210 tumor system with T/C values of 125%. Phenothiazines such as fluphenazine and butyrophenones such as triperidol were prominent among the confirmed active structural types. Dopamine, a beta-phenethylamine neurotrasmitter, was active. While reproducible B16 melanoma activity was not observed among the psychotropic drugs, most of the L1210 confirmed active agents were effective against the ip P388 tumor model and also were active in vitro against KB cells. Ic L1210 activity was not observed among the few compounds chosen for testing in that tumor system. The yield of ip L1210 confirmed actives from this group of psychotropic agents was 18 times that which would have been expected from the random screening of compounds. | Psychotropic drugs as potential antitumor agents: a selective screening study. Compounds with known psychotropic properties were tested for activity in murine ip L1210 leukemia and B 16 melanoma in a protocol designed to obtain leads for new antitumor agents which might also possess central nervous system (CNS) antitumor properties. Barbiturates and hallucinogenic compounds were the only compound types deliberately excluded. Representatives from most of the other known CNS agent classes were included among the 297 psychotropic drugs evaluated. Sixteen of these agents were reproducibly active against the L1210 tumor system with T/C values of 125%. Phenothiazines such as fluphenazine and butyrophenones such as triperidol were prominent among the confirmed active structural types. Dopamine, a beta-phenethylamine neurotrasmitter, was active. While reproducible B16 melanoma activity was not observed among the psychotropic drugs, most of the L1210 confirmed active agents were effective against the ip P388 tumor model and also were active in vitro against KB cells. Ic L1210 activity was not observed among the few compounds chosen for testing in that tumor system. The yield of ip L1210 confirmed actives from this group of psychotropic agents was 18 times that which would have been expected from the random screening of compounds. | [
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PMID:23899 | Lack of effect of hypercapnic acidosis on elasticity of cat papillary muscles. | Chages in external pH from 7.40 to 6.95 obtained by changing the pCO2 of the medium at constant bicarbonate concentration produced in cat papillary muscles a significant decrease in isometric tension with no changes in time to peak tension. Active and resting stiffness, as determined by two different methods, did not change under conditions of hypercapnic acidosis. | Lack of effect of hypercapnic acidosis on elasticity of cat papillary muscles. Chages in external pH from 7.40 to 6.95 obtained by changing the pCO2 of the medium at constant bicarbonate concentration produced in cat papillary muscles a significant decrease in isometric tension with no changes in time to peak tension. Active and resting stiffness, as determined by two different methods, did not change under conditions of hypercapnic acidosis. | [
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PMID:23901 | Species-specific aggregation factor in sponges. VII. Its effect on cyclic amp and cyclic gmp metabolism in cells of Geodia cydonium. | In dissociated single cells from the sponge Geodia cydonium, DNA synthesis is initiated after incubation with a homologous, soluble aggregation factor. During the DNA-initiation phase the cyclic AMP- and cyclic GMP levels vary drastically; the cyclic AMP content drops from 2.2 pmol/10(6) cells to 0.3 pmol/10(6) cells while the cyclic GMP content increases from 0.6 pmol to 3.7 pmol/10(6) cells. The activity of neither the adenylate cyclase nor of the guanylate cyclase isolated from cells which have been incubated for different periods of time with the aggregation factor, is changed. The soluble as well as the particulate enzyme activities were checked in vitro. The cyclic nucleotide receptors have been isolated from the sponge cells and characterized with respect to their molecular weight, dissociation constant for cyclic AMP or cyclic GMP and intracellular concentration. None of these parameters are altered during aggregation factor-mediated DNA initiation. From these data it is concluded that the regulation of cyclic nucleotide levels is a consequence of a changed activity of nucleotide cyclases or of phosphodiesterases, but this is presumably not caused by a changed rate of synthesis of nucleotide cyclases or of cyclic nucleotide receptors. | Species-specific aggregation factor in sponges. VII. Its effect on cyclic amp and cyclic gmp metabolism in cells of Geodia cydonium. In dissociated single cells from the sponge Geodia cydonium, DNA synthesis is initiated after incubation with a homologous, soluble aggregation factor. During the DNA-initiation phase the cyclic AMP- and cyclic GMP levels vary drastically; the cyclic AMP content drops from 2.2 pmol/10(6) cells to 0.3 pmol/10(6) cells while the cyclic GMP content increases from 0.6 pmol to 3.7 pmol/10(6) cells. The activity of neither the adenylate cyclase nor of the guanylate cyclase isolated from cells which have been incubated for different periods of time with the aggregation factor, is changed. The soluble as well as the particulate enzyme activities were checked in vitro. The cyclic nucleotide receptors have been isolated from the sponge cells and characterized with respect to their molecular weight, dissociation constant for cyclic AMP or cyclic GMP and intracellular concentration. None of these parameters are altered during aggregation factor-mediated DNA initiation. From these data it is concluded that the regulation of cyclic nucleotide levels is a consequence of a changed activity of nucleotide cyclases or of phosphodiesterases, but this is presumably not caused by a changed rate of synthesis of nucleotide cyclases or of cyclic nucleotide receptors. | [
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PMID:23902 | Enteroendocrine cells in the digestive tract of Barbus conchonius (teleostei, cyprinidae). | Just as in other cyprinids, three zones can be distinguished in the digestive tract of Barbus conchonius. A fat absorptive zone (65--75%), including the intestinal bulb, is followed by a protein absorptive zone (25--35%) and a small ion and water absorptive zone (less than 5%). The main characteristics of these zones are described. Four types of enteroendocrine cells can be distinguished between the intestinal epithelial cells. The number decreases in the caudal direction, and there are very few in the protein absorptive zone. All the enteroendocrine cells are argyrophilic and differ mainly in the size and shape of their secretory granules. Serotonin producing and hence argentaffin cells have not been found. Amine precursor uptake and decarboxylation (APUD) by the enteroendocrine cells of adult fishes has not been observed. The possible functions of the enteroendocrine cells are discussed. (Auto-)phagosomes, common in epithelial cells of the gut of B. conchonius, show similar staining characteristics as the enteroendocrine cells; their function is discussed. | Enteroendocrine cells in the digestive tract of Barbus conchonius (teleostei, cyprinidae). Just as in other cyprinids, three zones can be distinguished in the digestive tract of Barbus conchonius. A fat absorptive zone (65--75%), including the intestinal bulb, is followed by a protein absorptive zone (25--35%) and a small ion and water absorptive zone (less than 5%). The main characteristics of these zones are described. Four types of enteroendocrine cells can be distinguished between the intestinal epithelial cells. The number decreases in the caudal direction, and there are very few in the protein absorptive zone. All the enteroendocrine cells are argyrophilic and differ mainly in the size and shape of their secretory granules. Serotonin producing and hence argentaffin cells have not been found. Amine precursor uptake and decarboxylation (APUD) by the enteroendocrine cells of adult fishes has not been observed. The possible functions of the enteroendocrine cells are discussed. (Auto-)phagosomes, common in epithelial cells of the gut of B. conchonius, show similar staining characteristics as the enteroendocrine cells; their function is discussed. | [
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PMID:23903 | Ultrastructural appearance of neurosecretory granules in the sinus gland of the crab after different fixation procedures. | The appearance of neurosecretory granules in the crab sinus gland was studied after fixation at different pHs. Whereas at pH 7.0 the neurosecretory granules were pleomorphic with respect to electron density, at pH 5.0 or 6.0 all the granules remained electron dense. The possible role of maturation as an explanation of this observation is discussed. | Ultrastructural appearance of neurosecretory granules in the sinus gland of the crab after different fixation procedures. The appearance of neurosecretory granules in the crab sinus gland was studied after fixation at different pHs. Whereas at pH 7.0 the neurosecretory granules were pleomorphic with respect to electron density, at pH 5.0 or 6.0 all the granules remained electron dense. The possible role of maturation as an explanation of this observation is discussed. | [
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PMID:23904 | A radioautographic study of the neuroepithelial bodies of the lungs in fetal and neonatal rabbits. | The neuroepithelial bodies (NEB's) of the lung of 29-day-old fetuses and 1-day-old rabbits, under the conditions of this study, neither take up 3H-thymidine nor undergo mitosis. Also the NEB's are not derived at these times from proliferations of other kinds of epithelial cells in the intrapulmonary airways. It is, therefore, suggested that the difference in numbers of NEB's previously observed by us, between the 29-day fetus and the 1-day-old rabbit, is due either to regranulation or acquisition of argyrophilic material by the NEB's or differentiation of other epithelial types. It is concluded that the NEB's are composed of well differentiated cells, which have a greatly reduced capacity to undergo mitosis. | A radioautographic study of the neuroepithelial bodies of the lungs in fetal and neonatal rabbits. The neuroepithelial bodies (NEB's) of the lung of 29-day-old fetuses and 1-day-old rabbits, under the conditions of this study, neither take up 3H-thymidine nor undergo mitosis. Also the NEB's are not derived at these times from proliferations of other kinds of epithelial cells in the intrapulmonary airways. It is, therefore, suggested that the difference in numbers of NEB's previously observed by us, between the 29-day fetus and the 1-day-old rabbit, is due either to regranulation or acquisition of argyrophilic material by the NEB's or differentiation of other epithelial types. It is concluded that the NEB's are composed of well differentiated cells, which have a greatly reduced capacity to undergo mitosis. | [
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PMID:23909 | P-Nitrophenol-alpha-D-glucopyranoside as substrate for measurement of maltase activity in human semen. | Hitherto, seminal plasma maltase has been measured with maltose as substrate; this method is time consuming and lacks specificity. The use of a synthetic substrate, p-nitrophenol-alpha-D-glucopyranoside, allows accurate and rapid determination of this activity. When maltase is added to the incubation medium (the substrate and reduced glutathione in potassium phosphate buffer, pH 6.8), maintained at 37 degrees C, hydrolysis of the original substrate to p-nitrophenol goes at a constant rate during 4 h. Under optimal conditions of incubation, the Michaelis constant of the reaction, calculated by the Hanes method, was 2.92 +/- 0.84 (SD) X 10(-3) for six different semen samples. Isomaltase appeared to be absent from seminal plasma. The enzyme is stable to freezing and slow thawing and can be stored for at least 26 days at -80 degrees C. Its molecular weight is 259 000. Tris(hydroxymethyl)aminomethane (pH 6.8) exerts a noncompetitive inhibition on the enzyme activity. In 68 men 23 to 45 years old, whose semen analyses were normal, the seminal plasma maltase activity was 467 +/- 135 (SD) mU/g of protein. It was generally decreased in patients with infertility disorders. | P-Nitrophenol-alpha-D-glucopyranoside as substrate for measurement of maltase activity in human semen. Hitherto, seminal plasma maltase has been measured with maltose as substrate; this method is time consuming and lacks specificity. The use of a synthetic substrate, p-nitrophenol-alpha-D-glucopyranoside, allows accurate and rapid determination of this activity. When maltase is added to the incubation medium (the substrate and reduced glutathione in potassium phosphate buffer, pH 6.8), maintained at 37 degrees C, hydrolysis of the original substrate to p-nitrophenol goes at a constant rate during 4 h. Under optimal conditions of incubation, the Michaelis constant of the reaction, calculated by the Hanes method, was 2.92 +/- 0.84 (SD) X 10(-3) for six different semen samples. Isomaltase appeared to be absent from seminal plasma. The enzyme is stable to freezing and slow thawing and can be stored for at least 26 days at -80 degrees C. Its molecular weight is 259 000. Tris(hydroxymethyl)aminomethane (pH 6.8) exerts a noncompetitive inhibition on the enzyme activity. In 68 men 23 to 45 years old, whose semen analyses were normal, the seminal plasma maltase activity was 467 +/- 135 (SD) mU/g of protein. It was generally decreased in patients with infertility disorders. | [
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PMID:23910 | Optimal reaction conditions for assaying human lactate dehydrogenase pyruvate-to-lactate at 25, 30, and 37 degrees C. | Optimal reaction conditions for assaying human lactate dehydrogenase pyruvate-to-lactate were determined for isoenzymes 1 and 5 at 25, 30, and 37 degrees C. Three of the nine different buffers examined--imidazole, triethanolamine, and N-tris(hydroxymethyl)-methyl-2-aminoethane sulfonic acid--are satisfactory. Beta-NADH, pyruvate, and hydrogen ion concentrations were chosen to measure both isoenzymes with maximal-equal-sustainable efficiency at the lowest substrate concentrations. Approximately 95% of each isoenzyme is measured, for activities up to threefold the upper normal limit, if the measurements are made immediately after the reaction is initiated. The Arrhenius relationship for each isoenzyme is unique. Interconversion of results from one temperature to another is practical only with reservations. Results at 37 degrees C are not as reliable as those at 25 degrees C. | Optimal reaction conditions for assaying human lactate dehydrogenase pyruvate-to-lactate at 25, 30, and 37 degrees C. Optimal reaction conditions for assaying human lactate dehydrogenase pyruvate-to-lactate were determined for isoenzymes 1 and 5 at 25, 30, and 37 degrees C. Three of the nine different buffers examined--imidazole, triethanolamine, and N-tris(hydroxymethyl)-methyl-2-aminoethane sulfonic acid--are satisfactory. Beta-NADH, pyruvate, and hydrogen ion concentrations were chosen to measure both isoenzymes with maximal-equal-sustainable efficiency at the lowest substrate concentrations. Approximately 95% of each isoenzyme is measured, for activities up to threefold the upper normal limit, if the measurements are made immediately after the reaction is initiated. The Arrhenius relationship for each isoenzyme is unique. Interconversion of results from one temperature to another is practical only with reservations. Results at 37 degrees C are not as reliable as those at 25 degrees C. | [
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PMID:23911 | D(-)-N-Methylglucamine buffer for pH 8.5 to 10.5. | A new amine buffer for the pH range 8.5 to 10.5 based on D(-)-N-methylglucamine and its hydrochloride is described. Important physical and chemical constants characterizing this buffer system have been determined: thermodynamic pKa = 9.52 (25 degrees C), buffer capacity = 0.055, dilution value pH 1/2 = -0.02, temperature coefficient dpKa/dT = 0.023, the heat of ionization delta H degrees = 36.93 kj mol-1. D(-)-N-Methylglucamine, which is available in highly purified form, is highly soluble in water. Thus one can prepare either buffer solutions or stable solid mixtures when combined with D(-)-N-methylglucaminium chloride. D(-)-N-Methylglucamine buffers are suitable for both biochemistry and pH control. | D(-)-N-Methylglucamine buffer for pH 8.5 to 10.5. A new amine buffer for the pH range 8.5 to 10.5 based on D(-)-N-methylglucamine and its hydrochloride is described. Important physical and chemical constants characterizing this buffer system have been determined: thermodynamic pKa = 9.52 (25 degrees C), buffer capacity = 0.055, dilution value pH 1/2 = -0.02, temperature coefficient dpKa/dT = 0.023, the heat of ionization delta H degrees = 36.93 kj mol-1. D(-)-N-Methylglucamine, which is available in highly purified form, is highly soluble in water. Thus one can prepare either buffer solutions or stable solid mixtures when combined with D(-)-N-methylglucaminium chloride. D(-)-N-Methylglucamine buffers are suitable for both biochemistry and pH control. | [
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PMID:23912 | Synthetic substrate beta-glucosidase activity in leukocytes: a reproducible method for the identification of patients and carriers of Gaucher's disease. | A method is described for the identification of patients and carriers of Gaucher's disease, using leukocytes from a small volume of blood. The fluorogenic substrate, 4-methylumbelliferyl-beta-D-glucopyranoside, was assayed in the presence of pure sodium taurocholate (2.5 mg/ml) and Triton X-100 (2.0 mg/ml). Some commercial brands of pure sodium taurocholate were satisfactory for this purpose. The pH optimum for controls, Gaucher disease carriers and Gaucher disease patients was 5.4 using citrate-phosphate buffer. Although leukocytes prepared from only a small amount of blood (2-8 ml) are required, there is sufficient quantity for measuring other lysosomal enzymes as controls. Using this method, 12 patients with all types of Gaucher's disease and 12 obligate heterozygotes were identified. Carrier status was predicted in six other family members and ruled out in six others. Eight unaffected people married to Gaucher carriers or Gaucher patients were predicted to be non-carriers of Gaucher's disease, thereby ruling out children affected with Gaucher's disease in that mating. | Synthetic substrate beta-glucosidase activity in leukocytes: a reproducible method for the identification of patients and carriers of Gaucher's disease. A method is described for the identification of patients and carriers of Gaucher's disease, using leukocytes from a small volume of blood. The fluorogenic substrate, 4-methylumbelliferyl-beta-D-glucopyranoside, was assayed in the presence of pure sodium taurocholate (2.5 mg/ml) and Triton X-100 (2.0 mg/ml). Some commercial brands of pure sodium taurocholate were satisfactory for this purpose. The pH optimum for controls, Gaucher disease carriers and Gaucher disease patients was 5.4 using citrate-phosphate buffer. Although leukocytes prepared from only a small amount of blood (2-8 ml) are required, there is sufficient quantity for measuring other lysosomal enzymes as controls. Using this method, 12 patients with all types of Gaucher's disease and 12 obligate heterozygotes were identified. Carrier status was predicted in six other family members and ruled out in six others. Eight unaffected people married to Gaucher carriers or Gaucher patients were predicted to be non-carriers of Gaucher's disease, thereby ruling out children affected with Gaucher's disease in that mating. | [
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PMID:23913 | Cystic fibrosis liver sialyltransferase. | The activity of sialytransferase with regard to the glycoprotein substrates asialofetuin and asialo-ovine submaxillary mucin was determined in normal, pathological control, and cystic fibrosis liver homogenates. Cystic fibrosis and pathological livers have about 40% of the average normal specific activity for sialytransferase. Several properties of cystic fibrosis sialytransferase were investigated and compared to those of the normal liver enzyme (Alhadeff et al. 1977). The pH optima curves were similar, but cystic fibrosis sialyltransferase appears to be more thermolabile than the normal liver enzyme. Isoelectric focusing studies revealed that the three most basic forms of sialyltransferase which are found in normal livers are deficient or absent in most cystic fibrosis liver. The data suggest that altered glycoprotein-sialyltransferases may be present in cystic fibrosis livers, probably a secondary effect due to general liver pathology. | Cystic fibrosis liver sialyltransferase. The activity of sialytransferase with regard to the glycoprotein substrates asialofetuin and asialo-ovine submaxillary mucin was determined in normal, pathological control, and cystic fibrosis liver homogenates. Cystic fibrosis and pathological livers have about 40% of the average normal specific activity for sialytransferase. Several properties of cystic fibrosis sialytransferase were investigated and compared to those of the normal liver enzyme (Alhadeff et al. 1977). The pH optima curves were similar, but cystic fibrosis sialyltransferase appears to be more thermolabile than the normal liver enzyme. Isoelectric focusing studies revealed that the three most basic forms of sialyltransferase which are found in normal livers are deficient or absent in most cystic fibrosis liver. The data suggest that altered glycoprotein-sialyltransferases may be present in cystic fibrosis livers, probably a secondary effect due to general liver pathology. | [
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PMID:23936 | [Standardisation of obtaining blood samples: influence of tourniquet application on 33 constituents of blood and serum (author's transl)]. | The extent and dynamics of changes by short (1 min) or prolonged (6 min) tourniquet application while obtaining venous blood samples were analysed with respect ot 33 frequently measured constituents of blood and serum. After 6-minute tourniquet application the values for red cells, haemoglobin, packed cell volume, total protein, albumen, gamma-glutamyl transferase, alkaline phosphatase, lactate dehydrogenase, creatinekinase, bilirubin, cholesterol, total glycerol and calcium increased by an average of 4-9%. One-minute tourniquet application did not have a significant effect. Levels of sodium, potassium, carbon dioxide, creatinine, uric acid, ratio of electrophoretic fractions and the MCV, MCH and MCHC indices were not affected even by 6-minute tourniquet applications. The introduction of blood sampling under standardised conditions is proposed. | [Standardisation of obtaining blood samples: influence of tourniquet application on 33 constituents of blood and serum (author's transl)]. The extent and dynamics of changes by short (1 min) or prolonged (6 min) tourniquet application while obtaining venous blood samples were analysed with respect ot 33 frequently measured constituents of blood and serum. After 6-minute tourniquet application the values for red cells, haemoglobin, packed cell volume, total protein, albumen, gamma-glutamyl transferase, alkaline phosphatase, lactate dehydrogenase, creatinekinase, bilirubin, cholesterol, total glycerol and calcium increased by an average of 4-9%. One-minute tourniquet application did not have a significant effect. Levels of sodium, potassium, carbon dioxide, creatinine, uric acid, ratio of electrophoretic fractions and the MCV, MCH and MCHC indices were not affected even by 6-minute tourniquet applications. The introduction of blood sampling under standardised conditions is proposed. | [
