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PMID:27253 | Difficulties in comparing catecholamine-related enzymes from the brains of schizophrenics and controls. | The catecholamine-forming and metabolizing enzyme tyrosine hydroxylase, dopa decarboxylase, dopamine-beta-hydroxylase, phenylethanolamine N-methyltransferase, and catecholamine-O-methyltransferase, as well as the endogenous inhibitor of dopamine-beta-hydroxylase were compared in the brains of schizophrenics and controls. While there were no statistically significant differences in the enzyme or inhibitor activity between groups, tbre was a decided trend toward a decreased enzyme activity in the brains of the schizophrenics. From another set of control brains it was found that changes in human enzyme activity following death are variable and may be dependent on how the brains were handled. Thus, it is unclear whether the apparent differences between schizophrenics and controls were present when they were alive or occurred after death. | Difficulties in comparing catecholamine-related enzymes from the brains of schizophrenics and controls. The catecholamine-forming and metabolizing enzyme tyrosine hydroxylase, dopa decarboxylase, dopamine-beta-hydroxylase, phenylethanolamine N-methyltransferase, and catecholamine-O-methyltransferase, as well as the endogenous inhibitor of dopamine-beta-hydroxylase were compared in the brains of schizophrenics and controls. While there were no statistically significant differences in the enzyme or inhibitor activity between groups, tbre was a decided trend toward a decreased enzyme activity in the brains of the schizophrenics. From another set of control brains it was found that changes in human enzyme activity following death are variable and may be dependent on how the brains were handled. Thus, it is unclear whether the apparent differences between schizophrenics and controls were present when they were alive or occurred after death. | [
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PMID:27258 | Effect of anticonvulsant drugs on alkaline phosphatase and gamma-glutamyltransferase activity in human leukocytes. | The authors compared the alkaline phosphatase and gamma-glutamyl-transferase activities in leukocytes from presumably healthy subjects and from epileptic patients treated with anticonvulsants, matched for age and sex. Intraleukocyte alkaline phosphatase activity was higher in women than in men and was greatly reduced by the anticonvulsant drugs. Intraleukocyte gammaglutamyltransferase activity did not vary with sex and was unaffected by the anticonvulsant drugs contrary to findings for the activity of this enzyme in plasma and liver. | Effect of anticonvulsant drugs on alkaline phosphatase and gamma-glutamyltransferase activity in human leukocytes. The authors compared the alkaline phosphatase and gamma-glutamyl-transferase activities in leukocytes from presumably healthy subjects and from epileptic patients treated with anticonvulsants, matched for age and sex. Intraleukocyte alkaline phosphatase activity was higher in women than in men and was greatly reduced by the anticonvulsant drugs. Intraleukocyte gammaglutamyltransferase activity did not vary with sex and was unaffected by the anticonvulsant drugs contrary to findings for the activity of this enzyme in plasma and liver. | [
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PMID:27259 | Thermodynamic investigations of proteins. IV. Calcium binding protein parvalbumin. | The conformational transitions of calcium binding protein parvalbumin III from carp muscle were studied by scanning calorimetry, potentiometric titration and isothermal calorimetric titration. Changes of Gibbs energy, enthalpy and partial heat capacity were determined. The removal of calcium ions by EDTA is accompanied by 1) a heat absorption of 75 +/- 10 kJ per mole of the protein, 2) a decrease in the Gibbs energy of protein structure stabilisation of about 42 kJ mol-1 and 3) a decrease in thermostability by more than 50 K. The protonation of the acidic groups leads to a loss of calcium followed by denaturation, while the pH of the transition strongly depends on calcium activity. The enthalpy and heat capacity changes at denaturation are comparable with the values observed for other compact globular proteins. | Thermodynamic investigations of proteins. IV. Calcium binding protein parvalbumin. The conformational transitions of calcium binding protein parvalbumin III from carp muscle were studied by scanning calorimetry, potentiometric titration and isothermal calorimetric titration. Changes of Gibbs energy, enthalpy and partial heat capacity were determined. The removal of calcium ions by EDTA is accompanied by 1) a heat absorption of 75 +/- 10 kJ per mole of the protein, 2) a decrease in the Gibbs energy of protein structure stabilisation of about 42 kJ mol-1 and 3) a decrease in thermostability by more than 50 K. The protonation of the acidic groups leads to a loss of calcium followed by denaturation, while the pH of the transition strongly depends on calcium activity. The enthalpy and heat capacity changes at denaturation are comparable with the values observed for other compact globular proteins. | [
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PMID:27256 | Immunosuppressive activity of two plant steroidal lactones withaferin A and withanolide E. | Withaferin A and withanolide E, two steroidal lactones of plant origin were demonstrated to have specific immunosuppressive effects on human B and T lymphocytes as well as on mice thymocytes. E rosettes and EAC rosette formation by normal human T and B lymphocytes were inhibited by the two compounds at very low concentrations. The formation of mouse red blood rosettes by chronic lymphatic leukemic cells were only inhibited by withaferin A. The functional activity of normal human T lymphocytes as assessed by a local xenogeneic graft versus host reaction was also affected by these two plant steroidal lactones. These experiments demonstrate a specific action of the compound on antigen recognition as well as proliferative capacity of T lymphocytes as well as B lymphocytes. | Immunosuppressive activity of two plant steroidal lactones withaferin A and withanolide E. Withaferin A and withanolide E, two steroidal lactones of plant origin were demonstrated to have specific immunosuppressive effects on human B and T lymphocytes as well as on mice thymocytes. E rosettes and EAC rosette formation by normal human T and B lymphocytes were inhibited by the two compounds at very low concentrations. The formation of mouse red blood rosettes by chronic lymphatic leukemic cells were only inhibited by withaferin A. The functional activity of normal human T lymphocytes as assessed by a local xenogeneic graft versus host reaction was also affected by these two plant steroidal lactones. These experiments demonstrate a specific action of the compound on antigen recognition as well as proliferative capacity of T lymphocytes as well as B lymphocytes. | [
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PMID:27260 | Temperature dependence of the interfacial behavior of uracil derivatives. | The effect of temperature on the interfacial behavior of uracil, thymine and 1,5-dimethyluracil has been used to characterize the thermodynamics of adsorption of these compounds at a mercury electrode-aqueous electrolyte interface. Between 4.5 degrees C and 40 degrees C all three compounds exhibit two adsorption regions. The first, a dilute layer where the molecules adsorb in a flat orientation on the electrode surface. The second, a compact layer where the molecules adsorb in a perpendicular orientation. The thermodynamic constants characterizing formation of these surface layers have been deduced. | Temperature dependence of the interfacial behavior of uracil derivatives. The effect of temperature on the interfacial behavior of uracil, thymine and 1,5-dimethyluracil has been used to characterize the thermodynamics of adsorption of these compounds at a mercury electrode-aqueous electrolyte interface. Between 4.5 degrees C and 40 degrees C all three compounds exhibit two adsorption regions. The first, a dilute layer where the molecules adsorb in a flat orientation on the electrode surface. The second, a compact layer where the molecules adsorb in a perpendicular orientation. The thermodynamic constants characterizing formation of these surface layers have been deduced. | [
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PMID:27257 | Action of beta-adrenoceptor blocking agents on hemodynamic variables and on baroreceptor and postganglionic sympathetic activities in rabbits. | Fifty-six experiments were performed in rabbits anesthetized by urethane or sodium pentobarbital. Baroreceptor (aortic nerve) and post-ganglionic sympathetic (renal nerve) activities were recorded, and peripheral resistances were determined after infusion of beta-adrenoceptor blocking agents (propranolol, pindolol, timolol). A progressive vasodilation was observed after the onset of infusion of the 3 drugs; at the same time arterial pressure decreased, but aortic flow did not vary significantly. Baroreceptor activity was enhanced, whereas efferent renal nerve activity was decreased. | Action of beta-adrenoceptor blocking agents on hemodynamic variables and on baroreceptor and postganglionic sympathetic activities in rabbits. Fifty-six experiments were performed in rabbits anesthetized by urethane or sodium pentobarbital. Baroreceptor (aortic nerve) and post-ganglionic sympathetic (renal nerve) activities were recorded, and peripheral resistances were determined after infusion of beta-adrenoceptor blocking agents (propranolol, pindolol, timolol). A progressive vasodilation was observed after the onset of infusion of the 3 drugs; at the same time arterial pressure decreased, but aortic flow did not vary significantly. Baroreceptor activity was enhanced, whereas efferent renal nerve activity was decreased. | [
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PMID:27261 | Volume changes during enzyme reactions. The influence of pressure on the action of invertase, dextranase and dextransucrase. | The pressure dependence of the maximum velocities and the Michaelis constants for the enzymes invertase and dextranase was measured up to 1400 bar. The corresponding activation volumes deltaV not equal to c and deltaV not equal to Km proved to be independent of pressure. Together with data from other sources the meaning of deltaV not equal to c and deltaV not equal to Km is established and the volume profiles of the reactions are constructed. These profiles are similar in contour to the volume profile of the dextran formation catalyzed by the enzyme dextransucrase, but the amount of the volume changes is very much larger for dextransucrase. The evaluation of salt effects shows, that for all three enzymes solvent interactions are not important in explaining the results. The reaction mechanisms seem to be governed by conformation changes of the enzymes. The larger effects in dextransucrase are explained by the produced dextran chain remaining tightly bound to the enzyme and being transported relative to the enzymes position in each reaction cycle. | Volume changes during enzyme reactions. The influence of pressure on the action of invertase, dextranase and dextransucrase. The pressure dependence of the maximum velocities and the Michaelis constants for the enzymes invertase and dextranase was measured up to 1400 bar. The corresponding activation volumes deltaV not equal to c and deltaV not equal to Km proved to be independent of pressure. Together with data from other sources the meaning of deltaV not equal to c and deltaV not equal to Km is established and the volume profiles of the reactions are constructed. These profiles are similar in contour to the volume profile of the dextran formation catalyzed by the enzyme dextransucrase, but the amount of the volume changes is very much larger for dextransucrase. The evaluation of salt effects shows, that for all three enzymes solvent interactions are not important in explaining the results. The reaction mechanisms seem to be governed by conformation changes of the enzymes. The larger effects in dextransucrase are explained by the produced dextran chain remaining tightly bound to the enzyme and being transported relative to the enzymes position in each reaction cycle. | [
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PMID:27262 | [Effect of the alpha-adrenoblocker pyrroxan on systemic hemodynamics in puppies and dogs with vasorenal hypertension]. | Vasorenal hypertension was induced in 2--3-month-old puppies and adult dogs by stricture of both renal arteries. 1.5 mg/kg of pyrroxan was injected intravenously 3 and 14 days later. A reduction of increased arterial pressure was noted both in adult dogs and in puppies, to the subnormal level in the latter. Hypotensive effect of the preparation was connected in adult animals with diminution of the general peripheral vascular resistance and in puppies, besides, with reduction of cardiac output. Pyrroxan injection was accompanied in all the animals with tachycardia, reduction of the phase of isometric contraction and activation of myocardial contractility. | [Effect of the alpha-adrenoblocker pyrroxan on systemic hemodynamics in puppies and dogs with vasorenal hypertension]. Vasorenal hypertension was induced in 2--3-month-old puppies and adult dogs by stricture of both renal arteries. 1.5 mg/kg of pyrroxan was injected intravenously 3 and 14 days later. A reduction of increased arterial pressure was noted both in adult dogs and in puppies, to the subnormal level in the latter. Hypotensive effect of the preparation was connected in adult animals with diminution of the general peripheral vascular resistance and in puppies, besides, with reduction of cardiac output. Pyrroxan injection was accompanied in all the animals with tachycardia, reduction of the phase of isometric contraction and activation of myocardial contractility. | [
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PMID:27263 | [Pharmacology of azidomorphine]. | As revealed, in acute experiments on rabbits, cats, and rats, a new morphine-like synthetic analgesic drug azidomorphine exceeded 20--100-fold the morphine ability to inhibit synaptic transmission in the thalamic structures, cerebral cortex, and the spinal cord during the nociceptive stimulation, and also by its analgesic activity. | [Pharmacology of azidomorphine]. As revealed, in acute experiments on rabbits, cats, and rats, a new morphine-like synthetic analgesic drug azidomorphine exceeded 20--100-fold the morphine ability to inhibit synaptic transmission in the thalamic structures, cerebral cortex, and the spinal cord during the nociceptive stimulation, and also by its analgesic activity. | [
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] |
PMID:27264 | Platelet storage at 22 degrees C: effect of type of agitation on morphology, viability, and function in vitro. | Recovery in vivo after 51Cr labeling, platelet morphology, and platelet aggregation were studied with platelet concentrates (PC) stored for transfusion under carefully controlled conditions. PC were prepared to a final volume of 50 ml from whole blood anticoagulated with citrate-phosphate-dextrose (CPD). The platelet count was kept between 0.8 and 1.6 X 10(12) platelets/liter. The PC were stored in bags constructed of polyvinylchloride (PVC) or polyethylene (PE) at 22 degrees C for 72 hr. The bags were placed on a horizontal shaker or a ferris wheel for agitation during storage. No significant changes in pH or platelet count were observed during storage. PC stored on the wheel showed moderate loss of viability and a marked deterioration of platelet morphology and aggregation compared to the shaker. PC stored on the shaker in bags made of PE showed better aggregation with ADP and thrombin but had the same viability and morphology as PC in bags constructed of PVC. Maintenance of normal platelet morphology as determined by phase-contrast microscopy, extent of shape change response, and the size distribution according to the Coulter Counter correlated with recovery in vivo. | Platelet storage at 22 degrees C: effect of type of agitation on morphology, viability, and function in vitro. Recovery in vivo after 51Cr labeling, platelet morphology, and platelet aggregation were studied with platelet concentrates (PC) stored for transfusion under carefully controlled conditions. PC were prepared to a final volume of 50 ml from whole blood anticoagulated with citrate-phosphate-dextrose (CPD). The platelet count was kept between 0.8 and 1.6 X 10(12) platelets/liter. The PC were stored in bags constructed of polyvinylchloride (PVC) or polyethylene (PE) at 22 degrees C for 72 hr. The bags were placed on a horizontal shaker or a ferris wheel for agitation during storage. No significant changes in pH or platelet count were observed during storage. PC stored on the wheel showed moderate loss of viability and a marked deterioration of platelet morphology and aggregation compared to the shaker. PC stored on the shaker in bags made of PE showed better aggregation with ADP and thrombin but had the same viability and morphology as PC in bags constructed of PVC. Maintenance of normal platelet morphology as determined by phase-contrast microscopy, extent of shape change response, and the size distribution according to the Coulter Counter correlated with recovery in vivo. | [
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PMID:27266 | Neurochemical anatomy of the neuroendocrine hypothalamus. Neurochemical anatomy of the hypothalamus. | Microdissection techniques for isolated removal of the various regions of the hypothalamus as well as the individual hypothalamic nuclei are detailed. Recent development of biochemical microassays have made it possible that the concentrations of neurohormones, neuropeptides, neurotransmitters and their related enzymes could be detected in such a small volume of brain tissue than the hypothalamic nuclei. Data available of the hypothalamic distribution of above substances are summarized. The possible role and origin of intra- and extrahypothalamic neurohormones as well as the existence of the so-called "hypophysiotrophic area" are discussed. | Neurochemical anatomy of the neuroendocrine hypothalamus. Neurochemical anatomy of the hypothalamus. Microdissection techniques for isolated removal of the various regions of the hypothalamus as well as the individual hypothalamic nuclei are detailed. Recent development of biochemical microassays have made it possible that the concentrations of neurohormones, neuropeptides, neurotransmitters and their related enzymes could be detected in such a small volume of brain tissue than the hypothalamic nuclei. Data available of the hypothalamic distribution of above substances are summarized. The possible role and origin of intra- and extrahypothalamic neurohormones as well as the existence of the so-called "hypophysiotrophic area" are discussed. | [
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PMID:27267 | Dopaminergic control of prolactin secretion. | A brief summary is given about present-day knowledge and views on the control of prolactin secretion in mammals. This is followed by the presentation of the evidence that dopamine is the inhibitory hypothalamic transducer controlling hormone release by prolactin cells. Most of the evidence is pharmacological in nature. It is only very recently that dopamine has been found in a meaningful concentration in the blood flowing from the median eminence to the pituitary. Although the dopamine concept of an inhibitory control of prolactin secretion seems quite satisfactory, the possibility of another unrelated inhibitory control system is not excluded. | Dopaminergic control of prolactin secretion. A brief summary is given about present-day knowledge and views on the control of prolactin secretion in mammals. This is followed by the presentation of the evidence that dopamine is the inhibitory hypothalamic transducer controlling hormone release by prolactin cells. Most of the evidence is pharmacological in nature. It is only very recently that dopamine has been found in a meaningful concentration in the blood flowing from the median eminence to the pituitary. Although the dopamine concept of an inhibitory control of prolactin secretion seems quite satisfactory, the possibility of another unrelated inhibitory control system is not excluded. | [
