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PMID:25215 | Nasal absorption of insulin in dogs. | The intranasal application of an insulin solution in dogs resulted in the rise of plasma immunoreactive insulin and in dose-dependent hypoglycemia. The absorption of insulin from this site was found to be enhanced when insulin was dissolved in an acid medium. In addition, when an insulin preparation with some surfactant was used, the effectiveness of nasally administered insulin was 25 to 30 per cent of that achieved with intravenously administered insulin. | Nasal absorption of insulin in dogs. The intranasal application of an insulin solution in dogs resulted in the rise of plasma immunoreactive insulin and in dose-dependent hypoglycemia. The absorption of insulin from this site was found to be enhanced when insulin was dissolved in an acid medium. In addition, when an insulin preparation with some surfactant was used, the effectiveness of nasally administered insulin was 25 to 30 per cent of that achieved with intravenously administered insulin. | [
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PMID:25217 | Trial of an alpha-adrenolytic drug (indoramin) for nocturnal enuresis. | The investigation of methods of treatment known to be effective in controlling nocturnal enuresis may yield information on the pathophysiology of the condition. An attempt has been made to mimic the known alpha-adrenolytic action of imipramine by treating a group of 14 enuretic school-children with a competitive alpha-adrenoceptor blocking substance, indoramin. Treatment in the doses used had no significant effect on night-wetting frequency. | Trial of an alpha-adrenolytic drug (indoramin) for nocturnal enuresis. The investigation of methods of treatment known to be effective in controlling nocturnal enuresis may yield information on the pathophysiology of the condition. An attempt has been made to mimic the known alpha-adrenolytic action of imipramine by treating a group of 14 enuretic school-children with a competitive alpha-adrenoceptor blocking substance, indoramin. Treatment in the doses used had no significant effect on night-wetting frequency. | [
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PMID:25218 | Intestinal mucins in health and disease. | Intestinal mucins are complex glycoproteins which are secreted from goblet cells, and form a gel-like covering over the mucosal surface. They are assumed to provide lubrication and protection of the underlying epithelium against potentially injurious chemicals, enzymes, bacteria and dietary constituents. Recent advances in our understanding of mucin structure, secretion and functional properties are reviewed in this paper. Implications for diseases such as cystic fibrosis, peptic ulcer, malignancy and inflammatory bowel disease are briefly discussed. | Intestinal mucins in health and disease. Intestinal mucins are complex glycoproteins which are secreted from goblet cells, and form a gel-like covering over the mucosal surface. They are assumed to provide lubrication and protection of the underlying epithelium against potentially injurious chemicals, enzymes, bacteria and dietary constituents. Recent advances in our understanding of mucin structure, secretion and functional properties are reviewed in this paper. Implications for diseases such as cystic fibrosis, peptic ulcer, malignancy and inflammatory bowel disease are briefly discussed. | [
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PMID:25221 | Prognosis in children with Crohn's disease. | The course of 86 children with Crohn's disease was examined during a 10-year period between 1966 and 1976. Patients were classified according to the initial site of disease. Ileocolitis was the most (52%) and colitis the least (9%) common form of disease with diffuse small bowel or ileal disease each comprising nearly 20% of the study group. These figures show a reversal from those of a previous decade when 42% of the patients had only terminal ileal disease and 17% had ileocolitis. Children with ileocolitis had the highest number of extracolonic manifestations and operations and required steroid therapy the longest. Those with only small bowel disease (with the exception of duodenal involvement) had fewer extraintestinal symptoms and operations and showed a consistently good response to medical treatment. | Prognosis in children with Crohn's disease. The course of 86 children with Crohn's disease was examined during a 10-year period between 1966 and 1976. Patients were classified according to the initial site of disease. Ileocolitis was the most (52%) and colitis the least (9%) common form of disease with diffuse small bowel or ileal disease each comprising nearly 20% of the study group. These figures show a reversal from those of a previous decade when 42% of the patients had only terminal ileal disease and 17% had ileocolitis. Children with ileocolitis had the highest number of extracolonic manifestations and operations and required steroid therapy the longest. Those with only small bowel disease (with the exception of duodenal involvement) had fewer extraintestinal symptoms and operations and showed a consistently good response to medical treatment. | [
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PMID:25222 | Inhibition of pancreatic secretion by enkephalin and morphine in dogs. | The nature and extent of enkephalin- and morphine-induced inhibition of pancreatic bicarbonate and protein secretion were studied in dogs with chronic pancreatic fistulae after administering exogenous secretin or octapeptide of cholecystokinin and stimulants for the endogenous release of these hormones. Enkephalin and morphine competitively inhibited the pancreatic bicarbonate secretion induced either by exogenous secretin or duodenal acidification. This inhibition was partially reversed by naloxone, an opiate antagonist. Opiate substance also profoundly inhibited pancreatic protein response to octapeptide of cholecystokinin and to various stimulants of endogenous cholecystokinin release. We conclude that enkephalin and morphine strongly inhibit the pancreatic responses to exogenous and endogenous stimulants by a mechanism involving separate opiate receptors. | Inhibition of pancreatic secretion by enkephalin and morphine in dogs. The nature and extent of enkephalin- and morphine-induced inhibition of pancreatic bicarbonate and protein secretion were studied in dogs with chronic pancreatic fistulae after administering exogenous secretin or octapeptide of cholecystokinin and stimulants for the endogenous release of these hormones. Enkephalin and morphine competitively inhibited the pancreatic bicarbonate secretion induced either by exogenous secretin or duodenal acidification. This inhibition was partially reversed by naloxone, an opiate antagonist. Opiate substance also profoundly inhibited pancreatic protein response to octapeptide of cholecystokinin and to various stimulants of endogenous cholecystokinin release. We conclude that enkephalin and morphine strongly inhibit the pancreatic responses to exogenous and endogenous stimulants by a mechanism involving separate opiate receptors. | [
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PMID:25227 | Some further characteristics of the growth of Naegleria fowleri and N. gruberi in axenic culture. | The effects of pH, various viscosity of the medium, changed ratio between the concentrations of dissolved and corpuscular components in the medium, and dissolved inorganic salts on the growth of axenic cultures of Naegleria fowleri and N. gruberi have been studied. The cultures were grown in liquid CALYG and BCS media. The pH optimum was 6.5 for N. fowleri and 6.0--6.5 for N. gruberi. No negative influence on the growth of N. fowleri was observed even at 0.5% concentration of highly viscous methylcellulose, whereas the growth of N. gruberi was distinctly inhibited by more than 0.2% of methycellulose. N. fowleri preferred the osmotorphic and N. gruberi phagotrophic nutrition in the given system of cultivation. The growth of both Naegleria species was inhibited by 0.1 N concentration of sodium chloride and potassium chloride without any significant difference in the tolerance. The inhibitory effect of these salts correlated primarily with the concentration of chloride anion. The ability to grow in a medium with increased viscosity and the preference for osmotrophic nutrtion are, besides the higher temperature optimum determined earlier, further characteristics of the pathogenic species N. fowleri. | Some further characteristics of the growth of Naegleria fowleri and N. gruberi in axenic culture. The effects of pH, various viscosity of the medium, changed ratio between the concentrations of dissolved and corpuscular components in the medium, and dissolved inorganic salts on the growth of axenic cultures of Naegleria fowleri and N. gruberi have been studied. The cultures were grown in liquid CALYG and BCS media. The pH optimum was 6.5 for N. fowleri and 6.0--6.5 for N. gruberi. No negative influence on the growth of N. fowleri was observed even at 0.5% concentration of highly viscous methylcellulose, whereas the growth of N. gruberi was distinctly inhibited by more than 0.2% of methycellulose. N. fowleri preferred the osmotorphic and N. gruberi phagotrophic nutrition in the given system of cultivation. The growth of both Naegleria species was inhibited by 0.1 N concentration of sodium chloride and potassium chloride without any significant difference in the tolerance. The inhibitory effect of these salts correlated primarily with the concentration of chloride anion. The ability to grow in a medium with increased viscosity and the preference for osmotrophic nutrtion are, besides the higher temperature optimum determined earlier, further characteristics of the pathogenic species N. fowleri. | [
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PMID:25228 | Virological examination of mosquito larvae from southern Moravia. | 254 pools of 4,115 mosquito larvae belonging to nine species were examined by isolation experiments. The larvae were collected in breeding places in an inundated forest--a natural focus of Tahyna virus, in April, June and July 1974 and 1975. Tahyna virus was isolated from one pool of 10 Culiseta annulata larvae collected in July 1974. Ecological questions concerning this finding are discussed. | Virological examination of mosquito larvae from southern Moravia. 254 pools of 4,115 mosquito larvae belonging to nine species were examined by isolation experiments. The larvae were collected in breeding places in an inundated forest--a natural focus of Tahyna virus, in April, June and July 1974 and 1975. Tahyna virus was isolated from one pool of 10 Culiseta annulata larvae collected in July 1974. Ecological questions concerning this finding are discussed. | [
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PMID:25229 | [Electroencephalographic effects of flurazepam in rabbits (author's transl)]. | Electroencephalographic (EEG) effects of flurazepam were investigated in unanesthetized, unrestrained rabbits with chronic electrode implants and compared with those of diazepam. Flurazepam, at doses of 0.5 approximately 5 mg/kg i.v., induced a drowsy EEG pattern, i.e. high voltage slow waves in the cortex and amygdaloid complex and desynchronization of the hippocampal theta waves. In addition, low voltage fast waves were superimposed, especially on the cortical EEG. Flurazepam suppressed the EEG arousal responses induced not only by auditory stimulation but also by electrical stimulation of the mesencephalic reticular formation, posterior hypothalamus and centromedian thalamus. The EEG arousal response induced by i.v. injection of physostigmine was suppressed by flurazepam. Flurazepam depressed the photic driving response and the augmenting response. The recruiting response was slightly enhanced by flurazepam. The limbic afterdischarges elicited by either hippocampal or amygdaloid stimulation were suppressed by flurazepam. Flurazepam caused reductions of pressor responses to stimulation of the posterior hypothalamus and the mesencephalic reticular formation in anaesthetized rabbits. There was little or no effect on pressor responses to the injection of noradrenaline, carotid artery occulusion and asphyxia with flurazepam. In general, these effects of flurazepam were similar to those of diazepam, but the drug induced actions which differed from those of diazepam. | [Electroencephalographic effects of flurazepam in rabbits (author's transl)]. Electroencephalographic (EEG) effects of flurazepam were investigated in unanesthetized, unrestrained rabbits with chronic electrode implants and compared with those of diazepam. Flurazepam, at doses of 0.5 approximately 5 mg/kg i.v., induced a drowsy EEG pattern, i.e. high voltage slow waves in the cortex and amygdaloid complex and desynchronization of the hippocampal theta waves. In addition, low voltage fast waves were superimposed, especially on the cortical EEG. Flurazepam suppressed the EEG arousal responses induced not only by auditory stimulation but also by electrical stimulation of the mesencephalic reticular formation, posterior hypothalamus and centromedian thalamus. The EEG arousal response induced by i.v. injection of physostigmine was suppressed by flurazepam. Flurazepam depressed the photic driving response and the augmenting response. The recruiting response was slightly enhanced by flurazepam. The limbic afterdischarges elicited by either hippocampal or amygdaloid stimulation were suppressed by flurazepam. Flurazepam caused reductions of pressor responses to stimulation of the posterior hypothalamus and the mesencephalic reticular formation in anaesthetized rabbits. There was little or no effect on pressor responses to the injection of noradrenaline, carotid artery occulusion and asphyxia with flurazepam. In general, these effects of flurazepam were similar to those of diazepam, but the drug induced actions which differed from those of diazepam. | [
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PMID:25230 | [Comparative studies of antihypertensive effects of several beta-blocking agents in conscious rats (author's transl)]. | The present study was undertaken to compare the antihypertensive effects of beta-blocking agents and to clarify the relations between antihypertensive effects and beta-blocking actions. In this study, blood pressure was measured by a direct cannulation of the abdominal aorta. Subcutaneous administration of carteolol and pindolol caused a significant fall in mean blood pressure in both normotensive and spontaneously hypertensive rats, whereas propranolol produced a rise in blood pressure. Maximum fall in blood pressure was observed 3 approximately 7 hr after the administration of carteolol and pindolol. In order to determine the beta-blocking action, changes in heart rate and blood pressure in response to isoproterenol (3 microgram/kg i.v.) were observed during the experiment. beta-Blocking action was found as early as 1 hr after subcutaneous administration. Carteolol showed the most effective blocking action throughout the experiment. Although beta-blocking agents lowered the blood pressure in this experiment, there was no apparent parallel between antihypertensive effects and beta-blocking action on the cardiac function. | [Comparative studies of antihypertensive effects of several beta-blocking agents in conscious rats (author's transl)]. The present study was undertaken to compare the antihypertensive effects of beta-blocking agents and to clarify the relations between antihypertensive effects and beta-blocking actions. In this study, blood pressure was measured by a direct cannulation of the abdominal aorta. Subcutaneous administration of carteolol and pindolol caused a significant fall in mean blood pressure in both normotensive and spontaneously hypertensive rats, whereas propranolol produced a rise in blood pressure. Maximum fall in blood pressure was observed 3 approximately 7 hr after the administration of carteolol and pindolol. In order to determine the beta-blocking action, changes in heart rate and blood pressure in response to isoproterenol (3 microgram/kg i.v.) were observed during the experiment. beta-Blocking action was found as early as 1 hr after subcutaneous administration. Carteolol showed the most effective blocking action throughout the experiment. Although beta-blocking agents lowered the blood pressure in this experiment, there was no apparent parallel between antihypertensive effects and beta-blocking action on the cardiac function. | [
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PMID:25231 | [Pharmacological studies of loperamide, an anti-diarrheal agent. I. Effects on diarrhea induced by castor oil and prostaglandin E. (author's transl)]. | Effects of loperamide on diarrhea induced by castor oil and prostaglandin E1 were investigated in rats and mice and compared with those of narcotic analgesics, atropine, mecamylamine and local anesthetics. The following results were obtained. Loperamide markedly suppressed the appearance of diarrhea induced by oral administration of castor oil in rats and the ED50 values for 1 and 2 hr protection was 0.082 and 0.42 mg/kg p.o., respectively. Loperamide markedly suppressed the appearance of diarrhea induced by i.v. administration of prostaglandin E1 and the ED50 value for 2 hr protection was 0.24 mg/kg p.o. in rats. The ID120 min value of loperamide which was calculated on the basis of the dose producing a 20% or more inhibition of the charcoal transport in the small intestine for 120 min was 0.8 mg/kg p.o. in mice and this activity was 9.2 times more potent than that of morphine. The analgesic ED50 value (Haffner's method) and LD50 value of loperamide was 149 and 249 mg/kg p.o., respectively. These results suggest that loperamide has a potent anti-diarrheal activity and specificity to the gastrointestinal tract and inhibits the effect of prostaglandin E1 and ricinoleic acid on the intestinal tract in rats. | [Pharmacological studies of loperamide, an anti-diarrheal agent. I. Effects on diarrhea induced by castor oil and prostaglandin E. (author's transl)]. Effects of loperamide on diarrhea induced by castor oil and prostaglandin E1 were investigated in rats and mice and compared with those of narcotic analgesics, atropine, mecamylamine and local anesthetics. The following results were obtained. Loperamide markedly suppressed the appearance of diarrhea induced by oral administration of castor oil in rats and the ED50 values for 1 and 2 hr protection was 0.082 and 0.42 mg/kg p.o., respectively. Loperamide markedly suppressed the appearance of diarrhea induced by i.v. administration of prostaglandin E1 and the ED50 value for 2 hr protection was 0.24 mg/kg p.o. in rats. The ID120 min value of loperamide which was calculated on the basis of the dose producing a 20% or more inhibition of the charcoal transport in the small intestine for 120 min was 0.8 mg/kg p.o. in mice and this activity was 9.2 times more potent than that of morphine. The analgesic ED50 value (Haffner's method) and LD50 value of loperamide was 149 and 249 mg/kg p.o., respectively. These results suggest that loperamide has a potent anti-diarrheal activity and specificity to the gastrointestinal tract and inhibits the effect of prostaglandin E1 and ricinoleic acid on the intestinal tract in rats. | [
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PMID:25232 | [Mechanism of methamphetamine toxicity in grouped mice and the effects of centrally acting drugs on its toxicity (author's transl)]. | The mortality of ddK mice treated with 40 mg/kg i.p. of methamphetamine (MA) was 85% in grouped conditions (10 mice in a cage) and 3% in individually isolated conditions. This mortality was not altered by the social environments even when other mice in the cage were not treated with MA. The mortality of mice individually isolated in cages with transparent walls was significantly higher than that of completely isolated mice. Almost all neuroleptics dose-dependently antagonized the MA toxicity in grouped mice, in small doses. The antagonizing activity of clozapine was somewhat weak, and sulpiride potentiated MA toxicity. Phentolamine and propranolol antagonized the MA toxicity at higher doses than neuroleptics. Reserpine and tetrabenazine previously given to mice remarkably antagonized the MA toxicity. H44/68 (a tyrosine hydroxylase inhibitor) had a considerable effect in antagonizing the MA toxicity, but diethyldithiocarbamate, U-14, 624 and FLA 63 (dopamine-beta-hydroxylase inhibitors) prevented the MA toxicity to a lesser extent than did H44/68. Apomorphine had no effect on the MA toxicity. The present data show that the MA toxicity in grouped mice (the increase in mortality) was enhanced by the presence of other mice, and suggest that the norepinephrine neurons play an important role in promoting the MA toxicity. Neuroleptics antagonize MA toxicity probably by blocking alpha-receptors in the central nervous system. | [Mechanism of methamphetamine toxicity in grouped mice and the effects of centrally acting drugs on its toxicity (author's transl)]. The mortality of ddK mice treated with 40 mg/kg i.p. of methamphetamine (MA) was 85% in grouped conditions (10 mice in a cage) and 3% in individually isolated conditions. This mortality was not altered by the social environments even when other mice in the cage were not treated with MA. The mortality of mice individually isolated in cages with transparent walls was significantly higher than that of completely isolated mice. Almost all neuroleptics dose-dependently antagonized the MA toxicity in grouped mice, in small doses. The antagonizing activity of clozapine was somewhat weak, and sulpiride potentiated MA toxicity. Phentolamine and propranolol antagonized the MA toxicity at higher doses than neuroleptics. Reserpine and tetrabenazine previously given to mice remarkably antagonized the MA toxicity. H44/68 (a tyrosine hydroxylase inhibitor) had a considerable effect in antagonizing the MA toxicity, but diethyldithiocarbamate, U-14, 624 and FLA 63 (dopamine-beta-hydroxylase inhibitors) prevented the MA toxicity to a lesser extent than did H44/68. Apomorphine had no effect on the MA toxicity. The present data show that the MA toxicity in grouped mice (the increase in mortality) was enhanced by the presence of other mice, and suggest that the norepinephrine neurons play an important role in promoting the MA toxicity. Neuroleptics antagonize MA toxicity probably by blocking alpha-receptors in the central nervous system. | [
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PMID:25233 | [Digestant effects of a new digestive enzyme capsule, Excelase, on jejunectomized and pancreato-jejunectomized Beagle dogs (author's transl)]. | The digestant effects of a new digestive capsule, excelase containing sanactase, proctase, meicelase, olipase-2S and pancreatic digestive enzyme TA, were investigated in vivo. Jejunectomy and pancreato-jejunectomy were performed to cause an artificial disturbance of gastro-intestinal digestion and absorption in Beagle dogs. Absorption of protein and fat was measured using chromic oxide as an indicator. Excelase (3 capsules/dog/day) was given orally to Beagle dogs 1 week after each operation for 7 weeks. Changes in body weight and absorption of protein and fat were observed during the administration. The decrease in body weight of dogs treated with excelase fully recovered, however, that of controls remained even 8 weeks after the surgery. Absorption of protein and fat in the groups of dogs treated with excelase was greatly improved as compared with controls. The digestant effects of excelase on percent absorption of protein and fat were more manifest in pancreato-jejunectomized dogs than in jejunectomized dogs. These results indicate that excelase is effective for gastro-intestinal disturbances of digestion and absorption. The digestant effects of excelase on starch, protein and cellulose were also investigated in vitro using a gastro-intestinal model. | [Digestant effects of a new digestive enzyme capsule, Excelase, on jejunectomized and pancreato-jejunectomized Beagle dogs (author's transl)]. The digestant effects of a new digestive capsule, excelase containing sanactase, proctase, meicelase, olipase-2S and pancreatic digestive enzyme TA, were investigated in vivo. Jejunectomy and pancreato-jejunectomy were performed to cause an artificial disturbance of gastro-intestinal digestion and absorption in Beagle dogs. Absorption of protein and fat was measured using chromic oxide as an indicator. Excelase (3 capsules/dog/day) was given orally to Beagle dogs 1 week after each operation for 7 weeks. Changes in body weight and absorption of protein and fat were observed during the administration. The decrease in body weight of dogs treated with excelase fully recovered, however, that of controls remained even 8 weeks after the surgery. Absorption of protein and fat in the groups of dogs treated with excelase was greatly improved as compared with controls. The digestant effects of excelase on percent absorption of protein and fat were more manifest in pancreato-jejunectomized dogs than in jejunectomized dogs. These results indicate that excelase is effective for gastro-intestinal disturbances of digestion and absorption. The digestant effects of excelase on starch, protein and cellulose were also investigated in vitro using a gastro-intestinal model. | [
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PMID:25234 | [Relation between the drug-metabolizing activities of liver microsomes and multiplicity of cytochrome P-450 (author's transl)]. | Wada et al. found a hyperbolic curve in the Lineweaver-Burk plots of the drug-metabolizing activity and competitive inhibition of steroid hormones to drugs, and suggested that several enzymes acting on different sites of various steroid hormone molecules might metabolize drugs without having specificities for hydroxylation sites. Recently, several cytochrome P-450s were separated and purified from liver microsomes. We studied the relation between the drug-metabolizing activities and multiplicity of cytochrome P-450 with the pH curve and the Lineweaver-Burk plots. Phenobarbital- and methylcholanthrene-treatment had remarkable effects on the pH curve of aniline hydroxylation by mouse liver microsomes. Addition of cortisol inhibited aniline hydroxylatin by liver microsomes from phenobarbital-treated mice more effectively at higher pH range and the pH curve was similar to that seen for normal mice. Haugen and Coon separated preparations [A] [B] and [C] with hydroxylapatite chromatography and we obtained [A'] (eluted by 100 mM K-phosphate buffer, pH 7.4) between [A] (50 mM) and [B] (300 mM). The preparation [A'] had the same absorption maximum (451 nm) of reduced CO complex as [A], but Km and pH curve for [A'] were similar to those for [B]. Each separated preparation of cytochrome P-450 showed a hyperbolic curve in Lineweaver-Burk plots and a different pH curve from each other. | [Relation between the drug-metabolizing activities of liver microsomes and multiplicity of cytochrome P-450 (author's transl)]. Wada et al. found a hyperbolic curve in the Lineweaver-Burk plots of the drug-metabolizing activity and competitive inhibition of steroid hormones to drugs, and suggested that several enzymes acting on different sites of various steroid hormone molecules might metabolize drugs without having specificities for hydroxylation sites. Recently, several cytochrome P-450s were separated and purified from liver microsomes. We studied the relation between the drug-metabolizing activities and multiplicity of cytochrome P-450 with the pH curve and the Lineweaver-Burk plots. Phenobarbital- and methylcholanthrene-treatment had remarkable effects on the pH curve of aniline hydroxylation by mouse liver microsomes. Addition of cortisol inhibited aniline hydroxylatin by liver microsomes from phenobarbital-treated mice more effectively at higher pH range and the pH curve was similar to that seen for normal mice. Haugen and Coon separated preparations [A] [B] and [C] with hydroxylapatite chromatography and we obtained [A'] (eluted by 100 mM K-phosphate buffer, pH 7.4) between [A] (50 mM) and [B] (300 mM). The preparation [A'] had the same absorption maximum (451 nm) of reduced CO complex as [A], but Km and pH curve for [A'] were similar to those for [B]. Each separated preparation of cytochrome P-450 showed a hyperbolic curve in Lineweaver-Burk plots and a different pH curve from each other. | [