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PMID:23937 | Drugs and depression. | Moderate or severe depression is now one of the most common diseases of our time with a prevalence of nearly 3%. It seems likely that this prevalence has increased as a result of the wider use of drugs which have an effect on the neurotransmitters. Changes in the levels of these neurotransmitters in the central nervous system are thought to be the biochemical basis for the development of at least some depressive illnesses. Drug-induced depressions are more likely to occur in those individuals who are genetically predisposed to depression or who have had a previous depressive illness. Other groups who are particularly susceptible to these effects are the elderly. Many groups of drugs have a primary or secondary action on the central nervous system neurotransmitter function. Some 200 drugs have been claimed to cause depression in certain patients, but only a relatively small number precipitate depressive symptoms with any frequency. Those most commonly implicated are the long-acting antipsychotics, barbiturates, ethanol, oral contraceptives and antihypertensive agents. It is important to remember that some drugs, such as reserpine, cause depression as a side-effect during their therapeutic use whereas others, such as fenfluramine, cause depression mainly when they are withdrawn too rapidly. In those patients presenting with depression, it is important to review the current drug therapy in order to assess the part played by these drugs in the development of the depression. Following this assessment, drug therapy should be adjusted appropriately. However, a distinction must be made between the symptoms of depression, those physiological changes which occur during treatment with a variety of drugs, and the patient's reaction to the disease for which they are being treated. | Drugs and depression. Moderate or severe depression is now one of the most common diseases of our time with a prevalence of nearly 3%. It seems likely that this prevalence has increased as a result of the wider use of drugs which have an effect on the neurotransmitters. Changes in the levels of these neurotransmitters in the central nervous system are thought to be the biochemical basis for the development of at least some depressive illnesses. Drug-induced depressions are more likely to occur in those individuals who are genetically predisposed to depression or who have had a previous depressive illness. Other groups who are particularly susceptible to these effects are the elderly. Many groups of drugs have a primary or secondary action on the central nervous system neurotransmitter function. Some 200 drugs have been claimed to cause depression in certain patients, but only a relatively small number precipitate depressive symptoms with any frequency. Those most commonly implicated are the long-acting antipsychotics, barbiturates, ethanol, oral contraceptives and antihypertensive agents. It is important to remember that some drugs, such as reserpine, cause depression as a side-effect during their therapeutic use whereas others, such as fenfluramine, cause depression mainly when they are withdrawn too rapidly. In those patients presenting with depression, it is important to review the current drug therapy in order to assess the part played by these drugs in the development of the depression. Following this assessment, drug therapy should be adjusted appropriately. However, a distinction must be made between the symptoms of depression, those physiological changes which occur during treatment with a variety of drugs, and the patient's reaction to the disease for which they are being treated. | [
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PMID:23938 | Drug interactions with antihypertensive drugs. | Drug interactions with antihypertensive drugs can be either beneficial or hazardous. The hazardous interactions are relatively infrequent but must be shown so they can be avoided. Those of most importance involve interaction with guanethidine-type agents and tricyclic antidepressants, amphetamine-type anorexiants or phenolpropanolamine-type common cold remedies; combined use of potassium retaining diuretics with potassium supplements; and incautious use of diuretics with cardiac glycosides. The beneficial interactions are the basis for modern antihypertensive therapy and can be of major help if logically applied to therapeutic problems. | Drug interactions with antihypertensive drugs. Drug interactions with antihypertensive drugs can be either beneficial or hazardous. The hazardous interactions are relatively infrequent but must be shown so they can be avoided. Those of most importance involve interaction with guanethidine-type agents and tricyclic antidepressants, amphetamine-type anorexiants or phenolpropanolamine-type common cold remedies; combined use of potassium retaining diuretics with potassium supplements; and incautious use of diuretics with cardiac glycosides. The beneficial interactions are the basis for modern antihypertensive therapy and can be of major help if logically applied to therapeutic problems. | [
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PMID:23941 | The molluscicidal properties of natural products from Ambrosia maritima. | The molluscicidal properties of Damsin "I", Ambrosin "II", and tribromodamsin "III" were investigated against the intermediate hosts of schistosomiasis Biomphalaria alexandrina and Bulinus truncatus. Although Damsin was to some extent more toxic against B. alexandrina than the other two compounds, it was less toxic than them against B. truncatus after 24 hr exposure periods. There is a flex relationship between concentration of Damsin and exposure period to prodce 100% mortality. The molluscicidal potency of Damsin remained stable over a wide range of pH values, but was affected by storage, sunlight and river bed-mud. Structure-activity relationship is included. | The molluscicidal properties of natural products from Ambrosia maritima. The molluscicidal properties of Damsin "I", Ambrosin "II", and tribromodamsin "III" were investigated against the intermediate hosts of schistosomiasis Biomphalaria alexandrina and Bulinus truncatus. Although Damsin was to some extent more toxic against B. alexandrina than the other two compounds, it was less toxic than them against B. truncatus after 24 hr exposure periods. There is a flex relationship between concentration of Damsin and exposure period to prodce 100% mortality. The molluscicidal potency of Damsin remained stable over a wide range of pH values, but was affected by storage, sunlight and river bed-mud. Structure-activity relationship is included. | [
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PMID:23942 | Arylsulfatases in bilharziasis. | Arylsulfatase A and B in urine have been estimated in 18 normal subjects and 50 bilharziasis patients. The bilharziasis patients were divided into two groups according to the type of infeciton. Those with bilharziasis haematobian type of infection and those with the bilharziasis mansoni type. Each group was further subdivided into subgroups according to the severity and progress of the disease. The activities of arylsulfatase A and B were significantly elevated in all the groups of patients studied and it is evident that there is a progressive increase with the progress of the disease in both types of bilharziasis infections (the haematobian and mansoni types). Liver dysfunction consequent of bilharzial infestation appears to take part in the mechanism of induction of the bilharzial bladder cancer. | Arylsulfatases in bilharziasis. Arylsulfatase A and B in urine have been estimated in 18 normal subjects and 50 bilharziasis patients. The bilharziasis patients were divided into two groups according to the type of infeciton. Those with bilharziasis haematobian type of infection and those with the bilharziasis mansoni type. Each group was further subdivided into subgroups according to the severity and progress of the disease. The activities of arylsulfatase A and B were significantly elevated in all the groups of patients studied and it is evident that there is a progressive increase with the progress of the disease in both types of bilharziasis infections (the haematobian and mansoni types). Liver dysfunction consequent of bilharzial infestation appears to take part in the mechanism of induction of the bilharzial bladder cancer. | [
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PMID:23943 | Laboratory model ecosystem evaluation of the chemical and biological behavior of radiolabeled micropollutants. | A standardized laboratory model ecosystem has been used to evaluate the comparative behavior of radiolabeled micropollutants including organochlorine, organophosphorus, carbamate, and hormone-mimic insecticides; herbicides; important industrial organic compounds including phthalate esters and PCB's; and specific pollutants such as TCBD and hexachlorobenzene. The compounds have been studies for ecological effects over a terrestrial-aquatic interface and through several food chains, especially with regard to the development of quantitative information on ecological magnification and biodegradability index. The pathways of chemical degradation in the various organisms have been evaluated and the ecological effects of degradation products investigated. Illustrations are given of the use of the model ecosystem technology for screening new candidate pesticides for effects on environmental quality; for evaluating the hazards of environmental pollution by industrial waste effluents; and for fundamental studies of the principles fo biodegradability of organic chemicals in a variety of organisms. | Laboratory model ecosystem evaluation of the chemical and biological behavior of radiolabeled micropollutants. A standardized laboratory model ecosystem has been used to evaluate the comparative behavior of radiolabeled micropollutants including organochlorine, organophosphorus, carbamate, and hormone-mimic insecticides; herbicides; important industrial organic compounds including phthalate esters and PCB's; and specific pollutants such as TCBD and hexachlorobenzene. The compounds have been studies for ecological effects over a terrestrial-aquatic interface and through several food chains, especially with regard to the development of quantitative information on ecological magnification and biodegradability index. The pathways of chemical degradation in the various organisms have been evaluated and the ecological effects of degradation products investigated. Illustrations are given of the use of the model ecosystem technology for screening new candidate pesticides for effects on environmental quality; for evaluating the hazards of environmental pollution by industrial waste effluents; and for fundamental studies of the principles fo biodegradability of organic chemicals in a variety of organisms. | [
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PMID:23944 | Influence of denervation and reinnervation on autolytic activity and on protein composition of skeletal muscle in rat. | Changes in the proteolytic activity and in the relative content of protein in soluble, myofibrillar and insoluble fractions were investigated following denervation and reinnervation of the soleus and tibialis anterior muscles of the rat. After denervation an increase of autolysis in the acid and neutral pH range, but not in the alkaline one, was found in both muscles. An increased autolysis at the acid and neutral pH range was also observed in both muscles after reinnervation, when the weight of the muscles increased. The results indicate the lack of inverse relationship between the changes of proteolytic activity and the decrease or increase of the amount of muscle protein in the course of muscle atrophy and regeneration. | Influence of denervation and reinnervation on autolytic activity and on protein composition of skeletal muscle in rat. Changes in the proteolytic activity and in the relative content of protein in soluble, myofibrillar and insoluble fractions were investigated following denervation and reinnervation of the soleus and tibialis anterior muscles of the rat. After denervation an increase of autolysis in the acid and neutral pH range, but not in the alkaline one, was found in both muscles. An increased autolysis at the acid and neutral pH range was also observed in both muscles after reinnervation, when the weight of the muscles increased. The results indicate the lack of inverse relationship between the changes of proteolytic activity and the decrease or increase of the amount of muscle protein in the course of muscle atrophy and regeneration. | [
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PMID:23945 | Glucokinase and hexokinase in pig erythrocytes. | We have shown different enzymatic activities responsible for the phosphorylation of glucose in pig erythrocytes. These activities were observed after partial purification from hemolyzed red cells. One of the enzymes involved is the hexokinase which is present in all tissues; the other is similar to hepatic glucokinase. We have determined the kinetic properties of these activities in hemolysates and in partially purified preparations. Their electrophoretic-migration characteristics were studied too. | Glucokinase and hexokinase in pig erythrocytes. We have shown different enzymatic activities responsible for the phosphorylation of glucose in pig erythrocytes. These activities were observed after partial purification from hemolyzed red cells. One of the enzymes involved is the hexokinase which is present in all tissues; the other is similar to hepatic glucokinase. We have determined the kinetic properties of these activities in hemolysates and in partially purified preparations. Their electrophoretic-migration characteristics were studied too. | [
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PMID:23947 | Electrostatic interactions at charged lipid membranes. Measurement of surface pH with fluorescent lipoid pH indicators. | The 5-dimethylaminonapthalene-1-sulfonyl (dansyl) chromophore attached to the polar head groups of lipids has been used as a fluorescent lipoid pH indicator to evaluate the interfacial pH in lipid-water lamellar systems prepared from negatively charged lipids. The pH in the vicinity of the charged lipid bilayers is different from the pH of the bulk aqueous phase and the difference is a function of the electrolyte concentration in the aqueous phase and of the lipid packing in the bilayer. At a fixed electrolyte concentration in the aqueous phase, the observed interfacial pH is 0.6 to 0.7 pH units lower above the thermal phase transition of the lipid than it is below this temperature. A quantitative interpretation of the results is given on the basis of the Gouy-Chapman theory. The results indicate that the dansyl chromophore is located in front of the charged surface and its distance from this surface increases with a decrease in lipid packing. | Electrostatic interactions at charged lipid membranes. Measurement of surface pH with fluorescent lipoid pH indicators. The 5-dimethylaminonapthalene-1-sulfonyl (dansyl) chromophore attached to the polar head groups of lipids has been used as a fluorescent lipoid pH indicator to evaluate the interfacial pH in lipid-water lamellar systems prepared from negatively charged lipids. The pH in the vicinity of the charged lipid bilayers is different from the pH of the bulk aqueous phase and the difference is a function of the electrolyte concentration in the aqueous phase and of the lipid packing in the bilayer. At a fixed electrolyte concentration in the aqueous phase, the observed interfacial pH is 0.6 to 0.7 pH units lower above the thermal phase transition of the lipid than it is below this temperature. A quantitative interpretation of the results is given on the basis of the Gouy-Chapman theory. The results indicate that the dansyl chromophore is located in front of the charged surface and its distance from this surface increases with a decrease in lipid packing. | [
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PMID:23948 | Kinetics of carbon monoxide binding to fully and partially reduced human hemoglobin valency hybrids. | The kinetics of carbon monoxide binding following fast reduction of the valency hybrids alpha2+betaCO2 and alphaCO2beta+2 by hydrated electrons have been studied at different degrees of reduction. The results show that at pH 6.0 and 7.0 reduction of one heme group yields a species which reacts fast with carbon monoxide (rate constant of the order of 10(6) M-1S-1). At pH 6.0 the intermediates alphaCO2beta2 and alpha2betaCO2 bind carbon monoxide with a rate characteristic of the T state. At pH 7.0 alphaCO2beta2 is for the greater part in the T state, while in the case of alpha2betaCO2 the R and the T state are about equally populated. | Kinetics of carbon monoxide binding to fully and partially reduced human hemoglobin valency hybrids. The kinetics of carbon monoxide binding following fast reduction of the valency hybrids alpha2+betaCO2 and alphaCO2beta+2 by hydrated electrons have been studied at different degrees of reduction. The results show that at pH 6.0 and 7.0 reduction of one heme group yields a species which reacts fast with carbon monoxide (rate constant of the order of 10(6) M-1S-1). At pH 6.0 the intermediates alphaCO2beta2 and alpha2betaCO2 bind carbon monoxide with a rate characteristic of the T state. At pH 7.0 alphaCO2beta2 is for the greater part in the T state, while in the case of alpha2betaCO2 the R and the T state are about equally populated. | [
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PMID:23949 | Mg2+ translocation across the thylakoid membrane: studies using the ionophore A 23187. | 1. The degree of the light-dependent Mg2+ efflux across the thylakoid membrane is a function of pH. There is a considerable efflux at pH 7.2 which decreases to a negligible amount at pH 8.6. This conclusion is derived from studies using the divalent cation-specific ionophore A23187. The ionophore is active as an uncoupler at pH 7.2 even in the absence of added Mg2+, and completely inactive at pH 8.6 unless Mg2+ is added. The activity was assayed as the effect on both the rate of electron transport (stimulation at pH 7.2 and inhibition at pH 8.6) and deltapH. 2. Under conditions of maximal Mg2+ efflux, pH 7.2, and in the presence of EDTA, it is shown that Mg2+ is transported across the thylakoid membrane, but it is not diluted in the medium. 3. At high pH, light-induced proton influx is not compensated by Mg2+ efflux in the presence of a relatively permeable anion. However, in the presence of an impermeable anion, where cation efflux is necessary, Mg2+ efflux occurs, indicating its preference to K+ efflux. It is suggested that although light-induced Mg2+ efflux across the thylakoid membrane is evident, its magnitude is small and it is not diluted in the stroma. Thus, it seems hard to visualize how this transport is supposed to play an important role in CO2 fixation. | Mg2+ translocation across the thylakoid membrane: studies using the ionophore A 23187. 1. The degree of the light-dependent Mg2+ efflux across the thylakoid membrane is a function of pH. There is a considerable efflux at pH 7.2 which decreases to a negligible amount at pH 8.6. This conclusion is derived from studies using the divalent cation-specific ionophore A23187. The ionophore is active as an uncoupler at pH 7.2 even in the absence of added Mg2+, and completely inactive at pH 8.6 unless Mg2+ is added. The activity was assayed as the effect on both the rate of electron transport (stimulation at pH 7.2 and inhibition at pH 8.6) and deltapH. 2. Under conditions of maximal Mg2+ efflux, pH 7.2, and in the presence of EDTA, it is shown that Mg2+ is transported across the thylakoid membrane, but it is not diluted in the medium. 3. At high pH, light-induced proton influx is not compensated by Mg2+ efflux in the presence of a relatively permeable anion. However, in the presence of an impermeable anion, where cation efflux is necessary, Mg2+ efflux occurs, indicating its preference to K+ efflux. It is suggested that although light-induced Mg2+ efflux across the thylakoid membrane is evident, its magnitude is small and it is not diluted in the stroma. Thus, it seems hard to visualize how this transport is supposed to play an important role in CO2 fixation. | [
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PMID:23950 | Therapeutic and collateral effects of 25-hydroxycholecalciferol in vitamin D deficiency. | The clinical and biochemical response to 25-hydroxycholecalciferol (25-HCC) and vitamin D3, 150 microgram/day for 20 days has been compared in infants aged 3--18 months with nutritional rickets. The infants were allocated at random to Group I (11 infants) treated with 25HCC and Group II (9 infants) treated with vitamin D3. In addition 15 matched control children without rickets were allocated to Group III and received 25-HCC 75 microgram/day for 20 days. Preliminary studies showed that plasma calcium, phosphorus, alkaline phosphatase and urine pH all differed significantly between the rachitic and control groups. The biochemical parameters in both groups of rachitic children became normal after treatment with the exception of plasma alkaline phosphatase which remained elevated. The control group showed a significant increase in plasma and urine calcium values in spite of the low dose of 25-HCC. The findings suggest that 25-HCC is as effective as vitamin D3 in the treatment of rickets but did not demonstrate any therapeutic advantage. | Therapeutic and collateral effects of 25-hydroxycholecalciferol in vitamin D deficiency. The clinical and biochemical response to 25-hydroxycholecalciferol (25-HCC) and vitamin D3, 150 microgram/day for 20 days has been compared in infants aged 3--18 months with nutritional rickets. The infants were allocated at random to Group I (11 infants) treated with 25HCC and Group II (9 infants) treated with vitamin D3. In addition 15 matched control children without rickets were allocated to Group III and received 25-HCC 75 microgram/day for 20 days. Preliminary studies showed that plasma calcium, phosphorus, alkaline phosphatase and urine pH all differed significantly between the rachitic and control groups. The biochemical parameters in both groups of rachitic children became normal after treatment with the exception of plasma alkaline phosphatase which remained elevated. The control group showed a significant increase in plasma and urine calcium values in spite of the low dose of 25-HCC. The findings suggest that 25-HCC is as effective as vitamin D3 in the treatment of rickets but did not demonstrate any therapeutic advantage. | [
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PMID:23952 | The contribution of intestinal endotoxin to mortality in hosts with compromised resistance: a review. | Sepsis, particularly with endotoxin-containing Gram-negative bacilli, is a serious complication in hosts whose defenses are compromised. This review examines work from our laboratory and others concerning infectious processes which may be critical to the survival of compromised individuals. Several avenues for control of sepsis are proposed. Gram-negative bacteria and their endotoxins can escape from the intestines of compromised animals to contaminate normally sterile host tissues. Endotoxins are especially toxic to compromised hosts because essential components of their inflammatory responses are missing (i.e., leukocytes and platelets in irradiated animals). Therefore, regulation of host responses to endotoxin is no longer possible. It is recommended that sepsis be controlled in compromised individuals through elimination of endogenous microbial agents. Should infection occur in these individuals, they should be transfused with blood cells necessary for clearance of bacteria and endotoxin and restoration of homeostasis. | The contribution of intestinal endotoxin to mortality in hosts with compromised resistance: a review. Sepsis, particularly with endotoxin-containing Gram-negative bacilli, is a serious complication in hosts whose defenses are compromised. This review examines work from our laboratory and others concerning infectious processes which may be critical to the survival of compromised individuals. Several avenues for control of sepsis are proposed. Gram-negative bacteria and their endotoxins can escape from the intestines of compromised animals to contaminate normally sterile host tissues. Endotoxins are especially toxic to compromised hosts because essential components of their inflammatory responses are missing (i.e., leukocytes and platelets in irradiated animals). Therefore, regulation of host responses to endotoxin is no longer possible. It is recommended that sepsis be controlled in compromised individuals through elimination of endogenous microbial agents. Should infection occur in these individuals, they should be transfused with blood cells necessary for clearance of bacteria and endotoxin and restoration of homeostasis. | [
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PMID:23956 | Simultaneous monitoring by optical techniques of respiratory chain and intracellular pH in toad ventricle strip. | Intracellular pH and oxidative metabolism can be measured in toad ventricle strips simultaneously by the use of the pH indicator dye, neutral red, and a rapid scanning spectrophotometer. The effects of hypoxia and acidification on mechanical function are approximately additive. The decrease in tension due to slight acidification is probably through an effect on the portion of the twitch tension supported by anaerobic metabolism. | Simultaneous monitoring by optical techniques of respiratory chain and intracellular pH in toad ventricle strip. Intracellular pH and oxidative metabolism can be measured in toad ventricle strips simultaneously by the use of the pH indicator dye, neutral red, and a rapid scanning spectrophotometer. The effects of hypoxia and acidification on mechanical function are approximately additive. The decrease in tension due to slight acidification is probably through an effect on the portion of the twitch tension supported by anaerobic metabolism. | [