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PMID:27268 | Hypothalamic regulatory hormones: physiological and clinical implications. | The hypothalamic regulatory hormones used for clinical studies are TRH, Gn-RH and somatostatin. In addition, as dopamine appears to be a physiological PIF, the dopamine agonists such as bromocriptine, could be considered as functional analogues of PIF. Gn-RH can be used to study the hypothalamic-pituitary gonadal relationship and to test the secretory reserve capacity of the gonadotrophs in disease states. Unfortunately Gn-RH testing discrimulates between pituitary and hypothalamic diseases only poorly. However gonadotrophin deficient men or women may be successfully treated with long-term Gn-RH with induction of puberty, potency, spermatogenesis and ovulation. Somatostatin has multiple actions in inhibiting endocrine and exocrine secretion but its actions are still being explored in diabetes. Bromocriptine, a long acting dopamine agonist (a functional analogue of PIF), suppresses prolactin and is highly effective in treating many hypogonadal states since hyperprolactinaemia is common. It also lowers growth hormone in acromegaly. TRH has provided a major, accurate, sensitive and safe test of thyroid function. | Hypothalamic regulatory hormones: physiological and clinical implications. The hypothalamic regulatory hormones used for clinical studies are TRH, Gn-RH and somatostatin. In addition, as dopamine appears to be a physiological PIF, the dopamine agonists such as bromocriptine, could be considered as functional analogues of PIF. Gn-RH can be used to study the hypothalamic-pituitary gonadal relationship and to test the secretory reserve capacity of the gonadotrophs in disease states. Unfortunately Gn-RH testing discrimulates between pituitary and hypothalamic diseases only poorly. However gonadotrophin deficient men or women may be successfully treated with long-term Gn-RH with induction of puberty, potency, spermatogenesis and ovulation. Somatostatin has multiple actions in inhibiting endocrine and exocrine secretion but its actions are still being explored in diabetes. Bromocriptine, a long acting dopamine agonist (a functional analogue of PIF), suppresses prolactin and is highly effective in treating many hypogonadal states since hyperprolactinaemia is common. It also lowers growth hormone in acromegaly. TRH has provided a major, accurate, sensitive and safe test of thyroid function. | [
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PMID:27269 | [Pathophysiology of sulfatide metabolism in metachromatic leukodystrophy]. | Metachromatic leucodystrophies (MLD) comprise a small group of heredodegenerative disorders of the nervous system. Deficiency of sulfatide-sulfatase or arylsulfatase A is the common defect in all forms of MLD leading to lysosomal sulfatide storage in the nervous tissue and in the kidney. On the basis of animal experiments, experiments with cultured fibroblasts of the patients as well as ultrastructural studies in a case of prenatal MLD, the following pathomechanism is proposed: 1. Lysosomal degradation of a large portion newly synthetised sulfatide normally regulating the net synthesis and incorporation of sulfatide into myelin, causes early accumulation of sulfatide in lysosomes of myelinating cells and in neurons in the genetic deficiency of arylsulfatase A. 2. Early accumulation of sulfatide does not lead to disturbance in myelination. Demyelination occurs possibly by storage of a cytotoxic compound, psychosin sulfate, also a substrate for the missing enzyme. Prevention of MLD is possible by prenatal diagnosis of arylsulfatase A deficiency in cultured amniotic cells. Enzyme substitution of the missing arylsulfatase A is possible by exogenous uptake of the enzyme in cultured fibroblasts. Thereby the defect of sulfatide degradation can be corrected. Although principles of enzyme substitution have been demonstrated, the problems of treating patients with MLD with arylsulfatase A infusions have yet to be overcome. | [Pathophysiology of sulfatide metabolism in metachromatic leukodystrophy]. Metachromatic leucodystrophies (MLD) comprise a small group of heredodegenerative disorders of the nervous system. Deficiency of sulfatide-sulfatase or arylsulfatase A is the common defect in all forms of MLD leading to lysosomal sulfatide storage in the nervous tissue and in the kidney. On the basis of animal experiments, experiments with cultured fibroblasts of the patients as well as ultrastructural studies in a case of prenatal MLD, the following pathomechanism is proposed: 1. Lysosomal degradation of a large portion newly synthetised sulfatide normally regulating the net synthesis and incorporation of sulfatide into myelin, causes early accumulation of sulfatide in lysosomes of myelinating cells and in neurons in the genetic deficiency of arylsulfatase A. 2. Early accumulation of sulfatide does not lead to disturbance in myelination. Demyelination occurs possibly by storage of a cytotoxic compound, psychosin sulfate, also a substrate for the missing enzyme. Prevention of MLD is possible by prenatal diagnosis of arylsulfatase A deficiency in cultured amniotic cells. Enzyme substitution of the missing arylsulfatase A is possible by exogenous uptake of the enzyme in cultured fibroblasts. Thereby the defect of sulfatide degradation can be corrected. Although principles of enzyme substitution have been demonstrated, the problems of treating patients with MLD with arylsulfatase A infusions have yet to be overcome. | [
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PMID:27271 | Alteration of medullary respiratory unit discharge by iontophoretic application of putative neurotransmitters. | 1 Cats with midcollicular decerebration were vagotomized, paralyzed and artificially ventilated. Phrenic nerve activity was recorded as an index of central respiratory rhythm. Medullary respiratory neurones and non-respiratory cells located in approximation to the ventral respiratory nucleus were tested for their responsiveness to iontophoretically applied gamma-aminobutyric acid (GABA), acetylcholine (ACh) and glutamate. 2 GABA tended to inhibit, whereas ACh and glutamate excited activity both of respiratory and non-respiratory units. Some phase-spanning respiratory unit activities were converted to phasic discharge patterns linked to either inspiration or expiration concomitant with application of low GABA doses. Appropriate applications of GABA also resulted in a complete cessation of the respiratory or non-respiratory neuronal activities. 3 While application of ACh or glutamate induced continuous firing in phasic, phase-spanning respiratory neurones, the periodic discharge patterns of inspiratory or expiratory units was not altered by ACh or, in many instances, by glutamate. Only at high doses of glutamate was the phasic discharge of some inspiratory or expiratory units converted to tonic activity. 4 These observations suggest that strong inhibitory processes serve to maintain the phasic firing pattern of respiratory units. These data also support the concept that active-inhibitory phase-switching mechanisms serve to define respiratory rhythmicity. | Alteration of medullary respiratory unit discharge by iontophoretic application of putative neurotransmitters. 1 Cats with midcollicular decerebration were vagotomized, paralyzed and artificially ventilated. Phrenic nerve activity was recorded as an index of central respiratory rhythm. Medullary respiratory neurones and non-respiratory cells located in approximation to the ventral respiratory nucleus were tested for their responsiveness to iontophoretically applied gamma-aminobutyric acid (GABA), acetylcholine (ACh) and glutamate. 2 GABA tended to inhibit, whereas ACh and glutamate excited activity both of respiratory and non-respiratory units. Some phase-spanning respiratory unit activities were converted to phasic discharge patterns linked to either inspiration or expiration concomitant with application of low GABA doses. Appropriate applications of GABA also resulted in a complete cessation of the respiratory or non-respiratory neuronal activities. 3 While application of ACh or glutamate induced continuous firing in phasic, phase-spanning respiratory neurones, the periodic discharge patterns of inspiratory or expiratory units was not altered by ACh or, in many instances, by glutamate. Only at high doses of glutamate was the phasic discharge of some inspiratory or expiratory units converted to tonic activity. 4 These observations suggest that strong inhibitory processes serve to maintain the phasic firing pattern of respiratory units. These data also support the concept that active-inhibitory phase-switching mechanisms serve to define respiratory rhythmicity. | [
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PMID:27272 | Factors limiting the rate of termination of the neuromuscular blocking action of fazadinium dibromide. | 1 Reasons for the termination of the neuromuscular blocking action of fazadinium dibromide have been investigated. 2 In the anaesthetized cat, maximum neuromuscular block of tibialis anterior muscle twitches following rapid intravenous injection of fazadinium was obtained as the injection bolus passed through the muscle, indicating the fazadinium very readily crosses capillary membranes. 3. The half-life of plasma clearance of fazadinium was about 1 min in both cat and man, despite a 10 fold difference in duration of action in these species. Plasma samples were bioassayed for neuromuscular blocking activity on an isolated, superfused phrenic nerve-diaphragm preparation of the rat. 4 In the anaesthetized cat, occlusion of the renal or hepatic circulations did not prolong the neuromuscular blocking action of single doses of fazadinium. Repeated doses accumulated slowly when the hepatic circulation was occluded but not when the renal circulation was occluded. 5 Fazadinium is eliminated from the body by both the liver and kidneys but the rates of these processes are insufficient to account for the initial rapid plasma clearance. 6 The rate-limiting step for the termination of the neuromuscular blocking action of fazadinium is most likely to be the rate of drug-receptor dissociation | Factors limiting the rate of termination of the neuromuscular blocking action of fazadinium dibromide. 1 Reasons for the termination of the neuromuscular blocking action of fazadinium dibromide have been investigated. 2 In the anaesthetized cat, maximum neuromuscular block of tibialis anterior muscle twitches following rapid intravenous injection of fazadinium was obtained as the injection bolus passed through the muscle, indicating the fazadinium very readily crosses capillary membranes. 3. The half-life of plasma clearance of fazadinium was about 1 min in both cat and man, despite a 10 fold difference in duration of action in these species. Plasma samples were bioassayed for neuromuscular blocking activity on an isolated, superfused phrenic nerve-diaphragm preparation of the rat. 4 In the anaesthetized cat, occlusion of the renal or hepatic circulations did not prolong the neuromuscular blocking action of single doses of fazadinium. Repeated doses accumulated slowly when the hepatic circulation was occluded but not when the renal circulation was occluded. 5 Fazadinium is eliminated from the body by both the liver and kidneys but the rates of these processes are insufficient to account for the initial rapid plasma clearance. 6 The rate-limiting step for the termination of the neuromuscular blocking action of fazadinium is most likely to be the rate of drug-receptor dissociation | [
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PMID:27279 | Evidence for the participation of beta1-adrenoceptors in isoprenaline-induced renin release from rat kidny slices in vitro. | 1. The inhibitory effects were studied of 4 beta-adrenoceptor antagonists against renin release induced by isoprenaline (0.5 mumol/1) in rat kidney slices. Additionally the pA2 values of these 4 drugs were measured against isoprenaline in guinea-pig isolated atria and trachea (against beta1- and beta2-adrenoceptors respectively). 2 When employed at a concentration of 2 mumol/1 propranolol and atenolol significantly inhibited renin release (P less than 0.001 and P less than 0.01) whereas practolol and IPS 339 [t-butyl-amino-3 ol-2 propyl) oximino-9 fluorene] had little effect. 2 A positive correlation was shown between the degree of inhibition of renin release and the pA2 of the antagonists at the beta1-adrenoceptors. 4 When practolol and IPS 339 were used in equipotent molar concentrations to propranolol for the beta1-adrenoceptors they inhibited renin release. 5 The results suggest that the adrenoceptor involved in the renin release induced by isoprenaline in the rat kidney is of the beta1-type. | Evidence for the participation of beta1-adrenoceptors in isoprenaline-induced renin release from rat kidny slices in vitro. 1. The inhibitory effects were studied of 4 beta-adrenoceptor antagonists against renin release induced by isoprenaline (0.5 mumol/1) in rat kidney slices. Additionally the pA2 values of these 4 drugs were measured against isoprenaline in guinea-pig isolated atria and trachea (against beta1- and beta2-adrenoceptors respectively). 2 When employed at a concentration of 2 mumol/1 propranolol and atenolol significantly inhibited renin release (P less than 0.001 and P less than 0.01) whereas practolol and IPS 339 [t-butyl-amino-3 ol-2 propyl) oximino-9 fluorene] had little effect. 2 A positive correlation was shown between the degree of inhibition of renin release and the pA2 of the antagonists at the beta1-adrenoceptors. 4 When practolol and IPS 339 were used in equipotent molar concentrations to propranolol for the beta1-adrenoceptors they inhibited renin release. 5 The results suggest that the adrenoceptor involved in the renin release induced by isoprenaline in the rat kidney is of the beta1-type. | [
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PMID:27280 | The effects of isoprenaline and a new beta-sympathomimetic amine upon spontaneous activity, diastolic depolarization and plateau height in cardiac Purkinje fibres. | 1 In spontaneously active Purkinje fibres of young cows the dose-response curves of the action of isoprenaline upon different electrophysiological parameters were measured.2 The increase in slope of diastolic depolarization could roughly be described by a one-for-one binding curve with a half maximum effect near 10(-8) M and the increase in the height of the plateau level by a two-for-one binding curve with a half maximum effect near 10(-7) M (-)-isoprenaline.3 These dose-response curves were similar to those of two parameters measured under voltage clamp conditions by other authors. The increase in slope of diastolic depolarization behaved like the shift of the activation curve for the pacemaker potassium current towards positive potentials and the growth in plateau height like the increase in the slow inward current mainly carried by Ca ions. From this conformity we propose that the parameters evaluated by us from action potential records could be used for a qualitative analysis of the action of catecholamines on pacemaker potassium current and Ca influx.4 The effects of the isomers of a new drug, 1-isopropylamino-3(4'hydroxyphenoxy)-propan-2-ol (IHP), were evaluated in the same way as those of isoprenaline. The (-)-isomer was at optimal concentrations (10(-5) M) nearly half as effective as isoprenaline in increasing frequency and slope of diastolic depolarization but caused no increase in plateau height. An identical relationship, but at 5 to 10 times higher concentrations, was obtained with the (+)-isomer.5 When 10(-4) M(-)-IHP was added to a preparation equilibrated with a maximum dose of (-)-isoprenaline (10(-6) M), frequency and plateau height declined. This result together with the observation that the effects of IHP could be blocked by the specific beta-antagonist propranolol, revealed the beta-agonistic nature of the new drug. Its inefficiency in increasing the plateau height and thus the slow (Ca) inward current was explained by its relatively low potency and intrinsic activity. | The effects of isoprenaline and a new beta-sympathomimetic amine upon spontaneous activity, diastolic depolarization and plateau height in cardiac Purkinje fibres. 1 In spontaneously active Purkinje fibres of young cows the dose-response curves of the action of isoprenaline upon different electrophysiological parameters were measured.2 The increase in slope of diastolic depolarization could roughly be described by a one-for-one binding curve with a half maximum effect near 10(-8) M and the increase in the height of the plateau level by a two-for-one binding curve with a half maximum effect near 10(-7) M (-)-isoprenaline.3 These dose-response curves were similar to those of two parameters measured under voltage clamp conditions by other authors. The increase in slope of diastolic depolarization behaved like the shift of the activation curve for the pacemaker potassium current towards positive potentials and the growth in plateau height like the increase in the slow inward current mainly carried by Ca ions. From this conformity we propose that the parameters evaluated by us from action potential records could be used for a qualitative analysis of the action of catecholamines on pacemaker potassium current and Ca influx.4 The effects of the isomers of a new drug, 1-isopropylamino-3(4'hydroxyphenoxy)-propan-2-ol (IHP), were evaluated in the same way as those of isoprenaline. The (-)-isomer was at optimal concentrations (10(-5) M) nearly half as effective as isoprenaline in increasing frequency and slope of diastolic depolarization but caused no increase in plateau height. An identical relationship, but at 5 to 10 times higher concentrations, was obtained with the (+)-isomer.5 When 10(-4) M(-)-IHP was added to a preparation equilibrated with a maximum dose of (-)-isoprenaline (10(-6) M), frequency and plateau height declined. This result together with the observation that the effects of IHP could be blocked by the specific beta-antagonist propranolol, revealed the beta-agonistic nature of the new drug. Its inefficiency in increasing the plateau height and thus the slow (Ca) inward current was explained by its relatively low potency and intrinsic activity. | [
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PMID:27281 | Enhancement by an antagonist of transmitter release from frog motor nerve terminals. | 1 The effect of Ba2+ on the synchronous release of acetylcholine from frog motor nerve terminals was studied by conventional electrophysiological techniques. 2 When Ca2+ and Ba2+ were the only divalent cations in the bathing fluid, Ba2+ caused a presynaptic reduction in the amplitude of the endplate potential (e.p.p.). This effect was surmountable by increasing the Ca2+ concentration. 3 The affinity constant (KA) for Ba2+, calculated on the assumption that Ba2+ is a competitive inhibitor of the agonist, Ca2+, was 1.1 +/- 0.4 mM-1 (mean +/- s.e. mean, n = 8). 4 When e.p.ps were depressed by the addition of 1 mM Mg2+, addition of Ba2+ (1 to 3 mM) caused either a further presynaptic depression of moderate magnitude or had no additional effect. 5 When e.p.p.s were depressed with [Mg2+] greater than or equal to 2 mM, addition of Ba2+ greater than or equal to 0.9 mM enhanced the e.p.p. amplitude by a presynaptic mechanism. 6 The interaction of the divalent cation antagonists Mg2+ and Ba2+ with the agonist, Ca2+ is discussed. It is demonstrated that a model which considers the nonequilibrium, kinetic properties of binding can be used to describe interactions between divalent cations at the external surface of the motor nerve ending. | Enhancement by an antagonist of transmitter release from frog motor nerve terminals. 1 The effect of Ba2+ on the synchronous release of acetylcholine from frog motor nerve terminals was studied by conventional electrophysiological techniques. 2 When Ca2+ and Ba2+ were the only divalent cations in the bathing fluid, Ba2+ caused a presynaptic reduction in the amplitude of the endplate potential (e.p.p.). This effect was surmountable by increasing the Ca2+ concentration. 3 The affinity constant (KA) for Ba2+, calculated on the assumption that Ba2+ is a competitive inhibitor of the agonist, Ca2+, was 1.1 +/- 0.4 mM-1 (mean +/- s.e. mean, n = 8). 4 When e.p.ps were depressed by the addition of 1 mM Mg2+, addition of Ba2+ (1 to 3 mM) caused either a further presynaptic depression of moderate magnitude or had no additional effect. 5 When e.p.p.s were depressed with [Mg2+] greater than or equal to 2 mM, addition of Ba2+ greater than or equal to 0.9 mM enhanced the e.p.p. amplitude by a presynaptic mechanism. 6 The interaction of the divalent cation antagonists Mg2+ and Ba2+ with the agonist, Ca2+ is discussed. It is demonstrated that a model which considers the nonequilibrium, kinetic properties of binding can be used to describe interactions between divalent cations at the external surface of the motor nerve ending. | [
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PMID:27283 | Rat cortical neurons in cell culture: culture methods, cell morphology, electrophysiology, and synapse formation. | Rat cortical neurons from 15 day embryos are grown in dissociated cell culture and maintained in vitro for 8--12 weeks. The neurons develop into forms which resemble mature cortical neurons in situ, stain with silver and exhibit passive and active electrophysiological properties similar to those of cortical neurons. Extensive chemical excitatory and inhibitory synapses develop de novo. These cultures can provide a model for future studies of mammalian CNS neuronal physiology, transmitter pharmacology, pathophysiology and mechanisms of drug action. | Rat cortical neurons in cell culture: culture methods, cell morphology, electrophysiology, and synapse formation. Rat cortical neurons from 15 day embryos are grown in dissociated cell culture and maintained in vitro for 8--12 weeks. The neurons develop into forms which resemble mature cortical neurons in situ, stain with silver and exhibit passive and active electrophysiological properties similar to those of cortical neurons. Extensive chemical excitatory and inhibitory synapses develop de novo. These cultures can provide a model for future studies of mammalian CNS neuronal physiology, transmitter pharmacology, pathophysiology and mechanisms of drug action. | [