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PMID:25237 | [Comparative studies on the protective effect of etilefrin and dihydroergotamine in circulatory instability after blood donation]. | In 20 female and 40 male subjects with circulatory instability, the response of pulse rate and blood pressure to blood loss and orthostasis was studied under the condition of protective pretreatment with drugs known to improve venous return by increasing the tone of capacitance vessels. Before blood loss (420 ml), in a cross-over double-blind procedure, each subject obtained 10 mg etilefrin or 2 mg dihydroergotamine, both administered orally. The procedure was repeated after at least 8 weeks. At this time, the subject obtained the drug which was not given in the first examination. By comparison of pulse rate and blood pressure taken after pretreatment, the statistical evaluation did not reveal any differences in the effectiveness of one or the other drug on the response to hypovolemic and orthostatic conditions. Patients preferred etilefrin. | [Comparative studies on the protective effect of etilefrin and dihydroergotamine in circulatory instability after blood donation]. In 20 female and 40 male subjects with circulatory instability, the response of pulse rate and blood pressure to blood loss and orthostasis was studied under the condition of protective pretreatment with drugs known to improve venous return by increasing the tone of capacitance vessels. Before blood loss (420 ml), in a cross-over double-blind procedure, each subject obtained 10 mg etilefrin or 2 mg dihydroergotamine, both administered orally. The procedure was repeated after at least 8 weeks. At this time, the subject obtained the drug which was not given in the first examination. By comparison of pulse rate and blood pressure taken after pretreatment, the statistical evaluation did not reveal any differences in the effectiveness of one or the other drug on the response to hypovolemic and orthostatic conditions. Patients preferred etilefrin. | [
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PMID:25241 | Macrophage colony development: properties of colony stimulating factors from murine embryo and pregnant uterus. | Extracts from embryonic and uterine tissue of mice, operationally defined as colony stimulating factor (CSF), promoted the growth of macrophage-granulocyte colonies in vitro. Uterine CSF focusses from pH 5.15 to 6.00 and embryonic CSF from pH 3.60 to 5.20, although both forms have similar biological activity. CSF is relatively resistant to denaturation but it is inactivated by periodate and dithiothreitol. Gel filtration indicates a molecular weight of 45,000 which is unchanged following treatment with insolubilized trypsin, a procedure which affords a useful purification (240-fold). Trypsin-sensitive material in CSF preparations modifies colonial form under certain conditions of culture, probably by increasing the motility of macrophages. Diaminoethane derivatives of CSF were prepared and retained biological activity at isoelectric points above pH 9.0. These derivatives may be covalently linked to Sepharose providing an insolubilized form of CSF to study interactions of CSF with the cell surface. | Macrophage colony development: properties of colony stimulating factors from murine embryo and pregnant uterus. Extracts from embryonic and uterine tissue of mice, operationally defined as colony stimulating factor (CSF), promoted the growth of macrophage-granulocyte colonies in vitro. Uterine CSF focusses from pH 5.15 to 6.00 and embryonic CSF from pH 3.60 to 5.20, although both forms have similar biological activity. CSF is relatively resistant to denaturation but it is inactivated by periodate and dithiothreitol. Gel filtration indicates a molecular weight of 45,000 which is unchanged following treatment with insolubilized trypsin, a procedure which affords a useful purification (240-fold). Trypsin-sensitive material in CSF preparations modifies colonial form under certain conditions of culture, probably by increasing the motility of macrophages. Diaminoethane derivatives of CSF were prepared and retained biological activity at isoelectric points above pH 9.0. These derivatives may be covalently linked to Sepharose providing an insolubilized form of CSF to study interactions of CSF with the cell surface. | [
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PMID:25242 | Effect of complement fixation on the release of lysosomal enzymes from rabbit alveolar macrophages. | Macrophage lysosomal enzymes that have specificities for substrates found in mammalian tissue may contribute to the tissue damage observed in chronic inflammatory diseases. Although a variety of agents that stimulate the release of hydrolases from peritoneal macrophages have been identified, little work has been done to establish the conditions and specific stimuli responsible for enzyme release by alveolar macrophages (AM). To assess the effect of phagocytosis by AM on the release phenomenon, hydrolytic enzymes normally sequestered in AM lysosomes were quantified in the supernatant fluids of phagocytosing and non-phagocytosing AM monolayers in both the presence and absence of serum. Maximum release occurred under conditions known to favor complement fixation by the classical or alternative pathways. Thermal destruction or immune fixation of serum complement before use in culture reduced the magnitude of release to that observed in cultures incubated in the complete absence of serum. Phagocytosis was not essential for release, since cells exposed to particles but treated with a known inhibitor of phagocytosis (cytochalasin B) still showed maximal release in the presence of fresh serum. These data are interpreted to indicate that a heat-labile component of serum, probably a by-product of complement fixation, was primarily responsible for the hydrolase release by AM. On the basis of these findings, individuals suffering from chronic pulmonary disease may suffer acute episodes of lung destruction from endogenous AM enzymes upon the inhalation of materials capable of fixing complement. | Effect of complement fixation on the release of lysosomal enzymes from rabbit alveolar macrophages. Macrophage lysosomal enzymes that have specificities for substrates found in mammalian tissue may contribute to the tissue damage observed in chronic inflammatory diseases. Although a variety of agents that stimulate the release of hydrolases from peritoneal macrophages have been identified, little work has been done to establish the conditions and specific stimuli responsible for enzyme release by alveolar macrophages (AM). To assess the effect of phagocytosis by AM on the release phenomenon, hydrolytic enzymes normally sequestered in AM lysosomes were quantified in the supernatant fluids of phagocytosing and non-phagocytosing AM monolayers in both the presence and absence of serum. Maximum release occurred under conditions known to favor complement fixation by the classical or alternative pathways. Thermal destruction or immune fixation of serum complement before use in culture reduced the magnitude of release to that observed in cultures incubated in the complete absence of serum. Phagocytosis was not essential for release, since cells exposed to particles but treated with a known inhibitor of phagocytosis (cytochalasin B) still showed maximal release in the presence of fresh serum. These data are interpreted to indicate that a heat-labile component of serum, probably a by-product of complement fixation, was primarily responsible for the hydrolase release by AM. On the basis of these findings, individuals suffering from chronic pulmonary disease may suffer acute episodes of lung destruction from endogenous AM enzymes upon the inhalation of materials capable of fixing complement. | [
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PMID:25243 | Evidence for quantitative variability of bacterial opsonic requirements. | We studied human serum opsonins by using combinations of heat inactivation and chelation to inhibit complement, adsorption to remove antibody, and trypan blue to inactivate the C3 receptor of human polymorphonuclear leukocytes. Streptococcus pneumoniae, serotype 25, required both complement and immunoglobulin for opsonization, even though that strain activated the alternative complement pathway. Both strains of Escherichia coli required antibody and complement, but varied in the degree of dependence on the C3 opsonin, since trypan blue moderately inhibited the killing of E. coli-1 and markedly inhibited the killing of E. coli-2. Serratia marcescens was opsonized in heat-inactivated serum (limited complement) or serum absorbed at 0 degrees C with S. marcescens (limited antibody), but depended on the C3 receptor in absorbed serum. S. marcescens activated the alternative pathway. Thus, opsonic requirements varied with the availability of opsonins. Requirements for bacterial opsonization vary with species and strains within species, perhaps reflecting quantitative relationships among alternative and classical pathway activation of C3, efficiency of adsorption of C3 or immunoglobulin G to bacterial surfaces, and efficiency of attachment of these ligands to polymorphonuclear leukocyte receptors. Furthermore, although not always sufficient for opsonization, the C3 opsonin (activated through either the classical or alternative pathway) appears necessary for effective phagocytosis and killing of all strains studied. | Evidence for quantitative variability of bacterial opsonic requirements. We studied human serum opsonins by using combinations of heat inactivation and chelation to inhibit complement, adsorption to remove antibody, and trypan blue to inactivate the C3 receptor of human polymorphonuclear leukocytes. Streptococcus pneumoniae, serotype 25, required both complement and immunoglobulin for opsonization, even though that strain activated the alternative complement pathway. Both strains of Escherichia coli required antibody and complement, but varied in the degree of dependence on the C3 opsonin, since trypan blue moderately inhibited the killing of E. coli-1 and markedly inhibited the killing of E. coli-2. Serratia marcescens was opsonized in heat-inactivated serum (limited complement) or serum absorbed at 0 degrees C with S. marcescens (limited antibody), but depended on the C3 receptor in absorbed serum. S. marcescens activated the alternative pathway. Thus, opsonic requirements varied with the availability of opsonins. Requirements for bacterial opsonization vary with species and strains within species, perhaps reflecting quantitative relationships among alternative and classical pathway activation of C3, efficiency of adsorption of C3 or immunoglobulin G to bacterial surfaces, and efficiency of attachment of these ligands to polymorphonuclear leukocyte receptors. Furthermore, although not always sufficient for opsonization, the C3 opsonin (activated through either the classical or alternative pathway) appears necessary for effective phagocytosis and killing of all strains studied. | [
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PMID:25244 | Interactions between effector cell activity and lymphokines: implications for recovery from herpesvirus infections. | The destruction of herpesvirus-infected target cells by antibody-dependent and direct cell cytotoxicity was enhanced by the presence of bovine lymphokine-containing preparations. To relate these effects to possible in vivo mechanisms of recovery, several in vitro approaches were used to measure the effects of lymphokine-containing preparations on controlling viral spread. In the first approach it was shown that in the presence of lymphokines, virus-infected cells could be killed earlier in the replication cycle by the mechanism of antibody-dependent cell cytotoxicity, thus possibly limiting spread of virus. That this was indeed the case was demonstrated by a decrease in the area of viral-induced cytopathology as well as in the total number of infected cells present. Secondly, the amount of infectious virus released was also markedly reduced in cultures incubated with lymphokines and immune peripheral blood lymphocytes as compared to cultures treated with either component alone. Finally, lymphokines caused the activation of macrophages. These results are discussed in terms of how various immune parameters may interact in a positive way so as to aid in the recovery from virus infection. | Interactions between effector cell activity and lymphokines: implications for recovery from herpesvirus infections. The destruction of herpesvirus-infected target cells by antibody-dependent and direct cell cytotoxicity was enhanced by the presence of bovine lymphokine-containing preparations. To relate these effects to possible in vivo mechanisms of recovery, several in vitro approaches were used to measure the effects of lymphokine-containing preparations on controlling viral spread. In the first approach it was shown that in the presence of lymphokines, virus-infected cells could be killed earlier in the replication cycle by the mechanism of antibody-dependent cell cytotoxicity, thus possibly limiting spread of virus. That this was indeed the case was demonstrated by a decrease in the area of viral-induced cytopathology as well as in the total number of infected cells present. Secondly, the amount of infectious virus released was also markedly reduced in cultures incubated with lymphokines and immune peripheral blood lymphocytes as compared to cultures treated with either component alone. Finally, lymphokines caused the activation of macrophages. These results are discussed in terms of how various immune parameters may interact in a positive way so as to aid in the recovery from virus infection. | [
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PMID:25245 | Synthesis and biological activity of ovine beta-lipotropin-(41--91)-henkaipentekontapeptide. | The 51-residue peptide ovine beta-lipotropin-(41--91) has been synthesized by the solid-phase method in about 5% overall yield. The synthetic product was characterized by partition chromatography on agarose gel, thin-layer chromatography in two solvent systems, paper electrophoresis at two pH values, polyacrylamide gel electrophoresis, amino acid analyses of acid and enzymic hydrolysates, and bioassay for lipolytic and melanotropic activities. The synthetic peptide is about 5.4 times as active on a weight basis as ovine beta-lipotropin in the lipolytic assay. In the melanotropic assay, it was about 2.4 times more active than the beta-lipotropin but only 5% as active as bovine beta-melanotropin. It had negligible opiate activity in the guinea pig ileum assay. | Synthesis and biological activity of ovine beta-lipotropin-(41--91)-henkaipentekontapeptide. The 51-residue peptide ovine beta-lipotropin-(41--91) has been synthesized by the solid-phase method in about 5% overall yield. The synthetic product was characterized by partition chromatography on agarose gel, thin-layer chromatography in two solvent systems, paper electrophoresis at two pH values, polyacrylamide gel electrophoresis, amino acid analyses of acid and enzymic hydrolysates, and bioassay for lipolytic and melanotropic activities. The synthetic peptide is about 5.4 times as active on a weight basis as ovine beta-lipotropin in the lipolytic assay. In the melanotropic assay, it was about 2.4 times more active than the beta-lipotropin but only 5% as active as bovine beta-melanotropin. It had negligible opiate activity in the guinea pig ileum assay. | [
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PMID:25248 | Glucose oxidation in the chick cornea: effect of diamide on the pentose shunt. | Chick embryo corneas (stages 38 and 45) have been used to study variations in pentose shunt activity following the use of a glutathione-specific oxidizing agent, diamide, and a sulfydryl blocking agent, N-ethylmaleimide (NEM). Shunt activity was measured by the ratio of radiolabeled carbon 1 (14C-1) of glucose to radiolabeled carbon 6 (14C-6) of glucose derived as expired 14CO2. Diamide and NEM were both found to increase pentose shunt activity relative to glycolysis, although by different means. Diamide appeared to exert its effect by oxidizing glutathione and creating a demand for higher shunt activity to facilitate glutathione reduction by NADPH. Both C-1 and C-6 oxidation were increased, but C-1 oxidation was increased to a much greater extent. In contrast, NEM decreased both C-1 and C-6 oxidation, with C-6 preferentially affected. Thus NEM appears to preferentially inhibit the enzymatic machinery of the glycolytic-tricarboxylic acid cycle pathway and acts as an effective metabolic stress on the cornea. Our data suggest that the pentose shunt in the cornea may serve as an important alternative pathway under conditions of metabolic stress for glucose utilization and the production of energy (ATP) in the corneal cells. | Glucose oxidation in the chick cornea: effect of diamide on the pentose shunt. Chick embryo corneas (stages 38 and 45) have been used to study variations in pentose shunt activity following the use of a glutathione-specific oxidizing agent, diamide, and a sulfydryl blocking agent, N-ethylmaleimide (NEM). Shunt activity was measured by the ratio of radiolabeled carbon 1 (14C-1) of glucose to radiolabeled carbon 6 (14C-6) of glucose derived as expired 14CO2. Diamide and NEM were both found to increase pentose shunt activity relative to glycolysis, although by different means. Diamide appeared to exert its effect by oxidizing glutathione and creating a demand for higher shunt activity to facilitate glutathione reduction by NADPH. Both C-1 and C-6 oxidation were increased, but C-1 oxidation was increased to a much greater extent. In contrast, NEM decreased both C-1 and C-6 oxidation, with C-6 preferentially affected. Thus NEM appears to preferentially inhibit the enzymatic machinery of the glycolytic-tricarboxylic acid cycle pathway and acts as an effective metabolic stress on the cornea. Our data suggest that the pentose shunt in the cornea may serve as an important alternative pathway under conditions of metabolic stress for glucose utilization and the production of energy (ATP) in the corneal cells. | [
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PMID:25256 | Blood gas changes during panting in a small East African antelope, the dik-dik. | Five adult male dik-dik (Madoqua kirkii) were exposed in a climatic chamber to an air temperature of 45 degrees C. Measurements were made of rectal temperature (Tre) and respiratory frequency (f) and arterial blood samples taken before and during heat exposure were analyzed for pH, PCO2 and PO2. During exposure, Tre and f increased in all animals. In the first 80 min dik-dik displayed thermal tachypnea and minor changes in blood gases. Continued exposure lead to hyperpnea accompanied by a fall in PaCO2 and a rise in pH. PaCO2 at first fell and then increased toward or above control levels. The dik-dik did not display second phase breathing. This observation confirms that second phase breathing is not essential to the development of respiratory alkalosis. The main conclusion of the study is that the dik-dik, unlike another heat-adapted antelope, the wildebeest (Taylor, Robertshaw, and Hoffmann. Am. J. Physiol. 217:907-910, 1969), is unable to resist alkalosis during heat stress. | Blood gas changes during panting in a small East African antelope, the dik-dik. Five adult male dik-dik (Madoqua kirkii) were exposed in a climatic chamber to an air temperature of 45 degrees C. Measurements were made of rectal temperature (Tre) and respiratory frequency (f) and arterial blood samples taken before and during heat exposure were analyzed for pH, PCO2 and PO2. During exposure, Tre and f increased in all animals. In the first 80 min dik-dik displayed thermal tachypnea and minor changes in blood gases. Continued exposure lead to hyperpnea accompanied by a fall in PaCO2 and a rise in pH. PaCO2 at first fell and then increased toward or above control levels. The dik-dik did not display second phase breathing. This observation confirms that second phase breathing is not essential to the development of respiratory alkalosis. The main conclusion of the study is that the dik-dik, unlike another heat-adapted antelope, the wildebeest (Taylor, Robertshaw, and Hoffmann. Am. J. Physiol. 217:907-910, 1969), is unable to resist alkalosis during heat stress. | [
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PMID:25260 | Alpha-l-arabinofuranosidase from Rhodotorula flava. | An alpha-L-arabinofuranosidase (EC 3.2.1.55) from the culture fluid of Rhodotorula flava IFO 0407 grown on beet arabinan as a carbon source has been highly purified. The purified enzyme has a pH optimum of 2.0. The enzyme is unusually acid stable, retaining 82% of its activity after being maintained for 24 h at pH 1.5 and at 30 degrees C. The apparent Km and Vmax values of the enzyme for phenyl alpha-L-arabinofuranoside were determined to be 9.1 mM and 72.5 mumol per min per mg of protein, respectively. | Alpha-l-arabinofuranosidase from Rhodotorula flava. An alpha-L-arabinofuranosidase (EC 3.2.1.55) from the culture fluid of Rhodotorula flava IFO 0407 grown on beet arabinan as a carbon source has been highly purified. The purified enzyme has a pH optimum of 2.0. The enzyme is unusually acid stable, retaining 82% of its activity after being maintained for 24 h at pH 1.5 and at 30 degrees C. The apparent Km and Vmax values of the enzyme for phenyl alpha-L-arabinofuranoside were determined to be 9.1 mM and 72.5 mumol per min per mg of protein, respectively. | [
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PMID:25261 | Protonmotive force and motility of Bacillus subtilis. | Motility of Bacillus subtilis was inhibited within a few minutes by a combination of valinomycin and a high concentration of potassium ions in the medium at neutral pH. Motility was restored by lowering the concentration of valinomycin or potassium ions. The valinomycin concentration necessary for motility inhibition was determined at various concentrations of potassium ions and various pH's. At pH 7.5, valinomycin of any concentration did not inhibit the motility, when the potassium ion concentration was lower than 9 mM. In the presence of 230 mM potassium ion, the motility inhibition by valinomycin was not detected at pH lower than 6.1. These results are easily explained by the idea that the motility of B. subtilis is supported by the electrochemical potential difference of the proton across the membrane, or the protonmotive force. The electrochemical potential difference necessary for motility was estimated to be about -90 mV. | Protonmotive force and motility of Bacillus subtilis. Motility of Bacillus subtilis was inhibited within a few minutes by a combination of valinomycin and a high concentration of potassium ions in the medium at neutral pH. Motility was restored by lowering the concentration of valinomycin or potassium ions. The valinomycin concentration necessary for motility inhibition was determined at various concentrations of potassium ions and various pH's. At pH 7.5, valinomycin of any concentration did not inhibit the motility, when the potassium ion concentration was lower than 9 mM. In the presence of 230 mM potassium ion, the motility inhibition by valinomycin was not detected at pH lower than 6.1. These results are easily explained by the idea that the motility of B. subtilis is supported by the electrochemical potential difference of the proton across the membrane, or the protonmotive force. The electrochemical potential difference necessary for motility was estimated to be about -90 mV. | [
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PMID:25262 | Fate of heterospecific transforming DNA bound to Streptococcus sanguis. | The fate of 3H-labeled str-r fus-s DNA from Streptococcus pneumoniae, bound after a 1-min uptake to 14C-labeled str-s fus-r S. sanguis recipients, was followed by techniques previously developed for analyzing the fate of homospecific DNA. Heterospecific S. pneumoniae DNA was bound and formed complexes with recipient protein in a manner similar to that of homospecific DNA but transformed relatively poorly. The rate at which complexed heterospecific DNA becomes physically associated with recipient DNA, and at which donor markers are integrated into the chromosome, was slower than in the case of homospecific DNA. In addition, about half of the heterospecific donor counts initially bound in trichloracetic acid-insoluble form were gradually solubilized and released from the cell. The association of heterospecific DNA with the recipient chromosome was more unstable than that involving homospecific DNA, since only associations of the former type were largely dissociated by isolation and resedimentation. The donor DNA-containing material so dissociated had the same sedimentation properties as complexed heterospecific DNA before association, indicating that the complex of single-stranded donor DNA and recipient protein formed on uptake moves as a whole from its site of formation to synapse with the chromosome. | Fate of heterospecific transforming DNA bound to Streptococcus sanguis. The fate of 3H-labeled str-r fus-s DNA from Streptococcus pneumoniae, bound after a 1-min uptake to 14C-labeled str-s fus-r S. sanguis recipients, was followed by techniques previously developed for analyzing the fate of homospecific DNA. Heterospecific S. pneumoniae DNA was bound and formed complexes with recipient protein in a manner similar to that of homospecific DNA but transformed relatively poorly. The rate at which complexed heterospecific DNA becomes physically associated with recipient DNA, and at which donor markers are integrated into the chromosome, was slower than in the case of homospecific DNA. In addition, about half of the heterospecific donor counts initially bound in trichloracetic acid-insoluble form were gradually solubilized and released from the cell. The association of heterospecific DNA with the recipient chromosome was more unstable than that involving homospecific DNA, since only associations of the former type were largely dissociated by isolation and resedimentation. The donor DNA-containing material so dissociated had the same sedimentation properties as complexed heterospecific DNA before association, indicating that the complex of single-stranded donor DNA and recipient protein formed on uptake moves as a whole from its site of formation to synapse with the chromosome. | [