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PMID:23957 | [Nature of the melipramine inhibition of the K+-n-nitrophenyl phosphatase activity of a synaptosomal fraction]. | The influence of antidepressant melipramine (imipramine) on the K+-paranitrophenyl-phosphatase (K+-n-NFP-ase) activity of the synaptosomal fraction in the bull's brain cortex, treated with sodium iodide, was studied in in vitro experiments. Melipramine (5.10(-5)--10(-3)M) is shown to inhibit the K+-n-NFP-ase activity, competing with potassium ions. With pH changing from acid and neutral to alkaline values the inhibitory action of melipramine gains in intensity, whereas the protective effect of potassium ions declines. It is suggested that one of the mechanisms responsible for the inhibition of the K+-p-NFP-ase activity by melipramine is interaction of the inhibitor in the anionic sections of the enzyme complex. | [Nature of the melipramine inhibition of the K+-n-nitrophenyl phosphatase activity of a synaptosomal fraction]. The influence of antidepressant melipramine (imipramine) on the K+-paranitrophenyl-phosphatase (K+-n-NFP-ase) activity of the synaptosomal fraction in the bull's brain cortex, treated with sodium iodide, was studied in in vitro experiments. Melipramine (5.10(-5)--10(-3)M) is shown to inhibit the K+-n-NFP-ase activity, competing with potassium ions. With pH changing from acid and neutral to alkaline values the inhibitory action of melipramine gains in intensity, whereas the protective effect of potassium ions declines. It is suggested that one of the mechanisms responsible for the inhibition of the K+-p-NFP-ase activity by melipramine is interaction of the inhibitor in the anionic sections of the enzyme complex. | [
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PMID:23958 | [Effect of pharmacological substances on the development of hemorrhagic erosions and on the noradrenaline level in the stomach wall in rats]. | Hemorrhagic erosions of the gastric mucosa were produced through electric stimulation of immobilized rats via electrodes stuck into the muscles of the fore-paws. In the experiment there was applied square pulsed current of 5--7 v (per 10 rats) with frequency of 50 Hz and the pulse time of 10 ms. Prior to stimulation the rats received intraperitoneally one of the following drugs: clonidin (0.1 mg/kg), benactyzine (3 mg/kg), etherophen (20 mg/kg), benzohexonium (10 mg/kg), iprazid (100 mg/kg), amphethamine (4 mg/kg), atropine (1 mg/kg), tyrosine (300 mg/kg), aethimizol (10 mg/kg) and orotic acid (10 mg/kg). The animals were sacrificed directly after stimulation and the norepinephrine level in the gastric wall was determined after E. Sh. Matlina and T. B. Rakhmanova (1967). Introduction of clonidin, benacyzine, etherophen, benzohexonium and iprazid to immobilized rats after electric stimulation prevented the development of hemorrhagic erosions and a drop of the norepinephrine level in the gastric wall, whereas amphethamine and tyrosine intensified the process of the erosions development in the gastric mucosa and did not prevent a decline in the norepinephrine level. Atropine and aethimizol had no effect on the findings under study. | [Effect of pharmacological substances on the development of hemorrhagic erosions and on the noradrenaline level in the stomach wall in rats]. Hemorrhagic erosions of the gastric mucosa were produced through electric stimulation of immobilized rats via electrodes stuck into the muscles of the fore-paws. In the experiment there was applied square pulsed current of 5--7 v (per 10 rats) with frequency of 50 Hz and the pulse time of 10 ms. Prior to stimulation the rats received intraperitoneally one of the following drugs: clonidin (0.1 mg/kg), benactyzine (3 mg/kg), etherophen (20 mg/kg), benzohexonium (10 mg/kg), iprazid (100 mg/kg), amphethamine (4 mg/kg), atropine (1 mg/kg), tyrosine (300 mg/kg), aethimizol (10 mg/kg) and orotic acid (10 mg/kg). The animals were sacrificed directly after stimulation and the norepinephrine level in the gastric wall was determined after E. Sh. Matlina and T. B. Rakhmanova (1967). Introduction of clonidin, benacyzine, etherophen, benzohexonium and iprazid to immobilized rats after electric stimulation prevented the development of hemorrhagic erosions and a drop of the norepinephrine level in the gastric wall, whereas amphethamine and tyrosine intensified the process of the erosions development in the gastric mucosa and did not prevent a decline in the norepinephrine level. Atropine and aethimizol had no effect on the findings under study. | [
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PMID:23959 | [Rat placental barrier permeability for the new antihistaminic preparation, fenkarol]. | As established, following a single introduction of phencarol tagged with tritium in a quinuclidine nucleus perorally to rats on the 13th day of gestation its insignificant amounts gain access to the fetus through the placenta. The maximum concentration of the drug in the maternal blood was recorded 3 hours after its introduction, following 6 hours--in the placenta, with the radioactivity continuing to be constant during the first 6 hours in the embryonal tissues. In 48 hours after introduction the radioactive tag is still demonstrable in the study tissues. It is concluded that the drug is largely retained in the placenta, when introduced on the 13th day of pregnancy; on the 21st day of gestation the barrier functions of the placenta declines. The presence of phencarol and its transformation products in the embryonal tissues is confirmed by the method of thin-layer chromotography. | [Rat placental barrier permeability for the new antihistaminic preparation, fenkarol]. As established, following a single introduction of phencarol tagged with tritium in a quinuclidine nucleus perorally to rats on the 13th day of gestation its insignificant amounts gain access to the fetus through the placenta. The maximum concentration of the drug in the maternal blood was recorded 3 hours after its introduction, following 6 hours--in the placenta, with the radioactivity continuing to be constant during the first 6 hours in the embryonal tissues. In 48 hours after introduction the radioactive tag is still demonstrable in the study tissues. It is concluded that the drug is largely retained in the placenta, when introduced on the 13th day of pregnancy; on the 21st day of gestation the barrier functions of the placenta declines. The presence of phencarol and its transformation products in the embryonal tissues is confirmed by the method of thin-layer chromotography. | [
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PMID:23960 | [Effect of the structure of liposomal membrane surface layer on the binding of phenothiazines]. | By using a fluorescent 3-metoxybenzatrone probe the binding of the phenothiazine series drugs (chlorpromazine, triftazine, aethaperazine) with phospholipid vesicles--liposomes, was investigated. Cholesterol is shown not to affect the binding of these drugs with liposomes. The surface charge of liposomes influences only the combination with the chlorpromazine membranes. With a higher positive charge of the membranes the binding of chlorpromazine diminishes in force. | [Effect of the structure of liposomal membrane surface layer on the binding of phenothiazines]. By using a fluorescent 3-metoxybenzatrone probe the binding of the phenothiazine series drugs (chlorpromazine, triftazine, aethaperazine) with phospholipid vesicles--liposomes, was investigated. Cholesterol is shown not to affect the binding of these drugs with liposomes. The surface charge of liposomes influences only the combination with the chlorpromazine membranes. With a higher positive charge of the membranes the binding of chlorpromazine diminishes in force. | [
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PMID:23965 | Acetylcholine--inhibition of transmitter release from adrenergic nerve terminals mediated by muscarinic receptors. | The evidence is reviewed for the presence of muscarinic receptors on the sympathetic nerves to blood vessels. Activation of these receptors by acetylcholine in doses that are too small to affect the smooth muscle cells directly inhibits the release of norepinephrine evoked by electric impulses or potassium ions. This inhibitory action of acetylcholine is prevented by muscarinic blocking agents and is probably due to hyperpolarization of the adrenergic nerve terminals. | Acetylcholine--inhibition of transmitter release from adrenergic nerve terminals mediated by muscarinic receptors. The evidence is reviewed for the presence of muscarinic receptors on the sympathetic nerves to blood vessels. Activation of these receptors by acetylcholine in doses that are too small to affect the smooth muscle cells directly inhibits the release of norepinephrine evoked by electric impulses or potassium ions. This inhibitory action of acetylcholine is prevented by muscarinic blocking agents and is probably due to hyperpolarization of the adrenergic nerve terminals. | [
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PMID:23966 | Histamine and 5-hydroxytryptamine-inhibition of transmitter release mediated by H2- and 5-hydroxytryptamine receptors. | The vasodilatation caused by histamine and 5-hydroxytryptamine may be due, at least in part, to their inhibitory action on adrenergic neurotransmission. The evidence for this is as follows: 1) contractions of isolated canine vascular strips caused by sympathetic nerve stimulation are depressed by these substances whereas contractions caused by norepinephrine are either unchanged or augmented; 2) histamine and 5-hydroxytryptamine inhibit the release of norepinephrine evoked by sympathetic nerve stimulation of isolated vascular strips previously incubated with the labeled transmitter. This inhibitory effect can be demonstrated using concentrations of the substinces less than those required to affect the smooth muscle cells directly. By contrast, neither histamine nor 5-hydroxytryptamine inhibits the displacement of neuronal norepinephrine by tyramine, suggesting that these substances interfere with the exocytotic process. Additional studies have identified the histamine-H2 receptor as the probable mediator of this prejunctional action of histamine, whereas the nature of the receptor for 5-hydroxytryptamine remains to be clarified. | Histamine and 5-hydroxytryptamine-inhibition of transmitter release mediated by H2- and 5-hydroxytryptamine receptors. The vasodilatation caused by histamine and 5-hydroxytryptamine may be due, at least in part, to their inhibitory action on adrenergic neurotransmission. The evidence for this is as follows: 1) contractions of isolated canine vascular strips caused by sympathetic nerve stimulation are depressed by these substances whereas contractions caused by norepinephrine are either unchanged or augmented; 2) histamine and 5-hydroxytryptamine inhibit the release of norepinephrine evoked by sympathetic nerve stimulation of isolated vascular strips previously incubated with the labeled transmitter. This inhibitory effect can be demonstrated using concentrations of the substinces less than those required to affect the smooth muscle cells directly. By contrast, neither histamine nor 5-hydroxytryptamine inhibits the displacement of neuronal norepinephrine by tyramine, suggesting that these substances interfere with the exocytotic process. Additional studies have identified the histamine-H2 receptor as the probable mediator of this prejunctional action of histamine, whereas the nature of the receptor for 5-hydroxytryptamine remains to be clarified. | [
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PMID:23967 | Actions of angiotensin on adrenergic nerve endings. | In the perfused vascular bed, vasoconstrictor responses to adrenergic nerve stimulation are augmented to a greater degree by angiotensin II than are the responses to injected norepinephrine. Overflow of adrenergic transmitter is also greater during nerve stimulation in the presence of angiotensin than in its absence. The evidence indicates that facilitation of adrenergic transmitter release rather than uptake blockade accounts for these results. In addition, an increased responsiveness of isolated arterial strips to norepinephrine as well as other agonists appears to contribute to the adrenergic potentiating effect of angiotensin II as well as angiotensin III. This action, which appears to be a cell membrane effect, seems to participate in adrenergic potentiation mainly in the arterial segment of the intact vascular bed. Both of these effects of angiotensin, i.e., facilitation of release and increased smooth muscle responsiveness, appear to be mediated by angiotensin receptors. | Actions of angiotensin on adrenergic nerve endings. In the perfused vascular bed, vasoconstrictor responses to adrenergic nerve stimulation are augmented to a greater degree by angiotensin II than are the responses to injected norepinephrine. Overflow of adrenergic transmitter is also greater during nerve stimulation in the presence of angiotensin than in its absence. The evidence indicates that facilitation of adrenergic transmitter release rather than uptake blockade accounts for these results. In addition, an increased responsiveness of isolated arterial strips to norepinephrine as well as other agonists appears to contribute to the adrenergic potentiating effect of angiotensin II as well as angiotensin III. This action, which appears to be a cell membrane effect, seems to participate in adrenergic potentiation mainly in the arterial segment of the intact vascular bed. Both of these effects of angiotensin, i.e., facilitation of release and increased smooth muscle responsiveness, appear to be mediated by angiotensin receptors. | [
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PMID:23968 | Prostaglandins--modulation of adrenergic nervous system. | Prostaglandins (PGs) affect vascular tone by a direct action on the vascular smooth muscle and by influencing vascular reactivity to adrenergic simuli and several vasoactive substances. Thus, in the isolated Tyrode's perfused rabbit renal, mesenteric and splenic vasculature PGE2 inhibited adrenergically induced vasoconstriction. Since the vasoconstrictor responses to renal nerve stimulation were enhanced by the blockade of PG synthesis and were reduced by stimulation of PG synthesis with arachidonic acid, this suggests that PGE2 functions as an inhibitory modulator of the adrenergic nervous system. However, our demonstration that PGE2 enhanced adrenergically induced vasoconstriction in the renal and mesenteric vasculature of the rat, but had opposite effects in the rat splenic vasculature indicates that the modulatory-effect of PGE-compounds on the adrenergic neuromuscular junction is species dependent and varies in different vascular beds within the same species. Prostaglandins, the release of which is evoked by several vasoactive substances including angiotensins, kinins, and adenine nucleotides, may also contribute to the regulation of vascular tone by either opposing or amplifying the vascular actions of vasoactive substances. | Prostaglandins--modulation of adrenergic nervous system. Prostaglandins (PGs) affect vascular tone by a direct action on the vascular smooth muscle and by influencing vascular reactivity to adrenergic simuli and several vasoactive substances. Thus, in the isolated Tyrode's perfused rabbit renal, mesenteric and splenic vasculature PGE2 inhibited adrenergically induced vasoconstriction. Since the vasoconstrictor responses to renal nerve stimulation were enhanced by the blockade of PG synthesis and were reduced by stimulation of PG synthesis with arachidonic acid, this suggests that PGE2 functions as an inhibitory modulator of the adrenergic nervous system. However, our demonstration that PGE2 enhanced adrenergically induced vasoconstriction in the renal and mesenteric vasculature of the rat, but had opposite effects in the rat splenic vasculature indicates that the modulatory-effect of PGE-compounds on the adrenergic neuromuscular junction is species dependent and varies in different vascular beds within the same species. Prostaglandins, the release of which is evoked by several vasoactive substances including angiotensins, kinins, and adenine nucleotides, may also contribute to the regulation of vascular tone by either opposing or amplifying the vascular actions of vasoactive substances. | [
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PMID:23969 | Metabolic modulation of neurotransmitter release--adenosine, adenine nucleotides, potassium, hyperosmolarity, and hydrogen ion. | Evidence has accumulated that several factors, which have been proposed as mediators of exercise hyperemia, can modulate adrenergic neurotransmission in blood vessels. Adenosine and the adenine nucleotides depress the response of isolated blood vessels of the dog to nerve stimulation more than that to exogenous norepinephrine; this difference is explained by a decreased release of the neurotransmitter. Potassium, hyperosmolarity, and acidosis also depress adrenergic neurotransmission in isolated veins. These results are consistent with the hypothesis that metabolic changes in the vicinity of the adrenergic neuroeffector junction are capable of decreasing the output of neurotransmitter to the blood vessels in the exercising muscle. | Metabolic modulation of neurotransmitter release--adenosine, adenine nucleotides, potassium, hyperosmolarity, and hydrogen ion. Evidence has accumulated that several factors, which have been proposed as mediators of exercise hyperemia, can modulate adrenergic neurotransmission in blood vessels. Adenosine and the adenine nucleotides depress the response of isolated blood vessels of the dog to nerve stimulation more than that to exogenous norepinephrine; this difference is explained by a decreased release of the neurotransmitter. Potassium, hyperosmolarity, and acidosis also depress adrenergic neurotransmission in isolated veins. These results are consistent with the hypothesis that metabolic changes in the vicinity of the adrenergic neuroeffector junction are capable of decreasing the output of neurotransmitter to the blood vessels in the exercising muscle. | [
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PMID:23973 | Oxypertine in combination with imipramine: a controlled trial. | A method for the controlled assessment of two agents for possible interaction in the treatment of endogenous depression is described. This consisted of dose-ranging the supplementary agent (oxypertine 30 mg and 60 mg daily or matching placebo) during Week 2 to Week 6 having established all patients on a therapeutic dose of imipramine during Week 1. The reported enhancement of imipramine by oxypertine was not confirmed. | Oxypertine in combination with imipramine: a controlled trial. A method for the controlled assessment of two agents for possible interaction in the treatment of endogenous depression is described. This consisted of dose-ranging the supplementary agent (oxypertine 30 mg and 60 mg daily or matching placebo) during Week 2 to Week 6 having established all patients on a therapeutic dose of imipramine during Week 1. The reported enhancement of imipramine by oxypertine was not confirmed. | [
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PMID:23974 | Studies of the androgen binding protein in the rete testis fluid of the ram and its relation to sexual season. | An androgen binding protein (ABP) with an electrophoretic mobility (Rf) of 0.56 is present in the rete testis fluid of adult rams. Its steroid specificity was found to be in the following order: 5alpha-DHT, testosterone, oestradiol-17 beta, dehydroepiandrosterone 5beta-DHT, androstenedione, cyproterone, cyproterone acetate, cortisol and progesterone. The characteristics of the ABP are similar to those found for the ABP of the testis and the epididymis of the rat and the rabbit. The concentration of ABP, determined by the dextran-coated charcoal method and sometimes confirmed by the steady-state polyacrylamide gel electrophoresis method, was significantly higher in the breeding season than in the non-breeding season (4.40 +/- 0.98 X 10(-9) M vs. 2.60 +/- 0.62 X 10(-9) M; P less than 0.037). The affinity constant of the ABP was independent of the season (2.45 +/- 0.21 X 10(9) M-1 vs. 2.66 +/- 0.1 X 10(9) M-1; NS). In addition, ABP was positively correlated with 5alpha-DHT (r = 0.506; P less than 0.0009), testosterone (r = 0.445; P less than 0.0003), total protein (r = 0.329; P less than 0.02) and spermatozoa (r = 0.406; P less than 0.006) in the RTF and with blood plasma testosterone (r = 0.584; P less than 0.0001). Furthermore, testosterone and 5alpha-DHT in RTF were positively correlated (r = 0.582; P less than 0.0001). These androgens were also correlated with plasma testosterone (r = 0.262, P less than 0.052 for testosterone in RTF; r = 0.341, P less than 0.018 for 5 alpha-DHT). Total proteins and spermatozoa were found to be positively correlated in the RTF (r = 0.789; P less than 0.0001). | Studies of the androgen binding protein in the rete testis fluid of the ram and its relation to sexual season. An androgen binding protein (ABP) with an electrophoretic mobility (Rf) of 0.56 is present in the rete testis fluid of adult rams. Its steroid specificity was found to be in the following order: 5alpha-DHT, testosterone, oestradiol-17 beta, dehydroepiandrosterone 5beta-DHT, androstenedione, cyproterone, cyproterone acetate, cortisol and progesterone. The characteristics of the ABP are similar to those found for the ABP of the testis and the epididymis of the rat and the rabbit. The concentration of ABP, determined by the dextran-coated charcoal method and sometimes confirmed by the steady-state polyacrylamide gel electrophoresis method, was significantly higher in the breeding season than in the non-breeding season (4.40 +/- 0.98 X 10(-9) M vs. 2.60 +/- 0.62 X 10(-9) M; P less than 0.037). The affinity constant of the ABP was independent of the season (2.45 +/- 0.21 X 10(9) M-1 vs. 2.66 +/- 0.1 X 10(9) M-1; NS). In addition, ABP was positively correlated with 5alpha-DHT (r = 0.506; P less than 0.0009), testosterone (r = 0.445; P less than 0.0003), total protein (r = 0.329; P less than 0.02) and spermatozoa (r = 0.406; P less than 0.006) in the RTF and with blood plasma testosterone (r = 0.584; P less than 0.0001). Furthermore, testosterone and 5alpha-DHT in RTF were positively correlated (r = 0.582; P less than 0.0001). These androgens were also correlated with plasma testosterone (r = 0.262, P less than 0.052 for testosterone in RTF; r = 0.341, P less than 0.018 for 5 alpha-DHT). Total proteins and spermatozoa were found to be positively correlated in the RTF (r = 0.789; P less than 0.0001). | [
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PMID:23976 | Animal influenza virus neuraminidase: studies on dependence of some of its properties on its association with hemagglutinin. | Neuraminidase (Nase) thermostability and sensitivity to pH treatment as well as specific enzymatic activity (Nase activity per 1 HA unit) were determined in two groups of animal influenza virus strains containing equine 1 and equine 2 Nase subtypes, respectively (A/equine/Prague/56 (Heq1 Neq1), A/equine/Cambridge/63 (Heq1 Neq1), A/FPV/Dutch/34 (Hav1 Neq1), A/chicken/Germany "N" (Hav2 Neq1), in one group, and A/equine/Miami/1/63 (Heq2 Neq2), A/turkey/Canada/63 (Hav6 Neq2), A/duck/Ukraine/1/63 (Hav7 Neq2), in the other group). Nase of all the strains used was thermotabile when heated at pH 4.5. Nase of Neq1 subtype irrespective of strain containing it was thermolabilt when heated both at pH 6.5 and 8.1 and sensitive to pH 4.5 treatment as such (without heating). Inversely, Nase of Neq2 antigenic subtype irrespective of the strain containing it, was thermostable when heated at pH 6.5 AND 8.1 and resistant to the treatment of pH 4.5. Specific enzymatic activity was considerably higher in all the strains containing Neq2 as compared to Neq1-containing strains (4-6 times as much). The results suggest that thermostability and pH sensitivity of equine Nases of both antigenic subtypes, as well as their specific activities, do not depend on the sort of HA which is coupled with enzyme subunits at viral envelope, but attributed rather to properties of the subunits themselves, such as glycoprotein entities. The data concerning specific activities may suggest that in the case of various combinations of Nase subunits with different HA subunits the amount of enzyme per virion is of the same order. | Animal influenza virus neuraminidase: studies on dependence of some of its properties on its association with hemagglutinin. Neuraminidase (Nase) thermostability and sensitivity to pH treatment as well as specific enzymatic activity (Nase activity per 1 HA unit) were determined in two groups of animal influenza virus strains containing equine 1 and equine 2 Nase subtypes, respectively (A/equine/Prague/56 (Heq1 Neq1), A/equine/Cambridge/63 (Heq1 Neq1), A/FPV/Dutch/34 (Hav1 Neq1), A/chicken/Germany "N" (Hav2 Neq1), in one group, and A/equine/Miami/1/63 (Heq2 Neq2), A/turkey/Canada/63 (Hav6 Neq2), A/duck/Ukraine/1/63 (Hav7 Neq2), in the other group). Nase of all the strains used was thermotabile when heated at pH 4.5. Nase of Neq1 subtype irrespective of strain containing it was thermolabilt when heated both at pH 6.5 and 8.1 and sensitive to pH 4.5 treatment as such (without heating). Inversely, Nase of Neq2 antigenic subtype irrespective of the strain containing it, was thermostable when heated at pH 6.5 AND 8.1 and resistant to the treatment of pH 4.5. Specific enzymatic activity was considerably higher in all the strains containing Neq2 as compared to Neq1-containing strains (4-6 times as much). The results suggest that thermostability and pH sensitivity of equine Nases of both antigenic subtypes, as well as their specific activities, do not depend on the sort of HA which is coupled with enzyme subunits at viral envelope, but attributed rather to properties of the subunits themselves, such as glycoprotein entities. The data concerning specific activities may suggest that in the case of various combinations of Nase subunits with different HA subunits the amount of enzyme per virion is of the same order. | [