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PMID:27284 | The use of kainic acid in the localization of enzymes in the substantia nigra. | The selective destruction of neuronal perikarya via intracerebral injections of kainic acid was used to elucidate the cellular location of four neurotransmitter-related enzymes in the substantia nigra (SN). Two weeks after intranigral injections of kainic acid, dopamine-sensitive adenylate cyclase, glutamic acid decarboxylase (GAD), choline acetyltransferase (CAT) and acetylcholinesterase (AChE) were measured in the SN. Histological examination of the SN, and a reduction of striatal tyrosine hydroxylase (TH) activity by 94%, confirmed the extensive loss of neuronal cell bodies in the SN. Dopamine stimulation of adenylate cyclase was not reduced in the lesioned SN, supporting the view that dendritically-released dopamine can regulate cyclic AMP synthesis in afferent terminals to these dendrites. Nigral GAD activity was significantly reduced by the lesions, suggesting that there are GAD-containing perikarya in the SN. CAT activity was not affected by the kainic injections, indicating the absence of cholinergic perikarya in the SN. Nigral AChE activity was significantly decreased after kainic injections, thus confirming the presence of AChE within the nigral perikarya. The results suggest that dopamine-sensitive adenylate cyclase and CAT are located within afferents to the SN, while GAD and AChE are found, to some extent at least, in neuronal soma of the SN. The differentail effects of kainic acid on these enzymes suggest that this compound may be a useful neurochemical tool with which to determine the cellular distribution of enzyme systems in the central nervous system. | The use of kainic acid in the localization of enzymes in the substantia nigra. The selective destruction of neuronal perikarya via intracerebral injections of kainic acid was used to elucidate the cellular location of four neurotransmitter-related enzymes in the substantia nigra (SN). Two weeks after intranigral injections of kainic acid, dopamine-sensitive adenylate cyclase, glutamic acid decarboxylase (GAD), choline acetyltransferase (CAT) and acetylcholinesterase (AChE) were measured in the SN. Histological examination of the SN, and a reduction of striatal tyrosine hydroxylase (TH) activity by 94%, confirmed the extensive loss of neuronal cell bodies in the SN. Dopamine stimulation of adenylate cyclase was not reduced in the lesioned SN, supporting the view that dendritically-released dopamine can regulate cyclic AMP synthesis in afferent terminals to these dendrites. Nigral GAD activity was significantly reduced by the lesions, suggesting that there are GAD-containing perikarya in the SN. CAT activity was not affected by the kainic injections, indicating the absence of cholinergic perikarya in the SN. Nigral AChE activity was significantly decreased after kainic injections, thus confirming the presence of AChE within the nigral perikarya. The results suggest that dopamine-sensitive adenylate cyclase and CAT are located within afferents to the SN, while GAD and AChE are found, to some extent at least, in neuronal soma of the SN. The differentail effects of kainic acid on these enzymes suggest that this compound may be a useful neurochemical tool with which to determine the cellular distribution of enzyme systems in the central nervous system. | [
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PMID:27286 | Evidence for glutamate as a neurotransmitter in the corticofugal fibres to the dorsal lateral geniculate body and the superior colliculus in rats. | The high-affinity uptake of L-glutamate, D-aspartate and GABA were examined in homogenates from the dorsal lateral geniculate body and the superior colliculus after removal of the right visual cortex of adult rats. The high-affinity uptake of D-aspartate and L-glutamate were reduced by 53 to 75% respectively in the dorsal lateral geniculate body and by 46 and 53% respectively in the superior colliculus ipsilateral to the lesion. The uptake on the contralateral side was unaffected. The reductions were detected 3 days after the lesion and were maximally developed after 7 days. Subcellular fractionation showed that the main part of the uptake was confined to the synaptosomal fraction of both regions and that the reductions were most prominent in this fraction. The lesion was not accompanied by significant changes in high-affinity uptake of GABA nor in changes of choline acetyltransferase and glutamate decarboxylase activities. The high-affinity L-glutamate uptake on the contralateral visual cortex was unchanged from control values. After ablation of the visual cortex the level of L-glutamate was reduced by 32 and 17% in the ipsilateral dorsal lateral geniculate body and superior colliculus, respectively. The levels of the other amino acids examined, including L-aspartate, were unchanged. Enucleation had no effect on the uptake of L-glutamate and of GABA in the dorsal lateral geniculate body or in the superior colliculus. | Evidence for glutamate as a neurotransmitter in the corticofugal fibres to the dorsal lateral geniculate body and the superior colliculus in rats. The high-affinity uptake of L-glutamate, D-aspartate and GABA were examined in homogenates from the dorsal lateral geniculate body and the superior colliculus after removal of the right visual cortex of adult rats. The high-affinity uptake of D-aspartate and L-glutamate were reduced by 53 to 75% respectively in the dorsal lateral geniculate body and by 46 and 53% respectively in the superior colliculus ipsilateral to the lesion. The uptake on the contralateral side was unaffected. The reductions were detected 3 days after the lesion and were maximally developed after 7 days. Subcellular fractionation showed that the main part of the uptake was confined to the synaptosomal fraction of both regions and that the reductions were most prominent in this fraction. The lesion was not accompanied by significant changes in high-affinity uptake of GABA nor in changes of choline acetyltransferase and glutamate decarboxylase activities. The high-affinity L-glutamate uptake on the contralateral visual cortex was unchanged from control values. After ablation of the visual cortex the level of L-glutamate was reduced by 32 and 17% in the ipsilateral dorsal lateral geniculate body and superior colliculus, respectively. The levels of the other amino acids examined, including L-aspartate, were unchanged. Enucleation had no effect on the uptake of L-glutamate and of GABA in the dorsal lateral geniculate body or in the superior colliculus. | [
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PMID:27287 | Receptor-linked cyclic AMP systems in rat neostriatum: differential localization revealed by kainic acid injection. | Various receptor-linked cyclic AMP systems were measured in rat neostriatum 2--14 days after selective destruction of neuronal cell bodies and dendrites by micro-injection of 3 microgram of kainic acid. Basal adenylate cyclase activity was reduced by up to 56% in the injected side and the sensitivity to dopamine was abolished. Up to 84% of cyclic nucleotide phosphodiesterase activity, hydrolyzing either cyclic AMP or cyclic GMP, was destroyed by kainic acid injection. Specific binding of [3H]etorphine and [3H]spiroperidol was reduced by up to 62% in the injected side, while non-specific binding was unchanged. All of these changes were time-dependent, and were greatest 7--14 days after kainic acid treatment. On the other hand, intrastriatal kainic acid injection caused no change in the steady-state concentration of cyclic AMP in striatal slices, or in the in vivo cyclic AMP content in the striatum of rats killed by microwave irradiation. Receptor-mediated increases in cyclic AMP accumulation in striatal slices were either unchanged or markedly potentiated by kainic acid treatment. The maximum response to adenosine was unchanged, while the response to isoprenaline was increased up to 3.7-fold, the response to dopamine increased up to 6.7-fold, and the response to PGE1 increased up to 30-fold. The effect of dopamine in kainic acid-treated striatal slices was no longer blocked by fluphenazine, but was blocked by propranolol, suggesting an interaction of dopamine with a beta-adrenoceptor in kainic acid-treated slices. The results suggest differential cellular localizations of the various receptor-linked cyclic AMP systems in rat neostriatum. Some dopamine and opiate receptors, as well as most of the phosphodiesterase activity, are associated with local neuronal elements, while beta-adrenoceptor, adenosine and PGE1 alterations in cyclic AMP are not. The potentiation of the beta-adrenoceptor and PGE1 responses suggests that they may occur in glial cells. In addition, the pool of adenylate cyclase destroyed by kainic acid appears to make little contribution to normal levels of cyclic AMP in the tissue. | Receptor-linked cyclic AMP systems in rat neostriatum: differential localization revealed by kainic acid injection. Various receptor-linked cyclic AMP systems were measured in rat neostriatum 2--14 days after selective destruction of neuronal cell bodies and dendrites by micro-injection of 3 microgram of kainic acid. Basal adenylate cyclase activity was reduced by up to 56% in the injected side and the sensitivity to dopamine was abolished. Up to 84% of cyclic nucleotide phosphodiesterase activity, hydrolyzing either cyclic AMP or cyclic GMP, was destroyed by kainic acid injection. Specific binding of [3H]etorphine and [3H]spiroperidol was reduced by up to 62% in the injected side, while non-specific binding was unchanged. All of these changes were time-dependent, and were greatest 7--14 days after kainic acid treatment. On the other hand, intrastriatal kainic acid injection caused no change in the steady-state concentration of cyclic AMP in striatal slices, or in the in vivo cyclic AMP content in the striatum of rats killed by microwave irradiation. Receptor-mediated increases in cyclic AMP accumulation in striatal slices were either unchanged or markedly potentiated by kainic acid treatment. The maximum response to adenosine was unchanged, while the response to isoprenaline was increased up to 3.7-fold, the response to dopamine increased up to 6.7-fold, and the response to PGE1 increased up to 30-fold. The effect of dopamine in kainic acid-treated striatal slices was no longer blocked by fluphenazine, but was blocked by propranolol, suggesting an interaction of dopamine with a beta-adrenoceptor in kainic acid-treated slices. The results suggest differential cellular localizations of the various receptor-linked cyclic AMP systems in rat neostriatum. Some dopamine and opiate receptors, as well as most of the phosphodiesterase activity, are associated with local neuronal elements, while beta-adrenoceptor, adenosine and PGE1 alterations in cyclic AMP are not. The potentiation of the beta-adrenoceptor and PGE1 responses suggests that they may occur in glial cells. In addition, the pool of adenylate cyclase destroyed by kainic acid appears to make little contribution to normal levels of cyclic AMP in the tissue. | [
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PMID:27288 | Effects of estrogen and aminergic drugs on thresholds of medial basal hypothalamic axons in the median eminence of the rat. | Antidromic responses were induced in the medial basal hypothalamus (MBH) of female rats by median eminence (ME) stimulation, and thresholds were measured under various endocrine conditions. Two different types of antidromic response were recognized. In 83 out of 97 responses, repetitive shocks to the ME at 20--40 Hz induced a fractionation of antidromically-driven spikes into A- and B-components. In the remainder, the antidromically activated spikes were stable to stimulation at high frequency. The threshold tended to show higher values in proestrus than in diestrus-I in both types of response. However, only in the former type of response, application of conditioning electrical shocks to the ME was effective in decreasing the threshold for successive test pulses. A decrease in the threshold lasted for 150--200 msec following the conditioning, with the lowest threshold at about 60 msec after the shock. Effects of the conditioning persisted in animals with chronic complete deafferentation of the MBH but were abolished in intact rats by reserpine treatment. Dopamine, but not norepinephrine or serotonin, mimicked the threshold-decreasing effect of the conditioning when applied to the ME. The results suggest the possibility of a depolarizing effect of some dopaminergic neural mechanism projecting from the MBH to the ME and acting on the axons of a particular type of MBH neuron characterized by fractionating spikes. | Effects of estrogen and aminergic drugs on thresholds of medial basal hypothalamic axons in the median eminence of the rat. Antidromic responses were induced in the medial basal hypothalamus (MBH) of female rats by median eminence (ME) stimulation, and thresholds were measured under various endocrine conditions. Two different types of antidromic response were recognized. In 83 out of 97 responses, repetitive shocks to the ME at 20--40 Hz induced a fractionation of antidromically-driven spikes into A- and B-components. In the remainder, the antidromically activated spikes were stable to stimulation at high frequency. The threshold tended to show higher values in proestrus than in diestrus-I in both types of response. However, only in the former type of response, application of conditioning electrical shocks to the ME was effective in decreasing the threshold for successive test pulses. A decrease in the threshold lasted for 150--200 msec following the conditioning, with the lowest threshold at about 60 msec after the shock. Effects of the conditioning persisted in animals with chronic complete deafferentation of the MBH but were abolished in intact rats by reserpine treatment. Dopamine, but not norepinephrine or serotonin, mimicked the threshold-decreasing effect of the conditioning when applied to the ME. The results suggest the possibility of a depolarizing effect of some dopaminergic neural mechanism projecting from the MBH to the ME and acting on the axons of a particular type of MBH neuron characterized by fractionating spikes. | [
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] |
PMID:27291 | Calcium-binding properties of cardiac and skeletal troponin C as determined by circular dichroism and ultraviolet difference spectroscopy. | Calcium titration of the conformational change in cardiac and skeletal troponin C (TN-C) was followed by circular dichroism (CD) at pH values in the range from 5.2 to 7.4. Computer analysis was used to resolve the contributions from the different classes of Ca2+ -binding sites. At pH 6.94 in skeletal TN-C, apparent affinity constants for calcium of 1.8 x 10(7) and 4.5 x 10(5) M-1 were determined for the two classes of binding sites. The more sophisticated computer analysis of the data has revealed a substantial CD contribution from the low-affinity sites (approximately 30% of the high affinity contribution at pH 6.94) and suggests that skeletal TN-C with Ca2+ bound at the low-affinity sites is in a different conformation from that when just the high-affinity sites are occupied, in agreement with a recent nuclear magnetic resonance (NMR) study on this system (Seaman, K. B., Hartshorne, D. J. & Bothener-By, A. A. (1977) Biochemistry 16,4039-4046). With the cardiac protein at pH 7.07, an apparent affinity constant for calcium of 2.0 x 10(7) M-1 was calculated while no low-affinity site at this pH was detected by CD. On the other hand, at lower pH values, such as 6.05, a CD contribution from the cardiac low-affinity Ca2+ -binding site is detected with an apparent binding constant of 3.7 +/- 0.7 x 10(4) M-1. At the lower pH values, protonation of a class of carboxyl groups in each protein which possesses a high pKa (6.2-6.3) elicits the conformational change at the high-affinity sites with a corresponding decrease in the overall magnitude of the Ca2+ -evoked changes. The expression of a conformational change upon Ca2+ binding at the level of the low-affinity sites is enchanced by protonation of a class of carboxyls with a pKa of 6.3 in cardiac TN-C and 6.7-6.8 with the skeletal homologue. In both cases, this contribution is reduced upon protonation of carboxyls with pKa less than or equal to 5.5. It was also observed that the low-affinity sites of skeletal TN-C have a much larger role to play in the total conformational change than the low-affinity sites of cardiac TN-C, a finding probably related to the inability of site 1 in the cardiac protein to bind calcium. In the cardiac protein, the Ca2+ -induced tyrosine difference-spectrum maximum is reduced from deltaepsilonM,287nm =330M-1.cm-1 to 20M-1.cm-1 by protonation of a class of groups with a pKa of 6.4, presumably the same carboxyl groups as those invoved in the CD conformational contribution from the high-affinity binding sites. No such effect was observed for the skeletal protein where deltaepsilonM,287nm was constant at 110M-1 .cm-1 over the pH range studied. The dramatic alterations in the tyrosine environment of cardiac TN-C with pH are attributed to either or both of the tyrosines located in the two high-affinity Ca2+ -binding sites (sites 3 and 4)... | Calcium-binding properties of cardiac and skeletal troponin C as determined by circular dichroism and ultraviolet difference spectroscopy. Calcium titration of the conformational change in cardiac and skeletal troponin C (TN-C) was followed by circular dichroism (CD) at pH values in the range from 5.2 to 7.4. Computer analysis was used to resolve the contributions from the different classes of Ca2+ -binding sites. At pH 6.94 in skeletal TN-C, apparent affinity constants for calcium of 1.8 x 10(7) and 4.5 x 10(5) M-1 were determined for the two classes of binding sites. The more sophisticated computer analysis of the data has revealed a substantial CD contribution from the low-affinity sites (approximately 30% of the high affinity contribution at pH 6.94) and suggests that skeletal TN-C with Ca2+ bound at the low-affinity sites is in a different conformation from that when just the high-affinity sites are occupied, in agreement with a recent nuclear magnetic resonance (NMR) study on this system (Seaman, K. B., Hartshorne, D. J. & Bothener-By, A. A. (1977) Biochemistry 16,4039-4046). With the cardiac protein at pH 7.07, an apparent affinity constant for calcium of 2.0 x 10(7) M-1 was calculated while no low-affinity site at this pH was detected by CD. On the other hand, at lower pH values, such as 6.05, a CD contribution from the cardiac low-affinity Ca2+ -binding site is detected with an apparent binding constant of 3.7 +/- 0.7 x 10(4) M-1. At the lower pH values, protonation of a class of carboxyl groups in each protein which possesses a high pKa (6.2-6.3) elicits the conformational change at the high-affinity sites with a corresponding decrease in the overall magnitude of the Ca2+ -evoked changes. The expression of a conformational change upon Ca2+ binding at the level of the low-affinity sites is enchanced by protonation of a class of carboxyls with a pKa of 6.3 in cardiac TN-C and 6.7-6.8 with the skeletal homologue. In both cases, this contribution is reduced upon protonation of carboxyls with pKa less than or equal to 5.5. It was also observed that the low-affinity sites of skeletal TN-C have a much larger role to play in the total conformational change than the low-affinity sites of cardiac TN-C, a finding probably related to the inability of site 1 in the cardiac protein to bind calcium. In the cardiac protein, the Ca2+ -induced tyrosine difference-spectrum maximum is reduced from deltaepsilonM,287nm =330M-1.cm-1 to 20M-1.cm-1 by protonation of a class of groups with a pKa of 6.4, presumably the same carboxyl groups as those invoved in the CD conformational contribution from the high-affinity binding sites. No such effect was observed for the skeletal protein where deltaepsilonM,287nm was constant at 110M-1 .cm-1 over the pH range studied. The dramatic alterations in the tyrosine environment of cardiac TN-C with pH are attributed to either or both of the tyrosines located in the two high-affinity Ca2+ -binding sites (sites 3 and 4)... | [