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PMID:25263 | Cell-free biosynthesis of the O-acetylated N-acetylneuraminic acid capsular polysaccharide of group C meningococci. | A cell-free system was established to study the biosynthesis of group C meningococcal capsular polysaccharide, an alpha-2 leads to 9-linked N-acetylneuraminic acid (NeuAc) homopolymer containing O-acetyl groups at either C7 or C8. Sialyltransferase activity, isolated from group C meningococcus strain C-11, catalyzed incorporation of [14C]NeuAc from CMP (CMP--[14C]NeuAc) into polymeric form. This sialyltransferase was stimulated by addition of meningococcus group C and Escherichia coli K92 capsular polysaccharides, the latter being an alpha-2 leads to 8- and alpha-2 leads to 9-linked NeuAc heteropolymer. Group C meningococcal sialyltransferase did not require divalent ions but was stimulated by Mn2+. Attempts to demonstrate a lipid-soluble intermediate in the biosynthesis of this NeuAc polymer were unsuccessful. Meningococcal group C sialyltransferase incorporated NeuAc into a membrane-associated product. The polysaccharide can be extracted from the membrane-bound fraction with Triton X-100. The newly synthesized polysaccharide coprecipitates with authentic group C antigen in meningococcal group C antiserum and is degraded by sodium metaperiodate, indicating that the NeuAc polymer synthesized by the cell-free system consists of alpha-2 leads to 9 linkage. Meningococcal group C spheroplast membranes contain an O-acetylase that can catalyze the transfer of acetyl groups from acetyl coenzyme A to the in vitro-synthesized polysaccharide. | Cell-free biosynthesis of the O-acetylated N-acetylneuraminic acid capsular polysaccharide of group C meningococci. A cell-free system was established to study the biosynthesis of group C meningococcal capsular polysaccharide, an alpha-2 leads to 9-linked N-acetylneuraminic acid (NeuAc) homopolymer containing O-acetyl groups at either C7 or C8. Sialyltransferase activity, isolated from group C meningococcus strain C-11, catalyzed incorporation of [14C]NeuAc from CMP (CMP--[14C]NeuAc) into polymeric form. This sialyltransferase was stimulated by addition of meningococcus group C and Escherichia coli K92 capsular polysaccharides, the latter being an alpha-2 leads to 8- and alpha-2 leads to 9-linked NeuAc heteropolymer. Group C meningococcal sialyltransferase did not require divalent ions but was stimulated by Mn2+. Attempts to demonstrate a lipid-soluble intermediate in the biosynthesis of this NeuAc polymer were unsuccessful. Meningococcal group C sialyltransferase incorporated NeuAc into a membrane-associated product. The polysaccharide can be extracted from the membrane-bound fraction with Triton X-100. The newly synthesized polysaccharide coprecipitates with authentic group C antigen in meningococcal group C antiserum and is degraded by sodium metaperiodate, indicating that the NeuAc polymer synthesized by the cell-free system consists of alpha-2 leads to 9 linkage. Meningococcal group C spheroplast membranes contain an O-acetylase that can catalyze the transfer of acetyl groups from acetyl coenzyme A to the in vitro-synthesized polysaccharide. | [
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PMID:25264 | Involvement of the product of the glnF gene in the autogenous regulation of glutamine synthetase formation in Klebsiella aerogenes. | Mutations in a site, glnF, linked by P1-mediated transduction of argG on the chromosome of Klebsiella aerogenes, result in a requirement for glutamine. Mutants in this gene have in all media a level of glutamine synthetase (GS) corresponding to the level found in the wild-type strain grown in the medium producing the strongest repression of GS. The adenylylation and deadenylylation of GS in glnF mutants is normal. The glutamine requirement of glnF mutants could be suppressed by mutations in the structural gene for GS, glnA. These mutations result in altered regulation of GS synthesis, regardless of the presence or absence of the glnF mutation (GlnR phenotype). In GlnR mutants the GS level is higher than in the wild-type strain when the cells are cultured in strongly repressing medium, but lower than in the wild-type strain when cells are cultured in a derepressing medium. Heterozygous merodiploids carrying a normal glnA gene as well as a glnA gene responsible for the GlnR phenotype behave in every respect like merodiploids carrying two normal glnA genes. These results confirm autogenous regulation of GS synthesis and indicate that GS is both a repressor and an activator of GS synthesis. The mutation in glnA responsible for the GLnR phenotype has apparently resulted in the formation of a GS that is incompetent both as repressor and as activator of GS synthesis. According to this hypothesis, the product of the glnF gene is necessary for activation of the glnA gene by GS. | Involvement of the product of the glnF gene in the autogenous regulation of glutamine synthetase formation in Klebsiella aerogenes. Mutations in a site, glnF, linked by P1-mediated transduction of argG on the chromosome of Klebsiella aerogenes, result in a requirement for glutamine. Mutants in this gene have in all media a level of glutamine synthetase (GS) corresponding to the level found in the wild-type strain grown in the medium producing the strongest repression of GS. The adenylylation and deadenylylation of GS in glnF mutants is normal. The glutamine requirement of glnF mutants could be suppressed by mutations in the structural gene for GS, glnA. These mutations result in altered regulation of GS synthesis, regardless of the presence or absence of the glnF mutation (GlnR phenotype). In GlnR mutants the GS level is higher than in the wild-type strain when the cells are cultured in strongly repressing medium, but lower than in the wild-type strain when cells are cultured in a derepressing medium. Heterozygous merodiploids carrying a normal glnA gene as well as a glnA gene responsible for the GlnR phenotype behave in every respect like merodiploids carrying two normal glnA genes. These results confirm autogenous regulation of GS synthesis and indicate that GS is both a repressor and an activator of GS synthesis. The mutation in glnA responsible for the GLnR phenotype has apparently resulted in the formation of a GS that is incompetent both as repressor and as activator of GS synthesis. According to this hypothesis, the product of the glnF gene is necessary for activation of the glnA gene by GS. | [
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PMID:25265 | Glycerolipid biosynthesis in Saccharomyces cerevisiae: sn-glycerol-3-phosphate and dihydroxyacetone phosphate acyltransferase activities. | Yeast acyl-coenzyme A:dihydroxyacetone-phosphate O-acyltransferase (DHAP acyltransferase; EC 2.3.1.42) was investigated to (i) determine whether its activity and that of acyl-coenzyme A:sn-glycerol-3-phosphate O-acyltransferase (glycerol-P acyltransferase; EC 2.3.1.15) represent dual catalytic functions of a single membranous enzyme, (ii) estimate the relative contributions of the glycerol-P and DHAP pathways for yeast glycerolipid synthesis, and (iii) evaluate the suitability of yeast for future genetic investigations of the eucaryotic glycerol-P and DHAP acyltransferase activities. The membranous DHAP acyltransferase activity showed an apparent Km of 0.79 mM for DHAP, with a Vmax of 5.3 nmol/min per mg, whereas the glycerol-P acyltransferase activity showed an apparent Km of 0.05 mM for glycerol-P, with a Vmax of 3.4 nmol/min per mg. Glycerol-P was a competitive inhibitor (Ki, 0.07 mM) of the DHAP acyltransferase activity, and DHAP was a competitive inhibitor (Ki, 0.91 mM) of the glycerol-P acyltransferase activity. The two acyltransferase activities exhibited marked similarities in their pH dependence, acyl-coenzyme A chain length preference and substrate concentration dependencies, thermolability, and patterns of inactivation by N-ethylmaleimide, trypsin, and detergents. Thus, the data strongly suggest that yeast glycerol-P and DHAP acyltransferase activities represent dual catalytic functions of a single membrane-bound enzyme. Furthermore, since no acyl-DHAP oxidoreductase activity could be detected in yeast membranes, the DHAP pathway for glycerolipid synthesis may not operate in yeast. | Glycerolipid biosynthesis in Saccharomyces cerevisiae: sn-glycerol-3-phosphate and dihydroxyacetone phosphate acyltransferase activities. Yeast acyl-coenzyme A:dihydroxyacetone-phosphate O-acyltransferase (DHAP acyltransferase; EC 2.3.1.42) was investigated to (i) determine whether its activity and that of acyl-coenzyme A:sn-glycerol-3-phosphate O-acyltransferase (glycerol-P acyltransferase; EC 2.3.1.15) represent dual catalytic functions of a single membranous enzyme, (ii) estimate the relative contributions of the glycerol-P and DHAP pathways for yeast glycerolipid synthesis, and (iii) evaluate the suitability of yeast for future genetic investigations of the eucaryotic glycerol-P and DHAP acyltransferase activities. The membranous DHAP acyltransferase activity showed an apparent Km of 0.79 mM for DHAP, with a Vmax of 5.3 nmol/min per mg, whereas the glycerol-P acyltransferase activity showed an apparent Km of 0.05 mM for glycerol-P, with a Vmax of 3.4 nmol/min per mg. Glycerol-P was a competitive inhibitor (Ki, 0.07 mM) of the DHAP acyltransferase activity, and DHAP was a competitive inhibitor (Ki, 0.91 mM) of the glycerol-P acyltransferase activity. The two acyltransferase activities exhibited marked similarities in their pH dependence, acyl-coenzyme A chain length preference and substrate concentration dependencies, thermolability, and patterns of inactivation by N-ethylmaleimide, trypsin, and detergents. Thus, the data strongly suggest that yeast glycerol-P and DHAP acyltransferase activities represent dual catalytic functions of a single membrane-bound enzyme. Furthermore, since no acyl-DHAP oxidoreductase activity could be detected in yeast membranes, the DHAP pathway for glycerolipid synthesis may not operate in yeast. | [
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PMID:25266 | Intracellular serine protease of Bacillus subtilis: sequence homology with extracellular subtilisins. | Intracellular serine protease was isolated from stationary-grown Bacillus subtilis A-50 cells and purified to homogeneity. The molecular weight of the enzyme is 31,000 +/- 1,000, with an isoelectric point of 4.3. Its amino acid composition is characteristically enriched in glutamic acid content, differing from that of extra-cellular subtilisins. The enzyme is completely inhibited with phenylmethylsulfonyl fluoride and ethylenediaminetetraacetic acid. Intracellular protease possesses negligible activity towards bovine serum albumin and hemoglobin, but has 5- to 20-fold higher specific activity against p-nitroanilides of benzyloxycarbonyl tripeptides than subtilisin BPN'. Esterolytic activity of the enzyme is also higher than that of subtilisin BPN'. The enzyme is sequence homologous with secretory subtilisins throughout 50 determined NH2-terminal residues, indicating the presence of duplicated structural genes for serine proteases in the B. subtilis genome. The occurrence of two homologous genes in the cell might accelerate the evolution of serine protease not only by the loosening of selective constrainst, but also by creation of sequence variants by means of intragenic recombination. Three molecular forms of intracellular protease were found, two of them with NH2-terminal glutamic acid and one minor form, three residues longer, with asparagine as NH2 terminus. These data indicate the possible presence of an enzyme precursor proteolytically modified during cell growth. | Intracellular serine protease of Bacillus subtilis: sequence homology with extracellular subtilisins. Intracellular serine protease was isolated from stationary-grown Bacillus subtilis A-50 cells and purified to homogeneity. The molecular weight of the enzyme is 31,000 +/- 1,000, with an isoelectric point of 4.3. Its amino acid composition is characteristically enriched in glutamic acid content, differing from that of extra-cellular subtilisins. The enzyme is completely inhibited with phenylmethylsulfonyl fluoride and ethylenediaminetetraacetic acid. Intracellular protease possesses negligible activity towards bovine serum albumin and hemoglobin, but has 5- to 20-fold higher specific activity against p-nitroanilides of benzyloxycarbonyl tripeptides than subtilisin BPN'. Esterolytic activity of the enzyme is also higher than that of subtilisin BPN'. The enzyme is sequence homologous with secretory subtilisins throughout 50 determined NH2-terminal residues, indicating the presence of duplicated structural genes for serine proteases in the B. subtilis genome. The occurrence of two homologous genes in the cell might accelerate the evolution of serine protease not only by the loosening of selective constrainst, but also by creation of sequence variants by means of intragenic recombination. Three molecular forms of intracellular protease were found, two of them with NH2-terminal glutamic acid and one minor form, three residues longer, with asparagine as NH2 terminus. These data indicate the possible presence of an enzyme precursor proteolytically modified during cell growth. | [
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PMID:25267 | Acid proteases. II. Fluorescence study of the interaction of Cladosporium acid protease with glycyl-DL-norleucine methyl ester in the presence of cupric ions. | Glycyl-DL-norleucine methyl ester (GN), a diazoacetyl-DL-norleucine methyl ester (DAN) analog, in the presence of cupric ions was found to partially quench the protein fluorescence of acid protease from Cladosporium sp. No. 45-2, and cupric ions were also found to quench the fluorescence. These quenchings were pH-dependent. GN alone did not quench the fluorescence of the enzyme. The interaction between the enzyme and GN in the presence of cupric ions was studied statically at pH 5.4 in terms of fluorescence change. The dissociation constant, Kd, of the enzyme-GN complex in the presence of a 20-fold molar excess of cupric ions (0.08 mM) determined by fluorescence titration at 30 degrees C (Kd = 1.86 mM) was in good agreement with that obtained for GN from kinetics of inhibition of DAN-induced inactivation in the presence of a 20-fold molar excess of cupric ions at 30 degrees C (KA = 1.94 mM) (Kanazawa, H. (1977) J. Biochem. 81, 1739-1744). At various concentrations of cupric ions, no change of Kd was found. These results suggest that cupric ions are attracted to a negatively charged carboxyl group responsible for the formation of the enzyme-GN complex. | Acid proteases. II. Fluorescence study of the interaction of Cladosporium acid protease with glycyl-DL-norleucine methyl ester in the presence of cupric ions. Glycyl-DL-norleucine methyl ester (GN), a diazoacetyl-DL-norleucine methyl ester (DAN) analog, in the presence of cupric ions was found to partially quench the protein fluorescence of acid protease from Cladosporium sp. No. 45-2, and cupric ions were also found to quench the fluorescence. These quenchings were pH-dependent. GN alone did not quench the fluorescence of the enzyme. The interaction between the enzyme and GN in the presence of cupric ions was studied statically at pH 5.4 in terms of fluorescence change. The dissociation constant, Kd, of the enzyme-GN complex in the presence of a 20-fold molar excess of cupric ions (0.08 mM) determined by fluorescence titration at 30 degrees C (Kd = 1.86 mM) was in good agreement with that obtained for GN from kinetics of inhibition of DAN-induced inactivation in the presence of a 20-fold molar excess of cupric ions at 30 degrees C (KA = 1.94 mM) (Kanazawa, H. (1977) J. Biochem. 81, 1739-1744). At various concentrations of cupric ions, no change of Kd was found. These results suggest that cupric ions are attracted to a negatively charged carboxyl group responsible for the formation of the enzyme-GN complex. | [
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PMID:25268 | Purification and some properties of a neutral muscle pyrophosphatase. | In the water-soluble fraction of rabbit skeletal muscle, at least two types of inorganic pyro phosphatase (PPase) are distinguishable on ion exchange column chromatography. One of them, pyrophosphatase-A (PPase-A), was isolated in an electrophoretically homogeneous form. This enzyme catalyzed the hydrolysis of PPi but not that of other phosphate esters. Only Mg2+ was required for activity and stability. Other cations such as Ca2+, Co2+, Mn2+, and Zn2+ had no activating effect. The activity of this PPase was optimum at pH 7.4. ATP, ADP, sodium imidodiphosphate (PNP), p-chloromercuribenzoate, and Ca2+ inhibited its enzymic activity. The enzyme was protected by dithiothreitol (DTT) against heat denaturation. The molecular weight was estimated to be 67,000 by gel filtration and the molecular size of the subunit was found to be 35,000 by gel electrophoresis in the presence of sodium dodecyl sulfate (SDS). The enzyme probably consists of two identical subunits of 35,000 daltons. | Purification and some properties of a neutral muscle pyrophosphatase. In the water-soluble fraction of rabbit skeletal muscle, at least two types of inorganic pyro phosphatase (PPase) are distinguishable on ion exchange column chromatography. One of them, pyrophosphatase-A (PPase-A), was isolated in an electrophoretically homogeneous form. This enzyme catalyzed the hydrolysis of PPi but not that of other phosphate esters. Only Mg2+ was required for activity and stability. Other cations such as Ca2+, Co2+, Mn2+, and Zn2+ had no activating effect. The activity of this PPase was optimum at pH 7.4. ATP, ADP, sodium imidodiphosphate (PNP), p-chloromercuribenzoate, and Ca2+ inhibited its enzymic activity. The enzyme was protected by dithiothreitol (DTT) against heat denaturation. The molecular weight was estimated to be 67,000 by gel filtration and the molecular size of the subunit was found to be 35,000 by gel electrophoresis in the presence of sodium dodecyl sulfate (SDS). The enzyme probably consists of two identical subunits of 35,000 daltons. | [
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PMID:25269 | Bacteriolytic enzyme induced from pyocinogenic Pseudomonas aeruginosa. Purification and characterization of PR1-lysozyme. | A bacteriolytic enzyme, PR1-lysozyme, has been purified from the lysate of mitomycin C-induced pyocinogenic Pseudomonas aeruginosa, by acrinol treatment, Amberlite CG-50 chromatography, ammonium sulfate fractionation, Sephadex G-100 gel filtration and two cycles of SP-Sephadex C-50 chromatography. Homogeneity of the preparation was demonstrated by three electrophoretic techniques. PR1-lysozyme is a basic protein (pI, 9.4) and consists of a single polypeptide chain having a molecular weight of 24,000. The amino acid composition of the protein was analyzed, and no cystein residue was found among more than 210 amino acid residues. The optimum pH for enzymatic activity was 6.4 and the enzyme exhibited about 50 to 70 times greater specific activity than hen egg-white lysozyme when assayed with chloroform-killed P. aeruginosa as a substrate. By analyzing the products of enzymatic action on purified peptidoglycan of P. aeruginosa, the enzyme was identified as an N-acetylmuramidase, i.e., the same classification as hen-egg-white lysozyme. PR1-lysozyme did not show any activity towards intact cells of gram-positive and gram-negative bacteria tested. However, the enzyme was able to lyse chloroform-killed gram-negative and gram-positive bacteria. | Bacteriolytic enzyme induced from pyocinogenic Pseudomonas aeruginosa. Purification and characterization of PR1-lysozyme. A bacteriolytic enzyme, PR1-lysozyme, has been purified from the lysate of mitomycin C-induced pyocinogenic Pseudomonas aeruginosa, by acrinol treatment, Amberlite CG-50 chromatography, ammonium sulfate fractionation, Sephadex G-100 gel filtration and two cycles of SP-Sephadex C-50 chromatography. Homogeneity of the preparation was demonstrated by three electrophoretic techniques. PR1-lysozyme is a basic protein (pI, 9.4) and consists of a single polypeptide chain having a molecular weight of 24,000. The amino acid composition of the protein was analyzed, and no cystein residue was found among more than 210 amino acid residues. The optimum pH for enzymatic activity was 6.4 and the enzyme exhibited about 50 to 70 times greater specific activity than hen egg-white lysozyme when assayed with chloroform-killed P. aeruginosa as a substrate. By analyzing the products of enzymatic action on purified peptidoglycan of P. aeruginosa, the enzyme was identified as an N-acetylmuramidase, i.e., the same classification as hen-egg-white lysozyme. PR1-lysozyme did not show any activity towards intact cells of gram-positive and gram-negative bacteria tested. However, the enzyme was able to lyse chloroform-killed gram-negative and gram-positive bacteria. | [
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PMID:25270 | Studies on soybean trypsin inhibitors. XIII. Preparation and characterization of active fragments from Bowman-Birk proteinase inhibitor. | Soybean Bowman-Birk inhibitor, a double-headed inhibitor of trypsin and alpha-chymotrypsin, was treated with cyanogen bromide and then pepsin to yield two inhibitory active fragments. Structural investigation showed that one of the fragments was derived from the trypsin inhibitory domain and the other from the chymotrypsin inhibitory domain of the inhibitor. In contrast to the unusual stability of the native inhibitor, the separated domains were less stable and could be inactivated with excess proteinases. These results suggest that the legume double-headed inhibitors acquired their unusual stability by duplicating an ancestral single-headed structure. | Studies on soybean trypsin inhibitors. XIII. Preparation and characterization of active fragments from Bowman-Birk proteinase inhibitor. Soybean Bowman-Birk inhibitor, a double-headed inhibitor of trypsin and alpha-chymotrypsin, was treated with cyanogen bromide and then pepsin to yield two inhibitory active fragments. Structural investigation showed that one of the fragments was derived from the trypsin inhibitory domain and the other from the chymotrypsin inhibitory domain of the inhibitor. In contrast to the unusual stability of the native inhibitor, the separated domains were less stable and could be inactivated with excess proteinases. These results suggest that the legume double-headed inhibitors acquired their unusual stability by duplicating an ancestral single-headed structure. | [
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PMID:25271 | A new affinity adsorbent for guanyloribonuclease. Guanylyl-(2'-5')-guanosine coupled to aminohexyl-Sepharose. | Guanylyl-(2'-5')-guanosine binds to RNase T1 in 1:1 stoichiometry with a dissociation constant of 0.22 mM at pH 5.0 and 25 degrees C. This nucleotide, coupled to aminohexyl-Sepharose 4B, is able to serve as an affinity adsorbent for guanyloribonuclease [EC 3.1.4.8]. The strength of interaction between the adsorbent and various guanyloribonucleases at pH 5.0 was found to decrease in the following order: RNase N1 greater than RNase F1 greater than RNase T1 greater than RNase St. The bound enzymes can be released from the adsorbent either by increase of ionic strength or by increasing the pH from 5.0 to 7.5. The interaction between RNase T1 and the adsorbent is weakened by the presence of a low concentration of 2', 3'-, or 5'-GMP, which are competitive inhibitors of the enzyme. RNase F1 was purified to homogeneity by use of this affinity adsorbent. | A new affinity adsorbent for guanyloribonuclease. Guanylyl-(2'-5')-guanosine coupled to aminohexyl-Sepharose. Guanylyl-(2'-5')-guanosine binds to RNase T1 in 1:1 stoichiometry with a dissociation constant of 0.22 mM at pH 5.0 and 25 degrees C. This nucleotide, coupled to aminohexyl-Sepharose 4B, is able to serve as an affinity adsorbent for guanyloribonuclease [EC 3.1.4.8]. The strength of interaction between the adsorbent and various guanyloribonucleases at pH 5.0 was found to decrease in the following order: RNase N1 greater than RNase F1 greater than RNase T1 greater than RNase St. The bound enzymes can be released from the adsorbent either by increase of ionic strength or by increasing the pH from 5.0 to 7.5. The interaction between RNase T1 and the adsorbent is weakened by the presence of a low concentration of 2', 3'-, or 5'-GMP, which are competitive inhibitors of the enzyme. RNase F1 was purified to homogeneity by use of this affinity adsorbent. | [
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PMID:25274 | Purification and characterization of lysozyme produced by Streptomyces erythraeus. | A species of lysozyme (SE lysozyme) was purified from culture filtrate of Streptomyces erythraeus. The enzyme has a molecular weight of 18,500 as determined by ultracentrifugation. Its isoelectric point is 9.5, and it shows optimal activity at pH 4.0 with an optimal ionic strength of 0.1. Investigation of the substrate specificity showed SE lysozyme to be an N-acetyl-muramidase. The simplest product in the digest of cell walls of Micrococcus lysodeikticus was identified as a disaccharide, [GlcNAcbeta(1 leads to 4) MurNAc]. While S. aureus as well as M. lysodeikticus was lysed by this lysozyme, chitin and its derivatives were not. | Purification and characterization of lysozyme produced by Streptomyces erythraeus. A species of lysozyme (SE lysozyme) was purified from culture filtrate of Streptomyces erythraeus. The enzyme has a molecular weight of 18,500 as determined by ultracentrifugation. Its isoelectric point is 9.5, and it shows optimal activity at pH 4.0 with an optimal ionic strength of 0.1. Investigation of the substrate specificity showed SE lysozyme to be an N-acetyl-muramidase. The simplest product in the digest of cell walls of Micrococcus lysodeikticus was identified as a disaccharide, [GlcNAcbeta(1 leads to 4) MurNAc]. While S. aureus as well as M. lysodeikticus was lysed by this lysozyme, chitin and its derivatives were not. | [
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PMID:25275 | Induction of hepatic tyrosine aminotransferase mRNA by protein synthesis inhibitors. | Several protein synthesis inhibitors were as effective as the inducers hydrocortisone or cyclic AMP in elevating rat liver tyrosine aminotransferase mRNA levels when assayed in the wheat germ cell-free translational system. Cycloheximide, emetine, or puromycin increased this mRNA activity 6- to 7-fold within 4 h after in vivo administration. No increase in total hepatic mRNA levels or tryptophan oxygenase mRNA was found after treatment with these protein synthesis inhibitors. Furthermesults suggest that a short lived protein may specifically regulate the level of functional hepatic tyrosine aminotransferase mRNA or that ongoing translation of this mRNA is required for its degradation. | Induction of hepatic tyrosine aminotransferase mRNA by protein synthesis inhibitors. Several protein synthesis inhibitors were as effective as the inducers hydrocortisone or cyclic AMP in elevating rat liver tyrosine aminotransferase mRNA levels when assayed in the wheat germ cell-free translational system. Cycloheximide, emetine, or puromycin increased this mRNA activity 6- to 7-fold within 4 h after in vivo administration. No increase in total hepatic mRNA levels or tryptophan oxygenase mRNA was found after treatment with these protein synthesis inhibitors. Furthermesults suggest that a short lived protein may specifically regulate the level of functional hepatic tyrosine aminotransferase mRNA or that ongoing translation of this mRNA is required for its degradation. | [