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PMID:23982 | Assessment of the plasma disappearance of cholyl'l14C-glycine as a test of hepatocellular disease. | The plasma disappearance of a tracer dose of cholyl-l14C-glycine has been examined in 12 control subjects and in 32 patients with hepatocellular dysfunction. Simple analysis of the data did not detect hepatic dysfunction except in severe hepatocellular disease. The greatest degree of discrimination between normal subjects and patients with mild liver disease was obtained by taking the ratio of the plasma retention at 60 minutes to that at 10 minutes; it was similar to that obtained with serum gamma-glutamyl transferase. The two hour post-prandial plasma "total" bile acid concentration gave complete separation between the control subjects and patients with liver disease. | Assessment of the plasma disappearance of cholyl'l14C-glycine as a test of hepatocellular disease. The plasma disappearance of a tracer dose of cholyl-l14C-glycine has been examined in 12 control subjects and in 32 patients with hepatocellular dysfunction. Simple analysis of the data did not detect hepatic dysfunction except in severe hepatocellular disease. The greatest degree of discrimination between normal subjects and patients with mild liver disease was obtained by taking the ratio of the plasma retention at 60 minutes to that at 10 minutes; it was similar to that obtained with serum gamma-glutamyl transferase. The two hour post-prandial plasma "total" bile acid concentration gave complete separation between the control subjects and patients with liver disease. | [
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PMID:23983 | Postprandial gastro-oesophageal reflux in healthy people. | Distal oesophageal pH was monitored for three hours after a standard meal in 10 young healthy subjects without symptoms of gastro-oesophageal reflux. Episodes of reflux occurred in nine of these subjects; and, in five, oesophageal pH was less than 5 for between 11 and 75% of the first postprandial hour. Intermittent incompetence of the lower oesophageal sphincter after food must, therefore, be regarded as a normal phenomenon. The method described would be suitable for the evaluation of agents believed to weaken or to strengthen the lower oesophageal sphincter. | Postprandial gastro-oesophageal reflux in healthy people. Distal oesophageal pH was monitored for three hours after a standard meal in 10 young healthy subjects without symptoms of gastro-oesophageal reflux. Episodes of reflux occurred in nine of these subjects; and, in five, oesophageal pH was less than 5 for between 11 and 75% of the first postprandial hour. Intermittent incompetence of the lower oesophageal sphincter after food must, therefore, be regarded as a normal phenomenon. The method described would be suitable for the evaluation of agents believed to weaken or to strengthen the lower oesophageal sphincter. | [
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PMID:23984 | Transport kinetics of 6-deoxy-D-glucose in Candida parapsilosis. | The strictly aerobic yeast Candida parapsilosis transports the nonmetabolizable monosaccharide 6-deoxy-D-glucose by an active process (inhibition by 2.4-dinitrophenol and other uncouplers but not by iodoacetamide), the accumulation ratio decreasing with increasing substrate concentration. Measured accumulation ratios are in agreement with those predicted from kinetic constants for influx and efflux. Energy for transport is probably required in the translocation step. The maximum rate is temperature-dependent with a transition point at 21 degrees C. the accumulation ratio is not. The uptake is most active at pH 4.5--8.5. It appears not to involve stoichiometric proton symport. The transport system is shared by D-glucose, D-mannose, D-galactose and possibly maltose but not by fructose, sucrose or pentoses. The apparent half-life of the transport system was 3.5--4 h. | Transport kinetics of 6-deoxy-D-glucose in Candida parapsilosis. The strictly aerobic yeast Candida parapsilosis transports the nonmetabolizable monosaccharide 6-deoxy-D-glucose by an active process (inhibition by 2.4-dinitrophenol and other uncouplers but not by iodoacetamide), the accumulation ratio decreasing with increasing substrate concentration. Measured accumulation ratios are in agreement with those predicted from kinetic constants for influx and efflux. Energy for transport is probably required in the translocation step. The maximum rate is temperature-dependent with a transition point at 21 degrees C. the accumulation ratio is not. The uptake is most active at pH 4.5--8.5. It appears not to involve stoichiometric proton symport. The transport system is shared by D-glucose, D-mannose, D-galactose and possibly maltose but not by fructose, sucrose or pentoses. The apparent half-life of the transport system was 3.5--4 h. | [
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PMID:23985 | Production of amylase by a submerged culture of Aspergillus wentii. | Soluble starch was hydrolysed to maltose by Aspergillus wentii Wehmer (IMI 17295). Studies on nutritional requirements of Aspergillus wentii for production of amylase revealed that the optimum conditions were achieved in fermentation culture medium containing 1% starch, and incubated at 20 degrees C for 3 days at pH 6.0. Tryptophan was the best nitrogen source. The amylase activity was completely inhibited when 1 mM sodium iodoacetate was incorporated into the medium. With 10 mM sodium citrate the amylase activity was increased from 3.51 to 6.0 mg/ml. | Production of amylase by a submerged culture of Aspergillus wentii. Soluble starch was hydrolysed to maltose by Aspergillus wentii Wehmer (IMI 17295). Studies on nutritional requirements of Aspergillus wentii for production of amylase revealed that the optimum conditions were achieved in fermentation culture medium containing 1% starch, and incubated at 20 degrees C for 3 days at pH 6.0. Tryptophan was the best nitrogen source. The amylase activity was completely inhibited when 1 mM sodium iodoacetate was incorporated into the medium. With 10 mM sodium citrate the amylase activity was increased from 3.51 to 6.0 mg/ml. | [
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PMID:23988 | Parallel occurrence of oxidant-sensitivity and decreased inhibition by NADPH in G-6-PD Lublin and G-6-PD Poxnań. | In two studied variants of G-6-PD without chronic hemolysis in probands, sensitivity of enzymes to inhibition by NADPH was decreased. Ki for NADPH was 28 micronM in Gd Lublin and 19 micronM in Gd Poznań. Susceptibility to the oxidant-induced hemolysis was described in probands, as well as in patients hemizygous for two other variants of G-6-PD with increased Ki for NADPH. It is suggested that in these cases, the oxidant-induced hemolysis is aggravated by their inability to counteract the drop in NADPH concentration with an increase in G-6-PD activity. | Parallel occurrence of oxidant-sensitivity and decreased inhibition by NADPH in G-6-PD Lublin and G-6-PD Poxnań. In two studied variants of G-6-PD without chronic hemolysis in probands, sensitivity of enzymes to inhibition by NADPH was decreased. Ki for NADPH was 28 micronM in Gd Lublin and 19 micronM in Gd Poznań. Susceptibility to the oxidant-induced hemolysis was described in probands, as well as in patients hemizygous for two other variants of G-6-PD with increased Ki for NADPH. It is suggested that in these cases, the oxidant-induced hemolysis is aggravated by their inability to counteract the drop in NADPH concentration with an increase in G-6-PD activity. | [
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PMID:23989 | Studies of cyclic AMP action using mutant tissue culture cells. | S49 mouse lymphoma cell mutants, each with a specific defect in its ability to generate or respond to cyclic AMP, have been isolated. Analysis of the properties of these cells has begun to provide information on complex and significant biologic problems related to the cyclic AMP system. | Studies of cyclic AMP action using mutant tissue culture cells. S49 mouse lymphoma cell mutants, each with a specific defect in its ability to generate or respond to cyclic AMP, have been isolated. Analysis of the properties of these cells has begun to provide information on complex and significant biologic problems related to the cyclic AMP system. | [
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PMID:23990 | [Correlation of growth-phase and streptodornase-production of beta-hemolytic streptococci group A (author's transl)]. | 77 M- and 48 T-typed strains of A-streptococci were compared with respect to their extinction, acidification and nuclease-production as a function of the growth-time. Strains of the same serotype (M 12) showed a change in the distribution of the isoenzymes of nucleases with respect to the growth-time. Half of the strains produced nuclease activity after 4 h. Streptodornase B was produced by all strains. With the growth the number of the nucleases found at the same time diminished, after 96 h in 70% of the strains the nuclease activity was represented by streptodornase B only. | [Correlation of growth-phase and streptodornase-production of beta-hemolytic streptococci group A (author's transl)]. 77 M- and 48 T-typed strains of A-streptococci were compared with respect to their extinction, acidification and nuclease-production as a function of the growth-time. Strains of the same serotype (M 12) showed a change in the distribution of the isoenzymes of nucleases with respect to the growth-time. Half of the strains produced nuclease activity after 4 h. Streptodornase B was produced by all strains. With the growth the number of the nucleases found at the same time diminished, after 96 h in 70% of the strains the nuclease activity was represented by streptodornase B only. | [
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PMID:23994 | Depression of cell-mediated immunity following inoculation of Trichinella spiralis extract in the mouse. | Mice pretreated with Trichinella spiralis extract (TsE), or infected with the parasite, rejected primary skin allografts in 18-23 days and secondary allografts in 12-16 days. Mice pretreated with saline or with bovine serum albumin (BSA) rejected the primary allografts in 12-18 days and did not accept the secondary grafts. Inoculation of increasing doses of parental spleen cells from mice pretreated with saline or with BSA in F1 hybrids produced proportionately stronger graft-versus-host reactions (GvHR) whereas increasing doses of cells from TsE pretreated mice reduced proportionately the capacity of the inoculum to induce a GvHR. Immunodepression of the parental cells was obtained with 7 and with 4, but not with 2, daily injections of TsE. The depression waned rapidly after the treatment with TsE but a significant degree still remained after 3 days. Immunodepression by TsE cannot be solely explained by antigenic competition and although our results are consistent with the induction of suppressor cells, it is probable that other mechanisms are also involved. | Depression of cell-mediated immunity following inoculation of Trichinella spiralis extract in the mouse. Mice pretreated with Trichinella spiralis extract (TsE), or infected with the parasite, rejected primary skin allografts in 18-23 days and secondary allografts in 12-16 days. Mice pretreated with saline or with bovine serum albumin (BSA) rejected the primary allografts in 12-18 days and did not accept the secondary grafts. Inoculation of increasing doses of parental spleen cells from mice pretreated with saline or with BSA in F1 hybrids produced proportionately stronger graft-versus-host reactions (GvHR) whereas increasing doses of cells from TsE pretreated mice reduced proportionately the capacity of the inoculum to induce a GvHR. Immunodepression of the parental cells was obtained with 7 and with 4, but not with 2, daily injections of TsE. The depression waned rapidly after the treatment with TsE but a significant degree still remained after 3 days. Immunodepression by TsE cannot be solely explained by antigenic competition and although our results are consistent with the induction of suppressor cells, it is probable that other mechanisms are also involved. | [
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PMID:23995 | Responses to polyvinyl pyrrolidone and pneumococcal polysaccharide in protein-deficient mice. | Indirect haemagglutination titres induced by polyvinyl pyrrolidone (PVP) or pneumococcal polysaccharide Type III (S111) were determined in mice maintained on a 4% albumin diet from weaning and normally-fed littermates. Responses to PVP, given intravenously (i.v.) or intraperitoneally (i.p.), were elevated by protein-deficiency at low antigen doses and increasingly depressed at high doses. Increases in the duration of protein-deficiency generally improved these responses. The persistence of tolerance was reduced by protein-deficiency and priming was evident in both groups when tolerance was broken. The low protein diet depressed responses to moderate doses of S111 given i.p. to C57Bl mice, but such responses were normal in BALB/c mice and in C57Bl mice injected i.v. High doses of S111 (i.p., i.v.) elicited poor responses in deficient mice. These findings are discussed in relation to previous studies using other antigens, with a view to elucidating mechanisms responsible for the effects of protein-deficiency. | Responses to polyvinyl pyrrolidone and pneumococcal polysaccharide in protein-deficient mice. Indirect haemagglutination titres induced by polyvinyl pyrrolidone (PVP) or pneumococcal polysaccharide Type III (S111) were determined in mice maintained on a 4% albumin diet from weaning and normally-fed littermates. Responses to PVP, given intravenously (i.v.) or intraperitoneally (i.p.), were elevated by protein-deficiency at low antigen doses and increasingly depressed at high doses. Increases in the duration of protein-deficiency generally improved these responses. The persistence of tolerance was reduced by protein-deficiency and priming was evident in both groups when tolerance was broken. The low protein diet depressed responses to moderate doses of S111 given i.p. to C57Bl mice, but such responses were normal in BALB/c mice and in C57Bl mice injected i.v. High doses of S111 (i.p., i.v.) elicited poor responses in deficient mice. These findings are discussed in relation to previous studies using other antigens, with a view to elucidating mechanisms responsible for the effects of protein-deficiency. | [
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PMID:23996 | Restoration of suppressor function in athymic mice. | Antibody responses of normal and congenitally athymic (nu/nu) mice were measured to the phosphorylcholine determinant of a pneumococcal C-polysaccharide. The plaque-forming cell responses of athymic mice were approximately tenfold higher than for intact controls. This enhancement was equivalent to that obtained by treatment of controls with antithymocyte serum. In both cases, the enhanced responses presumably result from a lack of suppressor T cells. Reconstitution of athymic mice with normal thymocytes restored both suppressor and helper T cell functions. Whereas suppressor activity did not appear until 6 days after transplantation of T cells, helper activity was fully restored within 24 h and had diminished by 6 days. | Restoration of suppressor function in athymic mice. Antibody responses of normal and congenitally athymic (nu/nu) mice were measured to the phosphorylcholine determinant of a pneumococcal C-polysaccharide. The plaque-forming cell responses of athymic mice were approximately tenfold higher than for intact controls. This enhancement was equivalent to that obtained by treatment of controls with antithymocyte serum. In both cases, the enhanced responses presumably result from a lack of suppressor T cells. Reconstitution of athymic mice with normal thymocytes restored both suppressor and helper T cell functions. Whereas suppressor activity did not appear until 6 days after transplantation of T cells, helper activity was fully restored within 24 h and had diminished by 6 days. | [
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PMID:24000 | Bactericidal activity of specific and azurophil granules from human neutrophils: studies with outer-membrane mutants of Salmonella typhimurium LT-2. | Extracts of specific granules and azurophil granules from human neutrophils were tested for their bactericidal activity against various lipopolysaccharide mutants of Salmonella typhimurium LT-2. Three purified granule populations, one specific and two azurophil, were obtained by isopycnic centrifugation of homogenized neutrophils. Each was extracted with 0.2 M acetate buffer (pH 4), and the extracts were dialyzed against phosphate-buffered saline (pH 7) to remove acetate. These extracts contained >/=84% of the lysozyme, lactoferrin, or myeloperoxidase initially present in the whole granules. The S. typhimurium mutants possessed Ra, Rc, Rd(1), Rd(2), or Re lipopolysaccharide. As the carbohydrate content of the lipopolysaccharide decreased, the bacteria became increasingly more susceptible to the bactericidal activity of all granule extracts. Bactericidal activity of the extracts was in the order: mixed (azurophil + specific) >/= azurophil >> specific. Specific granules were bacteriostatic for S through Rd(2) bacteria. They were bactericidal only for the Re mutant. Both azurophil granule populations were equally bactericidal. Extracts boiled for 30 min retained none of their bactericidal activity for any of the bacteria; however, they remained bacteriostatic for the deep rough (Rd(2), Re) mutants. Bactericidal activity was dependent upon pH, in that mixed and azurophil granule contents killed the smooth parent and Ra mutant best at pH 5, the Rc and Rd(1) mutants to the same degree at pH 5 to 8, and the deep rough mutants (Rd(2) and Re) best at pH 8. Specific granule contents were most bacteriostatic for S through Rd(2) bacteria at pH 5 and killed the Re mutant only at pH 8. Thus, as the S. typhimurium lipopolysaccharide content decreased, the bactericidal pH optimum increased. Killing by all extracts was dependent upon incubation temperature, with almost no bactericidal or bacteriostatic activity observed when bacteria and granule fractions were incubated on ice (2 degrees C) and plated immediately. Intermediate killing was observed at 22 degrees C. If bacteria were incubated with granule extracts at 2 degrees C, washed free of extract, suspended in medium without extract, and reincubated at 37 degrees C, killing was observed. This suggested that a component(s) of the extracts was sticking to the bacteria at 2 degrees C but killing only at 37 degrees C. | Bactericidal activity of specific and azurophil granules from human neutrophils: studies with outer-membrane mutants of Salmonella typhimurium LT-2. Extracts of specific granules and azurophil granules from human neutrophils were tested for their bactericidal activity against various lipopolysaccharide mutants of Salmonella typhimurium LT-2. Three purified granule populations, one specific and two azurophil, were obtained by isopycnic centrifugation of homogenized neutrophils. Each was extracted with 0.2 M acetate buffer (pH 4), and the extracts were dialyzed against phosphate-buffered saline (pH 7) to remove acetate. These extracts contained >/=84% of the lysozyme, lactoferrin, or myeloperoxidase initially present in the whole granules. The S. typhimurium mutants possessed Ra, Rc, Rd(1), Rd(2), or Re lipopolysaccharide. As the carbohydrate content of the lipopolysaccharide decreased, the bacteria became increasingly more susceptible to the bactericidal activity of all granule extracts. Bactericidal activity of the extracts was in the order: mixed (azurophil + specific) >/= azurophil >> specific. Specific granules were bacteriostatic for S through Rd(2) bacteria. They were bactericidal only for the Re mutant. Both azurophil granule populations were equally bactericidal. Extracts boiled for 30 min retained none of their bactericidal activity for any of the bacteria; however, they remained bacteriostatic for the deep rough (Rd(2), Re) mutants. Bactericidal activity was dependent upon pH, in that mixed and azurophil granule contents killed the smooth parent and Ra mutant best at pH 5, the Rc and Rd(1) mutants to the same degree at pH 5 to 8, and the deep rough mutants (Rd(2) and Re) best at pH 8. Specific granule contents were most bacteriostatic for S through Rd(2) bacteria at pH 5 and killed the Re mutant only at pH 8. Thus, as the S. typhimurium lipopolysaccharide content decreased, the bactericidal pH optimum increased. Killing by all extracts was dependent upon incubation temperature, with almost no bactericidal or bacteriostatic activity observed when bacteria and granule fractions were incubated on ice (2 degrees C) and plated immediately. Intermediate killing was observed at 22 degrees C. If bacteria were incubated with granule extracts at 2 degrees C, washed free of extract, suspended in medium without extract, and reincubated at 37 degrees C, killing was observed. This suggested that a component(s) of the extracts was sticking to the bacteria at 2 degrees C but killing only at 37 degrees C. | [
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PMID:24001 | Role of complement in lethal bacterial lipopolysaccharide-induced hypotensive and coagulative changes. | The effect of C3 depletion on the multiple pathophysiological changes produced by a lethal dose of Serratia marcescens lipopolysaccharide (LPS) was evaluated. The injection of this LPS into rabbits resulted in biphasic hypotensive changes and thrombocytopenia. These changes were characterized by an acute, transient decrease occurring within minutes after injection followed by a second more gradual decrease beginning 30 to 60 min post-LPS. Prior depletion of C3, with the anticomplementary protein from cobra venom (CoF), did not alter the extent of either the gradual hypotensive and platelet changes or the coagulative and metabolic changes when normal and C3-depleted rabbits were compared. Importantly, the lethal effects of S. marcescens LPS were not reduced by prior depletion of C3. Only the immediate, reversible thrombocytopenia and hypotension were abrogated by C3 depletion. | Role of complement in lethal bacterial lipopolysaccharide-induced hypotensive and coagulative changes. The effect of C3 depletion on the multiple pathophysiological changes produced by a lethal dose of Serratia marcescens lipopolysaccharide (LPS) was evaluated. The injection of this LPS into rabbits resulted in biphasic hypotensive changes and thrombocytopenia. These changes were characterized by an acute, transient decrease occurring within minutes after injection followed by a second more gradual decrease beginning 30 to 60 min post-LPS. Prior depletion of C3, with the anticomplementary protein from cobra venom (CoF), did not alter the extent of either the gradual hypotensive and platelet changes or the coagulative and metabolic changes when normal and C3-depleted rabbits were compared. Importantly, the lethal effects of S. marcescens LPS were not reduced by prior depletion of C3. Only the immediate, reversible thrombocytopenia and hypotension were abrogated by C3 depletion. | [