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PMID:27292 | Guanosine triphosphate catabolism in human and rabbit erythrocytes: role of reductive deamination of guanylate to inosinate. | The reductive deamination of guanylate to inosinate was demonstrable but occurred at low rates in human and rabbit erythrocytes incubated in vitro with or without glucose. However, the process was considerably accelerated in erythrocytes incubated with deoxyglucose. In human erythrocytes incubated with deoxyglucose, deamination was the major pathway of catabolism of guanylate; little or no guanylate was dephosphorylated. In rabbit erythrocytes, guanylate was both deaminated and dephosphorylated. Inosinate formed from guanylate was metabolized only by dephosphorylation in human erythrocytes, but in rabbit erythrocytes, it was also converted to xanthylate. | Guanosine triphosphate catabolism in human and rabbit erythrocytes: role of reductive deamination of guanylate to inosinate. The reductive deamination of guanylate to inosinate was demonstrable but occurred at low rates in human and rabbit erythrocytes incubated in vitro with or without glucose. However, the process was considerably accelerated in erythrocytes incubated with deoxyglucose. In human erythrocytes incubated with deoxyglucose, deamination was the major pathway of catabolism of guanylate; little or no guanylate was dephosphorylated. In rabbit erythrocytes, guanylate was both deaminated and dephosphorylated. Inosinate formed from guanylate was metabolized only by dephosphorylation in human erythrocytes, but in rabbit erythrocytes, it was also converted to xanthylate. | [
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PMID:27293 | Some properties of a Ca2+- and (or) Mg2+-requiring nucleoside di- and tri-phosphatase(s) associated with the membranes of rat pancreatic zymogen granules. | Membranes of rat pancreatic zymogen granules have been purified and found to be essentially free of contamination by mitochondria, microsomes, and plasma membranes. They possessed an acid phosphatase activity which derived probably from lysed lysosomes contaminating the purified zymogen granules from which the membranes were prepared. These membranes were found to contain a strong Ca2+- and (or) Mg2+-requiring activity toward all nucleoside di- and tri-phosphates. Various data support the tentative conclusion that a single protein catalyzes the hydrolysis of the nucleoside di- and tri-phosphates. This protein appears to be intrinsic with its active site localized on the internal face of the membranes. | Some properties of a Ca2+- and (or) Mg2+-requiring nucleoside di- and tri-phosphatase(s) associated with the membranes of rat pancreatic zymogen granules. Membranes of rat pancreatic zymogen granules have been purified and found to be essentially free of contamination by mitochondria, microsomes, and plasma membranes. They possessed an acid phosphatase activity which derived probably from lysed lysosomes contaminating the purified zymogen granules from which the membranes were prepared. These membranes were found to contain a strong Ca2+- and (or) Mg2+-requiring activity toward all nucleoside di- and tri-phosphates. Various data support the tentative conclusion that a single protein catalyzes the hydrolysis of the nucleoside di- and tri-phosphates. This protein appears to be intrinsic with its active site localized on the internal face of the membranes. | [
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PMID:27294 | The control of pyruvate kinases of Escherichia coli: further studies of the enzyme activated by ribose-5-phosphate. | The pyruvate kinase activated by ribose-5-phosphate from Escherichia coli has been purified to homogeneity, taking advantage of the stabilization of the enzyme by its inhibitor phosphate and by thiol reagents. The native enzyme has a tetrameric quaternary structure which, while prone to dissociation under many conditions, remains intact in the presence of the above reagents. The enzyme was found to reactivate on dilution out of 8 M urea. Interestingly, the recovery of activity is greatly increased by phosphate, an allosteric inhibitor, but markedly reduced by the allosteric activator, ribose-5-phosphate, implying that it is harder for the enzyme to refold to a 'relaxed state.' Proteolysis studies indicate a more open structure in the presence of the activator. | The control of pyruvate kinases of Escherichia coli: further studies of the enzyme activated by ribose-5-phosphate. The pyruvate kinase activated by ribose-5-phosphate from Escherichia coli has been purified to homogeneity, taking advantage of the stabilization of the enzyme by its inhibitor phosphate and by thiol reagents. The native enzyme has a tetrameric quaternary structure which, while prone to dissociation under many conditions, remains intact in the presence of the above reagents. The enzyme was found to reactivate on dilution out of 8 M urea. Interestingly, the recovery of activity is greatly increased by phosphate, an allosteric inhibitor, but markedly reduced by the allosteric activator, ribose-5-phosphate, implying that it is harder for the enzyme to refold to a 'relaxed state.' Proteolysis studies indicate a more open structure in the presence of the activator. | [
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PMID:27297 | Effect of pH on the antimicrobial activity of some triphenylmethane dyes. | Four common dyes were tested as inhibitors of four types of bacteria over the pH range 5.0-9.0. Inhibition of the gram-negative types, Salmonella anatum and Enterobacter aerogenes, was markedly affected by the pH of the medium. These organisms tolerated concentrations of crystal violet and ethyl violet about 100-fold higher at pH 5.0 than at pH 9.0. Above pH 7.0 brilliant green (BG) and malachite green (MG) were precipitated as their respective carbinols and lost their inhibitory properties with these two organisms. Two gram-positive types, Staphylococcus aureus and Bacillus cereus, were more sensitive to dyes and results were less affected by pH. The carbinol forms of MG and BG were nearly as inhibitory to these organisms as the ionized forms. | Effect of pH on the antimicrobial activity of some triphenylmethane dyes. Four common dyes were tested as inhibitors of four types of bacteria over the pH range 5.0-9.0. Inhibition of the gram-negative types, Salmonella anatum and Enterobacter aerogenes, was markedly affected by the pH of the medium. These organisms tolerated concentrations of crystal violet and ethyl violet about 100-fold higher at pH 5.0 than at pH 9.0. Above pH 7.0 brilliant green (BG) and malachite green (MG) were precipitated as their respective carbinols and lost their inhibitory properties with these two organisms. Two gram-positive types, Staphylococcus aureus and Bacillus cereus, were more sensitive to dyes and results were less affected by pH. The carbinol forms of MG and BG were nearly as inhibitory to these organisms as the ionized forms. | [
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PMID:27299 | Reduced nicotinamide adenine dinucleotide phosphate-dependent formation of 2,3-dihydro-2,3-dihydroxyaflatoxin B1 from aflatoxin B1 by hepatic microsomes. | 2,3-Dihydro-2,3-dihydroxyaflatoxin B1 (dihydrodiol) was formed as a major metabolite in the incubation of aflatoxin B1 with rat and hamster liver microsomes. The yield of the dihydrodiol was maximal at pH 6.5, was reduced nicotinmide adenine dinucleotide phosphate- and cytochrome P-450-dependent, and was increased 2- to 4-fold by pretreatment of the animals with phenobarbital; pretreatment with 3-methylcholanthrene did not alter the activity of rat hepatic microsomes. Inhibitors of epoxide hydrase did not lower the yield of the dihydrodiol in these systems. Negligible yields of the dihydrodiol were formed from aflatoxin B1 and rat liver microsomes in the presence of DNA. Little or no formation of the dihydrodiol was noted with microsomes from rat intestinal mucosa, kidney, or lung. These results further support the formation of aflatoxin B1 2,3-oxide as a major electrophilic metabolite of aflatoxin B1 in rat and hamster liver microsomal systems, since this highly reactive epoxide would be expected to hydrolyze readily to form the dihydrodiol. | Reduced nicotinamide adenine dinucleotide phosphate-dependent formation of 2,3-dihydro-2,3-dihydroxyaflatoxin B1 from aflatoxin B1 by hepatic microsomes. 2,3-Dihydro-2,3-dihydroxyaflatoxin B1 (dihydrodiol) was formed as a major metabolite in the incubation of aflatoxin B1 with rat and hamster liver microsomes. The yield of the dihydrodiol was maximal at pH 6.5, was reduced nicotinmide adenine dinucleotide phosphate- and cytochrome P-450-dependent, and was increased 2- to 4-fold by pretreatment of the animals with phenobarbital; pretreatment with 3-methylcholanthrene did not alter the activity of rat hepatic microsomes. Inhibitors of epoxide hydrase did not lower the yield of the dihydrodiol in these systems. Negligible yields of the dihydrodiol were formed from aflatoxin B1 and rat liver microsomes in the presence of DNA. Little or no formation of the dihydrodiol was noted with microsomes from rat intestinal mucosa, kidney, or lung. These results further support the formation of aflatoxin B1 2,3-oxide as a major electrophilic metabolite of aflatoxin B1 in rat and hamster liver microsomal systems, since this highly reactive epoxide would be expected to hydrolyze readily to form the dihydrodiol. | [
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PMID:27304 | Quantitative characteristics of the Feyrter cells and neuroepithelial bodies of the fetal rabbit lung in normoxia and short term chronic hypoxia. | We report here quantitative data on the Feyrter (single) cells (APUD cells) and neuroepithelial bodies (grouped Feyrter cells), in the lungs of rabbit fetuses at 26, 27.5 and 29 days gestational age, during normoxia and short term chronic hypoxia. The apparent number of these cells declines during this period; we suggest that this might be due to increased hypoxemia. Moreover, the number of cells in the lungs of fetuses from short term chronically hypoxic mothers is lower than in the normoxic animals. These findings are in agreement with our previous studies in short term chronically hypoxic neonatal rabbits, and suggest that the increased hypoxemia in the fetus, caused by the induction of hypoxia in the mother, constitutes a stimulus for secretory activity of the Feyrter cells and neuroepithelial bodies (NEBs). This in turn could be part of the mechanism responsible for maintaining the pulmonary vasoconstriction due to hypoxemia. Our results from fetuses of normoxic does provide base line data on the chronological importance of the Feyrter cells and NEBs. | Quantitative characteristics of the Feyrter cells and neuroepithelial bodies of the fetal rabbit lung in normoxia and short term chronic hypoxia. We report here quantitative data on the Feyrter (single) cells (APUD cells) and neuroepithelial bodies (grouped Feyrter cells), in the lungs of rabbit fetuses at 26, 27.5 and 29 days gestational age, during normoxia and short term chronic hypoxia. The apparent number of these cells declines during this period; we suggest that this might be due to increased hypoxemia. Moreover, the number of cells in the lungs of fetuses from short term chronically hypoxic mothers is lower than in the normoxic animals. These findings are in agreement with our previous studies in short term chronically hypoxic neonatal rabbits, and suggest that the increased hypoxemia in the fetus, caused by the induction of hypoxia in the mother, constitutes a stimulus for secretory activity of the Feyrter cells and neuroepithelial bodies (NEBs). This in turn could be part of the mechanism responsible for maintaining the pulmonary vasoconstriction due to hypoxemia. Our results from fetuses of normoxic does provide base line data on the chronological importance of the Feyrter cells and NEBs. | [
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PMID:27305 | pH-dependent behaviour of erythrocyte membrane elevations. | Freeze-etch electron microscopy of human erythrocytes from heparinized normal blood reveals membrane elevations. These elevations disappear at low pH values and reappear under normal pH conditions in vitro, both in the absence and the presence of plasma. This pH-dependent behaviour is discussed as an intrinsic membrane event. | pH-dependent behaviour of erythrocyte membrane elevations. Freeze-etch electron microscopy of human erythrocytes from heparinized normal blood reveals membrane elevations. These elevations disappear at low pH values and reappear under normal pH conditions in vitro, both in the absence and the presence of plasma. This pH-dependent behaviour is discussed as an intrinsic membrane event. | [
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PMID:27306 | Kinetic studies on the interaction of gold (III) with nucleic acids. IV. RNA-Au (III) system. | The kinetics of the interaction of Au(III) with whole yeast RNA has been studied using UV-spectrophotometry. The reaction is second order with respect to the nucleotide unit of RNA and first order with respect to Au(III) in the respective stoichiometry of 2 : 1. The effects of initial composition, temperature, ionic strength, pH and chloride ion on the kinetics have been studied. Activation energy is found to be 11.5 kcal/mol. Effect of ionic strength indicates that both the positively charged and neutral species of Au(III) take part in the rate limiting step, the former being dominant at low ionic strength. A plausible mechanism has been proposed which involves the interaction of two nucleotide units of RNA with one species of Au(III) in the rate limiting step. | Kinetic studies on the interaction of gold (III) with nucleic acids. IV. RNA-Au (III) system. The kinetics of the interaction of Au(III) with whole yeast RNA has been studied using UV-spectrophotometry. The reaction is second order with respect to the nucleotide unit of RNA and first order with respect to Au(III) in the respective stoichiometry of 2 : 1. The effects of initial composition, temperature, ionic strength, pH and chloride ion on the kinetics have been studied. Activation energy is found to be 11.5 kcal/mol. Effect of ionic strength indicates that both the positively charged and neutral species of Au(III) take part in the rate limiting step, the former being dominant at low ionic strength. A plausible mechanism has been proposed which involves the interaction of two nucleotide units of RNA with one species of Au(III) in the rate limiting step. | [
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PMID:27308 | [Culicidae of the French Territory of Afars and Issas. The genera Aedes, Culiseta, Uranotaenia and Mimomyia]. | During an entomological survey conducted in French Territory of Afars and Issas from november 1973 to june 1975, the following species were collected: Aedes caspius, A. vittatus, Culiseta longeareolata, Uranotaenia balfouri, Mimomyia mimomyiaformis, M. mediolineata. To these mosquitoes, we must add the record of A. aegypti larvae on a dhow berthing in Djibuti while this mosquito does not exist actually in the Territory. The epidemiological importance of these culicids is discussed. | [Culicidae of the French Territory of Afars and Issas. The genera Aedes, Culiseta, Uranotaenia and Mimomyia]. During an entomological survey conducted in French Territory of Afars and Issas from november 1973 to june 1975, the following species were collected: Aedes caspius, A. vittatus, Culiseta longeareolata, Uranotaenia balfouri, Mimomyia mimomyiaformis, M. mediolineata. To these mosquitoes, we must add the record of A. aegypti larvae on a dhow berthing in Djibuti while this mosquito does not exist actually in the Territory. The epidemiological importance of these culicids is discussed. | [
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PMID:27313 | [Pharmacological study of para-aminoclonidine, an alpha sympathomimetic]. | Para-aminoclonidine is a more potent alpha-adrenergic agent than clonidine; in vitro on Rat aorta it is 150 times more active than NE, in vivo on pithed Rat it is still twice as active. The hypotensive action of paraaminoclondine is only manifested when given intraventricularly. In the conscious Rat, it has a diuretic action when given per os. Following IV injection in the anaesthetized Dog, the para-aminoclonidine induces an essentially hydric diuresis which is independet of its hypertensive action. | [Pharmacological study of para-aminoclonidine, an alpha sympathomimetic]. Para-aminoclonidine is a more potent alpha-adrenergic agent than clonidine; in vitro on Rat aorta it is 150 times more active than NE, in vivo on pithed Rat it is still twice as active. The hypotensive action of paraaminoclondine is only manifested when given intraventricularly. In the conscious Rat, it has a diuretic action when given per os. Following IV injection in the anaesthetized Dog, the para-aminoclonidine induces an essentially hydric diuresis which is independet of its hypertensive action. | [
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PMID:27309 | [Significance of the various toxins present on dead coral substrata in the origin of ciguatera]. | The authors look directly on the coral reefs the toxins carried by poisonous fish. Pointing out two major toxic substances, they discuss their respective role in ciguatera physiopathology and suggest a new hypothesis for the origin of the phenomenon. | [Significance of the various toxins present on dead coral substrata in the origin of ciguatera]. The authors look directly on the coral reefs the toxins carried by poisonous fish. Pointing out two major toxic substances, they discuss their respective role in ciguatera physiopathology and suggest a new hypothesis for the origin of the phenomenon. | [
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] |
PMID:27314 | [Respiratory function of blood and relative weight of certain organs in two species of shrew: Crocidura russula and Suncus etruscus (Soricidae mammals)]. | A high oxygen capacity combined with a low oxygen affinity and a large Bohr effect achieve a high oxygen transport capacity in small shrews. The high metabolic rate (Suncus estruscus: 100 to 350 ml O2/kg. min) can be explained by the combination of the favorable blood parameters with a high relative heart weight (1,2% of body weight) and heart frequencies of 1,000 to 1,350 per minute. | [Respiratory function of blood and relative weight of certain organs in two species of shrew: Crocidura russula and Suncus etruscus (Soricidae mammals)]. A high oxygen capacity combined with a low oxygen affinity and a large Bohr effect achieve a high oxygen transport capacity in small shrews. The high metabolic rate (Suncus estruscus: 100 to 350 ml O2/kg. min) can be explained by the combination of the favorable blood parameters with a high relative heart weight (1,2% of body weight) and heart frequencies of 1,000 to 1,350 per minute. | [
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PMID:27310 | [Examination of the enzymatic functions of the normal liver in black Africans (Apropos of 50 Senegalese cases)]. | In order to establish the hepatic enzymogram in healthy African black people, four enzymes have been studied in 50 apparently healthy male Africans: transaminases (GOT, GPT), alcaline phosphatases, ornithine carbamoyltransferase (OCT) and gamma-glutamyl transpeptidase (GGT). The findings do not show any difference with the usually admitted levels in European countries, except for alcaline phosphatases which are situated at the upper limit of the normal. | [Examination of the enzymatic functions of the normal liver in black Africans (Apropos of 50 Senegalese cases)]. In order to establish the hepatic enzymogram in healthy African black people, four enzymes have been studied in 50 apparently healthy male Africans: transaminases (GOT, GPT), alcaline phosphatases, ornithine carbamoyltransferase (OCT) and gamma-glutamyl transpeptidase (GGT). The findings do not show any difference with the usually admitted levels in European countries, except for alcaline phosphatases which are situated at the upper limit of the normal. | [
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PMID:27315 | [Effect of total and prolonged starvation on the acid soluble nucleotides in carp liver]. | The composition of the acid soluble nucleotide pool of Carp liver was determined by column chromatography, and its changes were followed during prolonged total fast. Total prolonged starvation caused substantial changes in the distribution of hepatocyte free nucleotides. Eleven months of starvation caused a reduction in the concentration of di- and triphosphate nucleosides as well as a diminution of the oxido-reduction of the hepatocyte. Only pyrimidine nucleoside monophosphates remained at concentration approaching those found in regularly fed Carp. | [Effect of total and prolonged starvation on the acid soluble nucleotides in carp liver]. The composition of the acid soluble nucleotide pool of Carp liver was determined by column chromatography, and its changes were followed during prolonged total fast. Total prolonged starvation caused substantial changes in the distribution of hepatocyte free nucleotides. Eleven months of starvation caused a reduction in the concentration of di- and triphosphate nucleosides as well as a diminution of the oxido-reduction of the hepatocyte. Only pyrimidine nucleoside monophosphates remained at concentration approaching those found in regularly fed Carp. | [