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PMID:25282 | The role of Ca2+ on pH-induced hydrodynamic changes of bovine pancreatic deoxyribonuclease A. | DNase A studied by gel filtration on Sephadex G-100 at pH 7.4 in 40 mM Tris-HCl buffer, behaves hydrodynamically as a spherical monomeric macromolecule of around 31,000 molecular weight, with a Stokes radius = 24.7 A, f/fo = 1.19, and D20,W = 8.69. Similar results were obtained by analytical dialysis using zinc chloride-modified cellophane membranes. The elution volume of DNase A decreases as the pH increases between pH 4.7 and pH 9.5. This effect has been attributed to a change in the tridimensional structure of the protein and interpreted as a modification in the axial ratio due to unfolding of the polypeptide chain with increase in the apparent Stokes radius. The addition of Ca2+ produce reversion of the pH-induced changes at pH 9.5. The transition occurs when Ca2+ binds to at least two binding sites (n = 1.66 in a Hill plot) with a Kd = 8.9 X 10(-5) M and the effect appears to be cooperative. These findings support the hypothesis that Ca2+-binding to DNase A causes a conformational change that maintains a more active structure of the enzyme, especially when the pH-induced unfolding reduces its activity. | The role of Ca2+ on pH-induced hydrodynamic changes of bovine pancreatic deoxyribonuclease A. DNase A studied by gel filtration on Sephadex G-100 at pH 7.4 in 40 mM Tris-HCl buffer, behaves hydrodynamically as a spherical monomeric macromolecule of around 31,000 molecular weight, with a Stokes radius = 24.7 A, f/fo = 1.19, and D20,W = 8.69. Similar results were obtained by analytical dialysis using zinc chloride-modified cellophane membranes. The elution volume of DNase A decreases as the pH increases between pH 4.7 and pH 9.5. This effect has been attributed to a change in the tridimensional structure of the protein and interpreted as a modification in the axial ratio due to unfolding of the polypeptide chain with increase in the apparent Stokes radius. The addition of Ca2+ produce reversion of the pH-induced changes at pH 9.5. The transition occurs when Ca2+ binds to at least two binding sites (n = 1.66 in a Hill plot) with a Kd = 8.9 X 10(-5) M and the effect appears to be cooperative. These findings support the hypothesis that Ca2+-binding to DNase A causes a conformational change that maintains a more active structure of the enzyme, especially when the pH-induced unfolding reduces its activity. | [
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PMID:25283 | Energy transduction in the photosynthetic membranes of the cyanobacterium (blue-green alga) P-lectonema boryanum. | Membrane preparations isolated from the photosynthetic lamellae of the cyanobacterium Plectonema boryanum generate upon illumination a transmembrane pH gradient of approximately 2 to 3 pH units (acid inside), as determined from the distribution of either fluorescent or radioactive amines (9 aminoacridine and [14C]methylamine, respectively). Using the distribution of permeant ions to measure the electrical potential across the membrane, it was found that the latter is practically nil under conditions in which the deltapH is formed and photophosphorylation takes place. In agreement with the above findings cyclic photophosphorylation in this membrane preparation is inhibited by agents shown to collapse the deltapH but not by agents which should collapse the electrical potential. It is deduced that the pattern of proton movement in the photosynthetic lamellae of intact Plectonema spheroplasts corresponds to that of the cell-free membrane system, as both preparations show similar light dependent accumulation of fluorescent amine. It is concluded that the pattern of energy transduction in Plectonema photosynthetic lamellae is similar to that of chloroplast thylakoid membranes and not to that of bacterial cytoplasmic membranes. The evolutionary implications of the findings are discussed and a model for the directionality of H+ movements in the whole cell is presented. | Energy transduction in the photosynthetic membranes of the cyanobacterium (blue-green alga) P-lectonema boryanum. Membrane preparations isolated from the photosynthetic lamellae of the cyanobacterium Plectonema boryanum generate upon illumination a transmembrane pH gradient of approximately 2 to 3 pH units (acid inside), as determined from the distribution of either fluorescent or radioactive amines (9 aminoacridine and [14C]methylamine, respectively). Using the distribution of permeant ions to measure the electrical potential across the membrane, it was found that the latter is practically nil under conditions in which the deltapH is formed and photophosphorylation takes place. In agreement with the above findings cyclic photophosphorylation in this membrane preparation is inhibited by agents shown to collapse the deltapH but not by agents which should collapse the electrical potential. It is deduced that the pattern of proton movement in the photosynthetic lamellae of intact Plectonema spheroplasts corresponds to that of the cell-free membrane system, as both preparations show similar light dependent accumulation of fluorescent amine. It is concluded that the pattern of energy transduction in Plectonema photosynthetic lamellae is similar to that of chloroplast thylakoid membranes and not to that of bacterial cytoplasmic membranes. The evolutionary implications of the findings are discussed and a model for the directionality of H+ movements in the whole cell is presented. | [
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PMID:25284 | The transport of L-alanine by the hamster kidney cell line BHK-21-C13. | The uptake of L-alanine into BHK21-C13 cells in culture has been studied. This amino acid appears to be transported essentially via a relatively low affinity, high capacity, sodium ion dependent transport system. Inhibition studies using other amino acids or their analogues provided information about the specificity of this system. This alanine transport system was shown to exhibit a broad substrate specificity and appeared to be capable of transporting most naturally occurring neutral alpha-amino acids. Kinetic studies of the inhibition of L-alanine uptake also indicated the presence of a second neutral amino acid transport system capable of transporting this amino acid. However, it is unlikely that this second uptake system contributes greatly to L-alanine uptake. Inhibition of the uptake of L-leucine indicated that this transport system has a similar specificity to the "L"-system initially described for Ehrlich ascites carcinoma cells. | The transport of L-alanine by the hamster kidney cell line BHK-21-C13. The uptake of L-alanine into BHK21-C13 cells in culture has been studied. This amino acid appears to be transported essentially via a relatively low affinity, high capacity, sodium ion dependent transport system. Inhibition studies using other amino acids or their analogues provided information about the specificity of this system. This alanine transport system was shown to exhibit a broad substrate specificity and appeared to be capable of transporting most naturally occurring neutral alpha-amino acids. Kinetic studies of the inhibition of L-alanine uptake also indicated the presence of a second neutral amino acid transport system capable of transporting this amino acid. However, it is unlikely that this second uptake system contributes greatly to L-alanine uptake. Inhibition of the uptake of L-leucine indicated that this transport system has a similar specificity to the "L"-system initially described for Ehrlich ascites carcinoma cells. | [
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PMID:25285 | Coordinacy of lysosomal enzyme excretion in human urine. | Assay conditions have been developed for the determination of urinary beta-glucuronidase, beta-galactosidase, alpha-galactosidase, and beta-hexosaminidase using fluorometric substrates. The assay conditions for beta-glucuronidase overcome interference by both low and high molecular weight inhibitors, a problem that has confused earlier studies of enzyme excretion. The four lysosomal enzymes are excreted corrdinately: although their absolute levels (in units per milligram of creatinine) vary during the day and from one day to the next, the ratio of one enzyme to another remains relatively constant. The lack of correlation betweem plasma and urine enzyme levels, together with the high molecular weights of these enzymes, suggests that the urinary enzymes are not derived by glomerular filtration. The lack of coordinacy with lactate dehydrogenase suggests they are not derived from exfoliated cells. by analogy with experimental animals, they may be derived from lysosomes extruded into the lumen of the proximal tubule by epithelial cells. There is considerable variation among a population of 125 healthy adult subjects for total enzyme excretion. Both total enzyme excretion and coordinacy ratios are log-normally distributed, suggesting that they are the resultants of many factors, each of which has a relative, or proportional, effect on enzyme excretion. About one-half the population variation resides in a process common to the excretion of all four enzymes (possibly the lysosome extrusion pathway), and about one-half resides in factors affecting each enzyme independently. | Coordinacy of lysosomal enzyme excretion in human urine. Assay conditions have been developed for the determination of urinary beta-glucuronidase, beta-galactosidase, alpha-galactosidase, and beta-hexosaminidase using fluorometric substrates. The assay conditions for beta-glucuronidase overcome interference by both low and high molecular weight inhibitors, a problem that has confused earlier studies of enzyme excretion. The four lysosomal enzymes are excreted corrdinately: although their absolute levels (in units per milligram of creatinine) vary during the day and from one day to the next, the ratio of one enzyme to another remains relatively constant. The lack of correlation betweem plasma and urine enzyme levels, together with the high molecular weights of these enzymes, suggests that the urinary enzymes are not derived by glomerular filtration. The lack of coordinacy with lactate dehydrogenase suggests they are not derived from exfoliated cells. by analogy with experimental animals, they may be derived from lysosomes extruded into the lumen of the proximal tubule by epithelial cells. There is considerable variation among a population of 125 healthy adult subjects for total enzyme excretion. Both total enzyme excretion and coordinacy ratios are log-normally distributed, suggesting that they are the resultants of many factors, each of which has a relative, or proportional, effect on enzyme excretion. About one-half the population variation resides in a process common to the excretion of all four enzymes (possibly the lysosome extrusion pathway), and about one-half resides in factors affecting each enzyme independently. | [
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PMID:25286 | Diminished spectrin extraction from ATP-depleted human erythrocytes. Evidence relating spectrin to changes in erythrocyte shape and deformability. | We measured spectrin "extractability" in erythrocytes which were metabolically depleted by incubation at 37 degrees C in plasma or glucose-free buffers. Membranes were extracted with 1 mM EDTA (pH 8, 40 h, 4 degrees C) and analyzed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. This procedure solubilized 85--90% of the spectrin, actin, and residual hemoglobin from ghosts of fresh erythrocytes. In incubated erythrocytes, inextractable spectrin rapidly accumulated when ATP concentrations fell below 0--15% of normal. In severely depleted cells, 60--90% of the total ghost spectrin became inextractable. Inextractability was not abolished by physically disrupting the ghost before extraction, but was reversed when erythrocyte ATP was replenished with adenosine. The accumulation of inextractable spectrin correlated temporally with the increase in apparent membrane deformability and the increases in erythrocyte vicosity, calcium content, sodium gain, and potassium loss characteristic of ATP-depleted erythrocytes. No change in integral membrane protein topography (assessed by the distribution of intramembranous particles and concanavalin A surface-binding sites) was detected in depleted cells. Analogous changes were observed in erythrocytes exposed to extremes of pH and temperature. When the pH in the erythrocyte interior fell below 5.5, a pH where spectrin was aggregated and isoelectrically precipitated, erythrocyte and ghost viscosity increased coincident with a marked decrease in spectrin extractability. Similarly above 49 degrees C, a temperature where spectrin was denatured and precipitated, erythrocyte viscosity rose as inextractable spectrin accumulated. These observations provide direct evidence of a change in the physical state of spectrin associated with a change in erythrocyte shape and deformability. They support the concept that erythrocyte shape and deformability are largely determined by the shape and deformability of the spectrin-actin protein meshwork which laminates the inner membrane surface. | Diminished spectrin extraction from ATP-depleted human erythrocytes. Evidence relating spectrin to changes in erythrocyte shape and deformability. We measured spectrin "extractability" in erythrocytes which were metabolically depleted by incubation at 37 degrees C in plasma or glucose-free buffers. Membranes were extracted with 1 mM EDTA (pH 8, 40 h, 4 degrees C) and analyzed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. This procedure solubilized 85--90% of the spectrin, actin, and residual hemoglobin from ghosts of fresh erythrocytes. In incubated erythrocytes, inextractable spectrin rapidly accumulated when ATP concentrations fell below 0--15% of normal. In severely depleted cells, 60--90% of the total ghost spectrin became inextractable. Inextractability was not abolished by physically disrupting the ghost before extraction, but was reversed when erythrocyte ATP was replenished with adenosine. The accumulation of inextractable spectrin correlated temporally with the increase in apparent membrane deformability and the increases in erythrocyte vicosity, calcium content, sodium gain, and potassium loss characteristic of ATP-depleted erythrocytes. No change in integral membrane protein topography (assessed by the distribution of intramembranous particles and concanavalin A surface-binding sites) was detected in depleted cells. Analogous changes were observed in erythrocytes exposed to extremes of pH and temperature. When the pH in the erythrocyte interior fell below 5.5, a pH where spectrin was aggregated and isoelectrically precipitated, erythrocyte and ghost viscosity increased coincident with a marked decrease in spectrin extractability. Similarly above 49 degrees C, a temperature where spectrin was denatured and precipitated, erythrocyte viscosity rose as inextractable spectrin accumulated. These observations provide direct evidence of a change in the physical state of spectrin associated with a change in erythrocyte shape and deformability. They support the concept that erythrocyte shape and deformability are largely determined by the shape and deformability of the spectrin-actin protein meshwork which laminates the inner membrane surface. | [
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PMID:25289 | Alpah-adrenergic receptor modulation of beta-adrenergic, adenosine and prostaglandin E1 increased adenosine 3':5'-cyclic monophosphate levels in primary cultures of glia. | Beta-adrenergic agonists, adenosine and prostaglandin E1 increased the level of adenosine 3':5'-monophosphate (cAMP) in glial cultures prepared from rat cerebral cortical tissue. In addition to these physiological effectors, cholera toxin also increased cAMP levels in these cultures. The accumulation of cAMP in response to each of these agen-s, including cholera toxin, was partially blocked (50--80%) by simultaneous alpha-adrenergic receptor stimulation. Basal levels of cAMP were not affected by alpha-adrenergic agonists. These results indicate that in glia, alpha-adrenergic receptors may serve to modulate the level of cAMP which normally accumulates in response to a number of neurohumoral substances. The modulatory effect of alpha-adrenergic agents does not appear to reduce cAMP accumulation by activating phosphodiesterase since the effect was not blocked by a potent inhibitor of this enzymemthe results suggest that the modulatory effect of alpha-adrenergic receptor activation results from an interaction which takes place at some point in between adenylate cyclase-associated-membrane receptors and the enzymatic degradation of cAMP. | Alpah-adrenergic receptor modulation of beta-adrenergic, adenosine and prostaglandin E1 increased adenosine 3':5'-cyclic monophosphate levels in primary cultures of glia. Beta-adrenergic agonists, adenosine and prostaglandin E1 increased the level of adenosine 3':5'-monophosphate (cAMP) in glial cultures prepared from rat cerebral cortical tissue. In addition to these physiological effectors, cholera toxin also increased cAMP levels in these cultures. The accumulation of cAMP in response to each of these agen-s, including cholera toxin, was partially blocked (50--80%) by simultaneous alpha-adrenergic receptor stimulation. Basal levels of cAMP were not affected by alpha-adrenergic agonists. These results indicate that in glia, alpha-adrenergic receptors may serve to modulate the level of cAMP which normally accumulates in response to a number of neurohumoral substances. The modulatory effect of alpha-adrenergic agents does not appear to reduce cAMP accumulation by activating phosphodiesterase since the effect was not blocked by a potent inhibitor of this enzymemthe results suggest that the modulatory effect of alpha-adrenergic receptor activation results from an interaction which takes place at some point in between adenylate cyclase-associated-membrane receptors and the enzymatic degradation of cAMP. | [
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PMID:25287 | Diazepam and lorazepam for intravenous surgical premedication. | Diazepam, 10 and 20 mg, and 2 and 4 mg lorazepam were studied as intravenous surgical premedicants in 120 patients. Relief of anxiety, sedation, patient acceptance, lack of recall, and side effects were the variables evaluated. Both diazepam and lorazepam proved to be excellent surgical premedicants. The basic difference between the two drugs is temporal. Both medications produce similar relief of anxiety, sedation, patient acceptance, and lack of recall. The clinical effects of intravenous diazepam peaks in 2 to 3 minutes and diminishes thereafter. Intravenous lorazepam has a latent period of 8 to 15 minutes, with increasing effects at 15 to 30 minutes. | Diazepam and lorazepam for intravenous surgical premedication. Diazepam, 10 and 20 mg, and 2 and 4 mg lorazepam were studied as intravenous surgical premedicants in 120 patients. Relief of anxiety, sedation, patient acceptance, lack of recall, and side effects were the variables evaluated. Both diazepam and lorazepam proved to be excellent surgical premedicants. The basic difference between the two drugs is temporal. Both medications produce similar relief of anxiety, sedation, patient acceptance, and lack of recall. The clinical effects of intravenous diazepam peaks in 2 to 3 minutes and diminishes thereafter. Intravenous lorazepam has a latent period of 8 to 15 minutes, with increasing effects at 15 to 30 minutes. | [
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PMID:25291 | Loss of carbofuran from rice paddy water: chemical and physical factors. | The loss of carbofuran was studied from rice paddy water treated with a granular formulation of the insecticide, and from ponds filled with drainage from the paddy. The average half-life (t 1/2) for carbofuran loss was 57 hr. Controlled experiments indicated that pH was the predominating factor governing carbofuran loss from water in the environment studied. The loss due to hydrolysis was over 700 times more rapid at pH (t 1/2 = 1.2 hr.) than at pH (t 1/2 = 864 hr.) in buffered deionized water. The average pH of the rice paddy was 8, but diurnal fluctuations of 7 to 9.5 are common in similar environments. Impurities in the water, sunlight, and temperature influence the rate of carbofuran loss but not nearly so much as pH. There was no evidence for significant loss due to evaporation or oxidation. The results have important implications for the duration of the insecticide's activity and the effect on fish within or downstream from treated paddies. | Loss of carbofuran from rice paddy water: chemical and physical factors. The loss of carbofuran was studied from rice paddy water treated with a granular formulation of the insecticide, and from ponds filled with drainage from the paddy. The average half-life (t 1/2) for carbofuran loss was 57 hr. Controlled experiments indicated that pH was the predominating factor governing carbofuran loss from water in the environment studied. The loss due to hydrolysis was over 700 times more rapid at pH (t 1/2 = 1.2 hr.) than at pH (t 1/2 = 864 hr.) in buffered deionized water. The average pH of the rice paddy was 8, but diurnal fluctuations of 7 to 9.5 are common in similar environments. Impurities in the water, sunlight, and temperature influence the rate of carbofuran loss but not nearly so much as pH. There was no evidence for significant loss due to evaporation or oxidation. The results have important implications for the duration of the insecticide's activity and the effect on fish within or downstream from treated paddies. | [
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PMID:25292 | Preservation of organophosphorous pesticides in water samples. | Four techniques were studied for capacity to preserve sixteen organophosphorous pesticides in distilled water and in creek water. A technique using chloroform effectively preserved all sixteen pesticides for the three weeks of the study and refrigeration was effective for fourteen of the pesticides, but buffers of pH 4 and pH 7 appeared undependable as preservatives. | Preservation of organophosphorous pesticides in water samples. Four techniques were studied for capacity to preserve sixteen organophosphorous pesticides in distilled water and in creek water. A technique using chloroform effectively preserved all sixteen pesticides for the three weeks of the study and refrigeration was effective for fourteen of the pesticides, but buffers of pH 4 and pH 7 appeared undependable as preservatives. | [
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PMID:25288 | Long-term hypnotic efficacy and safety of triazolam and flurazepam. | Both triazolam and flurazepam are effective hypnotics when administered nightly for 12 consecutive weeks. However, at the dosages tested, 0.6 mg triazolam had a significantly faster onset of activity than 30 mg flurazepam. Long-term administration of either treatment did not influence the patient's capability to recognize the difference between active drug and placebo. This supports the conclusion that there was no tolerance development on either treatment. There were no deleterious effects attributable to either treatment as measured by the 35-Item Hopkins Symptom Checklist or by physical examinations, laboratory tests, ECGs, and ophthalmologic examinations. Side effects occurred more often on flurazepam than on triazolam, and the number of patients experiencing side effects was significantly higher in the flurazepam group. Drowsiness and grogginess were reported most frequently on both treatments, and the number of patients reporting drowsiness or grogginess was also significantly higher in the flurazepam group. | Long-term hypnotic efficacy and safety of triazolam and flurazepam. Both triazolam and flurazepam are effective hypnotics when administered nightly for 12 consecutive weeks. However, at the dosages tested, 0.6 mg triazolam had a significantly faster onset of activity than 30 mg flurazepam. Long-term administration of either treatment did not influence the patient's capability to recognize the difference between active drug and placebo. This supports the conclusion that there was no tolerance development on either treatment. There were no deleterious effects attributable to either treatment as measured by the 35-Item Hopkins Symptom Checklist or by physical examinations, laboratory tests, ECGs, and ophthalmologic examinations. Side effects occurred more often on flurazepam than on triazolam, and the number of patients experiencing side effects was significantly higher in the flurazepam group. Drowsiness and grogginess were reported most frequently on both treatments, and the number of patients reporting drowsiness or grogginess was also significantly higher in the flurazepam group. | [
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PMID:25304 | Biological characteristics of peptidoglycans of group A streptococcus and some other bacterial species. I. Tolerance and effect of antibody in fever response, and heart damaging effect in rabbits. | Induced tolerance to the pyrogenic action of group A streptococcus peptidoglycan decreased after one week and was no longer detectable after the second week. However, one or two further doses of peptidoglycan rapidly restored the tolerance. The passive transfer of plasma from rabbits tolerant to streptococcus peptidoglycan to nontolerant animals failed to transfer tolerance. Antiserum to streptococcus peptidoglycan neutralized the pyrogenic effect of not only streptococcus but also staphylococcus and pneumococcus peptidoglycan; it did not influence the febrile response to endotoxin. Histopathologic changes in the rabbit heart produced by the intravenous injection of staphylococcus or pneumococcus peptidoglycans were similar and were characterized by various stages of degeneration and necrosis. The changes were less pronounced than after streptococcus peptidoglycan. Antiserum to streptococcus peptidoglycan had modest or no counteracting effect on the development of heart alterations after staphylococcus or pneumococcus peptidoglycan. | Biological characteristics of peptidoglycans of group A streptococcus and some other bacterial species. I. Tolerance and effect of antibody in fever response, and heart damaging effect in rabbits. Induced tolerance to the pyrogenic action of group A streptococcus peptidoglycan decreased after one week and was no longer detectable after the second week. However, one or two further doses of peptidoglycan rapidly restored the tolerance. The passive transfer of plasma from rabbits tolerant to streptococcus peptidoglycan to nontolerant animals failed to transfer tolerance. Antiserum to streptococcus peptidoglycan neutralized the pyrogenic effect of not only streptococcus but also staphylococcus and pneumococcus peptidoglycan; it did not influence the febrile response to endotoxin. Histopathologic changes in the rabbit heart produced by the intravenous injection of staphylococcus or pneumococcus peptidoglycans were similar and were characterized by various stages of degeneration and necrosis. The changes were less pronounced than after streptococcus peptidoglycan. Antiserum to streptococcus peptidoglycan had modest or no counteracting effect on the development of heart alterations after staphylococcus or pneumococcus peptidoglycan. | [