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PMID:24002 | Attachment of Bacteroides melaninogenicus subsp. asaccharolyticus to oral surfaces and its possible role in colonization of the mouth and of periodontal pockets. | This investigation examined the ability of cells of Bacteroides melaninogenicus subsp. asaccharolyticus 381 to adhere to surfaces that might be important for its initial colonization of the mouth and its subsequent colonization in periodontal pockets. Of 48 asaccharolytic strains of B. melaninogenicus, 47 agglutinated human erythrocytes, whereas none of 20 fermentative strains, which included reference cultures of the subspecies intermedius and melaninogenicus, were active. Electron microscopy indicated that both asaccharolytic and fermentative strains possessed pili; hence, the presence of pili did not correlate with the hemagglutinating activities of B. melaninogenicus strains. Both asaccharolytic and fermentative B. melaninogenicus strains suspended in phosphate-buffered saline adhered in high numbers to buccal epithelial cells and to the surfaces of several gram-positive bacteria tested, including Actinomyces viscosus, A. naeslundii, A. israelii, Streptococcus sanguis, and S. mitis. B. melaninogenicus subsp. asaccharolyticus 381 also attached, but in comparatively low numbers, to untreated and to saliva-treated hydroxyapatite. Addition of clarified whole saliva to suspensions of strain 381 almost completely eliminated adherence to buccal epithelial cells and to hydroxyapatite surfaces, but saliva had no detectable effect on attachment to gram-positive plaque bacteria. Both fermentative and nonfermentative strains of B. melaninogenicus also attached in high numbers to crevicular epithelial cells derived from human periodontal pockets, but normal human serum strongly inhibited attachment. Serum also inhibited attachment of strain 381 to saliva- and serum-treated hydroxyapatite, but it had little effect upon attachment to gram-positive bacteria. These observations suggested that salivary and serum components would strongly inhibit the attachment of B. melaninogenicus cells to several oral surfaces, but not to the surfaces of certain gram-positive bacteria commonly present in human dental plaque. This was confirmed by an in vivo experiment in which streptomycin-labeled cells of B. melaninogenicus 381-R were introduced into the mouths of two volunteers. After 10 min, several hundred-fold higher numbers of the organism were recovered from preformed bacterial plaque present on teeth than from clean tooth surfaces or from the buccal mucosa and tongue dorsum. High numbers of B. melaninogenicus cells were also recovered from preformed plaque after 150 min, but virtually no cells of the organism were recovered from the other surfaces studied. These data suggest that the presence of dental plaque containing Actinomyces and other gram-positive bacteria may be essential for the attachment and colonization of B. melaninogenicus cells after their initial introduction into the mouth. Similarly, the presence of subgingival plaque containing gram-positive bacteria may be necessary for its secondary colonization in periodontal pockets. | Attachment of Bacteroides melaninogenicus subsp. asaccharolyticus to oral surfaces and its possible role in colonization of the mouth and of periodontal pockets. This investigation examined the ability of cells of Bacteroides melaninogenicus subsp. asaccharolyticus 381 to adhere to surfaces that might be important for its initial colonization of the mouth and its subsequent colonization in periodontal pockets. Of 48 asaccharolytic strains of B. melaninogenicus, 47 agglutinated human erythrocytes, whereas none of 20 fermentative strains, which included reference cultures of the subspecies intermedius and melaninogenicus, were active. Electron microscopy indicated that both asaccharolytic and fermentative strains possessed pili; hence, the presence of pili did not correlate with the hemagglutinating activities of B. melaninogenicus strains. Both asaccharolytic and fermentative B. melaninogenicus strains suspended in phosphate-buffered saline adhered in high numbers to buccal epithelial cells and to the surfaces of several gram-positive bacteria tested, including Actinomyces viscosus, A. naeslundii, A. israelii, Streptococcus sanguis, and S. mitis. B. melaninogenicus subsp. asaccharolyticus 381 also attached, but in comparatively low numbers, to untreated and to saliva-treated hydroxyapatite. Addition of clarified whole saliva to suspensions of strain 381 almost completely eliminated adherence to buccal epithelial cells and to hydroxyapatite surfaces, but saliva had no detectable effect on attachment to gram-positive plaque bacteria. Both fermentative and nonfermentative strains of B. melaninogenicus also attached in high numbers to crevicular epithelial cells derived from human periodontal pockets, but normal human serum strongly inhibited attachment. Serum also inhibited attachment of strain 381 to saliva- and serum-treated hydroxyapatite, but it had little effect upon attachment to gram-positive bacteria. These observations suggested that salivary and serum components would strongly inhibit the attachment of B. melaninogenicus cells to several oral surfaces, but not to the surfaces of certain gram-positive bacteria commonly present in human dental plaque. This was confirmed by an in vivo experiment in which streptomycin-labeled cells of B. melaninogenicus 381-R were introduced into the mouths of two volunteers. After 10 min, several hundred-fold higher numbers of the organism were recovered from preformed bacterial plaque present on teeth than from clean tooth surfaces or from the buccal mucosa and tongue dorsum. High numbers of B. melaninogenicus cells were also recovered from preformed plaque after 150 min, but virtually no cells of the organism were recovered from the other surfaces studied. These data suggest that the presence of dental plaque containing Actinomyces and other gram-positive bacteria may be essential for the attachment and colonization of B. melaninogenicus cells after their initial introduction into the mouth. Similarly, the presence of subgingival plaque containing gram-positive bacteria may be necessary for its secondary colonization in periodontal pockets. | [
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PMID:24004 | Adoptive immunochemotherapy of a transplantable AKR leukemia (K36). | Adoptive immunotherapy of a transplantable AKR leukemia (K36) was carried out as an adjunct to cytoxan chemotherapy using normal allogeneic H-2-incompatible spleen cells as well as sensitized H-2-matched allogeneic spleen cells. A significant therapeutic effect was obtained with cytoxan and allogeneic C57BL/6 splenocytes, demonstrating the potential use of the graft-versus-host reaction. Utilizing specific adoptive immunochemotherapy, a maximum effect was found with splenocytes from allogeneic but H-2-compatible CBA/J mice immunized against an allogeneic Gross-virus-induced lymphoma (E female G2). This therapeutic effect was most likely the result of prior sensitization of donor lymphocytes to common virus-associated tumor antigens. | Adoptive immunochemotherapy of a transplantable AKR leukemia (K36). Adoptive immunotherapy of a transplantable AKR leukemia (K36) was carried out as an adjunct to cytoxan chemotherapy using normal allogeneic H-2-incompatible spleen cells as well as sensitized H-2-matched allogeneic spleen cells. A significant therapeutic effect was obtained with cytoxan and allogeneic C57BL/6 splenocytes, demonstrating the potential use of the graft-versus-host reaction. Utilizing specific adoptive immunochemotherapy, a maximum effect was found with splenocytes from allogeneic but H-2-compatible CBA/J mice immunized against an allogeneic Gross-virus-induced lymphoma (E female G2). This therapeutic effect was most likely the result of prior sensitization of donor lymphocytes to common virus-associated tumor antigens. | [
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PMID:24008 | Sperm-freezing and donor insemination. | A comparison between freezing of human sperm using glycerol or glycerol egg-yolk citrate is presented. Freezing was performed with a gradual lowering of the temperature using alchol-liquid nitrogen or by immersion into liquid nitrogen. Great variations were observed within the same donor sperm sample when the sperm was frozen according to different techniques and marked post-thaw, motility variations were also found between spermatozoa from different donors. No statistically significant differences were found in the number of pregnancies obtained following inseminations with sperm frozen according to the two methods. | Sperm-freezing and donor insemination. A comparison between freezing of human sperm using glycerol or glycerol egg-yolk citrate is presented. Freezing was performed with a gradual lowering of the temperature using alchol-liquid nitrogen or by immersion into liquid nitrogen. Great variations were observed within the same donor sperm sample when the sperm was frozen according to different techniques and marked post-thaw, motility variations were also found between spermatozoa from different donors. No statistically significant differences were found in the number of pregnancies obtained following inseminations with sperm frozen according to the two methods. | [
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PMID:24009 | Electrochemical effects of metals on IUDs. | Electrochemical effects within the uterus are being studied, particularly as they pertain to IUDs made in total or in part of metals. The intrauterine potential as measured with either metal or nonpolarizable salt electrodes is positive at the fundal with respect to the cervical end of the uterus. Placement of dissimilar metal electrodes within the uterus will induce voltage gradients that are roughly predictable from their cell potential in blood as an electrolyte. When copper and zinc electrodes are placed at opposite ends of the uterus, they induce voltage gradients that are typically greater than 100 mv per cm. External copper and zinc wires connected at one end, and with the other ends in contact with various positions on the skin of rats, were found to influence potential gradients within the uterus. When a length of zinc wire was wound around the upper portion of the stem of a small Cu-7 200 IUD, it took about 1 year from insertion for the zinc to disappear. When the zinc wire was wound around the lower portion of the stem, it disappeared in 1 month. The zinc reduced intrauterine corrosion of the copper and it was more effective at reducing copper corrosion when in the lower stem position. | Electrochemical effects of metals on IUDs. Electrochemical effects within the uterus are being studied, particularly as they pertain to IUDs made in total or in part of metals. The intrauterine potential as measured with either metal or nonpolarizable salt electrodes is positive at the fundal with respect to the cervical end of the uterus. Placement of dissimilar metal electrodes within the uterus will induce voltage gradients that are roughly predictable from their cell potential in blood as an electrolyte. When copper and zinc electrodes are placed at opposite ends of the uterus, they induce voltage gradients that are typically greater than 100 mv per cm. External copper and zinc wires connected at one end, and with the other ends in contact with various positions on the skin of rats, were found to influence potential gradients within the uterus. When a length of zinc wire was wound around the upper portion of the stem of a small Cu-7 200 IUD, it took about 1 year from insertion for the zinc to disappear. When the zinc wire was wound around the lower portion of the stem, it disappeared in 1 month. The zinc reduced intrauterine corrosion of the copper and it was more effective at reducing copper corrosion when in the lower stem position. | [
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PMID:24010 | Antigens from human seminal plasma. Part III: studies on two new antigens from the tricholoracetic acid soluble fraction. | Two new antigens were isolated from the trichloroacetic acid soluble fraction of human seminal plasma: a polysaccharide (G-3) and a glycopeptide (G-4). Both were homogeneous, and hand sedimentation constants at infinite dilution of 2.9 X 10(-13) and 2.0 X 10(-13) resectively. Chemical and physicochemical studies suggest that both are branched molecules composed mainly of alpha-D-glucopyranosidic residues. The peptide component of G-R contains a high proporiton of the basic amino acids arginine and lysine. Since both antigens are polydispered, the low percentage of sialic acids or of some amino acids determined might have antigenic significance. | Antigens from human seminal plasma. Part III: studies on two new antigens from the tricholoracetic acid soluble fraction. Two new antigens were isolated from the trichloroacetic acid soluble fraction of human seminal plasma: a polysaccharide (G-3) and a glycopeptide (G-4). Both were homogeneous, and hand sedimentation constants at infinite dilution of 2.9 X 10(-13) and 2.0 X 10(-13) resectively. Chemical and physicochemical studies suggest that both are branched molecules composed mainly of alpha-D-glucopyranosidic residues. The peptide component of G-R contains a high proporiton of the basic amino acids arginine and lysine. Since both antigens are polydispered, the low percentage of sialic acids or of some amino acids determined might have antigenic significance. | [
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PMID:24011 | Augmentative effect of ascorbic acid upon induction of human ovulation in clomiphene-ineffective anovulatory women. | The effect of the administration of 1-ascorbic acid, either alone or combined with clomiphene, upon induction of human ovulation was investigated in clomiphene-inffective anovulatory women. Oral administration of daily 400 mg of ascorbic acid induced ovulation in two out of five habitually anovulatory cycles and in one out of eight first-grade amenorrhea cases, and was ineffective in all six second-grade hypothalamic amenorrhea cases. Combined administration of ascorbic acid with 5 days of clomiphene induced ovulation in five out of five habitually anovulatory cycles, in 10 out of 17 first-grade hypothalamic amenorrhea cases, and in two out of nine second-grade hypothalamic amenorrhea cases. Pregnancy was established in eight out of 18 sterile, habitually-anovulatory or first-grade amenorrheic women with the combined ascorbic acid-clomiphene therapy, and in one out of five sterile, habitually anovulatory women with ascorbic acid therapy alone. Since administration of ascorbic acid induced no changed in blood FSH, LH, and amount of cervical mucus, and it is well established that LH decreases dose-dependency of the ascorbic acid content in the rat ovaries, the possible site of action of ascorbic acid seems to be at the ovarian level. | Augmentative effect of ascorbic acid upon induction of human ovulation in clomiphene-ineffective anovulatory women. The effect of the administration of 1-ascorbic acid, either alone or combined with clomiphene, upon induction of human ovulation was investigated in clomiphene-inffective anovulatory women. Oral administration of daily 400 mg of ascorbic acid induced ovulation in two out of five habitually anovulatory cycles and in one out of eight first-grade amenorrhea cases, and was ineffective in all six second-grade hypothalamic amenorrhea cases. Combined administration of ascorbic acid with 5 days of clomiphene induced ovulation in five out of five habitually anovulatory cycles, in 10 out of 17 first-grade hypothalamic amenorrhea cases, and in two out of nine second-grade hypothalamic amenorrhea cases. Pregnancy was established in eight out of 18 sterile, habitually-anovulatory or first-grade amenorrheic women with the combined ascorbic acid-clomiphene therapy, and in one out of five sterile, habitually anovulatory women with ascorbic acid therapy alone. Since administration of ascorbic acid induced no changed in blood FSH, LH, and amount of cervical mucus, and it is well established that LH decreases dose-dependency of the ascorbic acid content in the rat ovaries, the possible site of action of ascorbic acid seems to be at the ovarian level. | [
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PMID:24012 | Varicocele: relation between anoxia and hypospermatogenesis. | Three groups of patients were analysed: varicocele patients with abnormal seminal fluid (17 cases), variococele and normal sperm (seven cases) and normal males (17 cases). Blood gas was secured in the internal spermatic vein, perpheric vein and femural artery, in an attempt to evaluate the possibility of altered gas content as a cause of spermatogenic depression. No statistical differences were found when we compared the three groups. We concluded that anoxia was not the cause of hypospermatogenesis in varicocele patients. | Varicocele: relation between anoxia and hypospermatogenesis. Three groups of patients were analysed: varicocele patients with abnormal seminal fluid (17 cases), variococele and normal sperm (seven cases) and normal males (17 cases). Blood gas was secured in the internal spermatic vein, perpheric vein and femural artery, in an attempt to evaluate the possibility of altered gas content as a cause of spermatogenic depression. No statistical differences were found when we compared the three groups. We concluded that anoxia was not the cause of hypospermatogenesis in varicocele patients. | [
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PMID:24013 | A morphological and physiological study of mesotubarium ovarica in humans. | In order to clarify the mechanisms by which the egg is transported from the ruptured follicle into the fimbrial end of the Fallopian tube in the human being, the mesotubarium ovarica (MTO), the unique anatomical structure which connects the tubal fimbriae and the ovary, was studied in seven human adnexal specimens histochemically and electron microscopically. The results demonstrated clearly the presence of smooth muscle cells in the MTO, and failed to demonstrate the presence of cilia in the lining epithelial cells of the MTO. Based on these morphological results, contractility of the MTO was studied in vitro by using a muscle chamber and a pressure transducer with 26 human adnexal specimens. Spontaneous contractile activites of regular frequency and moderate intensity were observed in the MTOs of all specimens examined. A possible role of the MTO in the mechanisms of ovum pickup at the time of ovulation is discussed. | A morphological and physiological study of mesotubarium ovarica in humans. In order to clarify the mechanisms by which the egg is transported from the ruptured follicle into the fimbrial end of the Fallopian tube in the human being, the mesotubarium ovarica (MTO), the unique anatomical structure which connects the tubal fimbriae and the ovary, was studied in seven human adnexal specimens histochemically and electron microscopically. The results demonstrated clearly the presence of smooth muscle cells in the MTO, and failed to demonstrate the presence of cilia in the lining epithelial cells of the MTO. Based on these morphological results, contractility of the MTO was studied in vitro by using a muscle chamber and a pressure transducer with 26 human adnexal specimens. Spontaneous contractile activites of regular frequency and moderate intensity were observed in the MTOs of all specimens examined. A possible role of the MTO in the mechanisms of ovum pickup at the time of ovulation is discussed. | [
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PMID:24014 | Postnidatory effects of luteinizing hormone releasing hormone (LHRH) in hamsters. | Whereas the administration of LHRH to pregnant hamsters has no effect during the prenidatory period, the hormone is effective in terminating pregnancy when given after implantation (days 6-10). The ED50 for pregnancy termination over this period approximates a dose of 0.35-0.4 mg b.i.d. When given to pregnant females in a second study, the effects of LHRH at this dose were completely reverse by minute doses of progesterone (30 microgram and above). Finally, administration of LHRH at 1.5 mg b.i.d., from days 6-10 was followed by daily sacrifice through day 12; bloods were sampled at autopsy for progesterone evaluation. Autopsies on days 7 and 8 showed few differences between controls and LHRH-treated hamsters, although decreased weights of the uterine/conceptus units signaled the initation of resorption. Significant LHRH-induced decreases in circulating progesterone were seen by day 9. Fetal resorption continued and was essentially complete by day 11, while progesterone levels continued depressed through the end of the study. | Postnidatory effects of luteinizing hormone releasing hormone (LHRH) in hamsters. Whereas the administration of LHRH to pregnant hamsters has no effect during the prenidatory period, the hormone is effective in terminating pregnancy when given after implantation (days 6-10). The ED50 for pregnancy termination over this period approximates a dose of 0.35-0.4 mg b.i.d. When given to pregnant females in a second study, the effects of LHRH at this dose were completely reverse by minute doses of progesterone (30 microgram and above). Finally, administration of LHRH at 1.5 mg b.i.d., from days 6-10 was followed by daily sacrifice through day 12; bloods were sampled at autopsy for progesterone evaluation. Autopsies on days 7 and 8 showed few differences between controls and LHRH-treated hamsters, although decreased weights of the uterine/conceptus units signaled the initation of resorption. Significant LHRH-induced decreases in circulating progesterone were seen by day 9. Fetal resorption continued and was essentially complete by day 11, while progesterone levels continued depressed through the end of the study. | [
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PMID:24015 | Vascular changes in traumatic amenorrhea and hypomenorrhea. | Pelvic angiography was performed in 12 cases of amenorrhea and hypomenorrhea which developed following curettage of abortion and in the puerperium. Six cases of similar age and obstetric history with normal menstrual cycles served as control. Pelvic angiography revealed widespread vascular occlusion of myometrial arteries, in seven of the twelve cases. These findings account for the small amount of endometrium removed on diagnostic curettage in these cases as well as the greatly reduced menstrual loss. The poor obstetric history of the cases studied may well be due to this excessive vascular damage. | Vascular changes in traumatic amenorrhea and hypomenorrhea. Pelvic angiography was performed in 12 cases of amenorrhea and hypomenorrhea which developed following curettage of abortion and in the puerperium. Six cases of similar age and obstetric history with normal menstrual cycles served as control. Pelvic angiography revealed widespread vascular occlusion of myometrial arteries, in seven of the twelve cases. These findings account for the small amount of endometrium removed on diagnostic curettage in these cases as well as the greatly reduced menstrual loss. The poor obstetric history of the cases studied may well be due to this excessive vascular damage. | [
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PMID:24016 | Quenching of tryptophan fluorescence in human antithrombin III by iodide ion. | Iodide is an efficient quencher of antithrombin III intrinsic tryptophan fluorescence. The quenching pattern indicates that about 60% of the tryptophyl fluorescence originates from exposed residues in the multitryptophan-containing protein. In denaturing media all of the tryptophyls are solvent-exposed. The binding of heparin to antithrombin III influences the number of solvent-exposed tryptophan residues. By studying the dependence of the quenching on pH, information regarding the presence of charged residues adjacent to tryptophyls was obtained. | Quenching of tryptophan fluorescence in human antithrombin III by iodide ion. Iodide is an efficient quencher of antithrombin III intrinsic tryptophan fluorescence. The quenching pattern indicates that about 60% of the tryptophyl fluorescence originates from exposed residues in the multitryptophan-containing protein. In denaturing media all of the tryptophyls are solvent-exposed. The binding of heparin to antithrombin III influences the number of solvent-exposed tryptophan residues. By studying the dependence of the quenching on pH, information regarding the presence of charged residues adjacent to tryptophyls was obtained. | [
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PMID:24017 | Photochemical evidence for the presence of histidyl residue in the active site of alkaline mesentericopeptidase. | The photosensitized oxidation of alkaline mesentericopeptidase in the presence of methylene blue results in a first-order rate of inactivation. The loss of enzymatic activity towards casein and N-acetyl-L-tyrosine ethyl ester closely correlates with the destruction of one histidyl residue. A pK value of 6.8 is determined from the sigmoid pH-dependence of the photoinactivation rate. This suggests the involvement of a normal titrating imidazole group in the active site of mesentericopeptidase. The competitive inhibitor Na-benzoyl-L-arginine protects the enzyme from photoinactivation. A conclusion is made that the active site histidyl residue is modified. Circular dichroism spectra show no change in the protein conformation during the photodynamic treatment. | Photochemical evidence for the presence of histidyl residue in the active site of alkaline mesentericopeptidase. The photosensitized oxidation of alkaline mesentericopeptidase in the presence of methylene blue results in a first-order rate of inactivation. The loss of enzymatic activity towards casein and N-acetyl-L-tyrosine ethyl ester closely correlates with the destruction of one histidyl residue. A pK value of 6.8 is determined from the sigmoid pH-dependence of the photoinactivation rate. This suggests the involvement of a normal titrating imidazole group in the active site of mesentericopeptidase. The competitive inhibitor Na-benzoyl-L-arginine protects the enzyme from photoinactivation. A conclusion is made that the active site histidyl residue is modified. Circular dichroism spectra show no change in the protein conformation during the photodynamic treatment. | [