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PMID:27316 | [Modification of the effect of cardio-accelerator nerve stimulation in dogs by clonidine and several alpha-adrenolytics]. | In dogs anaesthetized with pentobarbital (30 mg. kg -1 i.v.), clonidine (0,01 mg.kg-1 i.v.) reduced the tachycardia induced at low frequencies by stimulation of the cardiac nerve. The effects of some alpha-adrenoceptor blocking agents on this effect have been studied. Small doses of yohimbine (0.3 mg. kg-1 i.v.) or piperoxan (0.3 mg. kg-1 i.v.) increased the effects of the stimulation and in addition antagonized the inhibitory effects of clonidine and reversed the pressor response to adrenaline. Thymoxamine (1 mg.kg-1 i.v.) and prazosin (1 mg.kg-1 i.v.) did not increase the effect of the stimulation of the cardiac nerve, but reduced the effect of clonidine. ARC239 (0.05 mg.kg-1) reversed the pressor response to adrenaline but even at high doses did not increase the effects of the stimulation of the cardiac nerve or the effects of clonidine. These observations afford further evidence for a dissimilarity between pre and post-synaptic alpha-adrenoceptors. | [Modification of the effect of cardio-accelerator nerve stimulation in dogs by clonidine and several alpha-adrenolytics]. In dogs anaesthetized with pentobarbital (30 mg. kg -1 i.v.), clonidine (0,01 mg.kg-1 i.v.) reduced the tachycardia induced at low frequencies by stimulation of the cardiac nerve. The effects of some alpha-adrenoceptor blocking agents on this effect have been studied. Small doses of yohimbine (0.3 mg. kg-1 i.v.) or piperoxan (0.3 mg. kg-1 i.v.) increased the effects of the stimulation and in addition antagonized the inhibitory effects of clonidine and reversed the pressor response to adrenaline. Thymoxamine (1 mg.kg-1 i.v.) and prazosin (1 mg.kg-1 i.v.) did not increase the effect of the stimulation of the cardiac nerve, but reduced the effect of clonidine. ARC239 (0.05 mg.kg-1) reversed the pressor response to adrenaline but even at high doses did not increase the effects of the stimulation of the cardiac nerve or the effects of clonidine. These observations afford further evidence for a dissimilarity between pre and post-synaptic alpha-adrenoceptors. | [
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PMID:27317 | [Changes in the distribution of gamma-glutamyl transferase in the organs of the mouse as a function of development]. | In the Mouse, gamma-glutamyl transferase distribution changes with development in the kidney and the liver; on the contrary, its activity remains almost equal and low in other organs. In the liver, a low activity is observed in the fetus and the new-born up to the 3rd day of life; then it is no more measurable. In the kidney, the low enzyme activity of the fetus is multiplied by 10 in the 10 first days of life and by 50 in the 6 first weeks. | [Changes in the distribution of gamma-glutamyl transferase in the organs of the mouse as a function of development]. In the Mouse, gamma-glutamyl transferase distribution changes with development in the kidney and the liver; on the contrary, its activity remains almost equal and low in other organs. In the liver, a low activity is observed in the fetus and the new-born up to the 3rd day of life; then it is no more measurable. In the kidney, the low enzyme activity of the fetus is multiplied by 10 in the 10 first days of life and by 50 in the 6 first weeks. | [
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PMID:27318 | [Comparative action of toxins isolated from the venom of the scorpion (Androctonus australis) and the tentacles of the sea anemone (Anemonia Sulcata) on an isolated frog neuromuscular preparation]. | We have shown that, if Scorpion venom is acting a skeletal muscle indirectly by releasing Acetycholine and directly by inducing an increase in intracellular free calcium, the main action of toxin II isolated from Anemonia Sulcata tentacles is presynaptic. | [Comparative action of toxins isolated from the venom of the scorpion (Androctonus australis) and the tentacles of the sea anemone (Anemonia Sulcata) on an isolated frog neuromuscular preparation]. We have shown that, if Scorpion venom is acting a skeletal muscle indirectly by releasing Acetycholine and directly by inducing an increase in intracellular free calcium, the main action of toxin II isolated from Anemonia Sulcata tentacles is presynaptic. | [
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] |
PMID:27319 | [Action of toxin II isolated from the tentacles of sea anemones on the neuromuscular transmission on normal and botulin toxin inhibited frogs]. | In our experimental conditions, toxin II from Anemonia sulcata restored Frog neuromuscular transmission blocked by botulinum toxin, type A. | [Action of toxin II isolated from the tentacles of sea anemones on the neuromuscular transmission on normal and botulin toxin inhibited frogs]. In our experimental conditions, toxin II from Anemonia sulcata restored Frog neuromuscular transmission blocked by botulinum toxin, type A. | [
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PMID:27322 | Observations on serum and urine alkaline ribonuclease activity and urate after burn injury in man. | In an investigation into the disturbances of body function associated with burn injury we have measured the activity of alkaline ribonuclease (EC 3.1.4.22) and the level of urate in the serum and urine of patients sustaining burn injury. Ribonuclease activity was elevated in all patients. The degree of elevation can be related to the percentage of the body surface area burned and to a predictive index of burn mortality. Increased serum ribonuclease activity was accompanied by increased urine ribonuclease output. The relationship between serum urea and ribonuclease activity has been investigated. A significant correlation between these two parameters was observed during the first week post burn. We suggest that this correlation shows as a result of increased protein catabolism and renal dysfunction. After the first week a significant correlation between serum urea and ribonuclease activity was not observed. It is possible that, at this stage, increased ribonuclease activity is perhaps a result of tissue repair. Serum urate was found to be decreased in all patients after burn injury. Serum urate decrease expressed as a percentage of initial value, correlated very strongly with the predictive index of burn mortality. In severely burned patients the decrease in serum urate was accompanied by increased urine urate output and may indicate a change in renal handling of urate after burn injury. | Observations on serum and urine alkaline ribonuclease activity and urate after burn injury in man. In an investigation into the disturbances of body function associated with burn injury we have measured the activity of alkaline ribonuclease (EC 3.1.4.22) and the level of urate in the serum and urine of patients sustaining burn injury. Ribonuclease activity was elevated in all patients. The degree of elevation can be related to the percentage of the body surface area burned and to a predictive index of burn mortality. Increased serum ribonuclease activity was accompanied by increased urine ribonuclease output. The relationship between serum urea and ribonuclease activity has been investigated. A significant correlation between these two parameters was observed during the first week post burn. We suggest that this correlation shows as a result of increased protein catabolism and renal dysfunction. After the first week a significant correlation between serum urea and ribonuclease activity was not observed. It is possible that, at this stage, increased ribonuclease activity is perhaps a result of tissue repair. Serum urate was found to be decreased in all patients after burn injury. Serum urate decrease expressed as a percentage of initial value, correlated very strongly with the predictive index of burn mortality. In severely burned patients the decrease in serum urate was accompanied by increased urine urate output and may indicate a change in renal handling of urate after burn injury. | [
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PMID:27323 | Some pitfalls in the quality control of alkaline phosphatase activity. | A study of the pH optimum of alkaline phosphatase activity in calibration and control serum preparations reveals a wide variation in pH optima. It is shown that as small an error of +/- 0.05 pH units in preparation of the buffered substrate results in an error in the order of +/- 5% in the response between the calibrating serum and patient specimens. It is recommended that a calibration serum should closely parallel the pH optimum of alkaline phosphatase in human sera and that a quality control serum containing alkaline phosphatase of alternative source to that present in the calibration serum be included with patient samples as a sensitive means of monitoring pH changes in the buffered substrate. | Some pitfalls in the quality control of alkaline phosphatase activity. A study of the pH optimum of alkaline phosphatase activity in calibration and control serum preparations reveals a wide variation in pH optima. It is shown that as small an error of +/- 0.05 pH units in preparation of the buffered substrate results in an error in the order of +/- 5% in the response between the calibrating serum and patient specimens. It is recommended that a calibration serum should closely parallel the pH optimum of alkaline phosphatase in human sera and that a quality control serum containing alkaline phosphatase of alternative source to that present in the calibration serum be included with patient samples as a sensitive means of monitoring pH changes in the buffered substrate. | [
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PMID:27327 | Characterization of beta-adrenergic receptor linked to adenylate cyclase in a human cancer cell line (COLO 16). | 1. A human cancer cell line (COLO 16) derived originally from an epidermal squamous cell carcinoma was found to possess adenylate cyclase responsiveness to beta-adrenergic agonists. 2. The adenylate cyclase response was characterized with respect to activation constants (KA) for various beta-adrenergic agonists and inhibition constants (Ki) for antagonists. 3. Intact cells responded with dose-dependent increases in production of cyclic adenosine 3':5'-monophosphate. 4. Properties of the beta-adrenergic receptor were evaluated by using the specific binding of [3H]propranolol to cell membranes. Specific binding was saturable, with KD 5.79 nmol/l and binding sites 0.68 pmol/mg of protein. 5. Competition for binding to cell membranes was shown by beta-adrenergic agonists and antagonists and was stereospecific. There was close agreement between the affinity of these various agents on adenylate cyclase and receptor binding. 6. It is likely that the beta-adrenergic receptor-linked adenylate cyclase in COLO 16 cells represents persistence in a cancer cell line of a receptor present normally in epidermal cells. | Characterization of beta-adrenergic receptor linked to adenylate cyclase in a human cancer cell line (COLO 16). 1. A human cancer cell line (COLO 16) derived originally from an epidermal squamous cell carcinoma was found to possess adenylate cyclase responsiveness to beta-adrenergic agonists. 2. The adenylate cyclase response was characterized with respect to activation constants (KA) for various beta-adrenergic agonists and inhibition constants (Ki) for antagonists. 3. Intact cells responded with dose-dependent increases in production of cyclic adenosine 3':5'-monophosphate. 4. Properties of the beta-adrenergic receptor were evaluated by using the specific binding of [3H]propranolol to cell membranes. Specific binding was saturable, with KD 5.79 nmol/l and binding sites 0.68 pmol/mg of protein. 5. Competition for binding to cell membranes was shown by beta-adrenergic agonists and antagonists and was stereospecific. There was close agreement between the affinity of these various agents on adenylate cyclase and receptor binding. 6. It is likely that the beta-adrenergic receptor-linked adenylate cyclase in COLO 16 cells represents persistence in a cancer cell line of a receptor present normally in epidermal cells. | [
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PMID:27328 | The nature of inactive renin in human plasma and amniotic fluid. | 1. The properties of inactive and active renin in human plasma and amniotic fluid were studied chromatographically. Activation was achieved at pH 3.3 with and without added pepsin. 2. Acid activation of renin was time- and temperature-dependent but was inhibited by dilution of the sample. The dilution effect was corrected by adding pepsin. Such characteristics indicate that activation at low pH is catalysed by intrinsic enzymes. 3. Separation and/or dilution of the activating enzyme during ion-exchange chromatography concealed the eluted position of inactive renin and reduced the amount recovered. Only after full activation of the eluted renin was achieved with added pepsin was a distinct peak of inactive renin exposed. 4. At pH 7.5 inactive renin carried a lower negative charge than the active enzyme. This charge difference was lost after activation. 5. No molecular-weight differences between active, inactive renin or the International Renin Standard were detected by gel filtration. No renin of larger molecular weight was present. 6. These findings will be helpful in purification studies of human inactive renin. | The nature of inactive renin in human plasma and amniotic fluid. 1. The properties of inactive and active renin in human plasma and amniotic fluid were studied chromatographically. Activation was achieved at pH 3.3 with and without added pepsin. 2. Acid activation of renin was time- and temperature-dependent but was inhibited by dilution of the sample. The dilution effect was corrected by adding pepsin. Such characteristics indicate that activation at low pH is catalysed by intrinsic enzymes. 3. Separation and/or dilution of the activating enzyme during ion-exchange chromatography concealed the eluted position of inactive renin and reduced the amount recovered. Only after full activation of the eluted renin was achieved with added pepsin was a distinct peak of inactive renin exposed. 4. At pH 7.5 inactive renin carried a lower negative charge than the active enzyme. This charge difference was lost after activation. 5. No molecular-weight differences between active, inactive renin or the International Renin Standard were detected by gel filtration. No renin of larger molecular weight was present. 6. These findings will be helpful in purification studies of human inactive renin. | [
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PMID:27331 | Immunologically mediated drug-induced acute renal failure. | (1) AIN is the most frequent pattern of drug-induced immunologically mediated renal injury. A number of drugs may be responsible for AIN, namely methicillin and other penicillin derivatives, rifampicin, phenindione and sulfonamides. Particular clinical and pathological features often suggest an immune pathogenetic mechanism. IgG anti-TBM and IgE antibodies have been found in only a few cases and it is likely that antibody-mediated and cell-mediated injury may operate in the same patient. (2) Only few examples of drug-induced vasculitis and glomerulonephritis are known, and the pathophysiology of this kind of renal damage is poorly understood. | Immunologically mediated drug-induced acute renal failure. (1) AIN is the most frequent pattern of drug-induced immunologically mediated renal injury. A number of drugs may be responsible for AIN, namely methicillin and other penicillin derivatives, rifampicin, phenindione and sulfonamides. Particular clinical and pathological features often suggest an immune pathogenetic mechanism. IgG anti-TBM and IgE antibodies have been found in only a few cases and it is likely that antibody-mediated and cell-mediated injury may operate in the same patient. (2) Only few examples of drug-induced vasculitis and glomerulonephritis are known, and the pathophysiology of this kind of renal damage is poorly understood. | [
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PMID:27337 | Coupling of cerebral metabolism and blood flow in epileptic seizures, hypoxia and hypoglycaemia. | This study examines the possibility that changes of cerebral extracellular pH (PH e) or adenosine concentration may provide coupling mechanisms of a general nautre, adjusting cerebral blood flow (CBF) to metabolic demands. Although there is considerable indirect evidence that CBF varies inversely with pHe, results obtained during the last few years indicate that large increases in flow may occur in the absence of a fall in pHe. Thus, induction of hypoxia or epileptic seizures leads to maximal increase in CBF before pHe falls or even when there is initial alkalosis due to concomitant hypocapnia. Furthermore, CBF increases in hypoglycaemia and after administration of amphetamine, two conditions unassociated with tissue acidosis. The possibility that adenosine may be a coupling factor was examined in hypoxia and during epileptic seizures in rats. In both conditions a four- to fivefold increase in CBF occurs in spite of the fact that tissue adenosine concentrations remain at or below 1 mumolkg-u. It is concluded that adenosine accumulates first when there is a perturbation of cerebral energy state with a rise in AMP concentration. It seems unlikely that adenosine, formed by breakdown of AMP, acts as a general coupling factor. | Coupling of cerebral metabolism and blood flow in epileptic seizures, hypoxia and hypoglycaemia. This study examines the possibility that changes of cerebral extracellular pH (PH e) or adenosine concentration may provide coupling mechanisms of a general nautre, adjusting cerebral blood flow (CBF) to metabolic demands. Although there is considerable indirect evidence that CBF varies inversely with pHe, results obtained during the last few years indicate that large increases in flow may occur in the absence of a fall in pHe. Thus, induction of hypoxia or epileptic seizures leads to maximal increase in CBF before pHe falls or even when there is initial alkalosis due to concomitant hypocapnia. Furthermore, CBF increases in hypoglycaemia and after administration of amphetamine, two conditions unassociated with tissue acidosis. The possibility that adenosine may be a coupling factor was examined in hypoxia and during epileptic seizures in rats. In both conditions a four- to fivefold increase in CBF occurs in spite of the fact that tissue adenosine concentrations remain at or below 1 mumolkg-u. It is concluded that adenosine accumulates first when there is a perturbation of cerebral energy state with a rise in AMP concentration. It seems unlikely that adenosine, formed by breakdown of AMP, acts as a general coupling factor. | [
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PMID:27339 | Control of cerebral vascular smooth muscle during general anaesthesia. | The effects of some general anaesthetics, for example thiopentone, Althesin (alphaxalone + alphadolone) and ketamine, on cerebral vascular smooth muscle are those which would be expected from their metabolic actions. With other anaesthetics, mainly those administered by inhalation, and especially the volatile agents, cerebral blood flow increases in excess of the metabolic activity, which is usually depressed to varying degrees. During general anaesthesia with any of these agents, responses to changes in arterial Pco2 or blood pressure are maintained. Furthermore, when seizure activity occurs during enflurane administration, there is a flow response to the associated metabolic stimulation. The time course of the flow response to the metabolically depressant drug Althesin has been measured in baboons and shown to be very rapid. Wtih this drug cerebrovascular resistance begins to increase within 2 s of its arrival in the brain. This rapid flow change occurs also after sympathetic denervation. Extracellular fluid pH of the cortex does not alter until after the initiation of the vascular smooth muscle response. | Control of cerebral vascular smooth muscle during general anaesthesia. The effects of some general anaesthetics, for example thiopentone, Althesin (alphaxalone + alphadolone) and ketamine, on cerebral vascular smooth muscle are those which would be expected from their metabolic actions. With other anaesthetics, mainly those administered by inhalation, and especially the volatile agents, cerebral blood flow increases in excess of the metabolic activity, which is usually depressed to varying degrees. During general anaesthesia with any of these agents, responses to changes in arterial Pco2 or blood pressure are maintained. Furthermore, when seizure activity occurs during enflurane administration, there is a flow response to the associated metabolic stimulation. The time course of the flow response to the metabolically depressant drug Althesin has been measured in baboons and shown to be very rapid. Wtih this drug cerebrovascular resistance begins to increase within 2 s of its arrival in the brain. This rapid flow change occurs also after sympathetic denervation. Extracellular fluid pH of the cortex does not alter until after the initiation of the vascular smooth muscle response. | [