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PMID:25305 | Studies on the mitogen responses of germfree allogeneic chimeras. II. Maturation of two cell types and partial restoration of responsiveness of the short-term chimeras. | Germfree allogeneic bone marrow chimeras (ABMC) were produced by the i.v. injection of approximately 10(7) bone marrow cells from germfree DBA/2 mice into lethally irradiated germfree C3H mice. In the germfree state, the short-term ABMC showed no histologic signs of graft-vs-host reactions (GVHR), yet splenic lymphocytes were unable to respond to PHA, Con A, or SRBC. Attempts to remove responsiveness by the implantation of a DBA/2 thymus under the host kidney capsule also resulted in failure. However, when the donor thymus was enclosed in a cell-impermeable chamber to eliminate a GVH reaction, responsiveness to Con A was restored. The PHA and SRBC responses were unaffected by this treatment. Daily injections of thymosin caused both an increased Con A response and increased numbers of PFC, although the PHA response was again unaffected. Thus, soluble substances from thymic tissue can be used to overcome partially the histocompatibility barrier present in the ABMC that affects at least two different functional cell populations. | Studies on the mitogen responses of germfree allogeneic chimeras. II. Maturation of two cell types and partial restoration of responsiveness of the short-term chimeras. Germfree allogeneic bone marrow chimeras (ABMC) were produced by the i.v. injection of approximately 10(7) bone marrow cells from germfree DBA/2 mice into lethally irradiated germfree C3H mice. In the germfree state, the short-term ABMC showed no histologic signs of graft-vs-host reactions (GVHR), yet splenic lymphocytes were unable to respond to PHA, Con A, or SRBC. Attempts to remove responsiveness by the implantation of a DBA/2 thymus under the host kidney capsule also resulted in failure. However, when the donor thymus was enclosed in a cell-impermeable chamber to eliminate a GVH reaction, responsiveness to Con A was restored. The PHA and SRBC responses were unaffected by this treatment. Daily injections of thymosin caused both an increased Con A response and increased numbers of PFC, although the PHA response was again unaffected. Thus, soluble substances from thymic tissue can be used to overcome partially the histocompatibility barrier present in the ABMC that affects at least two different functional cell populations. | [
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PMID:25309 | Tyrosine aminotransferase in AKR/J albino and C57BL/6J black mouse skin. | L-Tyrosine aminotransferase is present in a high speed supernatant fraction of skin homogenate of AKR/J albino and C57BL/6J black mice. The conversion of tyrosine to p-hydroxyphenylpyruvate was shown to be catalyzed by an aminotransferase by the following observations: the reaction was partially dependent on the presence of low concentrations of alpha-ketoglutarate; catalase was ineffective in increasing the yield of p-hydroxyphenylpyruvate; there was potent inhibition by typical inhibitors of pyridoxal phosphate enzymes and of rat liver tyrosine aminotransferase; there was no inhibition by inhibitors of L-amino acid oxidase; and there was no oxidation of L-leucine, the best substrate for rat kidney L-amino acid oxidase. The aminotransferase was stimulated by mercaptoethanol and was inhibited by high concentrations of alpha-ketoglutarate. The apparent Km for tyrosine was 5 X 10(-3) M and the molecular weight, determined by sucrose density gradient centrifugation, was 150-200,000. Dopa was also transaminated by the crude enzyme. No tyrosine aminotransferase could be detected in extracts of hamster melanoma. | Tyrosine aminotransferase in AKR/J albino and C57BL/6J black mouse skin. L-Tyrosine aminotransferase is present in a high speed supernatant fraction of skin homogenate of AKR/J albino and C57BL/6J black mice. The conversion of tyrosine to p-hydroxyphenylpyruvate was shown to be catalyzed by an aminotransferase by the following observations: the reaction was partially dependent on the presence of low concentrations of alpha-ketoglutarate; catalase was ineffective in increasing the yield of p-hydroxyphenylpyruvate; there was potent inhibition by typical inhibitors of pyridoxal phosphate enzymes and of rat liver tyrosine aminotransferase; there was no inhibition by inhibitors of L-amino acid oxidase; and there was no oxidation of L-leucine, the best substrate for rat kidney L-amino acid oxidase. The aminotransferase was stimulated by mercaptoethanol and was inhibited by high concentrations of alpha-ketoglutarate. The apparent Km for tyrosine was 5 X 10(-3) M and the molecular weight, determined by sucrose density gradient centrifugation, was 150-200,000. Dopa was also transaminated by the crude enzyme. No tyrosine aminotransferase could be detected in extracts of hamster melanoma. | [
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PMID:25310 | The relationships among arterial oxygen flow rate, oxygen binding by hemoglobin, and oxygen utilization in chronic cardiac decompensation. | We have examined the interrelationships among CaO2, blood flow, oxygen binding by hemoglobin, and VO2 in cardiac patients with and without chronic cardiac decompensation. We have quantified the role that decreased oxygen-binding to hemoglobin may play in maintaining VO2 in the presence of low systemic blood flow rates. The volume rate of oxygen delivery to tissues was expressed as the OFIa, the product of CO2 and blood flow. OFIa varied from 738 to 262 ml/min/m2, whereas VO2 varied from 170 to 117 ml/min/m2. Thus, in the patients with lowest OFIa (63% below the highest OFIa), VO2 was only down 19%. VO2 was maintained because the extraction of oxygen rose from about 20% to 50% in close association with the decrease in OFIa. Oxygen binding to hemoglobin was lower in patients with the lowest OFIa--and therefore, at in vivo conditions of pH, PCO2, and temperature, P50 in vivo was higher. The resulting facilitation of oxygen release at the PO2 of tissue capillaries could explain about one third of the observed increment in oxygen extraction in patients with low OFIa. An alternative interpretation is that a high P50 in vivo minimizes the reduction in PVO2 needed to maintain VO2 when increased proportional extraction of O2 compensates for decreased OFIa. | The relationships among arterial oxygen flow rate, oxygen binding by hemoglobin, and oxygen utilization in chronic cardiac decompensation. We have examined the interrelationships among CaO2, blood flow, oxygen binding by hemoglobin, and VO2 in cardiac patients with and without chronic cardiac decompensation. We have quantified the role that decreased oxygen-binding to hemoglobin may play in maintaining VO2 in the presence of low systemic blood flow rates. The volume rate of oxygen delivery to tissues was expressed as the OFIa, the product of CO2 and blood flow. OFIa varied from 738 to 262 ml/min/m2, whereas VO2 varied from 170 to 117 ml/min/m2. Thus, in the patients with lowest OFIa (63% below the highest OFIa), VO2 was only down 19%. VO2 was maintained because the extraction of oxygen rose from about 20% to 50% in close association with the decrease in OFIa. Oxygen binding to hemoglobin was lower in patients with the lowest OFIa--and therefore, at in vivo conditions of pH, PCO2, and temperature, P50 in vivo was higher. The resulting facilitation of oxygen release at the PO2 of tissue capillaries could explain about one third of the observed increment in oxygen extraction in patients with low OFIa. An alternative interpretation is that a high P50 in vivo minimizes the reduction in PVO2 needed to maintain VO2 when increased proportional extraction of O2 compensates for decreased OFIa. | [
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PMID:25311 | Allosteric modifiers of fish hemoglobins: in vitro and in vivo studies of the effect of ambient oxygen and pH on erythrocyte ATP concentrations. | Fundulus heteroclitus decreases erythrocyte adenosine triphosphate and increases blood hematocrit when acclimated to hypoxic conditions. A defined medium has been developed which allows isolated F. heteroclitus erythrocytes to be maintained for several hours without an appreciable loss of cellular ATP. The effect of oxygen tension, pH and metabolic inhibitors on the cellular concentration of ATP of fish red cells has been investigated as an in vitro model to explain in vivo responses to environmental changes. The isolated red cells significantly decrease their ATP/Hb molar ratio when exposed either to anaerobiosis or metabolic inhibitors. It is concluded that the in vivo response is mediated at the red cell level via decreased oxidative phosphorylation in the presence of low environmental oxygen. The length of time necessary to elicit the responce both in vivo and in vitro is also discussed. | Allosteric modifiers of fish hemoglobins: in vitro and in vivo studies of the effect of ambient oxygen and pH on erythrocyte ATP concentrations. Fundulus heteroclitus decreases erythrocyte adenosine triphosphate and increases blood hematocrit when acclimated to hypoxic conditions. A defined medium has been developed which allows isolated F. heteroclitus erythrocytes to be maintained for several hours without an appreciable loss of cellular ATP. The effect of oxygen tension, pH and metabolic inhibitors on the cellular concentration of ATP of fish red cells has been investigated as an in vitro model to explain in vivo responses to environmental changes. The isolated red cells significantly decrease their ATP/Hb molar ratio when exposed either to anaerobiosis or metabolic inhibitors. It is concluded that the in vivo response is mediated at the red cell level via decreased oxidative phosphorylation in the presence of low environmental oxygen. The length of time necessary to elicit the responce both in vivo and in vitro is also discussed. | [
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PMID:25317 | Stronglyoides ransomi: factors influencing the in vitro development of the free-living generation. | Among the progeny of parasitic females of Strongyloides ransomi, ransomi, males did not appear in significant numbers until the 7th week of infection in cases of simple infection, and until the 3rd week of infection in cases of multiple infection. The appearance of males was attributed to the effect of host immunity, the physiological ageing of the parasitic females, or both. Type of culture substrate and other cultural conditions did not influence the percent of larvae developing into males. Sex of larvae was determined prior to hatching, probably during oogenesis or embryogenesis. Culture conditions influenced the direction of development of female larvae. An initial pH below 5.9 or above 7.2 favored differentiation of larvae into infective larvae, whereas, intermediate initial pH levels favored development of free-living females. Baby pig substrate, autoclaved substrate, and substrate washed free of soluble chemicals (adverse cultural conditions) promoted differentiation toward infective larvae. Adult pig substrate, nonautoclaved substrate and unwashed substrate promoted differentiation toward free-living females. In general, adverse conditions inside the host and favor an indirect life cycle. | Stronglyoides ransomi: factors influencing the in vitro development of the free-living generation. Among the progeny of parasitic females of Strongyloides ransomi, ransomi, males did not appear in significant numbers until the 7th week of infection in cases of simple infection, and until the 3rd week of infection in cases of multiple infection. The appearance of males was attributed to the effect of host immunity, the physiological ageing of the parasitic females, or both. Type of culture substrate and other cultural conditions did not influence the percent of larvae developing into males. Sex of larvae was determined prior to hatching, probably during oogenesis or embryogenesis. Culture conditions influenced the direction of development of female larvae. An initial pH below 5.9 or above 7.2 favored differentiation of larvae into infective larvae, whereas, intermediate initial pH levels favored development of free-living females. Baby pig substrate, autoclaved substrate, and substrate washed free of soluble chemicals (adverse cultural conditions) promoted differentiation toward infective larvae. Adult pig substrate, nonautoclaved substrate and unwashed substrate promoted differentiation toward free-living females. In general, adverse conditions inside the host and favor an indirect life cycle. | [
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PMID:25318 | A simple quantitative technique for testing behavioral responses of Schistosoma mansoni miracidia to chemicals. | We describe a new technique for testing responses of Schistosoma mansoni miracidia to chemicals. Miracidia in spring water were placed in a chamber shaped like the Greek letter phi. Small volumes of test chemicals were inoculated into one side of the chamber. After 30 sec a dam was inserted to bisect the chamber and the percentage of miracidia on the inoculated side was calculated. Reproducible quantitative results were obtained using the known miracidial stimulants, snail-conditioned water (Biomphalaria glabrata) and Mg2+; up to 80% of the miracidia were recovered on the inoculated side of the chamber. Other substances also stimulated miracidia: several inorganic cations, 4 neurotransmitters, 3 acetycholine antagonists, and 1 acetycholine agonist. Modifying the technique by testing stimulants in altered chemical "backgrounds" allowed us to test for inhibitors of miracidial responses. Assays of the Mg2+ content of several of the test solutions indicated that their stimulatory or inhibitory effects could not be ascribed to Mg2+ contamination. However, results obtained with neurotransmitters and drugs were not sufficiently consistent to implicate specific neurotransmitters in the mechanism by which miracidia detect and respond to stimulants in snail-conditioned water. | A simple quantitative technique for testing behavioral responses of Schistosoma mansoni miracidia to chemicals. We describe a new technique for testing responses of Schistosoma mansoni miracidia to chemicals. Miracidia in spring water were placed in a chamber shaped like the Greek letter phi. Small volumes of test chemicals were inoculated into one side of the chamber. After 30 sec a dam was inserted to bisect the chamber and the percentage of miracidia on the inoculated side was calculated. Reproducible quantitative results were obtained using the known miracidial stimulants, snail-conditioned water (Biomphalaria glabrata) and Mg2+; up to 80% of the miracidia were recovered on the inoculated side of the chamber. Other substances also stimulated miracidia: several inorganic cations, 4 neurotransmitters, 3 acetycholine antagonists, and 1 acetycholine agonist. Modifying the technique by testing stimulants in altered chemical "backgrounds" allowed us to test for inhibitors of miracidial responses. Assays of the Mg2+ content of several of the test solutions indicated that their stimulatory or inhibitory effects could not be ascribed to Mg2+ contamination. However, results obtained with neurotransmitters and drugs were not sufficiently consistent to implicate specific neurotransmitters in the mechanism by which miracidia detect and respond to stimulants in snail-conditioned water. | [
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PMID:25319 | Effect of diamfenetide on experimental infections of Fasciola hepatica in lambs: anthelmintic and clinical investigations. | The activity of diamfenetide (N,N'-[oxybis(2,1-ethan diyloxy-4,1-phenylene)] bis acetamide) was studied in lambs experimentally inoculated with Fasciola hepatica. The drug was given orally at a dose level of 100 mg/kg either 1,3,5,7, or 9 weeks postinoculation. It was 100% effective 1, 3, and 5 weeks postinoculation, 73% effective 7 weeks postinoculation, and 57% effective 9 weeks postinoculation. Serum gamma-glutamyl transpeptidase activity remained normal in all lambs for 5 weeks after infection; it then began to increase in infected, untreated lambs at 6 weeks, and had increased 5- to 6-fold 9 weeks postinoculation in infected lambs. This enzyme activity was the most sensitive hematologic parameter used in this test to detect hepatobiliary damage by the parasite. The drug was well tolerated at the dose level used. | Effect of diamfenetide on experimental infections of Fasciola hepatica in lambs: anthelmintic and clinical investigations. The activity of diamfenetide (N,N'-[oxybis(2,1-ethan diyloxy-4,1-phenylene)] bis acetamide) was studied in lambs experimentally inoculated with Fasciola hepatica. The drug was given orally at a dose level of 100 mg/kg either 1,3,5,7, or 9 weeks postinoculation. It was 100% effective 1, 3, and 5 weeks postinoculation, 73% effective 7 weeks postinoculation, and 57% effective 9 weeks postinoculation. Serum gamma-glutamyl transpeptidase activity remained normal in all lambs for 5 weeks after infection; it then began to increase in infected, untreated lambs at 6 weeks, and had increased 5- to 6-fold 9 weeks postinoculation in infected lambs. This enzyme activity was the most sensitive hematologic parameter used in this test to detect hepatobiliary damage by the parasite. The drug was well tolerated at the dose level used. | [
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PMID:25321 | Effect of vehicles and other active ingredients on stability of hydrocortisone. | The stability of hydrocortisone in various types of vehicles, aqueous, water-washable (polyethylene glycol ointment base), and oil in water or water in oil-type emulsified vehicles, and in the presence of other ingredients, iodochlorhydroxyquin, menthol, and phenol, was studied under normal conditions (room temperature and weakly acidic pH). The study was conducted using a stability-indicating assay method, high-pressure liquid chromatography. The hydrocortisone was very unstable in water and water-washable ointment base. The addition of alcohol and glycerin to water had a stabilizing effect. Under drastic conditions (very acidic or very basic pH), hydrocortisone proved to be unstable only on the basic side. The data at higher temperatures confirmed that the decomposition in water and polyethylene glycol was pseudo-first order. The decomposition process appears to be different in the highly basic solution versus weakly acidic media or in water versus polyethylene glycol ointment base. | Effect of vehicles and other active ingredients on stability of hydrocortisone. The stability of hydrocortisone in various types of vehicles, aqueous, water-washable (polyethylene glycol ointment base), and oil in water or water in oil-type emulsified vehicles, and in the presence of other ingredients, iodochlorhydroxyquin, menthol, and phenol, was studied under normal conditions (room temperature and weakly acidic pH). The study was conducted using a stability-indicating assay method, high-pressure liquid chromatography. The hydrocortisone was very unstable in water and water-washable ointment base. The addition of alcohol and glycerin to water had a stabilizing effect. Under drastic conditions (very acidic or very basic pH), hydrocortisone proved to be unstable only on the basic side. The data at higher temperatures confirmed that the decomposition in water and polyethylene glycol was pseudo-first order. The decomposition process appears to be different in the highly basic solution versus weakly acidic media or in water versus polyethylene glycol ointment base. | [
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PMID:25322 | Demonstration of penicillamine as a product in benzylpenicillenic acid degradation in neutral media using differential pulse polarography. | During the study of the temporal changes of benzylpenicillenic acid in aqueous buffers using differential pulse polarography, penicillamine was found to be a degradation product at neutral pH. Since this result was not previously reported, the effects of pH and buffer concentration on penicillamine formation were investigated. The amount of penicillamine produced was greatest under conditions producing maximum benzylpenicillenic acid stability. Penicillamine was not obtained from benzylpenicilloic acid, the reported degradation product of benzylpenicillenic acid at neutral pH. Penicillamine also was detected in penicillin G solutions of neutral pH. Therefore, it is suggested that penicillamine found in penicillin G solutions arises from benzylpenicillenic acid degradation which, in turn, is produced from penicillin G isomerization. A pathway is proposed to show that penicillamine originates from the UV-absorbing isomer of benzylpenicillenic acid. | Demonstration of penicillamine as a product in benzylpenicillenic acid degradation in neutral media using differential pulse polarography. During the study of the temporal changes of benzylpenicillenic acid in aqueous buffers using differential pulse polarography, penicillamine was found to be a degradation product at neutral pH. Since this result was not previously reported, the effects of pH and buffer concentration on penicillamine formation were investigated. The amount of penicillamine produced was greatest under conditions producing maximum benzylpenicillenic acid stability. Penicillamine was not obtained from benzylpenicilloic acid, the reported degradation product of benzylpenicillenic acid at neutral pH. Penicillamine also was detected in penicillin G solutions of neutral pH. Therefore, it is suggested that penicillamine found in penicillin G solutions arises from benzylpenicillenic acid degradation which, in turn, is produced from penicillin G isomerization. A pathway is proposed to show that penicillamine originates from the UV-absorbing isomer of benzylpenicillenic acid. | [
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PMID:25323 | Effect of ionization on absorption of cephalosporins. | To explore the relative absorbabilities of different ionic forms of cephalosporins, the absorption rates of four compounds were measured in the pH 5-9 region using an in situ rat gut technique. Cephalexin, cephradine, and cephaloglycin have some oral activity, while 3-[(acetyloxy)methyl]-8-oxo-7-[[(4-oxo-1(4H-pyridinyl)acetyl]-amino]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid (I) has insignificant oral activity. The pH-species profiles calculated from their ionization constants showed that cephalexin, cephradine, and cephaloglycin have a large proportion of uncharged molecules plus zwitterions in the pH range of the small intestine, while I exists as the anion throughout this range. When the species profiles are compared with the pH-absorption rate profiles for cephalexin, cephradine, and I, the results are consistent with a model in which the zwitterionic and/or uncharged forms of the molecules are well absorbed, whereas the anions show little or no absorption. Although it has a pH profile for zwitterions plus uncharged molecules similar to cephalexin, cephaloglycin shows poor absorption, suggesting that the ratio of uncharged molecules to zwitterions may be important in absorption. | Effect of ionization on absorption of cephalosporins. To explore the relative absorbabilities of different ionic forms of cephalosporins, the absorption rates of four compounds were measured in the pH 5-9 region using an in situ rat gut technique. Cephalexin, cephradine, and cephaloglycin have some oral activity, while 3-[(acetyloxy)methyl]-8-oxo-7-[[(4-oxo-1(4H-pyridinyl)acetyl]-amino]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid (I) has insignificant oral activity. The pH-species profiles calculated from their ionization constants showed that cephalexin, cephradine, and cephaloglycin have a large proportion of uncharged molecules plus zwitterions in the pH range of the small intestine, while I exists as the anion throughout this range. When the species profiles are compared with the pH-absorption rate profiles for cephalexin, cephradine, and I, the results are consistent with a model in which the zwitterionic and/or uncharged forms of the molecules are well absorbed, whereas the anions show little or no absorption. Although it has a pH profile for zwitterions plus uncharged molecules similar to cephalexin, cephaloglycin shows poor absorption, suggesting that the ratio of uncharged molecules to zwitterions may be important in absorption. | [
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PMID:25324 | Kinetics of digoxin stability in aqueous solution. | Digoxin hydrolysis was studied as a function of pH. Conversion of digoxin to digoxigenin was followed by high-pressure liquid chromatography and shown to proceed by the initial loss of one, two, or three sugars. The hydrolysis rate was directly proportional to parent drug concentration and hydrogen-ion activity. The individual hydrolysis rate constants of digoxin, digoxigenin bisdigitoxoside, and digoxigenin monodigitoxoside were determined by a simplex fitting procedure. Data are presented suggesting that at least some variation in the bioavailability of orally administered digoxin arises from observed variations in gastric pH; these variations influence the extent to which hydrolysis occurs and, thus, modify the composition of digoxin species available for absorption. | Kinetics of digoxin stability in aqueous solution. Digoxin hydrolysis was studied as a function of pH. Conversion of digoxin to digoxigenin was followed by high-pressure liquid chromatography and shown to proceed by the initial loss of one, two, or three sugars. The hydrolysis rate was directly proportional to parent drug concentration and hydrogen-ion activity. The individual hydrolysis rate constants of digoxin, digoxigenin bisdigitoxoside, and digoxigenin monodigitoxoside were determined by a simplex fitting procedure. Data are presented suggesting that at least some variation in the bioavailability of orally administered digoxin arises from observed variations in gastric pH; these variations influence the extent to which hydrolysis occurs and, thus, modify the composition of digoxin species available for absorption. | [