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PMID:24018 | The peripheral and central neural actions of clonidine in normal and glaucomatous eyes. | The peripheral and central neural actions of clonidine on normal and glaucomatous eyes have been investigated. Threshold doses of clonidine applied topically induced a monotonic decrease of intraocular pressure in the treated eye and had no effect on the contralateral eye. With increased clonidine dose, a decrease of intraocular pressure occurred in the untreated eye, and there was a concomitant decrease of systemic arterial blood pressure. Analysis of aqueous humor dynamics showed that the ocular response to the peripheral and the central neural actions of clonidine were without effect on the tonographic coefficient of outflow facility. The episcleral venous pressure decreased in both the treated and the untreated eyes, but the changes were too small to account for the observed decrease of intraocular pressure. The results are consistent with the concept that both the peripheral and central ocular hypotensive actions of clonidine are mediated by an inhibition of adrenergic neurogenic vasoconstriction in the eye. | The peripheral and central neural actions of clonidine in normal and glaucomatous eyes. The peripheral and central neural actions of clonidine on normal and glaucomatous eyes have been investigated. Threshold doses of clonidine applied topically induced a monotonic decrease of intraocular pressure in the treated eye and had no effect on the contralateral eye. With increased clonidine dose, a decrease of intraocular pressure occurred in the untreated eye, and there was a concomitant decrease of systemic arterial blood pressure. Analysis of aqueous humor dynamics showed that the ocular response to the peripheral and the central neural actions of clonidine were without effect on the tonographic coefficient of outflow facility. The episcleral venous pressure decreased in both the treated and the untreated eyes, but the changes were too small to account for the observed decrease of intraocular pressure. The results are consistent with the concept that both the peripheral and central ocular hypotensive actions of clonidine are mediated by an inhibition of adrenergic neurogenic vasoconstriction in the eye. | [
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PMID:24019 | Reappraisal of the sympathetic role in the sphincteric urethra. Denervation supersensitivity of the urethra of the chronic neurogenic bladder to alpha-adrenergic drugs. | The response of urethral pressure to administration of an alpha-stimulant was compared between a group of eight patients with chronic neurogenic bladders as evidenced by positive denervation supersensitivity to parasympathomimetic bethanechol chloride and a group of ten control patients. A supersensitive response to administration of an alpha-stimulant with a rise of maximum urethral pressure, 10 mm Hg or more above the control urethral pressure, was uniformly observed in the urethra of patients with chronically denervated bladders. Our results appear to add pharmacologic evidence of alpha-adrenergic predominance over the parasympathetic in the urethra which is believed to be innervated dually in the recently envolving new concept. | Reappraisal of the sympathetic role in the sphincteric urethra. Denervation supersensitivity of the urethra of the chronic neurogenic bladder to alpha-adrenergic drugs. The response of urethral pressure to administration of an alpha-stimulant was compared between a group of eight patients with chronic neurogenic bladders as evidenced by positive denervation supersensitivity to parasympathomimetic bethanechol chloride and a group of ten control patients. A supersensitive response to administration of an alpha-stimulant with a rise of maximum urethral pressure, 10 mm Hg or more above the control urethral pressure, was uniformly observed in the urethra of patients with chronically denervated bladders. Our results appear to add pharmacologic evidence of alpha-adrenergic predominance over the parasympathetic in the urethra which is believed to be innervated dually in the recently envolving new concept. | [
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PMID:24021 | [Endocrine active pancreatic neoplasms]. | The tumor-forming endocrine cells of the pancreas belong to the APUD system. These cells are of neuroectodermal origin. The tumor can be diagnosed in most cases by a distinct clinical picture, and the diagnosis can be veryfied by direct hormone determination or/and by the biochemical disorders caused by the hormones. For localisation angiography, szintigrams, endoscopic pancreatography, and sonograms were used up to now without convincing results in many cases; computerized tomography promises to be the decisive examination in the future. Three hormones can, up to now, not yet be correlated with a distinct clinical picture specific for a pancreatic tumor. On the other hand, four tumors are responsible for a very typical clinical entity, the insulinoma, the glucagonoma, the gastrinoma, and the vipoma, as illustrated by our own cases. The surgical therapy consists mainly in enucleation of an adenoma or in partial pancreatic resection. Total pancreatectomy is indicated only in few cases. The Zollinger-Ellison syndrome is treated best by total gastrectomy. Malignant tumors are sensible to streptozotozin. | [Endocrine active pancreatic neoplasms]. The tumor-forming endocrine cells of the pancreas belong to the APUD system. These cells are of neuroectodermal origin. The tumor can be diagnosed in most cases by a distinct clinical picture, and the diagnosis can be veryfied by direct hormone determination or/and by the biochemical disorders caused by the hormones. For localisation angiography, szintigrams, endoscopic pancreatography, and sonograms were used up to now without convincing results in many cases; computerized tomography promises to be the decisive examination in the future. Three hormones can, up to now, not yet be correlated with a distinct clinical picture specific for a pancreatic tumor. On the other hand, four tumors are responsible for a very typical clinical entity, the insulinoma, the glucagonoma, the gastrinoma, and the vipoma, as illustrated by our own cases. The surgical therapy consists mainly in enucleation of an adenoma or in partial pancreatic resection. Total pancreatectomy is indicated only in few cases. The Zollinger-Ellison syndrome is treated best by total gastrectomy. Malignant tumors are sensible to streptozotozin. | [
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PMID:24022 | Hemoglobin Lufkin: beta 29 (B11) Gly replaced by Asp. An unstable hemoglobin variant involving an internal amino acid residue. | Hemoglobin Lufkin was found in a Black-American family. Structural analysis of the abnormal hemoglobin indicates a substitution of aspartic acid for glycine at position 29 in the beta chain. Marked instability of the variant hemoglobin is demonstrated by the rapid formation of inclusion bodies upon exposure of the red cells to redox dyes and by the large percentage of precipitated hemoglobin at 65 degrees C. The oxygen affinity, the Bohr effect, and the degree of cooperativity of Hb Lufkin and Hb A are similar over the physiologic pH range. However, at acid pH the oxygen affinity of the variant is increased. Unlike several other reported variants in the B helix, Hb Lufkin is not associated with methemoglobinemia. | Hemoglobin Lufkin: beta 29 (B11) Gly replaced by Asp. An unstable hemoglobin variant involving an internal amino acid residue. Hemoglobin Lufkin was found in a Black-American family. Structural analysis of the abnormal hemoglobin indicates a substitution of aspartic acid for glycine at position 29 in the beta chain. Marked instability of the variant hemoglobin is demonstrated by the rapid formation of inclusion bodies upon exposure of the red cells to redox dyes and by the large percentage of precipitated hemoglobin at 65 degrees C. The oxygen affinity, the Bohr effect, and the degree of cooperativity of Hb Lufkin and Hb A are similar over the physiologic pH range. However, at acid pH the oxygen affinity of the variant is increased. Unlike several other reported variants in the B helix, Hb Lufkin is not associated with methemoglobinemia. | [
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PMID:24030 | Contribution of a net transmembrane HCO3- flux to intracellular acid-base regulation. | Experiments were performed to determine the relative effects of a net extracellular-to-intracellular HCO3- flux and of elevated carbon dioxide tension (PCO2) on cellular acid-base regulation. Isolated rabbit hearts were perfused by recirculating a small volume of Ringer solution in which the PCO2 and the HCO3- concentration could be independently altered. Net HCO3- flux was assessed by the disappearance of HCO3- from perfusate. Between 40 and 100 Torr PCO2, a HCO3- flux into the cell occurs only when perfusate HCO3- concentration is increased. Therefore, by selective manipulation of perfusate HCO3- and PCO2 it is possible to induce hypercapnia with or without an accompanying HCO3- flux. When perfusate HCO3- concentration was increased from 20 to 36 mM, cellular HCO3- concentration increased from 22.5 +/- 0.8 to 26.1 +/- 1.0 mM at 40 Torr PCO2 and from 27.8 +/- 0.7 to 34.1 +/- 1.4 mM at 98 Torr PCO2. These increases can be accounted for by the amount of HCO3- that disappeared from the perfusate. The results suggest that most of the initial cell CO2 buffering is provided by the net HCO3- flux in addition to the passive physicochemical buffering. | Contribution of a net transmembrane HCO3- flux to intracellular acid-base regulation. Experiments were performed to determine the relative effects of a net extracellular-to-intracellular HCO3- flux and of elevated carbon dioxide tension (PCO2) on cellular acid-base regulation. Isolated rabbit hearts were perfused by recirculating a small volume of Ringer solution in which the PCO2 and the HCO3- concentration could be independently altered. Net HCO3- flux was assessed by the disappearance of HCO3- from perfusate. Between 40 and 100 Torr PCO2, a HCO3- flux into the cell occurs only when perfusate HCO3- concentration is increased. Therefore, by selective manipulation of perfusate HCO3- and PCO2 it is possible to induce hypercapnia with or without an accompanying HCO3- flux. When perfusate HCO3- concentration was increased from 20 to 36 mM, cellular HCO3- concentration increased from 22.5 +/- 0.8 to 26.1 +/- 1.0 mM at 40 Torr PCO2 and from 27.8 +/- 0.7 to 34.1 +/- 1.4 mM at 98 Torr PCO2. These increases can be accounted for by the amount of HCO3- that disappeared from the perfusate. The results suggest that most of the initial cell CO2 buffering is provided by the net HCO3- flux in addition to the passive physicochemical buffering. | [
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PMID:24031 | Effect of pH on cardiorespiratory and metabolic responses to exercise. | Five male subjects performed exercise at 33, 66, and 95% of their maximum power output on three occasions in random order. Each study was preceded by a 3-h period in which capsules were taken by mouth, containing either CaCO3 (control, NH4Cl (acidosis), or NaHCO3 (alkalosis) in a dose of 0.3 g/kg body wt; preexercise blood pH was 7.38 +/- 0.015, 7.21 +/- 0.033, and 7.43 +/- 0.029, respectively. Exercise was continuous and maintained for 20 min at the two lower power outputs and for as long as possible at the highest. Compared with control (270 +/- 13 s), endurance time at the highest power output was reduced in acidosis (160 +/- 22 s) and increased in alkalosis (438 +/- 120 s). No differences were observed for central cardiovascular changes in exercise (cardiac output, frequency, or stroke volume). The respiratory changes expected from changes in blood pH were observed, with a higher alveolar ventilation in acidosis. At all power outputs arterialized venous lactate was lowest in acidosis and highest in alkalosis. Plasma glycerol and free fatty acids were lowest in acidosis. Changes in blood [HCO3-] and pH were shown to have major effects on metabolism in exercise which presumably were responsible for impaired endurance. | Effect of pH on cardiorespiratory and metabolic responses to exercise. Five male subjects performed exercise at 33, 66, and 95% of their maximum power output on three occasions in random order. Each study was preceded by a 3-h period in which capsules were taken by mouth, containing either CaCO3 (control, NH4Cl (acidosis), or NaHCO3 (alkalosis) in a dose of 0.3 g/kg body wt; preexercise blood pH was 7.38 +/- 0.015, 7.21 +/- 0.033, and 7.43 +/- 0.029, respectively. Exercise was continuous and maintained for 20 min at the two lower power outputs and for as long as possible at the highest. Compared with control (270 +/- 13 s), endurance time at the highest power output was reduced in acidosis (160 +/- 22 s) and increased in alkalosis (438 +/- 120 s). No differences were observed for central cardiovascular changes in exercise (cardiac output, frequency, or stroke volume). The respiratory changes expected from changes in blood pH were observed, with a higher alveolar ventilation in acidosis. At all power outputs arterialized venous lactate was lowest in acidosis and highest in alkalosis. Plasma glycerol and free fatty acids were lowest in acidosis. Changes in blood [HCO3-] and pH were shown to have major effects on metabolism in exercise which presumably were responsible for impaired endurance. | [
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PMID:24032 | Catecholamine-synthesizing enzymes in adrenals of seasonally acclimatized voles. | Tyrosine hydroxylase (TH) and phenylethanolamine-N-methyltransferase (PNMT) activities were assayed in adrenal glands of the following groups of the Alaskan red-backed vole (Clethrionomys rutilus dawsoni): 1) laboratory reared at 20 degrees C and 2) exposed to 5 degrees C for 1, 3, 7, and 28 days; 3) wild, summer acclimatized; 4) wild, fall acclimatized; and 5) wild, winter acclimatized. TH activity in laboratory-acclimated voles exposed to 5 degrees C was increased by 2 times after 3 days and remained elevated after 28 days. PNMT activity in these same voles was increased after 7 days and also remained elevated after 28 days of cold exposure. In wild-acclimatized voles TH activity and PNMT activity in summer were equivalent to levels in 28-day cold-acclimated laboratory voles. In fall, TH activity was increased to 2.5 times the summer value. It decreased by midwinter, but remained elevated above the summer level. In contrast, PNMT activity appeared unchanged from summer through fall and winter. Pregnant summer voles had markedly increased TH activity. Adrenal norepinephrine and epinephrine did not change significantly with cold acclimation or seasonal acclimatization. Thus, acclimatization of wild voles to fall and winter conditions involved aquisition of a greater capacity to synthesize adrenal catecholamines than that produced by exposing laboratory-reared voles to an extended period of cold. | Catecholamine-synthesizing enzymes in adrenals of seasonally acclimatized voles. Tyrosine hydroxylase (TH) and phenylethanolamine-N-methyltransferase (PNMT) activities were assayed in adrenal glands of the following groups of the Alaskan red-backed vole (Clethrionomys rutilus dawsoni): 1) laboratory reared at 20 degrees C and 2) exposed to 5 degrees C for 1, 3, 7, and 28 days; 3) wild, summer acclimatized; 4) wild, fall acclimatized; and 5) wild, winter acclimatized. TH activity in laboratory-acclimated voles exposed to 5 degrees C was increased by 2 times after 3 days and remained elevated after 28 days. PNMT activity in these same voles was increased after 7 days and also remained elevated after 28 days of cold exposure. In wild-acclimatized voles TH activity and PNMT activity in summer were equivalent to levels in 28-day cold-acclimated laboratory voles. In fall, TH activity was increased to 2.5 times the summer value. It decreased by midwinter, but remained elevated above the summer level. In contrast, PNMT activity appeared unchanged from summer through fall and winter. Pregnant summer voles had markedly increased TH activity. Adrenal norepinephrine and epinephrine did not change significantly with cold acclimation or seasonal acclimatization. Thus, acclimatization of wild voles to fall and winter conditions involved aquisition of a greater capacity to synthesize adrenal catecholamines than that produced by exposing laboratory-reared voles to an extended period of cold. | [
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PMID:24033 | A32390A, a new biologically active metabolite. I. Discovery and fermentation studies. | A32390A is an isonitrile-containing derivative of diacyl D-mannitol. The compound is produced in fermentation as the major component of a metabolic complex known as A32390. A32390A inhibits dopamine-beta-hydroxylase reduces heart and adrenal norepinephrine levels, lowers blood pressure in hypertensive rats, and possesses antibiotic activity vs. Gram-positive bacteria and fungi, including Candida albicans. A32390 is produced in submerged culture by a mold, a species of Pyrenochaeta, NRRL-5786. Glucose and sucrose are among the best carbon sources for the biosynthesis of A32390. Mannitol, although a substituent of the A32390A molecule, supports little or no biosynthesis of the compound when employed as the major carbon source for the fermentation. The addition of crotonic acid derivatives. ethanol, or L-histidine to the fermentation medium enhances the level of A32390 produced. | A32390A, a new biologically active metabolite. I. Discovery and fermentation studies. A32390A is an isonitrile-containing derivative of diacyl D-mannitol. The compound is produced in fermentation as the major component of a metabolic complex known as A32390. A32390A inhibits dopamine-beta-hydroxylase reduces heart and adrenal norepinephrine levels, lowers blood pressure in hypertensive rats, and possesses antibiotic activity vs. Gram-positive bacteria and fungi, including Candida albicans. A32390 is produced in submerged culture by a mold, a species of Pyrenochaeta, NRRL-5786. Glucose and sucrose are among the best carbon sources for the biosynthesis of A32390. Mannitol, although a substituent of the A32390A molecule, supports little or no biosynthesis of the compound when employed as the major carbon source for the fermentation. The addition of crotonic acid derivatives. ethanol, or L-histidine to the fermentation medium enhances the level of A32390 produced. | [
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PMID:24037 | Regulation of gamma-aminobutyric acid degradation in Escherichia coli by nitrogen metabolism enzymes. | The possible role of glutamate dehydrogenase, glutamate synthase, and glutamine synthetase in the regulation of enzyme formation in the gamma-aminobutyric acid (GABA) catabolic pathway of Escherichia coli K-12 was investigated. Evidence is presented indicating that glutamine synthetase acts as a positive regulator in the E. coli GABA control system. Mutations impairing glutamate synthase activity prevent the depression of the enzymes of the GABA pathway in ammonia-limited glucose media. However, mutations resulting in constitutive synthesis of glutamine synthetase (GlnC) restore the ability of the glutamate synthase-less mutants to grow in glucose-GABA media and result in depressed synthesis of the GABA enzymes. It is suggested that the loss of glutamate synthesis activity affects the GABA control system indirectly by lowering glutamine synthetase levels. | Regulation of gamma-aminobutyric acid degradation in Escherichia coli by nitrogen metabolism enzymes. The possible role of glutamate dehydrogenase, glutamate synthase, and glutamine synthetase in the regulation of enzyme formation in the gamma-aminobutyric acid (GABA) catabolic pathway of Escherichia coli K-12 was investigated. Evidence is presented indicating that glutamine synthetase acts as a positive regulator in the E. coli GABA control system. Mutations impairing glutamate synthase activity prevent the depression of the enzymes of the GABA pathway in ammonia-limited glucose media. However, mutations resulting in constitutive synthesis of glutamine synthetase (GlnC) restore the ability of the glutamate synthase-less mutants to grow in glucose-GABA media and result in depressed synthesis of the GABA enzymes. It is suggested that the loss of glutamate synthesis activity affects the GABA control system indirectly by lowering glutamine synthetase levels. | [
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PMID:24038 | Mitochondrial adenosine triphosphatase of wild-type and poky Neurospora crassa. | We have compared the adenosine triphosphatase (ATPase) activity of mitochondria prepared from wild-type Neurospora crassa and from poky, a maternally inherited mutant known to possess defective mitochondrial ribosomes and reduced amounts of cytochromes aa3 and b. poky contains two distinct forms of mitochondrial ATPase. The first is normal in its Km for ATP, specificity for nucleotides and divalent cations, pH optimum, cold stability, and sensitivity to inhibitors (oligomycin, N,N-dicyclohexyl carbodiimide, and adenylyl imidodiphosphate). The fact that membrane-bound, cold-stable, oligomycin-sensitive ATPase activity is present in poky (with an activity of 1.93 +/- 0.03 mumol/min-mg of protein compared with 1.33 +/- 0.07 mumol/min-mg of protein in the wild-type strain) and also in chloramphenicol-grown wild-type cells suggests that products of mitochondrial protein synthesis play only a limited role in the attachment of the mitochondrial ATPase to the membrane in Neurospora. poky also contains a second form of mitochondrial ATPase, which has an activity of 1.5 +/- 0.2 mumol/min-mg of protein, is oligomycin sensitive but cold labile, and presumably is attached less firmly to the mitochondrial membrane. The two forms, added together, represent a substantial overproduction of mitochondrial ATPase by poky. | Mitochondrial adenosine triphosphatase of wild-type and poky Neurospora crassa. We have compared the adenosine triphosphatase (ATPase) activity of mitochondria prepared from wild-type Neurospora crassa and from poky, a maternally inherited mutant known to possess defective mitochondrial ribosomes and reduced amounts of cytochromes aa3 and b. poky contains two distinct forms of mitochondrial ATPase. The first is normal in its Km for ATP, specificity for nucleotides and divalent cations, pH optimum, cold stability, and sensitivity to inhibitors (oligomycin, N,N-dicyclohexyl carbodiimide, and adenylyl imidodiphosphate). The fact that membrane-bound, cold-stable, oligomycin-sensitive ATPase activity is present in poky (with an activity of 1.93 +/- 0.03 mumol/min-mg of protein compared with 1.33 +/- 0.07 mumol/min-mg of protein in the wild-type strain) and also in chloramphenicol-grown wild-type cells suggests that products of mitochondrial protein synthesis play only a limited role in the attachment of the mitochondrial ATPase to the membrane in Neurospora. poky also contains a second form of mitochondrial ATPase, which has an activity of 1.5 +/- 0.2 mumol/min-mg of protein, is oligomycin sensitive but cold labile, and presumably is attached less firmly to the mitochondrial membrane. The two forms, added together, represent a substantial overproduction of mitochondrial ATPase by poky. | [
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PMID:24039 | Catabolic N2-acetylornithine 5-aminotransferase of Klebsiella aerogenes: control of synthesis by induction, catabolite repression, and activation by glutamine synthetase. | Klebsiella aerogenes formed two N2-acetylornithine 5-aminotransferases (ACOAT) which were separable by diethylaminoethyl-cellulose chromatography. One ACOAT was repressed when the cells grew on arginine-containing medium, indicating its function in arginine biosynthesis. The second ACOAT was induced when arginine or ornithine was present in the medium as the sole source of carbon or nitrogen, suggesting its function in the catabolism of these compounds. The induced enzyme was purified almost to homogeneity. Its molecular weight is 59,000; it is a pyridoxal 5-phosphate-dependent enzyme and exhibits activity with N2-acetylornithine (Km = 1.1 mM) as well as with ornithine (Km = 5.4 mM). ACOAT did not catalyze the transamination of putrescine or 4-aminobutyrate. The best amino acceptor was 2-ketoglutarate (Km = 0.7 mM). ACOAT formation was subject to catabolite repression exerted by glucose when ammonia was present in excess. When the cells were deprived of nitrogen, ACOAT escaped from catabolite repression. This activation was mediated by glutamine synthetase as shown by the fact that mutants affected in the regulation or synthesis of glutamine synthetase were also affected in the control of ACOAT formation. | Catabolic N2-acetylornithine 5-aminotransferase of Klebsiella aerogenes: control of synthesis by induction, catabolite repression, and activation by glutamine synthetase. Klebsiella aerogenes formed two N2-acetylornithine 5-aminotransferases (ACOAT) which were separable by diethylaminoethyl-cellulose chromatography. One ACOAT was repressed when the cells grew on arginine-containing medium, indicating its function in arginine biosynthesis. The second ACOAT was induced when arginine or ornithine was present in the medium as the sole source of carbon or nitrogen, suggesting its function in the catabolism of these compounds. The induced enzyme was purified almost to homogeneity. Its molecular weight is 59,000; it is a pyridoxal 5-phosphate-dependent enzyme and exhibits activity with N2-acetylornithine (Km = 1.1 mM) as well as with ornithine (Km = 5.4 mM). ACOAT did not catalyze the transamination of putrescine or 4-aminobutyrate. The best amino acceptor was 2-ketoglutarate (Km = 0.7 mM). ACOAT formation was subject to catabolite repression exerted by glucose when ammonia was present in excess. When the cells were deprived of nitrogen, ACOAT escaped from catabolite repression. This activation was mediated by glutamine synthetase as shown by the fact that mutants affected in the regulation or synthesis of glutamine synthetase were also affected in the control of ACOAT formation. | [