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PMID:27343 | Production, metabolism and possible functions of adenosine in brain tissue in situ. | Adenosine and H+ may act synergistically to regulate cerebral blood flow because adenosine production is enhanced under various experimental conditions associated with an imbalance between oxygen supply and oxygen need. Direct application of adenosine dilates the pial vessels, but changes in cerebral vascular resistance are not observed when adenosine is infused intraarterially. This is because adenosine does not readily cross the blood-brain barrier. The studies reported here show that in dogs the adenosine released into the interstitium is partly reincorporated into adenine nucleotides via an adenosine kinase (EC 2.7.1.20) reaction (salvage pathway) and partly degraded to inosine and hypoxanthine. However, in contrast to other tissues, the accumulation of iosine and hypoxanthine in brain tissue proceeds at a rate slower than that of adenosine because one of the degradative enzymes, namely purine-nucleoside phosphorylase (EC 2.4.2.1) is located only in the vessel wall, which is not readily permeable to adenosine. Thus, the slow access of adenosine to its degradative enzymes delays the appearance of its products, inosine and hypoxanthine. | Production, metabolism and possible functions of adenosine in brain tissue in situ. Adenosine and H+ may act synergistically to regulate cerebral blood flow because adenosine production is enhanced under various experimental conditions associated with an imbalance between oxygen supply and oxygen need. Direct application of adenosine dilates the pial vessels, but changes in cerebral vascular resistance are not observed when adenosine is infused intraarterially. This is because adenosine does not readily cross the blood-brain barrier. The studies reported here show that in dogs the adenosine released into the interstitium is partly reincorporated into adenine nucleotides via an adenosine kinase (EC 2.7.1.20) reaction (salvage pathway) and partly degraded to inosine and hypoxanthine. However, in contrast to other tissues, the accumulation of iosine and hypoxanthine in brain tissue proceeds at a rate slower than that of adenosine because one of the degradative enzymes, namely purine-nucleoside phosphorylase (EC 2.4.2.1) is located only in the vessel wall, which is not readily permeable to adenosine. Thus, the slow access of adenosine to its degradative enzymes delays the appearance of its products, inosine and hypoxanthine. | [
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PMID:27344 | Cellular microenvironment in relation to local blood flow. | The investigation was designed to identify microenvironmental factors that might be important in the regulation of local blood flow. Cellular microenvironment in terms of Po2, pH, [K+], [Cl-], [Ca2+] and lactate was measured in rat brain by means of specific microelectrodes. Vascular endothelium was stained in vivo with Thioflavine S. Local blood flow was measured with micro hydrogen electrodes. Some intracellular measurements of pH were made in conjunction with extracellular measurements. The main findings were that local autoregulation responses and blood flow changes in response to imposed hypoxic changes were very rapid (1-1.5 s). Microflow responses to changes in local cell activity were limited to a region not more than 250 micron in diameter. Increased blood flow in acute hypoxia occurred within 1-2 s of the fall in tissue Po2 and was much more rapid than changes in either pH or potassium. Intracellular pH changed within 10 s of the onset of severe hypoxia but in all cases the blood flow followed the Po2 much more closely than any other parameter. It is suggested that changes in capillary endothelium and local membrane transmission may play a part in autoregulatory mechanisms. | Cellular microenvironment in relation to local blood flow. The investigation was designed to identify microenvironmental factors that might be important in the regulation of local blood flow. Cellular microenvironment in terms of Po2, pH, [K+], [Cl-], [Ca2+] and lactate was measured in rat brain by means of specific microelectrodes. Vascular endothelium was stained in vivo with Thioflavine S. Local blood flow was measured with micro hydrogen electrodes. Some intracellular measurements of pH were made in conjunction with extracellular measurements. The main findings were that local autoregulation responses and blood flow changes in response to imposed hypoxic changes were very rapid (1-1.5 s). Microflow responses to changes in local cell activity were limited to a region not more than 250 micron in diameter. Increased blood flow in acute hypoxia occurred within 1-2 s of the fall in tissue Po2 and was much more rapid than changes in either pH or potassium. Intracellular pH changed within 10 s of the onset of severe hypoxia but in all cases the blood flow followed the Po2 much more closely than any other parameter. It is suggested that changes in capillary endothelium and local membrane transmission may play a part in autoregulatory mechanisms. | [
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PMID:27346 | [Diethylpentenamide, a substitute for carbromal? (author's transl)]. | Bromcarbamide-containing sleeping pills are frequently used in suicide attempts and cause severe, often fatal, intoxication. Since 1975, the chemically related drug diethylpentenamide had been available (in the German Federal Republic) without prescription. The authors report four cases of attempted suicide with the drug. The signs were similar to those after carbromal intoxication. There was severe respiratory depression, successfully treated by extracorporeal detoxication with combined haemoperfusion and haemodialysis. One patient developed acute pancreatitis as a complication. | [Diethylpentenamide, a substitute for carbromal? (author's transl)]. Bromcarbamide-containing sleeping pills are frequently used in suicide attempts and cause severe, often fatal, intoxication. Since 1975, the chemically related drug diethylpentenamide had been available (in the German Federal Republic) without prescription. The authors report four cases of attempted suicide with the drug. The signs were similar to those after carbromal intoxication. There was severe respiratory depression, successfully treated by extracorporeal detoxication with combined haemoperfusion and haemodialysis. One patient developed acute pancreatitis as a complication. | [
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PMID:27347 | [Salazosulfapyridine-induced lupus erythematosus syndrome in ulcerative colitis (author's transl)]. | In a 25-year old woman with ulcerative colitis a lupus erythematosus syndrome occurred during treatment with salazosulfapyridine. The connection with medication was ascertained by reexposition. Some of the systemic complications of ulcerative colitis may possibly be due to the salazosulfapyridine medication. | [Salazosulfapyridine-induced lupus erythematosus syndrome in ulcerative colitis (author's transl)]. In a 25-year old woman with ulcerative colitis a lupus erythematosus syndrome occurred during treatment with salazosulfapyridine. The connection with medication was ascertained by reexposition. Some of the systemic complications of ulcerative colitis may possibly be due to the salazosulfapyridine medication. | [
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PMID:27348 | Pharmacological study of cholinergic mechanisms of compensatory ovarian hypertrophy in rats. | Treatment of hemicastrated adult female rats with nicotine increased the compensatory ovarian hypertrophy (COH) while central n-cholinolytic IEM-506 decreased COH and prevented the estrogen-induced suppression of COH. Administration of L-DOPA abolished the effect of IEM-506, and disulfiram blocked gonadotropic action of nicotine. Elevation of the dose of arecoline decreased COH and the sensitivity of hypothalamo-gonadotropic complex to estrogen suppression. Treatment of rats with L-DOPA and disulfiram abolished the late effect of arecoline. Central m-cholinolytic metamizyl decreased COH, potentiated the effect of estrogen and prevented the gonadotropic effect of L-DOPA. The regulatory role of m-cholinergic systems in noradrenaline mediation of gonadotropic function and n-cholinergic system in dopamine ones are suggested. | Pharmacological study of cholinergic mechanisms of compensatory ovarian hypertrophy in rats. Treatment of hemicastrated adult female rats with nicotine increased the compensatory ovarian hypertrophy (COH) while central n-cholinolytic IEM-506 decreased COH and prevented the estrogen-induced suppression of COH. Administration of L-DOPA abolished the effect of IEM-506, and disulfiram blocked gonadotropic action of nicotine. Elevation of the dose of arecoline decreased COH and the sensitivity of hypothalamo-gonadotropic complex to estrogen suppression. Treatment of rats with L-DOPA and disulfiram abolished the late effect of arecoline. Central m-cholinolytic metamizyl decreased COH, potentiated the effect of estrogen and prevented the gonadotropic effect of L-DOPA. The regulatory role of m-cholinergic systems in noradrenaline mediation of gonadotropic function and n-cholinergic system in dopamine ones are suggested. | [
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PMID:27353 | The stimulus-secretion coupling of glucose-induced insulin release. Metabolic effects of menadione in isolated islets. | Pancreatic islets contain an enzyme system which catalyzes the donation of hydrogen from NAD(P)H to menadione (2-methyl-1,4-naphthoquinone). In high concentrations (20 to 50 micrometer), menadione, in addition to lowering the concentration of reduced pyridine nucleotides in the islets, also impairs glycolysis and glucose oxidation, decreases ATP concentration, and inhibits proinsulin biosynthesis. However, at a 10 micrometer concentration, menadione fails to affect the concentration of adenine nucleotides, the utilization of glucose, the production of lactate and pyruvate, the oxidation of [6-14C]glucose and the synthesis of proinsulin; whereas the metabolism of glucose through the pentose shunt is markedly increased. The sole inhibitory effect of menadione 10 micrometer upon metabolic parameters is to reduce the concentration of both NADH and NADPH, such an effect being noticed in islets exposed to glucose 11.1 mM but not in those incubated at a higher glucose level (27.8 mM). Since, in the presence of glucose 11.1 mM, menadione 10 micrometer also severely decreases glucose-stimulated45 calcium net uptake and subsequent insulin release, it is concluded that the availability of reduced pyridine nucleotides may play an essential role in the secretory sequence by coupling metabolic to cationic events. Thus, when insulinotropic nutrients are oxidized in the B-cell, the increased availability of reduced pyridine nucleotides could modify the affinity for cations of native ionophoretic systems, eventually leading to the accumulation of calcium up to a level sufficient to trigger insulin release. | The stimulus-secretion coupling of glucose-induced insulin release. Metabolic effects of menadione in isolated islets. Pancreatic islets contain an enzyme system which catalyzes the donation of hydrogen from NAD(P)H to menadione (2-methyl-1,4-naphthoquinone). In high concentrations (20 to 50 micrometer), menadione, in addition to lowering the concentration of reduced pyridine nucleotides in the islets, also impairs glycolysis and glucose oxidation, decreases ATP concentration, and inhibits proinsulin biosynthesis. However, at a 10 micrometer concentration, menadione fails to affect the concentration of adenine nucleotides, the utilization of glucose, the production of lactate and pyruvate, the oxidation of [6-14C]glucose and the synthesis of proinsulin; whereas the metabolism of glucose through the pentose shunt is markedly increased. The sole inhibitory effect of menadione 10 micrometer upon metabolic parameters is to reduce the concentration of both NADH and NADPH, such an effect being noticed in islets exposed to glucose 11.1 mM but not in those incubated at a higher glucose level (27.8 mM). Since, in the presence of glucose 11.1 mM, menadione 10 micrometer also severely decreases glucose-stimulated45 calcium net uptake and subsequent insulin release, it is concluded that the availability of reduced pyridine nucleotides may play an essential role in the secretory sequence by coupling metabolic to cationic events. Thus, when insulinotropic nutrients are oxidized in the B-cell, the increased availability of reduced pyridine nucleotides could modify the affinity for cations of native ionophoretic systems, eventually leading to the accumulation of calcium up to a level sufficient to trigger insulin release. | [
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PMID:27354 | Characterization of deoxycytidylate methyltransferase in Xanthomonas oryzae infected with bacteriophage Xp12. | Three methods, chromatographic, spectrophotometric and tritium-release assay, were used and compared for the assay of deoxycytidylate methyltransferase. All three methods can be used for assay of this enzyme but the tritium-release assay appears to be the most simple and convenient. With the help of this assay the deoxycytidylate methyltransferase has been isolated and purified from sonically disrupted cells of Xp12-infected Xanthomonas oryzae. Using a procedure that involves fractionation with streptomycin sulfate and ammonium sulfate, filtration through Sephadex G-100 and chromatography on DEAE-cellulose, a 214-fold increase in specific activity was obtained. The enzyme displays a narrow pH optimum at 6.0 Among the buffers tested, 6-morpholinoethane sulfonate with the addition of Mg2 is the best. The enzyme can utilize dCMP as a substrate. The enzyme can also convert tetrahydrofolic acid into dihydrofolic acid. The Km value for dCMP is 31.3 micrometer and the Km value for tetrahydrofolic acid is 71.4 micrometer. There is no absolute requirement of ions for the activity of the enzyme; however, the presence of ions causes stimulating or inhibiting effects on enzyme activity that are dependent on the variety and concentration of ions used. | Characterization of deoxycytidylate methyltransferase in Xanthomonas oryzae infected with bacteriophage Xp12. Three methods, chromatographic, spectrophotometric and tritium-release assay, were used and compared for the assay of deoxycytidylate methyltransferase. All three methods can be used for assay of this enzyme but the tritium-release assay appears to be the most simple and convenient. With the help of this assay the deoxycytidylate methyltransferase has been isolated and purified from sonically disrupted cells of Xp12-infected Xanthomonas oryzae. Using a procedure that involves fractionation with streptomycin sulfate and ammonium sulfate, filtration through Sephadex G-100 and chromatography on DEAE-cellulose, a 214-fold increase in specific activity was obtained. The enzyme displays a narrow pH optimum at 6.0 Among the buffers tested, 6-morpholinoethane sulfonate with the addition of Mg2 is the best. The enzyme can utilize dCMP as a substrate. The enzyme can also convert tetrahydrofolic acid into dihydrofolic acid. The Km value for dCMP is 31.3 micrometer and the Km value for tetrahydrofolic acid is 71.4 micrometer. There is no absolute requirement of ions for the activity of the enzyme; however, the presence of ions causes stimulating or inhibiting effects on enzyme activity that are dependent on the variety and concentration of ions used. | [
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PMID:27358 | Dependence upon pH of steady-state parameters for the beta-galactosidase-catalysed hydrolyses of beta-D-galactopyranosyl derivatives of different chemical types. | The effect of pH upon the beta-galactosidase-catalyzed hydrolyses of aryl galactosides is essentially similar for each of the three steps of their hydrolysis. It differs markedly from that on the hydrolysis of galactosyl pyridinium salts; these proceed through a 'non-bottleneck' pathway. While pH increase abolishes the rate of every step of the reaction for aryl galactosides, it favors the first step of hydrolysis of the galactosyl pyridinium salts, which supports the hypothesis that catalysis of these compounds originates largely in non-covalent interactions. | Dependence upon pH of steady-state parameters for the beta-galactosidase-catalysed hydrolyses of beta-D-galactopyranosyl derivatives of different chemical types. The effect of pH upon the beta-galactosidase-catalyzed hydrolyses of aryl galactosides is essentially similar for each of the three steps of their hydrolysis. It differs markedly from that on the hydrolysis of galactosyl pyridinium salts; these proceed through a 'non-bottleneck' pathway. While pH increase abolishes the rate of every step of the reaction for aryl galactosides, it favors the first step of hydrolysis of the galactosyl pyridinium salts, which supports the hypothesis that catalysis of these compounds originates largely in non-covalent interactions. | [
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PMID:27359 | Effect of pH on the process of ternary-complex interconversion in the liver-alcohol-dehydrogenase reaction. | 1. Kinetic relationships referring to multiple-turnover conditions have been derived for the slowest exponential transient appearing in two-substrate enzyme reactions proceeding by an ordered ternary-complex mechanism. The validity of these and previously derived theoretical relationships for this mechanism has been tested by application to the liver alcohol dehydrogenase reaction. 2. All essential features of the transient-state kinetics of alcohol oxidation by NAD+ in the liver alcohol dehydrogenase system can be qualitatively and quantitatively explained in view of the compulsory-order mechanism in the proposed scheme. There is no kinetic evidence for any half-of-the-sites reactivity of the enzyme. A consistent set of rate constants is reported for the enzymic oxidation of benzyl alcohol at pH 8.75. 3. Transient-state rate parameters for benzyl alcohol/benzaldehyde catalysis by liver alcohol dehydrogenase have been determined at different pH. The interpretation of such rate parameters is critically discussed with reference to their informative value for the purpose of determination of rate constants (k and k') for the process of ternary-complex interconversion in the proposed scheme. It is concluded that the apparent rate constant (k') for hydride transfer from benzyl alcohol to NAD+ is dependent on a proton dissociation step with a pKa of 6.4, whereas the rate constant (k) for hydride transfer from NADH to benzaldehyde exhibits no corresponding dependence on proton association. 4. The asymmetric pH dependence of the forward and reverse rate of ternary-complex interconversion during liver alcohol dehydrogenase catalysis appears to reflect an obligatory step of alcohol/alcoholate ion equilibration occurring at the ternary-complex level. It is suggested that the observed pKa 6.4 dependence of the transient rate of alcohol oxidation can be attributed to a coupled acid-base system involving minimally the enzyme-bound alcohol and the protein residues Ser-48 and His-51. | Effect of pH on the process of ternary-complex interconversion in the liver-alcohol-dehydrogenase reaction. 1. Kinetic relationships referring to multiple-turnover conditions have been derived for the slowest exponential transient appearing in two-substrate enzyme reactions proceeding by an ordered ternary-complex mechanism. The validity of these and previously derived theoretical relationships for this mechanism has been tested by application to the liver alcohol dehydrogenase reaction. 2. All essential features of the transient-state kinetics of alcohol oxidation by NAD+ in the liver alcohol dehydrogenase system can be qualitatively and quantitatively explained in view of the compulsory-order mechanism in the proposed scheme. There is no kinetic evidence for any half-of-the-sites reactivity of the enzyme. A consistent set of rate constants is reported for the enzymic oxidation of benzyl alcohol at pH 8.75. 3. Transient-state rate parameters for benzyl alcohol/benzaldehyde catalysis by liver alcohol dehydrogenase have been determined at different pH. The interpretation of such rate parameters is critically discussed with reference to their informative value for the purpose of determination of rate constants (k and k') for the process of ternary-complex interconversion in the proposed scheme. It is concluded that the apparent rate constant (k') for hydride transfer from benzyl alcohol to NAD+ is dependent on a proton dissociation step with a pKa of 6.4, whereas the rate constant (k) for hydride transfer from NADH to benzaldehyde exhibits no corresponding dependence on proton association. 4. The asymmetric pH dependence of the forward and reverse rate of ternary-complex interconversion during liver alcohol dehydrogenase catalysis appears to reflect an obligatory step of alcohol/alcoholate ion equilibration occurring at the ternary-complex level. It is suggested that the observed pKa 6.4 dependence of the transient rate of alcohol oxidation can be attributed to a coupled acid-base system involving minimally the enzyme-bound alcohol and the protein residues Ser-48 and His-51. | [