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PMID:25325 | Spectroscopic approach to estimation of microequilibrium constants of prototropic reactions of aminobenzoic acids. | The microequilibrium constants of protolytic dissociation of diprotic acids, dihydric bases, or ampholytes such as the aminobenzoic acids, with dissimilar ionizing groups, can be estimated by spectrophotometric titration and measurement of the molar absorptivity at the long wavelength absorption maximum of simple alkylated derivatives. The method is applicable when the long wavelength absorption spectral bands of the tautomeric species are well resolved. Compared to the traditional method of estimating microequilibrium constants using the dissociation constants of alkylated derivatives, the proposed method is simpler, faster, and more accurate. | Spectroscopic approach to estimation of microequilibrium constants of prototropic reactions of aminobenzoic acids. The microequilibrium constants of protolytic dissociation of diprotic acids, dihydric bases, or ampholytes such as the aminobenzoic acids, with dissimilar ionizing groups, can be estimated by spectrophotometric titration and measurement of the molar absorptivity at the long wavelength absorption maximum of simple alkylated derivatives. The method is applicable when the long wavelength absorption spectral bands of the tautomeric species are well resolved. Compared to the traditional method of estimating microequilibrium constants using the dissociation constants of alkylated derivatives, the proposed method is simpler, faster, and more accurate. | [
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PMID:25326 | Characterization of impurities in sulfasalazine. | The chemical structures of four impurities isolated from sulfasalazine were determined. Three impurities are the by-products of the reaction process in drug synthesis, i.e., during diazotization of sulfapyridine and during coupling with salicylic acid. Only one contaminant was identified as a starting material, sulfapyridine, in the drug synthesis. The four impurities were characterized as 2-[[p-(2-pyridylsulfamoyl)-phenyl]azo]hydroxybenzene (I), 3-[[P-(2-pyridylsulfamoyl) phenyl]-azo]salicylic acid (II), 5-[[p-[4-(2-pyridylanilino)]-N-phenyl]azo]salicylic acid (III), and sulfapyridine (IV). Compounds I-III are novel molecules, and IV is the precursor of sulfasalazine. The isolation of the impurities was accomplished by TLC and liquid extraction procedures. The methods used to characterize the impurities were a combination of IR, UV, and NMR spectroscopy, mass spectrometry, and TLC. For I and III, comparisons also were made with the synthesized materials to supplement the evaluation. | Characterization of impurities in sulfasalazine. The chemical structures of four impurities isolated from sulfasalazine were determined. Three impurities are the by-products of the reaction process in drug synthesis, i.e., during diazotization of sulfapyridine and during coupling with salicylic acid. Only one contaminant was identified as a starting material, sulfapyridine, in the drug synthesis. The four impurities were characterized as 2-[[p-(2-pyridylsulfamoyl)-phenyl]azo]hydroxybenzene (I), 3-[[P-(2-pyridylsulfamoyl) phenyl]-azo]salicylic acid (II), 5-[[p-[4-(2-pyridylanilino)]-N-phenyl]azo]salicylic acid (III), and sulfapyridine (IV). Compounds I-III are novel molecules, and IV is the precursor of sulfasalazine. The isolation of the impurities was accomplished by TLC and liquid extraction procedures. The methods used to characterize the impurities were a combination of IR, UV, and NMR spectroscopy, mass spectrometry, and TLC. For I and III, comparisons also were made with the synthesized materials to supplement the evaluation. | [
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PMID:25327 | In vitro evidence for ipecac inactivation by activated charcoal. | The in vitro adsorption of the alkaloid emetine, a primary constituent of ipecac, on activated charcoal was studied. The results support the supposition that syrup of ipecac should not be given to counteract poisonings if activated charcoal is also to be administered. | In vitro evidence for ipecac inactivation by activated charcoal. The in vitro adsorption of the alkaloid emetine, a primary constituent of ipecac, on activated charcoal was studied. The results support the supposition that syrup of ipecac should not be given to counteract poisonings if activated charcoal is also to be administered. | [
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PMID:25328 | Determination of desmethyldiazepam in plasma by electron-capture GLC: application to pharmacokinetic studies of clorazepate. | Plasma desmethyldiazepam concentrations were quantitated by a rapid and sensitive technique using electron-capture GLC. Following addition of diazepam as the internal standard, plasma is extracted at physiological pH into benzene-isoamyl alcohol. The extract is evaporated to dryness and reconstituted with toluene-isoamyl alcohol prior to chromatography. Both diazepam and desmethyldiazepam are quantitatively extracted. The variation of identical samples is 5%, and the sensitivity is 5 ng of desmethyldiazepam/ml of original sample. The method is applicable to pharmacokinetic studies of clorazepate, a benzodiazepine derivative transformed to desmethyldiazepam prior to absorption. | Determination of desmethyldiazepam in plasma by electron-capture GLC: application to pharmacokinetic studies of clorazepate. Plasma desmethyldiazepam concentrations were quantitated by a rapid and sensitive technique using electron-capture GLC. Following addition of diazepam as the internal standard, plasma is extracted at physiological pH into benzene-isoamyl alcohol. The extract is evaporated to dryness and reconstituted with toluene-isoamyl alcohol prior to chromatography. Both diazepam and desmethyldiazepam are quantitatively extracted. The variation of identical samples is 5%, and the sensitivity is 5 ng of desmethyldiazepam/ml of original sample. The method is applicable to pharmacokinetic studies of clorazepate, a benzodiazepine derivative transformed to desmethyldiazepam prior to absorption. | [
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PMID:25329 | In vitro evaluation of three commercial sustained-release papaverine hydrochloride products. | Three commercial sustained-release papaverine hydrochloride products in the form of microencapsulated pellets were evaluated. Three different dissolution apparatuses were used: a continuous flow apparatus, the USP rotating basket apparatus, and a modified reciprocating basket apparatus. The frequency rate of the reciprocating basket apparatus could be varied from 0 to 31 strokes/min. Salicylic acid compacts were used as a standard to characterize each apparatus. A linear log--log correlation between dissolution rate and apparatus speed or flow rate was obtained. Release of papaverine hydrochloride from the commercial preparations was affected significantly by the pH of the dissolution media but not by the agitation intensity. | In vitro evaluation of three commercial sustained-release papaverine hydrochloride products. Three commercial sustained-release papaverine hydrochloride products in the form of microencapsulated pellets were evaluated. Three different dissolution apparatuses were used: a continuous flow apparatus, the USP rotating basket apparatus, and a modified reciprocating basket apparatus. The frequency rate of the reciprocating basket apparatus could be varied from 0 to 31 strokes/min. Salicylic acid compacts were used as a standard to characterize each apparatus. A linear log--log correlation between dissolution rate and apparatus speed or flow rate was obtained. Release of papaverine hydrochloride from the commercial preparations was affected significantly by the pH of the dissolution media but not by the agitation intensity. | [
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PMID:25330 | Prodrug approaches to enhancement of physicochemical properties of drugs IX: acetaminophen prodrug. | The synthesis, hydrolysis rate, and bioavailability of 1-(p-acetaminophenoxy)-1-ethoxyethane, an acetaminophen prodrug, are described. The prodrug is less soluble than acetaminophen and stable at neutral pH. However, in an acidic environment, the compound cleaves rapidly, generating acetaminophen. When both the prodrug and acetaminophen were administered to dogs in equivalent amounts, the blood acetaminophen levels were comparable. | Prodrug approaches to enhancement of physicochemical properties of drugs IX: acetaminophen prodrug. The synthesis, hydrolysis rate, and bioavailability of 1-(p-acetaminophenoxy)-1-ethoxyethane, an acetaminophen prodrug, are described. The prodrug is less soluble than acetaminophen and stable at neutral pH. However, in an acidic environment, the compound cleaves rapidly, generating acetaminophen. When both the prodrug and acetaminophen were administered to dogs in equivalent amounts, the blood acetaminophen levels were comparable. | [
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PMID:25331 | Hydrolysis kinetics of the monophosphate ester triclofos sodium. | The hydrolysis kinetics of the monophosphate ester triclofos sodium were studied in the pH 0.00--7.80 range, the 80.1--98.4 degrees range, and ionic strengths of 0.02--1.50. The pK1 and pK2 values for triclofos were 1.00 +/- 0.05 and 5.80 +/- 0.05, respectively. The pH dependency observed was consistent with a theory in which the hydrolysis rates of the triclofos species follow the order: monoanion greater than unionized greater than dianion. The apparent first-order rate constants at 80.1 degrees associated with each species were 6.4 +/- 0.6 X 10(-3), 28.0 +/- 0.5 X 10(-3), and 0.3 +/- 0.1 hr-1, respectively, Different Arrhenius activation energies were observed at pH 0.00, 3.50, and 7.80, at which values the unionized, monoanion, and dianion forms were the dominant species present, respectively. The reaction showed a modest positive kinetic salt effect at pH 3.50 and a modest negative effect at pH 7.80. Various mechanistic possibilities are discussed. | Hydrolysis kinetics of the monophosphate ester triclofos sodium. The hydrolysis kinetics of the monophosphate ester triclofos sodium were studied in the pH 0.00--7.80 range, the 80.1--98.4 degrees range, and ionic strengths of 0.02--1.50. The pK1 and pK2 values for triclofos were 1.00 +/- 0.05 and 5.80 +/- 0.05, respectively. The pH dependency observed was consistent with a theory in which the hydrolysis rates of the triclofos species follow the order: monoanion greater than unionized greater than dianion. The apparent first-order rate constants at 80.1 degrees associated with each species were 6.4 +/- 0.6 X 10(-3), 28.0 +/- 0.5 X 10(-3), and 0.3 +/- 0.1 hr-1, respectively, Different Arrhenius activation energies were observed at pH 0.00, 3.50, and 7.80, at which values the unionized, monoanion, and dianion forms were the dominant species present, respectively. The reaction showed a modest positive kinetic salt effect at pH 3.50 and a modest negative effect at pH 7.80. Various mechanistic possibilities are discussed. | [
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PMID:25332 | Determination of serum nadolol levels by GLC--selected ion monitoring mass spectrometry: comparison with a spectrofluorometric method. | A method to determine the serum concentration of the beta-adrenergic receptor blocking agent, nadolol, by GLC--selected ion monitoring mass spectrometry of the tri(trimethysilyl) ether derivative is described. A basic solution of serum was extracted, known amounts of internal standard were added to the extract, and the extract was back-extracted into acidic media and lyophilized. The resulting solids were reacted with N-trimethylsilylimidazole. Coded serum samples of 12 subjects, given nadolol alone or in combination with a second drug, were analyzed. The ions at m/e 86 and 100 were monitored to establish the relative concentration ratio of nadolol and the internal reference N-methylnadolol. No interferences from blood components or other administered drugs were observed. A detection level of 6.95 ng/ml of serum was found. | Determination of serum nadolol levels by GLC--selected ion monitoring mass spectrometry: comparison with a spectrofluorometric method. A method to determine the serum concentration of the beta-adrenergic receptor blocking agent, nadolol, by GLC--selected ion monitoring mass spectrometry of the tri(trimethysilyl) ether derivative is described. A basic solution of serum was extracted, known amounts of internal standard were added to the extract, and the extract was back-extracted into acidic media and lyophilized. The resulting solids were reacted with N-trimethylsilylimidazole. Coded serum samples of 12 subjects, given nadolol alone or in combination with a second drug, were analyzed. The ions at m/e 86 and 100 were monitored to establish the relative concentration ratio of nadolol and the internal reference N-methylnadolol. No interferences from blood components or other administered drugs were observed. A detection level of 6.95 ng/ml of serum was found. | [
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PMID:25333 | Microionization constants of commercial cephalosporins. | The equilibrium constants of five commercial cephalosporins were determined. Two are monoacidic and possess one Ka while three are amphoteric and have four microconstants. Although several assumptions were made in the calculations, good agreement was found between the compounds and with previously reported macroionization constants. By utilizing the microionization constants, the ratios of zwitterion to unchanged species were calculated to be in the 900-50,000 range and to have a maximum concentration between pH 3.5 and 5. | Microionization constants of commercial cephalosporins. The equilibrium constants of five commercial cephalosporins were determined. Two are monoacidic and possess one Ka while three are amphoteric and have four microconstants. Although several assumptions were made in the calculations, good agreement was found between the compounds and with previously reported macroionization constants. By utilizing the microionization constants, the ratios of zwitterion to unchanged species were calculated to be in the 900-50,000 range and to have a maximum concentration between pH 3.5 and 5. | [
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PMID:25334 | Pharmacokinetic studies on ditazole, a novel inhibitor of platelet aggregation. | The distribution of ditazole in blood and tissues of rats was determined by a simple GLC technique. Ditazole, after intravenous injection in rats (20 mg/kg), entered preferentially into the brain, the liver, and the heart in decreasing order. In the epididymal adipose tissue, the drug was present only in small amounts. Ditazole disappeared from the rat organs 4 hr after the treatment. The apparent ditazole half-life in rat blood was 41 min, the volume of distribution was 2.068 liters/kg, and the body clearance was 0.0345 liter/kg/min. | Pharmacokinetic studies on ditazole, a novel inhibitor of platelet aggregation. The distribution of ditazole in blood and tissues of rats was determined by a simple GLC technique. Ditazole, after intravenous injection in rats (20 mg/kg), entered preferentially into the brain, the liver, and the heart in decreasing order. In the epididymal adipose tissue, the drug was present only in small amounts. Ditazole disappeared from the rat organs 4 hr after the treatment. The apparent ditazole half-life in rat blood was 41 min, the volume of distribution was 2.068 liters/kg, and the body clearance was 0.0345 liter/kg/min. | [
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PMID:25335 | Differential spectrophotometric determination of tyrosine and tryptophan in pharmaceutical amino acid solutions. | A method is proposed for the simultaneous determination of tyrosine and tryptophan in solutions by differential spectrophotometry. The concentrations are calculated from the measurement of the absorbance of the amino acid mixture in an alkaline medium and from the differential absorbance of the alkaline solution against the acid solution at 294.4 nm. A comparison with three other well-known methods is discussed. | Differential spectrophotometric determination of tyrosine and tryptophan in pharmaceutical amino acid solutions. A method is proposed for the simultaneous determination of tyrosine and tryptophan in solutions by differential spectrophotometry. The concentrations are calculated from the measurement of the absorbance of the amino acid mixture in an alkaline medium and from the differential absorbance of the alkaline solution against the acid solution at 294.4 nm. A comparison with three other well-known methods is discussed. | [
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PMID:25337 | Rat jugular vein relaxes to norepinephrine, phenylephrine and histamine. | Circular muscle of the rat external jugular vein contracted to serotonin, angiotensin and potassium chloride but not to norepinephrine, phenylephrine, histamine or carbamylcholine. In contrast, rabbit and guinea-pig jugular veins contracted to norepinephrine, phenylephrine and histamine, although contractions to norepinephrine were small in guinea-pig jugular veins. Norepinephrine, phenylephrine and histamine produced a concentration-dependent sustained relaxation of serotonin-induced contractions in the rat jugular vein, as did isoproterenol, nitroglycerin and papaverine. Propranolol blocked relaxation to norepinephrine, phenylephrine and isoproterenol whereas metiamide, a H2 receptor antagonist blocked relaxation to histamine. alpha adrenergic receptor blockade with phentolamine or prazosin resulted in greater relaxation to norepinephrine whereas cocaine did not enhance norepinephrine-induced vasodilation. This study supports the premise that norepinephrine may exert prominent beta adrenergic receptor stimulation in some blood vessels and that this effect may be more apparent in veins than arteries. | Rat jugular vein relaxes to norepinephrine, phenylephrine and histamine. Circular muscle of the rat external jugular vein contracted to serotonin, angiotensin and potassium chloride but not to norepinephrine, phenylephrine, histamine or carbamylcholine. In contrast, rabbit and guinea-pig jugular veins contracted to norepinephrine, phenylephrine and histamine, although contractions to norepinephrine were small in guinea-pig jugular veins. Norepinephrine, phenylephrine and histamine produced a concentration-dependent sustained relaxation of serotonin-induced contractions in the rat jugular vein, as did isoproterenol, nitroglycerin and papaverine. Propranolol blocked relaxation to norepinephrine, phenylephrine and isoproterenol whereas metiamide, a H2 receptor antagonist blocked relaxation to histamine. alpha adrenergic receptor blockade with phentolamine or prazosin resulted in greater relaxation to norepinephrine whereas cocaine did not enhance norepinephrine-induced vasodilation. This study supports the premise that norepinephrine may exert prominent beta adrenergic receptor stimulation in some blood vessels and that this effect may be more apparent in veins than arteries. | [
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PMID:25341 | Nucleoside transport in mammalian cell membranes. IV. Organomercurials and organomercurial-mercaptonucleoside complexes as probes for nucleoside transport systems in hamster cells. | Organomercurials form stable stoichiometric complexes with thiolated nucleosides. The complexes inhibited uptake of ribonucleosides and cytosine arabinoside (CAR) in various types of normal and transformed cells. The inhibition was competitive and reversible (Ki = 3--6 micrometer). The interaction between complexes and transport system displayed a 1:1 stoichiometry. Chemical factors which contributed to the inhibitory power were evaluated with a series of S-alkylated derivatives and S--Hg--R complexes of mercaptonucleosides. The inhibitory potency was not determined exclusively by the hydrophobic nature of either the S-alkylated or the S--Hg--R moieties. Chemical modification of cells with penetrating and nonpenetrating organomercurials lead to stimulation of nucleoside uptake and to an increase in its susceptibility to inhibition by S--Hg--R complexes or S-aklylated derivatives of mercaptopurine ribosides. The kinetic and chemical data obtained with nucleoside analogs and with chemical modifiers suggested complex features of nucleoside transport systems. Four distinct classes of sites were implied: (i) a substrate binding site susceptible directly to competitive inhibition by organomercurial-mercaptonucleoside complexes, (ii) an additional site susceptible either to S-arylalkylated or S-mercuriated derivatives of 6-mercaptopurine ribosides, (iii) SH-containing modifier sites which stimulate uridine uptake upon binding of organomercurials, and (iv) SH-containing modifier sites which inhibit the function upon binding of organomercurials. From the observation that only SH sites related to stimulation were susceptible to modification by macromolecular-SH modifier probes, some conclusions can be drawn regarding the disposition of the various sites in the cell membrane in general and among membrane components in particular. | Nucleoside transport in mammalian cell membranes. IV. Organomercurials and organomercurial-mercaptonucleoside complexes as probes for nucleoside transport systems in hamster cells. Organomercurials form stable stoichiometric complexes with thiolated nucleosides. The complexes inhibited uptake of ribonucleosides and cytosine arabinoside (CAR) in various types of normal and transformed cells. The inhibition was competitive and reversible (Ki = 3--6 micrometer). The interaction between complexes and transport system displayed a 1:1 stoichiometry. Chemical factors which contributed to the inhibitory power were evaluated with a series of S-alkylated derivatives and S--Hg--R complexes of mercaptonucleosides. The inhibitory potency was not determined exclusively by the hydrophobic nature of either the S-alkylated or the S--Hg--R moieties. Chemical modification of cells with penetrating and nonpenetrating organomercurials lead to stimulation of nucleoside uptake and to an increase in its susceptibility to inhibition by S--Hg--R complexes or S-aklylated derivatives of mercaptopurine ribosides. The kinetic and chemical data obtained with nucleoside analogs and with chemical modifiers suggested complex features of nucleoside transport systems. Four distinct classes of sites were implied: (i) a substrate binding site susceptible directly to competitive inhibition by organomercurial-mercaptonucleoside complexes, (ii) an additional site susceptible either to S-arylalkylated or S-mercuriated derivatives of 6-mercaptopurine ribosides, (iii) SH-containing modifier sites which stimulate uridine uptake upon binding of organomercurials, and (iv) SH-containing modifier sites which inhibit the function upon binding of organomercurials. From the observation that only SH sites related to stimulation were susceptible to modification by macromolecular-SH modifier probes, some conclusions can be drawn regarding the disposition of the various sites in the cell membrane in general and among membrane components in particular. | [
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PMID:25342 | Hypertonic cryohemolysis: ionophore and pH effects. | Human erythrocytes suspended at 37 degrees C in hypertonic solution of either electrolytes or nonelectrolytes undergo hemolysis when the temperature is lowered toward 0 degrees C (Green, F.A., Jung, C.Y. 1977 J. Membrane Biol. 33:249). In the present studies this hypertonic cryohemolysis was profoundly affected by the pH of incubation, and was completely abolished at ph 5. In hypertonic NaCl, there was an apparent pH optimum at 6--6.5. In hypertonic sucrose, on the other hand, hemolysis increased progressively with increasing pH between 6 and 9. Amphotericin B inhibited hypertonic cryohemolysis in NaCl or KCl solution. No inhibiting effect of amphotericin B was observed when hypertonicity was due to sodium sulfate or sucrose. Valinomycin also inhibited hypertonic cryohemolysis in KCl, but did not affect the process in NaCl or sucrose solution. SITS (4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonate) and phloretin interfered with this valinomycin effect, whereas phlorizin did not. These results indicate that dissipation of an osmotic gradient across membranes may be responsible for the inhibition of the hemolysis by these inophores. Iso-osmotic cell shrinkage induced by valinomycin in 150 mM NaCl solution did not result in cryohemolysis. | Hypertonic cryohemolysis: ionophore and pH effects. Human erythrocytes suspended at 37 degrees C in hypertonic solution of either electrolytes or nonelectrolytes undergo hemolysis when the temperature is lowered toward 0 degrees C (Green, F.A., Jung, C.Y. 1977 J. Membrane Biol. 33:249). In the present studies this hypertonic cryohemolysis was profoundly affected by the pH of incubation, and was completely abolished at ph 5. In hypertonic NaCl, there was an apparent pH optimum at 6--6.5. In hypertonic sucrose, on the other hand, hemolysis increased progressively with increasing pH between 6 and 9. Amphotericin B inhibited hypertonic cryohemolysis in NaCl or KCl solution. No inhibiting effect of amphotericin B was observed when hypertonicity was due to sodium sulfate or sucrose. Valinomycin also inhibited hypertonic cryohemolysis in KCl, but did not affect the process in NaCl or sucrose solution. SITS (4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonate) and phloretin interfered with this valinomycin effect, whereas phlorizin did not. These results indicate that dissipation of an osmotic gradient across membranes may be responsible for the inhibition of the hemolysis by these inophores. Iso-osmotic cell shrinkage induced by valinomycin in 150 mM NaCl solution did not result in cryohemolysis. | [
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PMID:25343 | Current-voltage relationships for the plasma membrane and its principal electrogenic pump in Neurospora crassa: I. Steady-state conditions. | The nonlinear membrane current-voltage relationship (I-V curve) for intact hyphae of Neurospora crassa has been determined by means of a 3-electrode voltage-clamp technique, plus "quasi-linear" cable theory. Under normal conditions of growth and respiration, the membrane I-V curve is best described as a parabolic segment convex in the direction of depolarizing current. At the average resting potential of - 174 mV, the membrane conductance is approximately 190 micronhos/cm2; conductance increase to approximately 240 micronhos/cm2 at -300 mV, and decreases to approximately 130 micronhos/cm2 at 0 mV. Irreversible membrane breakdown occurs at potentials beyond this range. Inhibition of the primary electrogenic pump in Neurospora by ATP withdrawal (with 1 mM KCN) depolarizes the membrane to the range of -40 to -70 mV and reduces the slope of the I-V curve by a fixed scaling factor of approximately 0.8. For wild-type Neurospora, compared under control conditions and during steady-state inhibition by cyanide, the I-V difference curve--presumed to define the current-voltage curve for the electrogenic pump--is a saturation function with maximal current of approximately 20 muA/cm2, a half saturation potential near -300 mV, and a projected reversal potential of ca. -400 mV. This value is close to the maximal free energy available to the pump from ATP hydrolysis, so that pump stoichiometry must be close to 1 H+ extruded:1 ATP split. The time-courses of change in membrane potential and resistance with cyanide are compatible with the steady-state I-V curves, under the assumption the cyanide has no major effects other than ATP withdrawal. Other inhibitors, uncouplers, and lowered temperature all have more complicated effects. The detailed temporal analysis of voltage-clamp data showed three time-constants in the clamping currents: one of 10 msec, for charging the membrane capacitance (0.9 muF/cm/2); a second of 50-75 msec; and a third of 20-30 sec, perhaps representing changes of intracellular composition. | Current-voltage relationships for the plasma membrane and its principal electrogenic pump in Neurospora crassa: I. Steady-state conditions. The nonlinear membrane current-voltage relationship (I-V curve) for intact hyphae of Neurospora crassa has been determined by means of a 3-electrode voltage-clamp technique, plus "quasi-linear" cable theory. Under normal conditions of growth and respiration, the membrane I-V curve is best described as a parabolic segment convex in the direction of depolarizing current. At the average resting potential of - 174 mV, the membrane conductance is approximately 190 micronhos/cm2; conductance increase to approximately 240 micronhos/cm2 at -300 mV, and decreases to approximately 130 micronhos/cm2 at 0 mV. Irreversible membrane breakdown occurs at potentials beyond this range. Inhibition of the primary electrogenic pump in Neurospora by ATP withdrawal (with 1 mM KCN) depolarizes the membrane to the range of -40 to -70 mV and reduces the slope of the I-V curve by a fixed scaling factor of approximately 0.8. For wild-type Neurospora, compared under control conditions and during steady-state inhibition by cyanide, the I-V difference curve--presumed to define the current-voltage curve for the electrogenic pump--is a saturation function with maximal current of approximately 20 muA/cm2, a half saturation potential near -300 mV, and a projected reversal potential of ca. -400 mV. This value is close to the maximal free energy available to the pump from ATP hydrolysis, so that pump stoichiometry must be close to 1 H+ extruded:1 ATP split. The time-courses of change in membrane potential and resistance with cyanide are compatible with the steady-state I-V curves, under the assumption the cyanide has no major effects other than ATP withdrawal. Other inhibitors, uncouplers, and lowered temperature all have more complicated effects. The detailed temporal analysis of voltage-clamp data showed three time-constants in the clamping currents: one of 10 msec, for charging the membrane capacitance (0.9 muF/cm/2); a second of 50-75 msec; and a third of 20-30 sec, perhaps representing changes of intracellular composition. | [