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PMID:24040 | Effect of glucose starvation on the nicotinamide adenine dinucleotide phosphate-dependent glutamate dehydrogenase of yeast. | Yeast cells growing on mineral medium plus ammonia and glucose contained high levels of nicotinamide adenine dinucleotide phosphate-dependent glutamate dehydrogenase activity, as measured in crude extracts. After suspension of cells in fresh medium lacking glucose, there was a loss of the glutamate dehydrogenase activity. Loss of activity was inhibited by 2,4-dinitrophenol, sodium azide, iodoacetic acid, and cycloheximide. The enzyme activity was restored when glucose was added back to the medium, and this recovery was fully prevented in the presence of cycloheximide. | Effect of glucose starvation on the nicotinamide adenine dinucleotide phosphate-dependent glutamate dehydrogenase of yeast. Yeast cells growing on mineral medium plus ammonia and glucose contained high levels of nicotinamide adenine dinucleotide phosphate-dependent glutamate dehydrogenase activity, as measured in crude extracts. After suspension of cells in fresh medium lacking glucose, there was a loss of the glutamate dehydrogenase activity. Loss of activity was inhibited by 2,4-dinitrophenol, sodium azide, iodoacetic acid, and cycloheximide. The enzyme activity was restored when glucose was added back to the medium, and this recovery was fully prevented in the presence of cycloheximide. | [
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PMID:24041 | Evidence for the degradation of nicotinamide adenine dinucleotide phosphate-dependent glutamate dehydrogenase of Candida utilis during rapid enzyme inactivation. | The nicotinamide adenine dinucleotide phosphate-dependent glutamate dehydrogenase (NADP-GDH) from the food yeast Candida utilis was found to be rapidly inactivated when cultures were starved of a carbon source. The addition of glutamate or alanine to the starvation medium stimulated the rate of inactivation. Loss of enzyme activity was irreversible since the reappearance of enzyme activity, following the addition of glucose to carbon-starved cultures, was blocked by cycloheximide. A specific rabbit antibody was prepared against the NADP-GDH from C. utilis and used to quantitate the enzyme during inactivation promoted by carbon starvation. The amount of precipitable antigenic material paralleled the rapid decrease of enzyme activity observed after transition of cells from NH(4) (+)-glucose to glutamate medium. No additional small-molecular-weight protein was precipitated by the antibody as a result of the inactivation, suggesting that the enzyme is considerably altered during the primary steps of the inactivation process. Analysis by immunoprecipitation of the reappearance of enzyme activity after enzyme inactivation showed that increase of NADP-GDH activity was almost totally due to de novo synthesis, ruling out the possibility that enzyme activity modulation is achieved by reversible covalent modification. Enzyme degradation was also measured during steady-state growth and other changes in nitrogen and carbon status of the culture media. In all instances so far estimated, the enzyme was found to be very stable and not normally subject to high rates of degradation. Therefore, the possibility that inactivation was caused by a change in the ratio of synthesis to degradation can be excluded. | Evidence for the degradation of nicotinamide adenine dinucleotide phosphate-dependent glutamate dehydrogenase of Candida utilis during rapid enzyme inactivation. The nicotinamide adenine dinucleotide phosphate-dependent glutamate dehydrogenase (NADP-GDH) from the food yeast Candida utilis was found to be rapidly inactivated when cultures were starved of a carbon source. The addition of glutamate or alanine to the starvation medium stimulated the rate of inactivation. Loss of enzyme activity was irreversible since the reappearance of enzyme activity, following the addition of glucose to carbon-starved cultures, was blocked by cycloheximide. A specific rabbit antibody was prepared against the NADP-GDH from C. utilis and used to quantitate the enzyme during inactivation promoted by carbon starvation. The amount of precipitable antigenic material paralleled the rapid decrease of enzyme activity observed after transition of cells from NH(4) (+)-glucose to glutamate medium. No additional small-molecular-weight protein was precipitated by the antibody as a result of the inactivation, suggesting that the enzyme is considerably altered during the primary steps of the inactivation process. Analysis by immunoprecipitation of the reappearance of enzyme activity after enzyme inactivation showed that increase of NADP-GDH activity was almost totally due to de novo synthesis, ruling out the possibility that enzyme activity modulation is achieved by reversible covalent modification. Enzyme degradation was also measured during steady-state growth and other changes in nitrogen and carbon status of the culture media. In all instances so far estimated, the enzyme was found to be very stable and not normally subject to high rates of degradation. Therefore, the possibility that inactivation was caused by a change in the ratio of synthesis to degradation can be excluded. | [
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PMID:24042 | Effect of external environmental factors on the morphology of Spiroplasma citri. | Spiroplasma citri was examined by electron microscopy for morphological changes when maintained under a variety of conditions. PPLO serum fraction maintained spiral and helical morphology of S. citri at pH values of 8.0, 7.5, and 7.0, but only partially at pH 6.0 and 5.0. The absence of PPLO serum fraction resulted in round, deteriorated cells at all pH values tested. Bovine serum albumin (BSA), Phytone, soluble starch, potato starch, spermine, lipid-extracted PPLO serum fraction, and lipid-extracted BSA could substitute for PPLO serum fraction in maintaining spiral and helical morphology at pH 7.5. At pH 5.0, only BSA, lipid-extracted BSA, and lipid-extracted PPLO serum fraction were effective. Only BSA supported growth of S. citri for more than two transfers, whereas all other substitutes could not support growth longer than two transfers. | Effect of external environmental factors on the morphology of Spiroplasma citri. Spiroplasma citri was examined by electron microscopy for morphological changes when maintained under a variety of conditions. PPLO serum fraction maintained spiral and helical morphology of S. citri at pH values of 8.0, 7.5, and 7.0, but only partially at pH 6.0 and 5.0. The absence of PPLO serum fraction resulted in round, deteriorated cells at all pH values tested. Bovine serum albumin (BSA), Phytone, soluble starch, potato starch, spermine, lipid-extracted PPLO serum fraction, and lipid-extracted BSA could substitute for PPLO serum fraction in maintaining spiral and helical morphology at pH 7.5. At pH 5.0, only BSA, lipid-extracted BSA, and lipid-extracted PPLO serum fraction were effective. Only BSA supported growth of S. citri for more than two transfers, whereas all other substitutes could not support growth longer than two transfers. | [
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PMID:24043 | The ATPase reaction in the steady state and in the initial burst catalyzed by chicken gizzard myosin in 0.6 M KCl1. | On studying the steady-state activity in 0.6 M KCl, it was found that Mg-ATPase of chicken gizzard myosin was identical with that of rabbit skeletal myosin in the pH-activity profile, Michaelis-Menten constant, and maximum velocity. As regards the "initial burst" of ATP splitting in the presence of Mg (0.6 M KCl), it was found that gizzard and skeletal myosins were identical both in the size of the initial burst and in the velocity-ATP concentration relationship. The only difference we observed was that the Ca- and EDTA-ATPase activities of gizzard myosin were, as reported by other investigators, approximately one-half to one-third of those of skeletal myosin, although the pH-activity profiles for the ATPase of gizzard myosin was essentially the same as that of skeletal myosin. | The ATPase reaction in the steady state and in the initial burst catalyzed by chicken gizzard myosin in 0.6 M KCl1. On studying the steady-state activity in 0.6 M KCl, it was found that Mg-ATPase of chicken gizzard myosin was identical with that of rabbit skeletal myosin in the pH-activity profile, Michaelis-Menten constant, and maximum velocity. As regards the "initial burst" of ATP splitting in the presence of Mg (0.6 M KCl), it was found that gizzard and skeletal myosins were identical both in the size of the initial burst and in the velocity-ATP concentration relationship. The only difference we observed was that the Ca- and EDTA-ATPase activities of gizzard myosin were, as reported by other investigators, approximately one-half to one-third of those of skeletal myosin, although the pH-activity profiles for the ATPase of gizzard myosin was essentially the same as that of skeletal myosin. | [
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PMID:24045 | Modification of rabbit muscle phosphorylase b by a water-soluble carbodiimide. | Glycogen phosphorylase b from rabbit muscle was rapidly inactivated by incubation with 1-cyclohexyl-3-(2-morpholinyl-(4)-ethyl)carbodiimide metho-p-toluenesulfonate (CMC) at pH 5.1. The inactivation was pH-dependent and was not restored by treatment with hydroxylamine. The addition of glycine ethyl ester or N-(2,4-dinitrophenyl)-ethylenediamine (DNP-EDA) markedly increased the rate of inactivation. Of the various amino analogs of glucose tested, only glucosyl amine accelerated the inactivation, although they are all bound to the glucose 1-phosphate site of the enzyme. In the absence of amines, incorporation of about 3 mol of [metho-14C]CMC per protein monomer was observed on complete inactivation. In the presence of DNP-EDA, however, only 2 mol of [metho-14C]CMC and 1 mol of DNP-EDA were incorporated before the activity was completely lost. The treatment of phosphorylase b with CMC did not change the Km values of the enzyme for glucose 1-phosphate and AMP, in spite of the 56% inactivation. It is suggested that, in the phosphorylase-catalyzed reaction, an essential carboxyl group of the enzyme plays a role in the protonation of the glucosidic oxygen of glucose 1-phosphate. | Modification of rabbit muscle phosphorylase b by a water-soluble carbodiimide. Glycogen phosphorylase b from rabbit muscle was rapidly inactivated by incubation with 1-cyclohexyl-3-(2-morpholinyl-(4)-ethyl)carbodiimide metho-p-toluenesulfonate (CMC) at pH 5.1. The inactivation was pH-dependent and was not restored by treatment with hydroxylamine. The addition of glycine ethyl ester or N-(2,4-dinitrophenyl)-ethylenediamine (DNP-EDA) markedly increased the rate of inactivation. Of the various amino analogs of glucose tested, only glucosyl amine accelerated the inactivation, although they are all bound to the glucose 1-phosphate site of the enzyme. In the absence of amines, incorporation of about 3 mol of [metho-14C]CMC per protein monomer was observed on complete inactivation. In the presence of DNP-EDA, however, only 2 mol of [metho-14C]CMC and 1 mol of DNP-EDA were incorporated before the activity was completely lost. The treatment of phosphorylase b with CMC did not change the Km values of the enzyme for glucose 1-phosphate and AMP, in spite of the 56% inactivation. It is suggested that, in the phosphorylase-catalyzed reaction, an essential carboxyl group of the enzyme plays a role in the protonation of the glucosidic oxygen of glucose 1-phosphate. | [
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PMID:24047 | Kinetic studies on redox reactions of hemoproteins. II. Reduction of thermoresistant cytochrome c-552 and horse heart cytochrome c by ascorbic acid. | The reductions of thermoresistant cytochrome c-552 and horse heart cytochrome c by ascorbic acid were studied by the stopped-flow method between pH 4 and 10. The results were as follows (1) The reduction of horse heart cytochrome c showed two relaxation decays above pH 8.5, one of which was pseudo-first order, as was the case below pH 8, while the other was nearly concentration-independent. These results were consistent with those reported by Greenwood and Palmer (J. Biol. Chem. (1965) 240, 3660-3663). (2) For the reduction of cytochrome c-552, only a single relaxational decay that obeyed pseudo-first order kinetics was observed. (3) It seems most reasonable to assume that the concentration-independent relaxation process can be attributed to the isomerization reaction accompanying ligand exchange, since it is known that only horse heart cytochrome c exhibits ligand exchange, involving a residue with pK 9.3. | Kinetic studies on redox reactions of hemoproteins. II. Reduction of thermoresistant cytochrome c-552 and horse heart cytochrome c by ascorbic acid. The reductions of thermoresistant cytochrome c-552 and horse heart cytochrome c by ascorbic acid were studied by the stopped-flow method between pH 4 and 10. The results were as follows (1) The reduction of horse heart cytochrome c showed two relaxation decays above pH 8.5, one of which was pseudo-first order, as was the case below pH 8, while the other was nearly concentration-independent. These results were consistent with those reported by Greenwood and Palmer (J. Biol. Chem. (1965) 240, 3660-3663). (2) For the reduction of cytochrome c-552, only a single relaxational decay that obeyed pseudo-first order kinetics was observed. (3) It seems most reasonable to assume that the concentration-independent relaxation process can be attributed to the isomerization reaction accompanying ligand exchange, since it is known that only horse heart cytochrome c exhibits ligand exchange, involving a residue with pK 9.3. | [
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PMID:24049 | Genetic expression of microsomal electron transport in mice. Different patterns of dependence of constitutive cytochrome P-450 reductase activity of pyridine nucleotide concentrations. | Cytochrome P-450 reductase and aryl hydrocarbon hydroxylase activities were investigated in hepatic microsomes from untreated C57BL/6J, DBA/2J, B6D2F1, and (B6D2) D2 mice. The dependence of the rate of P-450 reduction on the concentration of added pyridine nucleotide (NADPH or NADH) was biphasic in DBA/2J microsomes but monophasic in C57BL/6J microsomes. Analogous strain-specific patterns were observed when the dependence of the rate of benzpyrene hydroxylation on NADPH concentration was examined. In crosses between the two inbred strains and between B6D2F1 mice and DBA/2J mice, the biphasic pattern for both the reductase and the hydroxylase activities was found to co-segregate with the recessive allele for aromatic hydrocarbon responsiveness. These results might reflect an architectural difference between the microsomal electron transport systems of responsive and nonresponsive mice. | Genetic expression of microsomal electron transport in mice. Different patterns of dependence of constitutive cytochrome P-450 reductase activity of pyridine nucleotide concentrations. Cytochrome P-450 reductase and aryl hydrocarbon hydroxylase activities were investigated in hepatic microsomes from untreated C57BL/6J, DBA/2J, B6D2F1, and (B6D2) D2 mice. The dependence of the rate of P-450 reduction on the concentration of added pyridine nucleotide (NADPH or NADH) was biphasic in DBA/2J microsomes but monophasic in C57BL/6J microsomes. Analogous strain-specific patterns were observed when the dependence of the rate of benzpyrene hydroxylation on NADPH concentration was examined. In crosses between the two inbred strains and between B6D2F1 mice and DBA/2J mice, the biphasic pattern for both the reductase and the hydroxylase activities was found to co-segregate with the recessive allele for aromatic hydrocarbon responsiveness. These results might reflect an architectural difference between the microsomal electron transport systems of responsive and nonresponsive mice. | [
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PMID:24050 | Oxygen equilibria of hybrid-heme hemoglobins containing proto- and mesoheme groups. On the nonequivalence of alpha and beta chains. | Hybrid-heme hemoglobins, alpha(meso)2beta(proto)2 and alpha(proto)2beta(meso)2, were prepared, and the O2 equilibria of their alpha and beta chains were measured separately at the isosbestic points of the partner chains at different pH values and in the presence and absence of inositol hexaphosphate. The Adair equation was extended to distinguish between the O2 saturations of the alpha and beta chains, and the seven equilibrium parameters were obtained by curve fitting to those equations. The results showed that the beta chains have an affinity slightly higher than the alpha chains in the binding of the first O2 molecule. For the second O2 molecule, the molecular species that has been oxygenated on the alpha chain has a higher affinity than that carrying O2 on the beta chain. The slopes of the Hill plots were higher for the alpha chain. The O2 saturation curves for the alpha and beta chains were calculated from the parameters averaged for the hybrids alpha(meso)2beta(proto)2 and alpha(proto)2beta(meso)2 in order to cancel the effects of the heme replacement. The curves showed that the difference in O2 saturation between the two kinds of chains depends on the conditions and on the degree of O2 saturation. It was concluded that the functional difference between the chains is small enough so that it is not required to modify the models already accepted for the cooperativity of hemoglobin. | Oxygen equilibria of hybrid-heme hemoglobins containing proto- and mesoheme groups. On the nonequivalence of alpha and beta chains. Hybrid-heme hemoglobins, alpha(meso)2beta(proto)2 and alpha(proto)2beta(meso)2, were prepared, and the O2 equilibria of their alpha and beta chains were measured separately at the isosbestic points of the partner chains at different pH values and in the presence and absence of inositol hexaphosphate. The Adair equation was extended to distinguish between the O2 saturations of the alpha and beta chains, and the seven equilibrium parameters were obtained by curve fitting to those equations. The results showed that the beta chains have an affinity slightly higher than the alpha chains in the binding of the first O2 molecule. For the second O2 molecule, the molecular species that has been oxygenated on the alpha chain has a higher affinity than that carrying O2 on the beta chain. The slopes of the Hill plots were higher for the alpha chain. The O2 saturation curves for the alpha and beta chains were calculated from the parameters averaged for the hybrids alpha(meso)2beta(proto)2 and alpha(proto)2beta(meso)2 in order to cancel the effects of the heme replacement. The curves showed that the difference in O2 saturation between the two kinds of chains depends on the conditions and on the degree of O2 saturation. It was concluded that the functional difference between the chains is small enough so that it is not required to modify the models already accepted for the cooperativity of hemoglobin. | [
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PMID:24052 | Dibutyryl cyclic AMP increases the amount of functional messenger RNA coding for tyrosine aminotransferase in rat liver. | The administration of N6,O2-dibutyryl cyclic AMP and theophylline to adrenalectomized rats results in an increase in the amount of functional mRNA coding for tyrosine aminotransferase that can be isolated from liver. The induction of this specific mRNA, as quantitated in a mRNA-dependent reticulocyte lysate system, and using poly(A)+ mRNA extracted from total tissue and polysomes, is very rapid. Within an hour after the intraperitoneal injection of the cyclic AMP derivative there is a 5- to 7-fold elevation of functional mRNA coding for tyrosine aminotransferase (mRNATAT), and by 3 h this has returned to basal levels. In contrast, the 4- to 5-fold induction of tyrosine aminotransferase catalytic activity is maximal at 2 h and is still significantly greater than the basal level at 5 h. In the basal state, tyrosine aminotransferase mRNA codes for 0.019 +/- 0.003% of the protein synthesized in the in vitro system, whereas after cyclic nucleotide treatment this value 0.115 +/- 0.015%, hence the increase in mRNATAT activity is relatively specific. Cordycepin, at a concentration which prevents the accumulation in cytoplasm of poly(A)+ mRNA, completely blocks the increase in both the catalytic and mRNA activity of this enzyme. The marked increase in functional mRNA, the requirement for continued synthesis of poly(A)+ RNA, and the rapid induction and deinduction suggest that the cyclic nucleotide is enhancing specific mRNA synthesis and/or, processing, however an effect on mRNA degradation cannot be excluded. | Dibutyryl cyclic AMP increases the amount of functional messenger RNA coding for tyrosine aminotransferase in rat liver. The administration of N6,O2-dibutyryl cyclic AMP and theophylline to adrenalectomized rats results in an increase in the amount of functional mRNA coding for tyrosine aminotransferase that can be isolated from liver. The induction of this specific mRNA, as quantitated in a mRNA-dependent reticulocyte lysate system, and using poly(A)+ mRNA extracted from total tissue and polysomes, is very rapid. Within an hour after the intraperitoneal injection of the cyclic AMP derivative there is a 5- to 7-fold elevation of functional mRNA coding for tyrosine aminotransferase (mRNATAT), and by 3 h this has returned to basal levels. In contrast, the 4- to 5-fold induction of tyrosine aminotransferase catalytic activity is maximal at 2 h and is still significantly greater than the basal level at 5 h. In the basal state, tyrosine aminotransferase mRNA codes for 0.019 +/- 0.003% of the protein synthesized in the in vitro system, whereas after cyclic nucleotide treatment this value 0.115 +/- 0.015%, hence the increase in mRNATAT activity is relatively specific. Cordycepin, at a concentration which prevents the accumulation in cytoplasm of poly(A)+ mRNA, completely blocks the increase in both the catalytic and mRNA activity of this enzyme. The marked increase in functional mRNA, the requirement for continued synthesis of poly(A)+ RNA, and the rapid induction and deinduction suggest that the cyclic nucleotide is enhancing specific mRNA synthesis and/or, processing, however an effect on mRNA degradation cannot be excluded. | [
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PMID:24055 | Purification and subunit structure of rat mammary gland acetyl coenzyme A carboxylase. | 1. Acetyl coenzyme A carboxylase from lactating rat mammary gland has been purified to apparent homogeneity. 2. The purified enzyme has the following characteristics: (a) its specific activity approaches 15 units/mg of protein, (b) the sedimentation constants of the protomeric and polymeric forms of the enzyme are 12 to 13 S and greater than or equal to 40 S, respectively, (c) the polymeric form of the enzyme shows filamentous structures in the electron microscope, and (d) the polypeptide(s) arising from its dissociation reveals a single major component of Mr = 240,000 to 260,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 3. The enzyme contains 1 mol of biotin and approximately 6 mol of phosphate/240,000 g of protein. | Purification and subunit structure of rat mammary gland acetyl coenzyme A carboxylase. 1. Acetyl coenzyme A carboxylase from lactating rat mammary gland has been purified to apparent homogeneity. 2. The purified enzyme has the following characteristics: (a) its specific activity approaches 15 units/mg of protein, (b) the sedimentation constants of the protomeric and polymeric forms of the enzyme are 12 to 13 S and greater than or equal to 40 S, respectively, (c) the polymeric form of the enzyme shows filamentous structures in the electron microscope, and (d) the polypeptide(s) arising from its dissociation reveals a single major component of Mr = 240,000 to 260,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 3. The enzyme contains 1 mol of biotin and approximately 6 mol of phosphate/240,000 g of protein. | [
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PMID:24057 | Prune-bell syndrome: a report of twelve cases and review of the literature. | Prune-belly syndrome is a rare congenital disease involving a deficiency of abdominal muscles, genitourinary abnormalities, and cryptorchidism. About half of the patients with the disease have associated musculoskeletal abnormalities, including club foot and dislocation of the hip. We saw twelve patients with this disease, four of whom required extensive orthopaedic treatment. | Prune-bell syndrome: a report of twelve cases and review of the literature. Prune-belly syndrome is a rare congenital disease involving a deficiency of abdominal muscles, genitourinary abnormalities, and cryptorchidism. About half of the patients with the disease have associated musculoskeletal abnormalities, including club foot and dislocation of the hip. We saw twelve patients with this disease, four of whom required extensive orthopaedic treatment. | [