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PMID:27360 | Isolation of a multiprotein complex containing cytochrome b and c1 from Neurospora crassa mitochondria by affinity chromatography on immobilized cytochrome c. Difference in the binding between ferricytochrome c and ferrocytochrome c to the multiprotein complex. | A multiprotein complex which contains in equimolar amounts two cytochromes b (Mr each about 27,000), one cytochrome c1 (Mr 31,000) and six subunits without known prosthetic groups (Mr 8000, 12,000, 14,000, 45,000, 45,000, and 50,000) has been isolated from the mitochondrial membranes of Neurospora crassa by affinity chromatography on immobilized cytochrome c. The chromatographic separation was based upon the specific binding of the complex to ferricytochrome c coupled to Sepharose and its specific release upon conversion of the coupled ferricytochrome c into ferrocytochrome c using ascorbate as a reductant. The chromatography was performed in the presence of the nonionic detergent Triton X-100 at low ionic strengths. A monodisperse preparation of the multiprotein complex was obtained which was used for binding studies with cytochrome c from Neurospora crassa, horse heart and Saccaromyces cerevisiae. At low ionic strength (20 mM Trisacetate) and slightly alkaline pH (pH 7 to 8), more than one molecule of ferricytochrome c were bound to the isolated multiprotein complex with dissociation constants below 1 x 10(-7) M. One of these bindings appeared different from the others, since its high affinity was preserved at an ionic strength at which the affinities of the other bindings decreased. Furthermore, the affinity of only this binding decreased upon reduction of cytochrome c. It is suggested that this binding is at or near the functionally active site(s) of the mulipprotein complex. | Isolation of a multiprotein complex containing cytochrome b and c1 from Neurospora crassa mitochondria by affinity chromatography on immobilized cytochrome c. Difference in the binding between ferricytochrome c and ferrocytochrome c to the multiprotein complex. A multiprotein complex which contains in equimolar amounts two cytochromes b (Mr each about 27,000), one cytochrome c1 (Mr 31,000) and six subunits without known prosthetic groups (Mr 8000, 12,000, 14,000, 45,000, 45,000, and 50,000) has been isolated from the mitochondrial membranes of Neurospora crassa by affinity chromatography on immobilized cytochrome c. The chromatographic separation was based upon the specific binding of the complex to ferricytochrome c coupled to Sepharose and its specific release upon conversion of the coupled ferricytochrome c into ferrocytochrome c using ascorbate as a reductant. The chromatography was performed in the presence of the nonionic detergent Triton X-100 at low ionic strengths. A monodisperse preparation of the multiprotein complex was obtained which was used for binding studies with cytochrome c from Neurospora crassa, horse heart and Saccaromyces cerevisiae. At low ionic strength (20 mM Trisacetate) and slightly alkaline pH (pH 7 to 8), more than one molecule of ferricytochrome c were bound to the isolated multiprotein complex with dissociation constants below 1 x 10(-7) M. One of these bindings appeared different from the others, since its high affinity was preserved at an ionic strength at which the affinities of the other bindings decreased. Furthermore, the affinity of only this binding decreased upon reduction of cytochrome c. It is suggested that this binding is at or near the functionally active site(s) of the mulipprotein complex. | [
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PMID:27361 | Reduction of methemoglobin by cobaltocytochrome c catalyzed by mediators. | The reduction of methemoglobin by cobaltocytochrome c (Cocyt c) has been measured using nine mediators of different half-reduction potentials, Em, 7. The rate increases with the increase of Em, 7 for the mediator but dropped precipitously when it becomes more positive than the Em, 7 for the methemoglobin/hemoglobin couple. The reaction is most efficient with phenzaine methosulfate, therefore it was studied in detail. The reaction is first order in the concentrations of Cocyt c and phenazine methosulfate. The average second-order rate constant for Cocyt c + phenazine methosulfate (M) k1 leads to Cocyt c+ M-. is 2.9 x 10(4) M-1 s-1 at 25 degrees C, 0.1 M phosphate pH 7.0. There is a slight negative temperature dependence of k1 at low temperature; at higher temperatures the process has deltaH not equal to approximately 27 kJ mol-1 and deltaS not equal to approxmately - 75 J mol-1 K-1. The effect of anions reflects the dependence of Em, 7 for the methemoglobin/hemoglobin couple with various anions. There is no significant effect on k1 by the addition of inositol hexakisphosphate. The variation of k1 with pH is complicated. The experimental rate constants are compared with values calculated with the theory of nonadiabatic multiphonon process of electron tunneling. | Reduction of methemoglobin by cobaltocytochrome c catalyzed by mediators. The reduction of methemoglobin by cobaltocytochrome c (Cocyt c) has been measured using nine mediators of different half-reduction potentials, Em, 7. The rate increases with the increase of Em, 7 for the mediator but dropped precipitously when it becomes more positive than the Em, 7 for the methemoglobin/hemoglobin couple. The reaction is most efficient with phenzaine methosulfate, therefore it was studied in detail. The reaction is first order in the concentrations of Cocyt c and phenazine methosulfate. The average second-order rate constant for Cocyt c + phenazine methosulfate (M) k1 leads to Cocyt c+ M-. is 2.9 x 10(4) M-1 s-1 at 25 degrees C, 0.1 M phosphate pH 7.0. There is a slight negative temperature dependence of k1 at low temperature; at higher temperatures the process has deltaH not equal to approximately 27 kJ mol-1 and deltaS not equal to approxmately - 75 J mol-1 K-1. The effect of anions reflects the dependence of Em, 7 for the methemoglobin/hemoglobin couple with various anions. There is no significant effect on k1 by the addition of inositol hexakisphosphate. The variation of k1 with pH is complicated. The experimental rate constants are compared with values calculated with the theory of nonadiabatic multiphonon process of electron tunneling. | [
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PMID:27362 | Nuclear-magnetic-resonance studies of 5'-ribonucleotide and 5'-deoxyribonucleotide conformations in solution using the lanthanide probe method. | The conformations of the metal-bound 5'-ribonucleotides and 5'-deoxyribonucleotides in aqueous solution at different pH values have been studied using the lanthanide probe method. The conformational analysis, based on mixing different conformations in fast exchange within the nuclear magnetic resonance time scale, agrees well with the results from coupling constants, nuclear Over-hauser effects and spin-lattice relaxation times, obtained for the metal-fixed systems. The equilibrium between the two basic conformational combinations for the 5'-nucleotides, anti-(N in equalibrium S)-gg-g'g' and syn-(N in equalibrium S)-gt-g'g' depends on the nature of the furanose ring, the base and also on the state of base protonation and phosphate ionization. The effect of base protonation is particularly strong for the guanine nucleotides. | Nuclear-magnetic-resonance studies of 5'-ribonucleotide and 5'-deoxyribonucleotide conformations in solution using the lanthanide probe method. The conformations of the metal-bound 5'-ribonucleotides and 5'-deoxyribonucleotides in aqueous solution at different pH values have been studied using the lanthanide probe method. The conformational analysis, based on mixing different conformations in fast exchange within the nuclear magnetic resonance time scale, agrees well with the results from coupling constants, nuclear Over-hauser effects and spin-lattice relaxation times, obtained for the metal-fixed systems. The equilibrium between the two basic conformational combinations for the 5'-nucleotides, anti-(N in equalibrium S)-gg-g'g' and syn-(N in equalibrium S)-gt-g'g' depends on the nature of the furanose ring, the base and also on the state of base protonation and phosphate ionization. The effect of base protonation is particularly strong for the guanine nucleotides. | [
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PMID:27363 | Ecdysterone biosynthesis: a microsomal cytochrome-P-450-linked ecdysone 20-monooxygenase from tissues of the African migratory locust. | Ecdysone 20-monooxygenase, an enzyme which converts ecdysone to ecdysterone (the major moulting hormone of insects) has been characterized in cell-free preparations of tissues from African migratory locust. The product of the reaction has been identified as ecdysterone on the basis of several microchemical derivatization and chromatographic methods. Ecdysone 20-monooxygenase activity is located primarily in the microsomal fraction which also carries NADPH cytochrome c reductase and cytochrome P-450, as shown by sucrose density gradient centrifugation. Optimal conditions for the ecdysone 20-monooxygenase assay have been determined. The enzyme has a Km for ecdysone of 2.7 x 10(-7) M and is competitvely inhibited by ecdysterone (Ki = 7.5 x 10(-7) M). Ecdysone 20-monooxygenase is a typical cytochrome P-450 linked monooxygenase: the reaction requires O2 and is inhibited by CO, an effect partially reversed by white light. The enzyme is effectively inhibited by several specific monooxygenase inhibitors and by sulfhydryl reagents, but not by cyanide ions. Ecdysone elicits a type I difference spectrum when added to oxidized microsomes. NADPH acts as preferential electron donor. The transfer of reducing equivalents proceeds through NADPH cytochrome c (P-450) reductase: ecdysone 20-monooxygenase is inhibited by cytochrome c. Both NADPH cytochrome c reductase and ecdysone 20-monooxygenase are inhibited by NADP+ and show a similar Km for NADPH. The Malpighian tubules have the highest specific activity of ecdysone 20-monooxygenase, while fat body contain most of the cytochrome P-450 and NADPH cytochrome c reductase. | Ecdysterone biosynthesis: a microsomal cytochrome-P-450-linked ecdysone 20-monooxygenase from tissues of the African migratory locust. Ecdysone 20-monooxygenase, an enzyme which converts ecdysone to ecdysterone (the major moulting hormone of insects) has been characterized in cell-free preparations of tissues from African migratory locust. The product of the reaction has been identified as ecdysterone on the basis of several microchemical derivatization and chromatographic methods. Ecdysone 20-monooxygenase activity is located primarily in the microsomal fraction which also carries NADPH cytochrome c reductase and cytochrome P-450, as shown by sucrose density gradient centrifugation. Optimal conditions for the ecdysone 20-monooxygenase assay have been determined. The enzyme has a Km for ecdysone of 2.7 x 10(-7) M and is competitvely inhibited by ecdysterone (Ki = 7.5 x 10(-7) M). Ecdysone 20-monooxygenase is a typical cytochrome P-450 linked monooxygenase: the reaction requires O2 and is inhibited by CO, an effect partially reversed by white light. The enzyme is effectively inhibited by several specific monooxygenase inhibitors and by sulfhydryl reagents, but not by cyanide ions. Ecdysone elicits a type I difference spectrum when added to oxidized microsomes. NADPH acts as preferential electron donor. The transfer of reducing equivalents proceeds through NADPH cytochrome c (P-450) reductase: ecdysone 20-monooxygenase is inhibited by cytochrome c. Both NADPH cytochrome c reductase and ecdysone 20-monooxygenase are inhibited by NADP+ and show a similar Km for NADPH. The Malpighian tubules have the highest specific activity of ecdysone 20-monooxygenase, while fat body contain most of the cytochrome P-450 and NADPH cytochrome c reductase. | [
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PMID:27365 | Ultramicro-electrophoresis of protein in sweat from patients with cystic fibrosis of the pancreas and controls. | 46 sweat-samples from 32 children with cystic fibrosis of the pancreas (C.F.) and 35 samples from 23 control-children were collected with glass micro-capillaries. Protein was determined by ultramicro-electrophoresis. 1. Protein was detected in 48% of the samples (C.F. and controls). 2. At pH 2.3, C.F.-sweat showed at least one more band than control-sweat. 3. At pH 8.9, C.F.-sweat occasionally showed one more band than the control-group. 4. At pH 2.3, more protein was found at the electrophoresis start point using C.F.-sweat, whereas at pH 8.9 the opposite was found. 5. At pH 2.3, protein was found in fewer samples than at pH 8.9. The "C.F.-factor" is postulated to represent a basic polyelectrolyte which induces the following pathogenic mechanisms: a) Aggregation of proteins giving rise to a high viscosity of secretions, such as saliva. b) Binding to the cell-membrane of the glandular epithelium, thus inducing a disturbance of active NaCl-reabsorption (e.g. by reduction of luminal passive Na+-influx). | Ultramicro-electrophoresis of protein in sweat from patients with cystic fibrosis of the pancreas and controls. 46 sweat-samples from 32 children with cystic fibrosis of the pancreas (C.F.) and 35 samples from 23 control-children were collected with glass micro-capillaries. Protein was determined by ultramicro-electrophoresis. 1. Protein was detected in 48% of the samples (C.F. and controls). 2. At pH 2.3, C.F.-sweat showed at least one more band than control-sweat. 3. At pH 8.9, C.F.-sweat occasionally showed one more band than the control-group. 4. At pH 2.3, more protein was found at the electrophoresis start point using C.F.-sweat, whereas at pH 8.9 the opposite was found. 5. At pH 2.3, protein was found in fewer samples than at pH 8.9. The "C.F.-factor" is postulated to represent a basic polyelectrolyte which induces the following pathogenic mechanisms: a) Aggregation of proteins giving rise to a high viscosity of secretions, such as saliva. b) Binding to the cell-membrane of the glandular epithelium, thus inducing a disturbance of active NaCl-reabsorption (e.g. by reduction of luminal passive Na+-influx). | [
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PMID:27366 | Erythrocyte organic phosphates in the anemia of renal failure in childhood. | Erythrocyte 2,3-diphosphoglycerate (2,3-DPG ) and adenosinetriphosphate (ATP) levels were determined in 43 children with chronic renal failure on conservative treatment (CT), and 12 children on regular hemodialysis (HD) immediately before and after a HD session. The results were compared to non-anemic and anemic controls. In spite of anemia, erythrocyte 2,3-DPG in renal failure was similar to non-anemic controls at normal blood pH, but rose during dialysis as a result of alkalosis. In contrast, ATP levels were high already at a normal blood pH. 2,3-DPG correlated with packed cell volume (PCV) in children with renal failure but at lower concentrations compared to controls. Both organic phosphates in the erythrocytes showed a significant correlation with blood pH. The poor increase of 2,3-DPG, in combination with elevated ATP levels, suggests uremia-induced inhibition of 2,3-DPG synthesis. | Erythrocyte organic phosphates in the anemia of renal failure in childhood. Erythrocyte 2,3-diphosphoglycerate (2,3-DPG ) and adenosinetriphosphate (ATP) levels were determined in 43 children with chronic renal failure on conservative treatment (CT), and 12 children on regular hemodialysis (HD) immediately before and after a HD session. The results were compared to non-anemic and anemic controls. In spite of anemia, erythrocyte 2,3-DPG in renal failure was similar to non-anemic controls at normal blood pH, but rose during dialysis as a result of alkalosis. In contrast, ATP levels were high already at a normal blood pH. 2,3-DPG correlated with packed cell volume (PCV) in children with renal failure but at lower concentrations compared to controls. Both organic phosphates in the erythrocytes showed a significant correlation with blood pH. The poor increase of 2,3-DPG, in combination with elevated ATP levels, suggests uremia-induced inhibition of 2,3-DPG synthesis. | [
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PMID:27367 | Methylmalonic acidemia. | A patient presenting with severe metabolic acidosis accompanied by hyperglycinemia, hyperuricemia, hypoglycemia and hypertammonemia is described. Metabolic acidosis was found to be due to accumulated methylmalonic acid and did not respond to vitamin B12 administration. The patient showed favorable growth and development when kept on a low isoleucine, methionine, threonine and valine diet. In vitro studies using a lymphoid cell line derived from the patient showed a deficiency of methylmalonyl-CoA carbonyl-mutase. | Methylmalonic acidemia. A patient presenting with severe metabolic acidosis accompanied by hyperglycinemia, hyperuricemia, hypoglycemia and hypertammonemia is described. Metabolic acidosis was found to be due to accumulated methylmalonic acid and did not respond to vitamin B12 administration. The patient showed favorable growth and development when kept on a low isoleucine, methionine, threonine and valine diet. In vitro studies using a lymphoid cell line derived from the patient showed a deficiency of methylmalonyl-CoA carbonyl-mutase. | [
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PMID:27372 | Controlled double-blind trial of camazepam, a new benzodiazepine derivative, versus chlordiazepoxide. | 31 in-patients with anxiety entered at random in a double-blind clinical trial of chlordiazepoxide and camazepam, a benzodiazepine derivative. They were assessed by three different rating scales. Statistical evaluation of the results showed similar psychopharmacological activity of both drugs. It is concluded that camazepam is suitable for treatment of mild and severe anxiety states. | Controlled double-blind trial of camazepam, a new benzodiazepine derivative, versus chlordiazepoxide. 31 in-patients with anxiety entered at random in a double-blind clinical trial of chlordiazepoxide and camazepam, a benzodiazepine derivative. They were assessed by three different rating scales. Statistical evaluation of the results showed similar psychopharmacological activity of both drugs. It is concluded that camazepam is suitable for treatment of mild and severe anxiety states. | [
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PMID:27373 | Phenprocoumon requirement, whole blood coagulation time, bleeding time and plasma gamma-GT in patients receiving mianserin. | A possible interaction between the tetracyclic antidepressant mianserin and a coumarin derivative has been investigated. Sixty-three subjects, 61 of whom required anticoagulant therapy for a variety of medical conditions, were treated for 5 consecutive weeks with phenprocoumon, in a dose adjusted to reduce the prothrombin time to 15%-25%. After an initial control period of one week, subjects were randomly treated under double-blind conditions with mianserin 3 X 10 mg daily or 3 X 20 mg daily, or with a matching placebo. The dose of mianserin was gradually increased to reach the maximum by the 6th day. Three subjects dropped out and 60 completed the trial. The dose of phenprocoumon and the prothrombin, bleeding, and coagulation times were not significantly affected by administration of mianserin. It can be concluded that there is no clinically important interaction between phenprocoumon and doses of mianserin effective in depression. Sedation was more frequent in patients taking mianserin than in those given placebo. | Phenprocoumon requirement, whole blood coagulation time, bleeding time and plasma gamma-GT in patients receiving mianserin. A possible interaction between the tetracyclic antidepressant mianserin and a coumarin derivative has been investigated. Sixty-three subjects, 61 of whom required anticoagulant therapy for a variety of medical conditions, were treated for 5 consecutive weeks with phenprocoumon, in a dose adjusted to reduce the prothrombin time to 15%-25%. After an initial control period of one week, subjects were randomly treated under double-blind conditions with mianserin 3 X 10 mg daily or 3 X 20 mg daily, or with a matching placebo. The dose of mianserin was gradually increased to reach the maximum by the 6th day. Three subjects dropped out and 60 completed the trial. The dose of phenprocoumon and the prothrombin, bleeding, and coagulation times were not significantly affected by administration of mianserin. It can be concluded that there is no clinically important interaction between phenprocoumon and doses of mianserin effective in depression. Sedation was more frequent in patients taking mianserin than in those given placebo. | [