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PMID:25347 | Bloody pericardial fluid. The value of blood gas measurements. | Clinically notable pericardial effusions developed in three patients with renal failure. Pericardiocentesis showed hemorrhagic fluid, the source of which was not apparent. Simultaneous determinations of PCO2, PO2, and pH values showed a substantial increase in PCO2 levels and decrease in PO2, pH, and bicarbonate levels in the pericardial compared with the intracardial aspirates. This was true when pericardial fluid PCO2, PO2, and pH values were compared with mixed venous samples. Determination of PO2, PCO2, pH, and bicarbonate values in pericardial aspirates may determine the source of the fluid. | Bloody pericardial fluid. The value of blood gas measurements. Clinically notable pericardial effusions developed in three patients with renal failure. Pericardiocentesis showed hemorrhagic fluid, the source of which was not apparent. Simultaneous determinations of PCO2, PO2, and pH values showed a substantial increase in PCO2 levels and decrease in PO2, pH, and bicarbonate levels in the pericardial compared with the intracardial aspirates. This was true when pericardial fluid PCO2, PO2, and pH values were compared with mixed venous samples. Determination of PO2, PCO2, pH, and bicarbonate values in pericardial aspirates may determine the source of the fluid. | [
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PMID:25348 | Some new or poorly recognized physical and graphic findings in various forms of pericardial disease. | The eight new items concerning pericarditis (acute pericarditis, acute or chronic pericardial effusion, and constrictive or sero-constrictive pericarditis) were demonstrated based on the clinical observation of 115 patients. Pericarditis, a disease of remote antiquity, still has many fascinating and unrecognized aspects, which will be uncovered by the meticulous observation of the patients. | Some new or poorly recognized physical and graphic findings in various forms of pericardial disease. The eight new items concerning pericarditis (acute pericarditis, acute or chronic pericardial effusion, and constrictive or sero-constrictive pericarditis) were demonstrated based on the clinical observation of 115 patients. Pericarditis, a disease of remote antiquity, still has many fascinating and unrecognized aspects, which will be uncovered by the meticulous observation of the patients. | [
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PMID:25351 | [Use of beta-adrenergic blockaders in the treatment of arterial hypertension]. | The results of treatment of over 300 patients suffering from hypertensive disease with beta-blocking agents Obsidan and Visken are analysed and the place of these agents among other hypotensive drugs is discussed. The therapeutic effect was most evident in patients with the early stages of the disease and hyperkinetic type of circulation. The daily doses were from 60 to 240 mg. The hemodynamic shifts were displayed by a decrease in the cardiac output and rate of cardiac contractions and reflex, ususlly moderate, increase in general peripheral resistance, as a result the arterial pressure decreased gradually. In 1 to 6 months general peripheral resistance diminished in half of the patients, evidently due to the gradual adaptation of the vascular tone to the chronic reduction of the cardiac output. Treatment with beta-blocking agents led to non-uniform changes in regional hemodynamics: the tone of the cerebral arteries significantly reduced, the tone of the arteries of the lower extremities had a tendency to increase. This gives rise to the conclusion that alpha-adrenergic receptors predominate in the vessels of the lower extremities. In the treatment of patients with high and stable hypertension, it is advisable to combine beta-blocking agents with saluretics, vasodilators, hemiton, and other hypotensive drugs. | [Use of beta-adrenergic blockaders in the treatment of arterial hypertension]. The results of treatment of over 300 patients suffering from hypertensive disease with beta-blocking agents Obsidan and Visken are analysed and the place of these agents among other hypotensive drugs is discussed. The therapeutic effect was most evident in patients with the early stages of the disease and hyperkinetic type of circulation. The daily doses were from 60 to 240 mg. The hemodynamic shifts were displayed by a decrease in the cardiac output and rate of cardiac contractions and reflex, ususlly moderate, increase in general peripheral resistance, as a result the arterial pressure decreased gradually. In 1 to 6 months general peripheral resistance diminished in half of the patients, evidently due to the gradual adaptation of the vascular tone to the chronic reduction of the cardiac output. Treatment with beta-blocking agents led to non-uniform changes in regional hemodynamics: the tone of the cerebral arteries significantly reduced, the tone of the arteries of the lower extremities had a tendency to increase. This gives rise to the conclusion that alpha-adrenergic receptors predominate in the vessels of the lower extremities. In the treatment of patients with high and stable hypertension, it is advisable to combine beta-blocking agents with saluretics, vasodilators, hemiton, and other hypotensive drugs. | [
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PMID:25352 | [Coronary circulation under alpha-adrenergic blockade]. | In canine experiments with coronary catheterization and perfusion, catheterization of the heart cavities and great vessles, and with thermodilution it was demonstrated that the alpha-adrenergic blocker Phentolamine causes a regular flow resistance in the coronary vessels. The maximal depressor effect was observed with 0.3 mg/kg (intravenously), and it did not grow under higher doses of the drug. The reduction of coronary resistance under the effect of Phentolamine was realized, firstly, by way of a moderate activation of myocardial beta-receptors, and secondly, by way of a specific block of alpha-adrenergic receptors of the heart vessels. Phentolamine inhibited the pressor reactions of the coronaries to mesatone, adrenalin, noradrenalin, and limited the manifestations of the pressor effects in the coronary vessels during systemic sinocarotid reflexes. | [Coronary circulation under alpha-adrenergic blockade]. In canine experiments with coronary catheterization and perfusion, catheterization of the heart cavities and great vessles, and with thermodilution it was demonstrated that the alpha-adrenergic blocker Phentolamine causes a regular flow resistance in the coronary vessels. The maximal depressor effect was observed with 0.3 mg/kg (intravenously), and it did not grow under higher doses of the drug. The reduction of coronary resistance under the effect of Phentolamine was realized, firstly, by way of a moderate activation of myocardial beta-receptors, and secondly, by way of a specific block of alpha-adrenergic receptors of the heart vessels. Phentolamine inhibited the pressor reactions of the coronaries to mesatone, adrenalin, noradrenalin, and limited the manifestations of the pressor effects in the coronary vessels during systemic sinocarotid reflexes. | [
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PMID:25355 | [The different effects of fe(II)- and fe(III)-ions on the hyaluronic acid of the vitreous body (author's transl)]. | Fe(II)-ions reduce the viscosity of a hyaluronic acid solution but do not form a precipitate. Fe(III)-ions only slightly lower the viscosity of hyaluronic acid solutions but cause a brown, water-insoluble iron-hyaluronic acid precipitate. After adding ascorbic acid--concentrate the pH-value as in vitreous body--to the reaction solution, Fe(III)-ions reduce the viscosity too. The iron-hyalurnic acid complex is also destroyed by ascorbate. Therefore, the ascorbate of the vitreous humor plays an important role in the generation of siderosis bulbi. | [The different effects of fe(II)- and fe(III)-ions on the hyaluronic acid of the vitreous body (author's transl)]. Fe(II)-ions reduce the viscosity of a hyaluronic acid solution but do not form a precipitate. Fe(III)-ions only slightly lower the viscosity of hyaluronic acid solutions but cause a brown, water-insoluble iron-hyaluronic acid precipitate. After adding ascorbic acid--concentrate the pH-value as in vitreous body--to the reaction solution, Fe(III)-ions reduce the viscosity too. The iron-hyalurnic acid complex is also destroyed by ascorbate. Therefore, the ascorbate of the vitreous humor plays an important role in the generation of siderosis bulbi. | [
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PMID:25356 | [Possibility of conserving the heat balance of the human organism in an extremely rarefied atmosphere by vacuum evaporation of sweat from the surface of the body]. | As a result of 64 experiments carried out on 24 test subjects, it has been shown that extreme overwarming due to moderate and heavy thermal loads can be prevented with the aid of heat release through sweat evaporation from the surface of the body under the action of a rarefied atmosphere (termed vacuum sweat evaporation). The test subjects wearing altitude suits remained in a thermal altitude chamber at 40 and 55 degrees C for 8 to 3 hours at rest and doing light work at altitudes of 7 to 25 km without marked signs of overwarming. The entire heat release occurred only through vacuum sweat evaporation from the surface of the body that was under the altitude suit during a gradual decrease of the chamber pressure to 20 mm Hg (equivalent to an altitude of 25 km). The findings give support to the hypothesis that heart release via vacuum sweat evaporation from the surface of the body can be used as a method of controlling thermal homeostasis during high altitude and space flights. | [Possibility of conserving the heat balance of the human organism in an extremely rarefied atmosphere by vacuum evaporation of sweat from the surface of the body]. As a result of 64 experiments carried out on 24 test subjects, it has been shown that extreme overwarming due to moderate and heavy thermal loads can be prevented with the aid of heat release through sweat evaporation from the surface of the body under the action of a rarefied atmosphere (termed vacuum sweat evaporation). The test subjects wearing altitude suits remained in a thermal altitude chamber at 40 and 55 degrees C for 8 to 3 hours at rest and doing light work at altitudes of 7 to 25 km without marked signs of overwarming. The entire heat release occurred only through vacuum sweat evaporation from the surface of the body that was under the altitude suit during a gradual decrease of the chamber pressure to 20 mm Hg (equivalent to an altitude of 25 km). The findings give support to the hypothesis that heart release via vacuum sweat evaporation from the surface of the body can be used as a method of controlling thermal homeostasis during high altitude and space flights. | [
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PMID:25357 | [Experimental substantiation of the maximum permissible concentrations of acetone and acetaldehyde in regenerated drinking water]. | The paper presents experimental data indicating maximum permissible concentrations of acetone and acetaldehyde in reclaimed potable water used for different periods of time--up to 1, 3 or 12 months. The recommended concentrations for reclaimed water used for a year are 14.0 mg/l and 0.2 mg/l for acetone and acetaldehyde, respectively. The recommended levels for reclaimed water used for 1--3 months are 39.6 mg/l and 0.2 mg/l for acetone and acetaldehyde, respectively. | [Experimental substantiation of the maximum permissible concentrations of acetone and acetaldehyde in regenerated drinking water]. The paper presents experimental data indicating maximum permissible concentrations of acetone and acetaldehyde in reclaimed potable water used for different periods of time--up to 1, 3 or 12 months. The recommended concentrations for reclaimed water used for a year are 14.0 mg/l and 0.2 mg/l for acetone and acetaldehyde, respectively. The recommended levels for reclaimed water used for 1--3 months are 39.6 mg/l and 0.2 mg/l for acetone and acetaldehyde, respectively. | [
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PMID:25361 | Submandibular salivary pH as a diagnostic aid for prognosis of facial palsy. | The present paper aims at introducing a new method of testing the function of the chorda tympani with the aid of submandibular salivary pH. This method has several advantages: (1.) it enables the measurement of the function of only the paralyzed side; (2.) it accurately predicts the outcome of facial palsy at an early stage; and (3.) the method is simple and inexpensive. | Submandibular salivary pH as a diagnostic aid for prognosis of facial palsy. The present paper aims at introducing a new method of testing the function of the chorda tympani with the aid of submandibular salivary pH. This method has several advantages: (1.) it enables the measurement of the function of only the paralyzed side; (2.) it accurately predicts the outcome of facial palsy at an early stage; and (3.) the method is simple and inexpensive. | [
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PMID:25368 | [Effect of soap and detergents on the restitution of acidity and lipids of the skin surface]. | A comparative evaluation of the effects of soaps and detergents on pH behaviour and lipids level on the skin surface and duration of their restitution was carried out. The studies were conducted in two groups. The first group of 32 persons used 1% solution of "BHP" soap, whereas the other one, composed of 31 persons, 0,5% solution of detergentpowder "POLLENA". PH measurements of skin were performed using pH--meter of Radelkis firm with a combined glass and calomel electrode. Lipids on the skin surface were determined by the Hodgson--Jones and Wheatly's gravimetric method. The evaluation of soap and detergent's effects on skin was based on the mean output and direct pH values of the skin surface, accidental and residual level of lipids, and comparison of the duration of these parameters restitution. The soap was found to alkalyse stronger than the detergent, and duration of pH restitution is longer after using the soap, whereas the detergent degreases the skin more intensively than the soap. Also restitution of lipid's jacket is slower after using the detergent. | [Effect of soap and detergents on the restitution of acidity and lipids of the skin surface]. A comparative evaluation of the effects of soaps and detergents on pH behaviour and lipids level on the skin surface and duration of their restitution was carried out. The studies were conducted in two groups. The first group of 32 persons used 1% solution of "BHP" soap, whereas the other one, composed of 31 persons, 0,5% solution of detergentpowder "POLLENA". PH measurements of skin were performed using pH--meter of Radelkis firm with a combined glass and calomel electrode. Lipids on the skin surface were determined by the Hodgson--Jones and Wheatly's gravimetric method. The evaluation of soap and detergent's effects on skin was based on the mean output and direct pH values of the skin surface, accidental and residual level of lipids, and comparison of the duration of these parameters restitution. The soap was found to alkalyse stronger than the detergent, and duration of pH restitution is longer after using the soap, whereas the detergent degreases the skin more intensively than the soap. Also restitution of lipid's jacket is slower after using the detergent. | [
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PMID:25369 | An affinity-column procedure for the purification of veratrate O-demethylase from fungi. | An affinity column procedure is reported for purifying veratrate O-demethylase from higher fungi. The procedure is based on the affinity of the fungal demethylases for veratrate, which was coupled to AH-Sepharose 4B. An over 300-fold purification of the enzyme from an Ascomycete (Chaetomium piluliferum), and a lower degree of purification (20-fold) from a Basidiomycete (Xerocomus badius), were obtained. The O-demethylases from higher fungi require NADH and oxygen. The enzyme activity is sensitive to exposure to oxygen. The pH optima are 5 for enzyme from Chaetomium, and 7 for demethylase from Xerocomus, respectively. The enzymes are not specific for veratrate. They also demethylate p- and m-anisate and 3,4-dimethoxycinnamate, but to a lower degree. | An affinity-column procedure for the purification of veratrate O-demethylase from fungi. An affinity column procedure is reported for purifying veratrate O-demethylase from higher fungi. The procedure is based on the affinity of the fungal demethylases for veratrate, which was coupled to AH-Sepharose 4B. An over 300-fold purification of the enzyme from an Ascomycete (Chaetomium piluliferum), and a lower degree of purification (20-fold) from a Basidiomycete (Xerocomus badius), were obtained. The O-demethylases from higher fungi require NADH and oxygen. The enzyme activity is sensitive to exposure to oxygen. The pH optima are 5 for enzyme from Chaetomium, and 7 for demethylase from Xerocomus, respectively. The enzymes are not specific for veratrate. They also demethylate p- and m-anisate and 3,4-dimethoxycinnamate, but to a lower degree. | [
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PMID:25370 | Specific agglutination of Escherichia coli O128B12 by the mannose-binding proteins of Pseudomonas aeruginosa. | The mannosephilic haemagglutinins of Pseudomonas aeruginosa were found to agglutinate cells of Escherichia coli O128B12, to be adsorbed onto them and to attach peroxidase to them. These reactions were specifically inhibited by D-mannose. No agglutination by this Pseudomonas haemagglutinin was obtained when several other enteropathogenic types of Escherichia coli and some other Gram-negative bacteria were examined. Concanavalin A, which also reacted with Escherichia coli O128B12 cells, interacted with some of the other bacteria examined, too. Escherichia coli O128B12 was not agglutinated by the Pseudomonas galactosephilic haemagglutinins and those of the plant Phaseolus vulgaris. Its maximal agglutination by the Pseudomonas mannosephilic haemagglutinins was obtained employing cells grown for 4-6 h in conventional media. The growth temperature, aeration and presence of certain amino acids, but not D-mannose, in the culture medium had some effect on the agglutination in tensity; pH 6-8 was optimal for it and only at pH 3.0-3.2 no agglutination was observed. Treatment of the bacteria by proteolytic enzymes, ethanol or formaldehyde did not alter their agglutinability by either the Pseudomonas lectin or by antibodies produced against them in rabbits. Heating of the bacteria to 100 degrees C prevented their agglutination by the Pseudomonas lectin and lowered their ability to adsorb it, but did not significantly affect their reactions with the rabbit antibodies. | Specific agglutination of Escherichia coli O128B12 by the mannose-binding proteins of Pseudomonas aeruginosa. The mannosephilic haemagglutinins of Pseudomonas aeruginosa were found to agglutinate cells of Escherichia coli O128B12, to be adsorbed onto them and to attach peroxidase to them. These reactions were specifically inhibited by D-mannose. No agglutination by this Pseudomonas haemagglutinin was obtained when several other enteropathogenic types of Escherichia coli and some other Gram-negative bacteria were examined. Concanavalin A, which also reacted with Escherichia coli O128B12 cells, interacted with some of the other bacteria examined, too. Escherichia coli O128B12 was not agglutinated by the Pseudomonas galactosephilic haemagglutinins and those of the plant Phaseolus vulgaris. Its maximal agglutination by the Pseudomonas mannosephilic haemagglutinins was obtained employing cells grown for 4-6 h in conventional media. The growth temperature, aeration and presence of certain amino acids, but not D-mannose, in the culture medium had some effect on the agglutination in tensity; pH 6-8 was optimal for it and only at pH 3.0-3.2 no agglutination was observed. Treatment of the bacteria by proteolytic enzymes, ethanol or formaldehyde did not alter their agglutinability by either the Pseudomonas lectin or by antibodies produced against them in rabbits. Heating of the bacteria to 100 degrees C prevented their agglutination by the Pseudomonas lectin and lowered their ability to adsorb it, but did not significantly affect their reactions with the rabbit antibodies. | [
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PMID:25373 | [X-ray examination of the abdomen in hypnotic-sedative drug poisoning--casuistical contribution (author's transl)]. | By means of an impressive example the significance of abdominal X-ray examination in cases of intoxication with hypnotic-sedative drugs is pointed out. Apart from the qualitative diagnosis (drug containing bromide) the radiological proof of shadow-giving substances also permits a quantitative clinical assessment and has prognostical as well as therapeutical consequences. | [X-ray examination of the abdomen in hypnotic-sedative drug poisoning--casuistical contribution (author's transl)]. By means of an impressive example the significance of abdominal X-ray examination in cases of intoxication with hypnotic-sedative drugs is pointed out. Apart from the qualitative diagnosis (drug containing bromide) the radiological proof of shadow-giving substances also permits a quantitative clinical assessment and has prognostical as well as therapeutical consequences. | [
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PMID:25372 | A solution to the graft-versus-host problem in bone marrow transplantation to humans. | Many clinical situations arise where it would be desirable to transplant bone marrow to a marrow function deficient patient. However, bone marrow is immunologically competent by virtue of its content of lymphocyte precursors. Marrow transplantation in these patients is followed by the graft's immunological rejection of the patient - a fatal disease. A system for specific abrogation of this graft-versus-host disease after xenogeneic bone marrow transplantation to humans is presented. In this system, prospective patient cells are removed by skin biopsy for example, and injected into a fetal chimpanzee. The fetal chimpanzee becomes tolerant to this patient's antigens and will not attack or reject them when its bone marrow is removed after birth and injected into the specific patient from whom the tolerogenic cells were obtained. A simple and straightforward experimental test of this system's clinical applicability is also presented. | A solution to the graft-versus-host problem in bone marrow transplantation to humans. Many clinical situations arise where it would be desirable to transplant bone marrow to a marrow function deficient patient. However, bone marrow is immunologically competent by virtue of its content of lymphocyte precursors. Marrow transplantation in these patients is followed by the graft's immunological rejection of the patient - a fatal disease. A system for specific abrogation of this graft-versus-host disease after xenogeneic bone marrow transplantation to humans is presented. In this system, prospective patient cells are removed by skin biopsy for example, and injected into a fetal chimpanzee. The fetal chimpanzee becomes tolerant to this patient's antigens and will not attack or reject them when its bone marrow is removed after birth and injected into the specific patient from whom the tolerogenic cells were obtained. A simple and straightforward experimental test of this system's clinical applicability is also presented. | [
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PMID:25379 | Release of [14C]tyrosine from tubulinyl-[14C]tyrosine by brain extract. Separation of a carboxypeptidase from tubulin-tyrosine ligase. | The carboxypeptidase previously described that releases tyrosine from tubulinyl-tyrosine was obtained from rat brain preparation free of tubulin-tyrosine ligase. The enzyme was purified 24-fold. Its activity was increased by 2 mM MgCl2 or 30 mM KCl. Mercaptoethanol (50 mM), colchicine (0.2 mM) and tyrosine (0.2 mM) showed practically no effect on the release of tyrosine whereas iodoacetate (2 mM), deoxycholate (0.5%), CuCl2 (0.1 mM), ZnCl2 (0.1 mM) and NaCl or KCl (240 mM) had a strong inhibitory effect. The optimal pH of this enzyme was 6.3--7. A preparation containing tubulin-tyrosine ligase free of carboxypeptidase was also obtained. This preparation catalyzed the release of tyrosine from tyrosinated tubulin in the presence of ADP, Mg2+, K+ and Pi and the incorporation of tyrosine into tubulin. For the releasing activity the optimal concentration of MgCl2 was 3--20 mM and of KCl was 10--30 mM. For ADP the maximal act;vity was at 0.3 mM or higher. An important difference between the activities of the carboxypeptidase and the ligase was that the former was active on denatured tubulin whereas the latter was not. | Release of [14C]tyrosine from tubulinyl-[14C]tyrosine by brain extract. Separation of a carboxypeptidase from tubulin-tyrosine ligase. The carboxypeptidase previously described that releases tyrosine from tubulinyl-tyrosine was obtained from rat brain preparation free of tubulin-tyrosine ligase. The enzyme was purified 24-fold. Its activity was increased by 2 mM MgCl2 or 30 mM KCl. Mercaptoethanol (50 mM), colchicine (0.2 mM) and tyrosine (0.2 mM) showed practically no effect on the release of tyrosine whereas iodoacetate (2 mM), deoxycholate (0.5%), CuCl2 (0.1 mM), ZnCl2 (0.1 mM) and NaCl or KCl (240 mM) had a strong inhibitory effect. The optimal pH of this enzyme was 6.3--7. A preparation containing tubulin-tyrosine ligase free of carboxypeptidase was also obtained. This preparation catalyzed the release of tyrosine from tyrosinated tubulin in the presence of ADP, Mg2+, K+ and Pi and the incorporation of tyrosine into tubulin. For the releasing activity the optimal concentration of MgCl2 was 3--20 mM and of KCl was 10--30 mM. For ADP the maximal act;vity was at 0.3 mM or higher. An important difference between the activities of the carboxypeptidase and the ligase was that the former was active on denatured tubulin whereas the latter was not. | [