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PMID:24058 | Survival of Streptococcus pneumoniae in sputum from patients with pneumonia. | The isolation rate of Streptococcus pneumoniae in sputum cultures from patients with pneumococcal pneumonia is low. An investigation was made to determine whether this low yield might be due to loss of pneumocci and/or overgrowth by pharyngeal flora before the specimen is plated. Pneumococcal survival times and pharyngeal overgrowth at 4 degrees C and at room temperature were determined in sputum obtained from 42 patients with pneumococcal pneumonia. It was found that pneumococci survived for long periods in sputum--2.2 +/- 1.4 days at room temperature and 9.5 +/- 3.6 days at 4 degrees C. Overgrowth by pharyngeal flora occurred in only 6 of 42 specimens kept at 4 degrees C and 31 of 42 specimens kept at room temperature. The low yield of S. pneumoniae in sputum from patients with pneumococcal pneumonia is not explained by decreased viability of the organism. | Survival of Streptococcus pneumoniae in sputum from patients with pneumonia. The isolation rate of Streptococcus pneumoniae in sputum cultures from patients with pneumococcal pneumonia is low. An investigation was made to determine whether this low yield might be due to loss of pneumocci and/or overgrowth by pharyngeal flora before the specimen is plated. Pneumococcal survival times and pharyngeal overgrowth at 4 degrees C and at room temperature were determined in sputum obtained from 42 patients with pneumococcal pneumonia. It was found that pneumococci survived for long periods in sputum--2.2 +/- 1.4 days at room temperature and 9.5 +/- 3.6 days at 4 degrees C. Overgrowth by pharyngeal flora occurred in only 6 of 42 specimens kept at 4 degrees C and 31 of 42 specimens kept at room temperature. The low yield of S. pneumoniae in sputum from patients with pneumococcal pneumonia is not explained by decreased viability of the organism. | [
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PMID:24059 | Quantitative evaluation of three commercial blood culture media for growth of anaerobic organisms. | The ability of three different commercial blood culture media--brain heart infusion broth (Pfizer), thiol broth (Difco), and PRS-peptone broth (Becton, Dickinson & Co.)--to support the growth of five different anaerobes is described. Inocula of 100 and 1,000 colony-forming units per ml were used to evaluate potential differences in survival, lag time, growth rate, and doubling times of each anaerobe in each medium. In addition, each medium was evaluated for its ability to neutralize the antibacterial effects of whole blood. The results of this study indicate that the PRS-peptone broth is superior to brain heart infusion and thiol broths. Shorter lag times and accelerated generation times and growth rates were noted for more different anaerobes in the PRS-peptone broth. Neither the size of inoculum nor the addition of normal whole blood appeared to alter the survival or growth characteristics of the anaerobes in any medium. However, the addition of normal whole blood did extend the lag time of each anaerobe by approximately 1 to 2 h in each medium. | Quantitative evaluation of three commercial blood culture media for growth of anaerobic organisms. The ability of three different commercial blood culture media--brain heart infusion broth (Pfizer), thiol broth (Difco), and PRS-peptone broth (Becton, Dickinson & Co.)--to support the growth of five different anaerobes is described. Inocula of 100 and 1,000 colony-forming units per ml were used to evaluate potential differences in survival, lag time, growth rate, and doubling times of each anaerobe in each medium. In addition, each medium was evaluated for its ability to neutralize the antibacterial effects of whole blood. The results of this study indicate that the PRS-peptone broth is superior to brain heart infusion and thiol broths. Shorter lag times and accelerated generation times and growth rates were noted for more different anaerobes in the PRS-peptone broth. Neither the size of inoculum nor the addition of normal whole blood appeared to alter the survival or growth characteristics of the anaerobes in any medium. However, the addition of normal whole blood did extend the lag time of each anaerobe by approximately 1 to 2 h in each medium. | [
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PMID:24061 | A double-blind controlled study of pipothiazine palmitate in the maintenance treatment of schizophrenic outpatients. | In a double-blind controlled study lasting 9 months, pipothiazine palmitate, a long-acting neuroleptic that can be administered intramuscularly once a month, was compared with fluphenazine enanthate in the maintenance treatment of 32 schizophrenic outpatients. Before the trial commenced, all patients were being treated with fluphenazine enanthate. The results indicate that pipothiazine palmitate was not as potent a neuroleptic as fluphenazine enanthate. Pipothiazine appears to resemble fluphenazine enanthate in its capacity to induce parkinsonism and tardive dyskinesia. | A double-blind controlled study of pipothiazine palmitate in the maintenance treatment of schizophrenic outpatients. In a double-blind controlled study lasting 9 months, pipothiazine palmitate, a long-acting neuroleptic that can be administered intramuscularly once a month, was compared with fluphenazine enanthate in the maintenance treatment of 32 schizophrenic outpatients. Before the trial commenced, all patients were being treated with fluphenazine enanthate. The results indicate that pipothiazine palmitate was not as potent a neuroleptic as fluphenazine enanthate. Pipothiazine appears to resemble fluphenazine enanthate in its capacity to induce parkinsonism and tardive dyskinesia. | [
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PMID:24060 | Effects of antianxiety drug and personality on stress-inducing psychomotor performance test. | The present study was carried out to clarify the effects of an antianxiety drug and of personality characteristics on a psychomotor performance test. Forty-eight healthy women college students were chosen from 64 volunteers as having either high or low levels of trait anxiety, neuroticism, or extroversion. Subjects with high trait anxiety and/or neuroticism tended to show a decrease in both speed and accuracy of the mirror drawing test (MDT) in the initial nondrug trials. Bromazepam, 5 mg, a benzodiazepine derivative, decreased this decrement in highly anxious subjects but worsened the speed in less anxious subjects. The personality traits of subjects, as well as the degree to which a performance test will induce stress, must be considered when evaluating the effects of antianxiety drugs on the performance of normal volunteers. The clinical anxiety-reducing efficacy of drugs may be predicted by using the MDT in subjects with high levels of anxiety and/or neuroticism. | Effects of antianxiety drug and personality on stress-inducing psychomotor performance test. The present study was carried out to clarify the effects of an antianxiety drug and of personality characteristics on a psychomotor performance test. Forty-eight healthy women college students were chosen from 64 volunteers as having either high or low levels of trait anxiety, neuroticism, or extroversion. Subjects with high trait anxiety and/or neuroticism tended to show a decrease in both speed and accuracy of the mirror drawing test (MDT) in the initial nondrug trials. Bromazepam, 5 mg, a benzodiazepine derivative, decreased this decrement in highly anxious subjects but worsened the speed in less anxious subjects. The personality traits of subjects, as well as the degree to which a performance test will induce stress, must be considered when evaluating the effects of antianxiety drugs on the performance of normal volunteers. The clinical anxiety-reducing efficacy of drugs may be predicted by using the MDT in subjects with high levels of anxiety and/or neuroticism. | [
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PMID:24067 | The effect of Skylab on the chemical composition of saliva. | The levels of specific proteins and electrolytes in stimulated whole saliva were monitored in Skylab crew members before and after each mission. With few exceptions, mission-associated compositional changes in saliva were relatively minimal. There were no changes in(formula see text), Cl-, albumin, or IgG concentrations. There were slight decreases in total protein coinciding with moderate saliva flow rate increases immediately before and after each flight. Other changes included diminutions in Na+ and lysozyme, and elevations in Mg++ and IgA. The IgA increase was the most pronounced mission-associated change observed. | The effect of Skylab on the chemical composition of saliva. The levels of specific proteins and electrolytes in stimulated whole saliva were monitored in Skylab crew members before and after each mission. With few exceptions, mission-associated compositional changes in saliva were relatively minimal. There were no changes in(formula see text), Cl-, albumin, or IgG concentrations. There were slight decreases in total protein coinciding with moderate saliva flow rate increases immediately before and after each flight. Other changes included diminutions in Na+ and lysozyme, and elevations in Mg++ and IgA. The IgA increase was the most pronounced mission-associated change observed. | [
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PMID:24068 | Tetramethyl benzidine for horseradish peroxidase neurohistochemistry: a non-carcinogenic blue reaction product with superior sensitivity for visualizing neural afferents and efferents. | Tetramethyl benzidine (TMB) is a presumptively non-carcinogenic chromogen which yields a blue reaction-product at sites of horseradish peroxidase activity. Sixty-six distinct procedures were performed in rats and monkeys in order to determine the optimal incubation parameters for TMB. As a result, a procedure is recommended whose sensitivity greatly surpasses that of a previously described benzidine dihydrochloride method. Indeed, the sensitivity of this new method in demonstrating retrograde transport is markedly superior to that of the previously described benzidine dihydrochloride method. Furthermore, as a consequence of this enhanced sensitivity, many efferent connections of the injection site are also visualized. The injection site demonstrated by this TMB procedure is significantly larger than the one demonstrated when benzidine dihydrochloride or diaminobenzidine is used as a chromogen. Finally, this TMB procedure has been compared to two other TMB procedures and found to provide superior morphology and sensitivity. | Tetramethyl benzidine for horseradish peroxidase neurohistochemistry: a non-carcinogenic blue reaction product with superior sensitivity for visualizing neural afferents and efferents. Tetramethyl benzidine (TMB) is a presumptively non-carcinogenic chromogen which yields a blue reaction-product at sites of horseradish peroxidase activity. Sixty-six distinct procedures were performed in rats and monkeys in order to determine the optimal incubation parameters for TMB. As a result, a procedure is recommended whose sensitivity greatly surpasses that of a previously described benzidine dihydrochloride method. Indeed, the sensitivity of this new method in demonstrating retrograde transport is markedly superior to that of the previously described benzidine dihydrochloride method. Furthermore, as a consequence of this enhanced sensitivity, many efferent connections of the injection site are also visualized. The injection site demonstrated by this TMB procedure is significantly larger than the one demonstrated when benzidine dihydrochloride or diaminobenzidine is used as a chromogen. Finally, this TMB procedure has been compared to two other TMB procedures and found to provide superior morphology and sensitivity. | [
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PMID:24069 | The pH of the hamster sperm acrosome. | The pH of the hamster sperm acrosome was estimated by a method based on the distribution of monoamines between membrane enclosed volumes maintaining pH gradients. A fluorescent amine, 9-aminoacridine, was used to permit both microscopic and fluorometric measurements of amine distribution. Cauda epididymal hamster sperm incubated with 9-aminoacridine accumulated the amine in the acrosomal volume. In the presence of NH4Cl or the ionophore Nigericin (compounds which discharge pH gradients) 9-aminoacridine fluorescence disappeared from the acrosome. Amine distribution between the acrosome and external volume was estimated by fluorometric measurement of sperm filtrates in the presence and absence of NH4Cl and Nigericin. These values, together with an estimated acrosomal volume of 0.4mu3 were used to calculate an acrosomal pH of less than 5. In addition, an acrosomal pH of 5 or less was obtained with 14C-methylamine. We suggest that such an acidic acrosomal pH of 5 or less could serve to inhibit the activation or autoactivation of the acrosomal zymogen proacrosin to acrosin, a trypsin-like enzyme involved in fertilization. | The pH of the hamster sperm acrosome. The pH of the hamster sperm acrosome was estimated by a method based on the distribution of monoamines between membrane enclosed volumes maintaining pH gradients. A fluorescent amine, 9-aminoacridine, was used to permit both microscopic and fluorometric measurements of amine distribution. Cauda epididymal hamster sperm incubated with 9-aminoacridine accumulated the amine in the acrosomal volume. In the presence of NH4Cl or the ionophore Nigericin (compounds which discharge pH gradients) 9-aminoacridine fluorescence disappeared from the acrosome. Amine distribution between the acrosome and external volume was estimated by fluorometric measurement of sperm filtrates in the presence and absence of NH4Cl and Nigericin. These values, together with an estimated acrosomal volume of 0.4mu3 were used to calculate an acrosomal pH of less than 5. In addition, an acrosomal pH of 5 or less was obtained with 14C-methylamine. We suggest that such an acidic acrosomal pH of 5 or less could serve to inhibit the activation or autoactivation of the acrosomal zymogen proacrosin to acrosin, a trypsin-like enzyme involved in fertilization. | [
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PMID:24077 | Effects of calcium on the absorption and retention of lead. | An inverse relationship between lead retention and dietary calcium content has been known to exist for many years, but the reasons for this association remained unknown. In rats, the manipulation of dietary calcium had no significant effect upon the absorption of lead, but calcium-deprived animals had decreased excretion and thus increased body retention of lead. Intraluminal calcium decreased the absorption of test doses of lead from the small intestine in a dose-related manner. We postulated that this occurred because the two metals competed for similar binding sites on intestinal mucosal proteins which were important in the absorptive process. In vivo, lead bound to two heat-stable intestinal mucosal fractions which have been shown to bind calcium. Although more lead bound to the higher molecular weight fraction and more calcium bound to the lower molecular weight vitamin D-induced CaBP, substantial amounts of lead and calcium were found in both fractions. Further, the addition of calcium to test doses of lead markedly diminished the amount of lead bound by both fractions. Shared binding sites on absorptive proteins would explain why dietary calcium decreases lead absorption. | Effects of calcium on the absorption and retention of lead. An inverse relationship between lead retention and dietary calcium content has been known to exist for many years, but the reasons for this association remained unknown. In rats, the manipulation of dietary calcium had no significant effect upon the absorption of lead, but calcium-deprived animals had decreased excretion and thus increased body retention of lead. Intraluminal calcium decreased the absorption of test doses of lead from the small intestine in a dose-related manner. We postulated that this occurred because the two metals competed for similar binding sites on intestinal mucosal proteins which were important in the absorptive process. In vivo, lead bound to two heat-stable intestinal mucosal fractions which have been shown to bind calcium. Although more lead bound to the higher molecular weight fraction and more calcium bound to the lower molecular weight vitamin D-induced CaBP, substantial amounts of lead and calcium were found in both fractions. Further, the addition of calcium to test doses of lead markedly diminished the amount of lead bound by both fractions. Shared binding sites on absorptive proteins would explain why dietary calcium decreases lead absorption. | [
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PMID:24080 | [Enzyme-immunological tests: determination of the activity of peroxidase with the aid of the "Trinder reagent" (author's transl)]. | Most of the currently used enzyme-immunological tests employ horse radish peroxidase as a marker enzyme. A new method is described for the determination of extremely small quantities of peroxidase. This largely prevents the inactivation of the peroxidase by H2O2 and thereby permits a much longer incubation time. In the presence of 25 mmol/l phenol, 2 mmol/l 4-amino-antipyrin and 0.8 mmol/l H2O2, peroxidase catalyses the formation of a red quinonimine, whose increase in adsorption is directly proportional to the enzyme concentration. | [Enzyme-immunological tests: determination of the activity of peroxidase with the aid of the "Trinder reagent" (author's transl)]. Most of the currently used enzyme-immunological tests employ horse radish peroxidase as a marker enzyme. A new method is described for the determination of extremely small quantities of peroxidase. This largely prevents the inactivation of the peroxidase by H2O2 and thereby permits a much longer incubation time. In the presence of 25 mmol/l phenol, 2 mmol/l 4-amino-antipyrin and 0.8 mmol/l H2O2, peroxidase catalyses the formation of a red quinonimine, whose increase in adsorption is directly proportional to the enzyme concentration. | [
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PMID:24081 | Thin layer chromatographic screening for methaqualone, phenothiazines, opiates and benzodiazepines. | A method is described which permits the simultaneous detection of methaqualone, phenothiazines, opiates and benzodiazepines in urine. Its diagnostic application is discussed. After cleavage of conjugates with hydrochloric acid, the substances are extracted and identified by thin-layer chromatography. In most cases analysis can be carried out using 2 solvent systems, phenothiazines, methaqualone and opiates being visualised using a three stage spray sequence. Since phenothiazines can interfere with the detection of methaqualone, a specific eluant is used to ensure reliable detection of the latter. Methaqualone can be positively identified by its characteristic metabolite pattern, whereas phenothiazines can only be detected as a group. | Thin layer chromatographic screening for methaqualone, phenothiazines, opiates and benzodiazepines. A method is described which permits the simultaneous detection of methaqualone, phenothiazines, opiates and benzodiazepines in urine. Its diagnostic application is discussed. After cleavage of conjugates with hydrochloric acid, the substances are extracted and identified by thin-layer chromatography. In most cases analysis can be carried out using 2 solvent systems, phenothiazines, methaqualone and opiates being visualised using a three stage spray sequence. Since phenothiazines can interfere with the detection of methaqualone, a specific eluant is used to ensure reliable detection of the latter. Methaqualone can be positively identified by its characteristic metabolite pattern, whereas phenothiazines can only be detected as a group. | [
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PMID:24082 | Glutamate dehydrogenase from coelenterates is NADP specific. | Glutamate dehydrogenases detected in tissue extracts of a broad sample of coelenterate species all require NADP(H) as a co-substrate, rather than being capable of using either NAD(H) or NADP(H). In this respect, the coelenterate phyla appear to be unique in the animal kingdom. | Glutamate dehydrogenase from coelenterates is NADP specific. Glutamate dehydrogenases detected in tissue extracts of a broad sample of coelenterate species all require NADP(H) as a co-substrate, rather than being capable of using either NAD(H) or NADP(H). In this respect, the coelenterate phyla appear to be unique in the animal kingdom. | [
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PMID:24083 | Purification and properties of beta-hydroxybutyrate dehydrogenase from Mycobacterium phlei ATCC354. | beta-Hydroxybutyrate dehydrogenase (EC 1.1.1.30) was purified 145-fold from Mycobacterium phlei ATCC354 by ammonium sulphate fractionation and DEAE-cellulose chromatography. The pH optima for oxidation and reduction reactions were 8.4 and 6.8 respectively. The purified enzyme was specific for NAD, NADH, acetoacetate and D(-)-beta-hydroxybutyrate. Km values for DL-beta-hydroxybutyrate and NAD were 7.4 mM and 0.66 mM respectively. The enzyme was inactivated by mercurial thiol inhibitors and by heat, but could be protected by NADH, Ca2+ and partially by Mn2+. The enzyme did not require metal ions and was insensitive to EDTA, glutathione, dithiothreitol, beta-mercaptoethanol and cysteine. | Purification and properties of beta-hydroxybutyrate dehydrogenase from Mycobacterium phlei ATCC354. beta-Hydroxybutyrate dehydrogenase (EC 1.1.1.30) was purified 145-fold from Mycobacterium phlei ATCC354 by ammonium sulphate fractionation and DEAE-cellulose chromatography. The pH optima for oxidation and reduction reactions were 8.4 and 6.8 respectively. The purified enzyme was specific for NAD, NADH, acetoacetate and D(-)-beta-hydroxybutyrate. Km values for DL-beta-hydroxybutyrate and NAD were 7.4 mM and 0.66 mM respectively. The enzyme was inactivated by mercurial thiol inhibitors and by heat, but could be protected by NADH, Ca2+ and partially by Mn2+. The enzyme did not require metal ions and was insensitive to EDTA, glutathione, dithiothreitol, beta-mercaptoethanol and cysteine. | [
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PMID:24084 | Virulence and resistance to superoxide, low pH and hydrogen peroxide among strains of Mycobacterium tuberculosis. | Six strains of Mycobacterium tuberculosis of different virulence in guinea-pigs were compared with regard to their resistance to low pH, to hydrogen peroxide (H2O2) at different pH values and to superoxide (O2-). Low virulence was associated with susceptibility to H2O2 in native and isoniazid-resistant strains but not in laboratory-attenuated strain H37Ra. H2O2 resistance was only partly related to catalase content. Low virulence was not associated with susceptibility to an acid environment but the tuberculocidal effect of H2O2 was significantly increased at low pH. The strains were uniformly resistant to O2- and contained similar amounts of superoxide dismutase. The implications of these observations are discussed in the context of mechanisms of host defence in tuberculosis. | Virulence and resistance to superoxide, low pH and hydrogen peroxide among strains of Mycobacterium tuberculosis. Six strains of Mycobacterium tuberculosis of different virulence in guinea-pigs were compared with regard to their resistance to low pH, to hydrogen peroxide (H2O2) at different pH values and to superoxide (O2-). Low virulence was associated with susceptibility to H2O2 in native and isoniazid-resistant strains but not in laboratory-attenuated strain H37Ra. H2O2 resistance was only partly related to catalase content. Low virulence was not associated with susceptibility to an acid environment but the tuberculocidal effect of H2O2 was significantly increased at low pH. The strains were uniformly resistant to O2- and contained similar amounts of superoxide dismutase. The implications of these observations are discussed in the context of mechanisms of host defence in tuberculosis. | [
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PMID:24085 | Exopolysaccharide production by Pseudomonas NCIB11264 grown in continuous culture. | Exopolysaccharide formation by Pseudomonas NCIB11264 in a single-stage continuous culture was maximal under nitrogen limitation with excess carbohydrate substrate at 30 +/- 1 degrees C and pH 7.0 +/- 0.1. Polysaccharide production was not enhanced by phosphate limitation but was dependent on the dilution rate. Steady states were maintained for up to 500 h without deterioration of the culture or the development of mutant strains. The efficiency of conversion of the glucose substrate utilized into exopolysaccharide by the chemostat cultures was as high as 73%. | Exopolysaccharide production by Pseudomonas NCIB11264 grown in continuous culture. Exopolysaccharide formation by Pseudomonas NCIB11264 in a single-stage continuous culture was maximal under nitrogen limitation with excess carbohydrate substrate at 30 +/- 1 degrees C and pH 7.0 +/- 0.1. Polysaccharide production was not enhanced by phosphate limitation but was dependent on the dilution rate. Steady states were maintained for up to 500 h without deterioration of the culture or the development of mutant strains. The efficiency of conversion of the glucose substrate utilized into exopolysaccharide by the chemostat cultures was as high as 73%. | [
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