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PMID:25380 | Reversible inhibition of cyclic AMP-dependent protein kinase by insulin. | Extracts of fasted rat diaphragms, previously treated with or without insulin were assayed for glycogen synthase, protein kinase and cyclic [3H]-AMP binding. Treatment with insulin produced an elevation in the % of glycogen synthase I and a concurrent decrease in cyclic AMP-dependent protein kinase activity and cyclic [3H]-AMP binding. Analysis of extracts by disc gel electrophoresis demonstrated the inhibition of cyclic [3H]-AMP binding to involve the Type I protein kinase holoenzyme. Inhibition of protein kinase activity was most apparent in the presence of 0.2 micrometer cyclic AMP, with enzymatic activity of the insulin-treated extracts typically 60--65% of control. Higher assay concentrations diminished the difference between control and insulin-treated extracts and concentrations greater than 20 micrometer abolished it. The inhibition of cyclic AMP-dependent protein kinase activity after insulin was a transient and labile phenomenon. The effect was independent of ATP concentration in the assay, but was sensitive to the pH of tissue extraction, requiring a pH of 7.0 to 8.4 to be observed. Insulin-mediated inhibition of protein kinase activity was reversed upon preincubation of extracts at 0--2 degrees. Relatively concentrated homogenates (less than 4 microliter buffer/mg tissue) yielded extracts which exhibited little or no inhibition of protein kinase activity compared to extracts prepared from more dilute (6--10 microliter/mg) homogenates. A model for the inhibition of the cyclic-AMP dependent protein kinase by an insulin-generated inhibitor which becomes directly associated with the Type 1 holoenzyme is proposed. | Reversible inhibition of cyclic AMP-dependent protein kinase by insulin. Extracts of fasted rat diaphragms, previously treated with or without insulin were assayed for glycogen synthase, protein kinase and cyclic [3H]-AMP binding. Treatment with insulin produced an elevation in the % of glycogen synthase I and a concurrent decrease in cyclic AMP-dependent protein kinase activity and cyclic [3H]-AMP binding. Analysis of extracts by disc gel electrophoresis demonstrated the inhibition of cyclic [3H]-AMP binding to involve the Type I protein kinase holoenzyme. Inhibition of protein kinase activity was most apparent in the presence of 0.2 micrometer cyclic AMP, with enzymatic activity of the insulin-treated extracts typically 60--65% of control. Higher assay concentrations diminished the difference between control and insulin-treated extracts and concentrations greater than 20 micrometer abolished it. The inhibition of cyclic AMP-dependent protein kinase activity after insulin was a transient and labile phenomenon. The effect was independent of ATP concentration in the assay, but was sensitive to the pH of tissue extraction, requiring a pH of 7.0 to 8.4 to be observed. Insulin-mediated inhibition of protein kinase activity was reversed upon preincubation of extracts at 0--2 degrees. Relatively concentrated homogenates (less than 4 microliter buffer/mg tissue) yielded extracts which exhibited little or no inhibition of protein kinase activity compared to extracts prepared from more dilute (6--10 microliter/mg) homogenates. A model for the inhibition of the cyclic-AMP dependent protein kinase by an insulin-generated inhibitor which becomes directly associated with the Type 1 holoenzyme is proposed. | [
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PMID:25384 | Antacid titration in the prevention of acute gastrointestinal bleeding: a controlled, randomized trial in 100 critically ill patients. | We randomized 100 critically ill patients at risk of developing acute gastrointestinal ulceration and bleeding into two groups. One (51 patients) received antacid prophylaxis, and the other (49 patients) received no specific form of prophylaxis. Hourly antacid titration kept the pH of the gastric contents above 3.5. Two of the 51 patients who received antacid prophylaxis and gastrointestinal bleeding. Twelve of the 49 control patients bled (P less than 0.005). Of the 12 patients in the control group who bled, seven were placed on antacid medication, and all seven apparently stopped bleeding. Analysis of all the patients showed that an increasing prevalence of respiratory, failure, sepsis, peritonitis, jaundice, renal failure and hypotension was correlated with a greater frequency of bleeding. No patients required operative treatment to control bleeding. These data indicate that the occurrence of acute gastrointestinal bleeding in critically ill patients can be reduced by antacid titration. | Antacid titration in the prevention of acute gastrointestinal bleeding: a controlled, randomized trial in 100 critically ill patients. We randomized 100 critically ill patients at risk of developing acute gastrointestinal ulceration and bleeding into two groups. One (51 patients) received antacid prophylaxis, and the other (49 patients) received no specific form of prophylaxis. Hourly antacid titration kept the pH of the gastric contents above 3.5. Two of the 51 patients who received antacid prophylaxis and gastrointestinal bleeding. Twelve of the 49 control patients bled (P less than 0.005). Of the 12 patients in the control group who bled, seven were placed on antacid medication, and all seven apparently stopped bleeding. Analysis of all the patients showed that an increasing prevalence of respiratory, failure, sepsis, peritonitis, jaundice, renal failure and hypotension was correlated with a greater frequency of bleeding. No patients required operative treatment to control bleeding. These data indicate that the occurrence of acute gastrointestinal bleeding in critically ill patients can be reduced by antacid titration. | [
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PMID:25385 | A manpower policy for primary health care. | A National Academy of Sciences study of policy options for the supply of primary health-care manpower has produced a comprehensive set of recommendations. The study finds an adequate overall supply of physicians, but a shortage of primary health-care practitioners. It recommends maintaining current enrollment levels in medical schools and training programs for nurse practitioners and physician assistants and increasing the proportion of primary-care residents. To enhance the availability of primary care, the report advocates reimbursement for all physicians within a state at the same payment level for the same primary-care service, a reduction in payment differentials between primary-care services and nonprimary-care services, and reimbursement for educational and preventive services and for new health-practitioner services. The report supports a team approach in primary-care training and recommends that all medical students obtain clinical experience in a primary-care setting and some instruction in epidemiology and behavioral and social sciences. | A manpower policy for primary health care. A National Academy of Sciences study of policy options for the supply of primary health-care manpower has produced a comprehensive set of recommendations. The study finds an adequate overall supply of physicians, but a shortage of primary health-care practitioners. It recommends maintaining current enrollment levels in medical schools and training programs for nurse practitioners and physician assistants and increasing the proportion of primary-care residents. To enhance the availability of primary care, the report advocates reimbursement for all physicians within a state at the same payment level for the same primary-care service, a reduction in payment differentials between primary-care services and nonprimary-care services, and reimbursement for educational and preventive services and for new health-practitioner services. The report supports a team approach in primary-care training and recommends that all medical students obtain clinical experience in a primary-care setting and some instruction in epidemiology and behavioral and social sciences. | [
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PMID:25387 | The structure of the flavoenzyme glutathione reductase. | The three-dimensional structure of the dimeric flavoenzyme glutathione reductase from human erythrocytes has been elucidated by an X-ray diffraction analysis at 0.3 nm resolution. The polypeptide chain has been traced, and the binding positions of FAD, NADP and glutathione have been determined. A mechanism for the electron transfer is discussed. | The structure of the flavoenzyme glutathione reductase. The three-dimensional structure of the dimeric flavoenzyme glutathione reductase from human erythrocytes has been elucidated by an X-ray diffraction analysis at 0.3 nm resolution. The polypeptide chain has been traced, and the binding positions of FAD, NADP and glutathione have been determined. A mechanism for the electron transfer is discussed. | [
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PMID:25400 | [Effect of antimediator preparations on the intercellular relations in early embryos of the sea urchin]. | The dated treatment of the early embryos of an irregular (flat) sea urchin Scaphechinus mirabilis by neuropharmacological drugs (anti-neurotransmitters) during one of the first four cleavage divisions results in the impairment of intercellular connections and leads to the formation of twin embryos, dwarf embryos, embryos of the dumb-bell shape etc. In the experiments with some of the drugs under study such developmental abnormalities were not seen or were expressed much more weakly when serotonin or bufotenin (N,N-dimethylserotonin) were added to the medium. A suggestion is put forward that the early embryos possess an intracellular mechanism participating in the interaction between the cells and operating via endogenous monoamines, primarily serotonin. | [Effect of antimediator preparations on the intercellular relations in early embryos of the sea urchin]. The dated treatment of the early embryos of an irregular (flat) sea urchin Scaphechinus mirabilis by neuropharmacological drugs (anti-neurotransmitters) during one of the first four cleavage divisions results in the impairment of intercellular connections and leads to the formation of twin embryos, dwarf embryos, embryos of the dumb-bell shape etc. In the experiments with some of the drugs under study such developmental abnormalities were not seen or were expressed much more weakly when serotonin or bufotenin (N,N-dimethylserotonin) were added to the medium. A suggestion is put forward that the early embryos possess an intracellular mechanism participating in the interaction between the cells and operating via endogenous monoamines, primarily serotonin. | [
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PMID:25401 | [Effect of cortisol administration and stress actions in early ontogeny on hormonal induction in adult rats]. | The influence of cortisole injections and stress ("handling") in the early ontogenesis (during the first 9 or 16 days of life) on the process of thyrosine aminotransferase induction by cortisol in the adult rats has been studied. It was shown that both the injection of the hormone in the young rats and "handling" led to the manifested trace effects: (1) stable increase of thyrosine aminotransferase activity in the liver of adult (5-6 months) rats and (2) appearance of peculiar tolerance to cortisol in the form of decrease in the ability of cells to induce thyrosine aminotransferase in response to the hormone injection. The data obtained suggest that the sensitive period of postnatal ontogenesis when cortisol or "handling" exert such a stable effect is limited by the 3rd and 9th days after birth. The causes of such "biochemical imprinting" are considered with respect to the increased sensitivity of the genetical system of cells-targets to the transcription inducers during the early postnatal development. | [Effect of cortisol administration and stress actions in early ontogeny on hormonal induction in adult rats]. The influence of cortisole injections and stress ("handling") in the early ontogenesis (during the first 9 or 16 days of life) on the process of thyrosine aminotransferase induction by cortisol in the adult rats has been studied. It was shown that both the injection of the hormone in the young rats and "handling" led to the manifested trace effects: (1) stable increase of thyrosine aminotransferase activity in the liver of adult (5-6 months) rats and (2) appearance of peculiar tolerance to cortisol in the form of decrease in the ability of cells to induce thyrosine aminotransferase in response to the hormone injection. The data obtained suggest that the sensitive period of postnatal ontogenesis when cortisol or "handling" exert such a stable effect is limited by the 3rd and 9th days after birth. The causes of such "biochemical imprinting" are considered with respect to the increased sensitivity of the genetical system of cells-targets to the transcription inducers during the early postnatal development. | [
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PMID:25407 | Cystic fibrosis: isolation and physical properties of a salivary cystic fibrosis factor. | A ciliostatic factor has been isolated from cystic fibrosis (CF) saliva by dialyzing it from purified alpha-amylase prepared by a glycogen-complex method. This method of isolating the CF factor is an improvement over the previously employed heparin procedure. The activity of the isolated factor is proportional with concentration using the oyster gill ciliostatic assay and in its inhibition of mammalian glycogen debranching enzyme. The ciliostatic action of the factor can be reversed by heparin under certain conditions. The type of inhibition of the debranching enzyme by the isolated CF factor indicates that its chemical structure is similar to that observed with hydroxyalkylamines and polyamine metabolites. Physical properties of the isolated factor indicate that it is of low molecular weight and is labile as a function of pH and temperature. At neutral pH the conditions under which it is maintained have a direct effect on the length of time that it is stable. | Cystic fibrosis: isolation and physical properties of a salivary cystic fibrosis factor. A ciliostatic factor has been isolated from cystic fibrosis (CF) saliva by dialyzing it from purified alpha-amylase prepared by a glycogen-complex method. This method of isolating the CF factor is an improvement over the previously employed heparin procedure. The activity of the isolated factor is proportional with concentration using the oyster gill ciliostatic assay and in its inhibition of mammalian glycogen debranching enzyme. The ciliostatic action of the factor can be reversed by heparin under certain conditions. The type of inhibition of the debranching enzyme by the isolated CF factor indicates that its chemical structure is similar to that observed with hydroxyalkylamines and polyamine metabolites. Physical properties of the isolated factor indicate that it is of low molecular weight and is labile as a function of pH and temperature. At neutral pH the conditions under which it is maintained have a direct effect on the length of time that it is stable. | [
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PMID:25410 | Immunologic studies of arylsulfatase A in normal and metachromatic leukodystrophy liver. | Purified human liver arylsulfatase A on polyacrylamide gel electrophoresis at pH 4.0 is separated into two protein forms with enzymatic activity and two distinct inactive subunits. All of these components were immunologically distinguishable using different antisera preparations. In late infantile metachromatic leukodystrophy, only one of the two inactive subunits was immunologically detected, whereas in the juvenile form of metachromatic leukodystrophy, both inactive subunits were antigenically present. | Immunologic studies of arylsulfatase A in normal and metachromatic leukodystrophy liver. Purified human liver arylsulfatase A on polyacrylamide gel electrophoresis at pH 4.0 is separated into two protein forms with enzymatic activity and two distinct inactive subunits. All of these components were immunologically distinguishable using different antisera preparations. In late infantile metachromatic leukodystrophy, only one of the two inactive subunits was immunologically detected, whereas in the juvenile form of metachromatic leukodystrophy, both inactive subunits were antigenically present. | [
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PMID:25411 | Hypophosphatemic vitamin D-resistant rickets: metabolic balance studies in a child receiving 1,25 dihydroxyvitamin D3, phosphate, and ascorbic acid. | A child with hypophosphatemic vitamin D-resistant rickets was treated for three years with the conventional vitamin D-inorganic phosphate supplementation followed by a new therapeutic regimen consisting of 1,25 dihydroxyvitamin D3 (1,25 (OH)2D3) and half of the previous phosphate supplementation. The effectiveness of the two treatment regimens was compared by calcium, phosphate, and magnesium balance techniques and by serial radiological examinations as well as careful height measurements. In addition, the lowering of the urinary pH with ascorbic acid supplementation seems to be associated with improvement in the renal tubular reabsorption of phosphate, but its distinct effect, separate from the rest of the treatment modalities, was not tested in this study. The conventional treatment did not correct the hypophosphatemia and alkaline phosphatase elevation, whereas the 1,25 (OH)2 D3-inorganic phosphate regimen is well tolerated and effective in achieving a sustained normalization of these variables. In addition, the improved growth and healing of rickets further attest to the efficacy of the new treatment. | Hypophosphatemic vitamin D-resistant rickets: metabolic balance studies in a child receiving 1,25 dihydroxyvitamin D3, phosphate, and ascorbic acid. A child with hypophosphatemic vitamin D-resistant rickets was treated for three years with the conventional vitamin D-inorganic phosphate supplementation followed by a new therapeutic regimen consisting of 1,25 dihydroxyvitamin D3 (1,25 (OH)2D3) and half of the previous phosphate supplementation. The effectiveness of the two treatment regimens was compared by calcium, phosphate, and magnesium balance techniques and by serial radiological examinations as well as careful height measurements. In addition, the lowering of the urinary pH with ascorbic acid supplementation seems to be associated with improvement in the renal tubular reabsorption of phosphate, but its distinct effect, separate from the rest of the treatment modalities, was not tested in this study. The conventional treatment did not correct the hypophosphatemia and alkaline phosphatase elevation, whereas the 1,25 (OH)2 D3-inorganic phosphate regimen is well tolerated and effective in achieving a sustained normalization of these variables. In addition, the improved growth and healing of rickets further attest to the efficacy of the new treatment. | [
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PMID:25412 | Arterial blood gases in Pranayama practice. | Pranayama is a Yogic breathing practice which is known experientially to produce a profound calming effect on the mind. In an experiment designed to determine whether the mental effects of this practice were accompanied by changes in the arterial blood gases, arterial blood was drawn from 10 trained individuals prior to and immediately after Pranayama practice. No significance changes in arterial blood gases were noted after Pranayama. A neural mechanism for the mental effects of this practice is proposed. | Arterial blood gases in Pranayama practice. Pranayama is a Yogic breathing practice which is known experientially to produce a profound calming effect on the mind. In an experiment designed to determine whether the mental effects of this practice were accompanied by changes in the arterial blood gases, arterial blood was drawn from 10 trained individuals prior to and immediately after Pranayama practice. No significance changes in arterial blood gases were noted after Pranayama. A neural mechanism for the mental effects of this practice is proposed. | [
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PMID:25413 | [Giant cell arteritis and takayasu's disease: histopathological criteria (author's transl)]. | Both of these arterial diseases may involve the aorta and the major arterial trunks. Two cases of subclavian involvement are used to contrast them from a histopathological standpoint. In giant cell arteritis, the lesions affect above all the internal elastic layer and the inner part of the media, destroyed by an inflammatory infiltrate with giant cells. In Takayasu's disease, the lesions involve the adventitia, the site of fibrosis and of inflammatory islets with the vasa vasorum at the centres. Involvement of the media is predominantly in its outer part, the internal elastic layer being intact. A histopathological definition of these arterial diseases may be envisaged on the basis of these facts. | [Giant cell arteritis and takayasu's disease: histopathological criteria (author's transl)]. Both of these arterial diseases may involve the aorta and the major arterial trunks. Two cases of subclavian involvement are used to contrast them from a histopathological standpoint. In giant cell arteritis, the lesions affect above all the internal elastic layer and the inner part of the media, destroyed by an inflammatory infiltrate with giant cells. In Takayasu's disease, the lesions involve the adventitia, the site of fibrosis and of inflammatory islets with the vasa vasorum at the centres. Involvement of the media is predominantly in its outer part, the internal elastic layer being intact. A histopathological definition of these arterial diseases may be envisaged on the basis of these facts. | [
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PMID:25414 | Preparative separation of the complementary strands of DNA restriction fragments by alkaline RPC-5 chromatography. | High pressure reversed phase chromatography (RPC-5) at pH 12 was used for preparative separation of the complementary strands of the smaller DNA fragments which are generated by the Hae III restriction endonuclease digestion of Col El DNA. A single high pressure RPC-5 chromatographic step at neutral pH served to purify duplex fragments 70, 172, 250 and 440 base pairs long; each of these yielded two elution peaks upon chromatography under alkaline denaturing conditions. | Preparative separation of the complementary strands of DNA restriction fragments by alkaline RPC-5 chromatography. High pressure reversed phase chromatography (RPC-5) at pH 12 was used for preparative separation of the complementary strands of the smaller DNA fragments which are generated by the Hae III restriction endonuclease digestion of Col El DNA. A single high pressure RPC-5 chromatographic step at neutral pH served to purify duplex fragments 70, 172, 250 and 440 base pairs long; each of these yielded two elution peaks upon chromatography under alkaline denaturing conditions. | [
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PMID:25415 | Turbidimetric and potentiometric studies of ribosomal subunits from an erythromycin resistant mutant of Escherichia coli. | Turbidimetric and potentiometric techniques were applied to the analysis of an EryR mutant. Results show that in the mutant, the 30S subunits are drastically altered, as indicated by a higher Mg2+ requirement for subunit association and by an important difference in the titratable groups. Replacement of parental 50S by mutant 50S subunits does not decrease the association capacity with 30S parental subunits, but a structural difference is detected in the mutant 50S with potentiometric measurements. The mutation results in decreased ribosomal in vitro activities at 22 degrees C including lowered polyphenylalanine synthesis, drastic altered initiation step and the loss of erythromycin binding to the ribosomes. The results extend previous observation of a gene eryC part in the maturation of both subunits. | Turbidimetric and potentiometric studies of ribosomal subunits from an erythromycin resistant mutant of Escherichia coli. Turbidimetric and potentiometric techniques were applied to the analysis of an EryR mutant. Results show that in the mutant, the 30S subunits are drastically altered, as indicated by a higher Mg2+ requirement for subunit association and by an important difference in the titratable groups. Replacement of parental 50S by mutant 50S subunits does not decrease the association capacity with 30S parental subunits, but a structural difference is detected in the mutant 50S with potentiometric measurements. The mutation results in decreased ribosomal in vitro activities at 22 degrees C including lowered polyphenylalanine synthesis, drastic altered initiation step and the loss of erythromycin binding to the ribosomes. The results extend previous observation of a gene eryC part in the maturation of both subunits. | [
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PMID:25416 | Affinity adsorbents consisting of nucleic acids immobilized via bisoxirane activated polysaccharides. | An easy and efficient procedure for the immobilization of polynucleotide ligands to bisoxirane activated insoluble polysaccharides has been elaborated and is described in this paper. The resulting materials have been applied to the chromatography of DNA polymerase I, and RNA polymerase from E.coli. Because of their extraordinary stability to temperature, formamide, and alkaline conditions they seem to be particularly useful adsorbents for nucleic acid hybridization. | Affinity adsorbents consisting of nucleic acids immobilized via bisoxirane activated polysaccharides. An easy and efficient procedure for the immobilization of polynucleotide ligands to bisoxirane activated insoluble polysaccharides has been elaborated and is described in this paper. The resulting materials have been applied to the chromatography of DNA polymerase I, and RNA polymerase from E.coli. Because of their extraordinary stability to temperature, formamide, and alkaline conditions they seem to be particularly useful adsorbents for nucleic acid hybridization. | [
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