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PMID:27369 | Long-term blood pressure control with pindolol given once daily. | The antihypertensive effect of beta-adrenoceptor blocking drugs has been documented in a number of papers [10--13] and over the last 10 yr beta-receptor blockade has become a more and more frequent primary therapy for hypertension. During this time, a number of different beta-blockers with various degrees of selectivity, potency and intrinsic sympathomimetic effect have come into use. There have also been attempts to reduce the number of daily doses in beta-adrenoceptor blocking therapy while maintaining the antihypertensive effect [5]. Pindolol (Viskén) is a beta-adrenoceptor blocking agent with intrinsic sympathomimetic effect and is characterized by high potency and long duration. | Long-term blood pressure control with pindolol given once daily. The antihypertensive effect of beta-adrenoceptor blocking drugs has been documented in a number of papers [10--13] and over the last 10 yr beta-receptor blockade has become a more and more frequent primary therapy for hypertension. During this time, a number of different beta-blockers with various degrees of selectivity, potency and intrinsic sympathomimetic effect have come into use. There have also been attempts to reduce the number of daily doses in beta-adrenoceptor blocking therapy while maintaining the antihypertensive effect [5]. Pindolol (Viskén) is a beta-adrenoceptor blocking agent with intrinsic sympathomimetic effect and is characterized by high potency and long duration. | [
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PMID:27375 | Participation of the sympathetic and the renin--angiotensin systems in blood pressure control during hypercapnia in the anaesthetized dog. | The effects of hypercapnia on plasma renin concentration and blood pressure were studied in anaesthetized dogs, untreated and after pretreatment with guanethidine, propranolol or prazosin. An increase in plasma renin concentration which accompanied hypercapnia in untreated dogs was completely suppressed by pretreatment with guanethidine or propranolol. Prazosin significantly reduced but did not abolish renin release during hypercapnia. The pressor response normally occurring during hypercapnia was abolished by propranolol and reversed by guanethidine and prazosin. | Participation of the sympathetic and the renin--angiotensin systems in blood pressure control during hypercapnia in the anaesthetized dog. The effects of hypercapnia on plasma renin concentration and blood pressure were studied in anaesthetized dogs, untreated and after pretreatment with guanethidine, propranolol or prazosin. An increase in plasma renin concentration which accompanied hypercapnia in untreated dogs was completely suppressed by pretreatment with guanethidine or propranolol. Prazosin significantly reduced but did not abolish renin release during hypercapnia. The pressor response normally occurring during hypercapnia was abolished by propranolol and reversed by guanethidine and prazosin. | [
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PMID:27376 | Isoelectric focusing study of radiation damage to dry conalbumin. | The effect of gamma-rays on iron-free conalbumin was studied by isoelectric focusing technique. The results obtained can be interpreted on the basis of radiation-induced conformational changes of the macromolecules through a cleavage of disulphide bridges. Treatments with reagents acting on disulphide bridges lead to isoelectric focusing patterns that confirm this hypothesis. | Isoelectric focusing study of radiation damage to dry conalbumin. The effect of gamma-rays on iron-free conalbumin was studied by isoelectric focusing technique. The results obtained can be interpreted on the basis of radiation-induced conformational changes of the macromolecules through a cleavage of disulphide bridges. Treatments with reagents acting on disulphide bridges lead to isoelectric focusing patterns that confirm this hypothesis. | [
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PMID:27378 | [Influence of isolanide and trasicor on the content of nicotinamide coenzymes in the myocardium of growing rats in hemic hypoxia]. | Hemic hypoxia was produced in 7,30-day and 3--5-month old (adult) rats through introduction of sodium nitrite (100 mg/kg) with subsequent determination of the nicotinamide co-enzymes content in the myocardium. The effect of combined and separate application of the cardiac glycoside isolanide and beta-adrenoblocking agent trasicor under hypoxia at early stages of post-natal ontogenesis is shown. | [Influence of isolanide and trasicor on the content of nicotinamide coenzymes in the myocardium of growing rats in hemic hypoxia]. Hemic hypoxia was produced in 7,30-day and 3--5-month old (adult) rats through introduction of sodium nitrite (100 mg/kg) with subsequent determination of the nicotinamide co-enzymes content in the myocardium. The effect of combined and separate application of the cardiac glycoside isolanide and beta-adrenoblocking agent trasicor under hypoxia at early stages of post-natal ontogenesis is shown. | [
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PMID:27390 | The cation composition of the seminal plasma and prostatic fluid and its correlation to semen quality. | The concentrations of calcium, magnesium, and zinc were measured by atomic absorption spectrometry. The concentrations of all three cations were found to be strongly correlated with one another and with acidity (pH). Analysis of the relative concentrations in prostatic fluid, split ejaculates, and whole seminal plasma confirmed an almost exclusively prostatic origin of these cations. Semen quality, as judged by motility, vitality, concentration, and morphology of spermatozoa, was inversely related to cation concentrations. Therefore, we recommend adoption of the measurement of seminal divalent cations as part of the routine andrologic examination. | The cation composition of the seminal plasma and prostatic fluid and its correlation to semen quality. The concentrations of calcium, magnesium, and zinc were measured by atomic absorption spectrometry. The concentrations of all three cations were found to be strongly correlated with one another and with acidity (pH). Analysis of the relative concentrations in prostatic fluid, split ejaculates, and whole seminal plasma confirmed an almost exclusively prostatic origin of these cations. Semen quality, as judged by motility, vitality, concentration, and morphology of spermatozoa, was inversely related to cation concentrations. Therefore, we recommend adoption of the measurement of seminal divalent cations as part of the routine andrologic examination. | [
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PMID:27391 | Treatment of cryptorchidism with pernasal gonadotropin-releasing hormone therapy. | Twenty-five boys between 1 and 10 years of age with unilateral or bilateral cryptorchidism were treated with 200 microgram of gonadotropin-releasing hormone (GnRH) pernasally six times daily until descensus was completed, or for 10 weeks at most. Complete descent of the tests occurred in 16 patients, usually after 2 to 5 weeks of treatment. No adverse side effects have been observed. Radioimmunologic measurements of serum luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone, and prolactin, carried out before treatment, at the end of treatment, and 6 months after treatment, showed a transitory increase of the LH responsiveness to GnRH in only four of the patients and in the group as a whole a slight but significant decrease of FSH responsiveness. There were no signs of precocious puberty. GnRH antibodies were not found. | Treatment of cryptorchidism with pernasal gonadotropin-releasing hormone therapy. Twenty-five boys between 1 and 10 years of age with unilateral or bilateral cryptorchidism were treated with 200 microgram of gonadotropin-releasing hormone (GnRH) pernasally six times daily until descensus was completed, or for 10 weeks at most. Complete descent of the tests occurred in 16 patients, usually after 2 to 5 weeks of treatment. No adverse side effects have been observed. Radioimmunologic measurements of serum luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone, and prolactin, carried out before treatment, at the end of treatment, and 6 months after treatment, showed a transitory increase of the LH responsiveness to GnRH in only four of the patients and in the group as a whole a slight but significant decrease of FSH responsiveness. There were no signs of precocious puberty. GnRH antibodies were not found. | [
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PMID:27392 | Treatment of cryptorchidism with a potent analog of gonadotropin-releasing hormone. | Pernasal therapy of cryptorchidism with D-Leu6-des-Gly10-gonadotropin-releasing hormone ethylamide (D-Leu6-des-Gly10-GnRH-EA), a potent, long-acting GnRH analog, was attempted. Eleven prepubertal cryptorchid boys received between 25 microgram once daily and 25 to 50 microgram twice daily for 5 to 12 weeks. Complete testicular descent was achieved in 4 of the 11 boys. GnRH tests (1.5 microgram/kg intravenously), conducted in six boys before treatment, after 4 weeks of treatment, and in 2 boys 3 months after treatment, did not reveal changes in gonadotropin secretion indicative of precocious puberty or of decreased hypophyseal sensitivity to GnRH. Antibodies to the GnRH analog or to GnRH could not be detected. | Treatment of cryptorchidism with a potent analog of gonadotropin-releasing hormone. Pernasal therapy of cryptorchidism with D-Leu6-des-Gly10-gonadotropin-releasing hormone ethylamide (D-Leu6-des-Gly10-GnRH-EA), a potent, long-acting GnRH analog, was attempted. Eleven prepubertal cryptorchid boys received between 25 microgram once daily and 25 to 50 microgram twice daily for 5 to 12 weeks. Complete testicular descent was achieved in 4 of the 11 boys. GnRH tests (1.5 microgram/kg intravenously), conducted in six boys before treatment, after 4 weeks of treatment, and in 2 boys 3 months after treatment, did not reveal changes in gonadotropin secretion indicative of precocious puberty or of decreased hypophyseal sensitivity to GnRH. Antibodies to the GnRH analog or to GnRH could not be detected. | [
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PMID:27393 | Histochemical demonstration of glucose-6-phosphate dehydrogenase, D-sorbitol dehydrogenase, and alkaline phosphatase in human ampulla ductus deferentis. | A cytochemical study on the localization of the enzymes glucose-6-phosphate dehydrogenase, sorbitol dehydrogenase, and alkaline phosphatase was carried out in human ampullary glands. Although the former two enzymes showed a marked reactivity, alkaline phosphatase was absent from epithelial cells. On the basis of these results it is concluded that, as in the human seminal vesicle, in the ampullary gland fructose is probably secreted via the oxidative mechanism proposed by Hers (Le Metabolisme du Fructose. Bruxelles, Arscia, 1957). | Histochemical demonstration of glucose-6-phosphate dehydrogenase, D-sorbitol dehydrogenase, and alkaline phosphatase in human ampulla ductus deferentis. A cytochemical study on the localization of the enzymes glucose-6-phosphate dehydrogenase, sorbitol dehydrogenase, and alkaline phosphatase was carried out in human ampullary glands. Although the former two enzymes showed a marked reactivity, alkaline phosphatase was absent from epithelial cells. On the basis of these results it is concluded that, as in the human seminal vesicle, in the ampullary gland fructose is probably secreted via the oxidative mechanism proposed by Hers (Le Metabolisme du Fructose. Bruxelles, Arscia, 1957). | [
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PMID:27396 | Cytological analysis of bone marrow present in the bone nodules induced by human FL cells in mice. | Bone marrow cells recovered from bone nodules induced in mice by intramuscular transplantation of human FL cells were compared with the host femoral bone marrow. The precursors and mature forms of all types of blood cells were found in the induced bone marrow. The concentration of lymphocytes was significantly higher, and that of erythropoietic cells significantly lower in the induced bone marrow as compared with heterotopic femoral bone marrow. | Cytological analysis of bone marrow present in the bone nodules induced by human FL cells in mice. Bone marrow cells recovered from bone nodules induced in mice by intramuscular transplantation of human FL cells were compared with the host femoral bone marrow. The precursors and mature forms of all types of blood cells were found in the induced bone marrow. The concentration of lymphocytes was significantly higher, and that of erythropoietic cells significantly lower in the induced bone marrow as compared with heterotopic femoral bone marrow. | [
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PMID:27397 | Evaluation of the local antihistaminic activity of dimetindene maleate by a quantitative method. | The topical application of a 0.1% dimetindene maleate gel on the forearms of 11 subjects reduces the areas of both erythema and wheal produced by the intradermal injection of 0.025 mg of histamine hydrochloride to a statistically greater degree than 1% d-chlorpheniramine maleate cream (p less than 0.005 and p less than 0.0005, respectively). The quantitative method employed in this study has proved to be an excellent means for the evaluation of the antihistaminic activity of topical preparations. | Evaluation of the local antihistaminic activity of dimetindene maleate by a quantitative method. The topical application of a 0.1% dimetindene maleate gel on the forearms of 11 subjects reduces the areas of both erythema and wheal produced by the intradermal injection of 0.025 mg of histamine hydrochloride to a statistically greater degree than 1% d-chlorpheniramine maleate cream (p less than 0.005 and p less than 0.0005, respectively). The quantitative method employed in this study has proved to be an excellent means for the evaluation of the antihistaminic activity of topical preparations. | [
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PMID:27398 | [Sodium dichloracetate: its application to the therapeutics of experimental hyperiactatemia (author's transl)]. | Different types of experimental hyperlactatemia and hyperpyruvicemia with or without lowering of blood pH were induced in anesthetized dogs. The initially studied experimental model was the hyperlactatemia and hyperpyruvicemia with lowering of blood pH induced by the intraduodenal administration of high doses of phenformin. Intravenous perfusion of sodium dichloroacetate (150 mg/kg infused during 20 minutes), which acts on the enzymatic complex of pyruvate dehydrogenase, reduced the hyperiactatemia and hyperpyruvicemia with or without acidosis provoked by phenformin injected intraduodenally (30 mg/kg), by intense muscular work, by hypoxia or by continous perfusion or adrenaline. Exogenous or endogenous insulin combined with sodium dichloracetate reduced the hyperlactatemia and hyperpyruvicemia as well as the changes in blood pH provoked by phenformin, more strongly than did sodium dichloroacetate alone. These findings have been confirmed in the conscious dog. | [Sodium dichloracetate: its application to the therapeutics of experimental hyperiactatemia (author's transl)]. Different types of experimental hyperlactatemia and hyperpyruvicemia with or without lowering of blood pH were induced in anesthetized dogs. The initially studied experimental model was the hyperlactatemia and hyperpyruvicemia with lowering of blood pH induced by the intraduodenal administration of high doses of phenformin. Intravenous perfusion of sodium dichloroacetate (150 mg/kg infused during 20 minutes), which acts on the enzymatic complex of pyruvate dehydrogenase, reduced the hyperiactatemia and hyperpyruvicemia with or without acidosis provoked by phenformin injected intraduodenally (30 mg/kg), by intense muscular work, by hypoxia or by continous perfusion or adrenaline. Exogenous or endogenous insulin combined with sodium dichloracetate reduced the hyperlactatemia and hyperpyruvicemia as well as the changes in blood pH provoked by phenformin, more strongly than did sodium dichloroacetate alone. These findings have been confirmed in the conscious dog. | [
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PMID:27399 | Increased trypsin sensitivity of cell surface macromolecules after malignant transformation. | The effect of treatment with 0.04% (w/v) trypsin (EC 3.4.4.4) for 3 h on the electrophoretic mobility (EPM) of polyoma-virus malignantly transformed BHK21 cells (Py6) and their normal counterparts has been investigated. These particular conditions were chosen because an earlier study had shown that such treatment released material from the Py6 cells which was not obtained from the BHK21 cells. The negative EPM of the Py6 cells at pH 7.5 was greatly increased by this treatment; whereas the EPM of the BHK21 cells remained unchanged. Active enzyme was required to produce the change. No evidence was obtained for cytolysis, cytotoxicity or uptake of the enzyme by the treated Py6 cells. Measurement of the EPM of the Py6 cells at different pH levels before and after trypsin treatment suggested that the enzyme was removing cationic groupings from the cell surface. | Increased trypsin sensitivity of cell surface macromolecules after malignant transformation. The effect of treatment with 0.04% (w/v) trypsin (EC 3.4.4.4) for 3 h on the electrophoretic mobility (EPM) of polyoma-virus malignantly transformed BHK21 cells (Py6) and their normal counterparts has been investigated. These particular conditions were chosen because an earlier study had shown that such treatment released material from the Py6 cells which was not obtained from the BHK21 cells. The negative EPM of the Py6 cells at pH 7.5 was greatly increased by this treatment; whereas the EPM of the BHK21 cells remained unchanged. Active enzyme was required to produce the change. No evidence was obtained for cytolysis, cytotoxicity or uptake of the enzyme by the treated Py6 cells. Measurement of the EPM of the Py6 cells at different pH levels before and after trypsin treatment suggested that the enzyme was removing cationic groupings from the cell surface. | [
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PMID:27412 | Relationship between site of peptic ulceration and gastric acid-peptic activity: new evaluation of gastric analysis in patients with acute gastric bleeding and chronic peptic ulcer. | In an attempt to elucidate the etiology of acute gastric bleeding and/or erosion and chronic peptic ulcer, a measurement of gastric juice and mucosal pepsin was carried out in surgically-treated patients. Patients with massive gastric mucosal bleeding in the fundic gland area showed high levels of fundic mucosal pepsin without acid-pepsin appearance in the gastric contents. In these patients, a significantly high value of the peptic activity ratio of gastric mucosa to gastric juice (MJPR, 36.4 +/- 6.7) was observed. It can be suggested that transient blockage of pepsin output from peptic cells with occur in the course of the acute mucosal bleeding, while acid-peptic digestion could be carried out within the fundic gland mucosa. On the other hand, a close correlation between relatively high acid-and-pepsin concentration of the gastric contents and a low level of MJPR (5.6 +/- 1.2) was observed in patients with chronic gastric ulcer. Patients who had a gastric ulcer within the pyloric gland mucosa had a highest acid-peptic activity among three groups with ulcers in fundic gland area, border zone and pyloric gland area. There is a rule that acid-peptic activity becomes low when the site of gastric ulcer moves from pylorus to fundus. A marked increase in acid-and-pepsin secretion into the gastric cavity was observed in patients suffering from chronic duodenal ulcer showing the lowest level of MJPR (3.40 +/- 0.50). | Relationship between site of peptic ulceration and gastric acid-peptic activity: new evaluation of gastric analysis in patients with acute gastric bleeding and chronic peptic ulcer. In an attempt to elucidate the etiology of acute gastric bleeding and/or erosion and chronic peptic ulcer, a measurement of gastric juice and mucosal pepsin was carried out in surgically-treated patients. Patients with massive gastric mucosal bleeding in the fundic gland area showed high levels of fundic mucosal pepsin without acid-pepsin appearance in the gastric contents. In these patients, a significantly high value of the peptic activity ratio of gastric mucosa to gastric juice (MJPR, 36.4 +/- 6.7) was observed. It can be suggested that transient blockage of pepsin output from peptic cells with occur in the course of the acute mucosal bleeding, while acid-peptic digestion could be carried out within the fundic gland mucosa. On the other hand, a close correlation between relatively high acid-and-pepsin concentration of the gastric contents and a low level of MJPR (5.6 +/- 1.2) was observed in patients with chronic gastric ulcer. Patients who had a gastric ulcer within the pyloric gland mucosa had a highest acid-peptic activity among three groups with ulcers in fundic gland area, border zone and pyloric gland area. There is a rule that acid-peptic activity becomes low when the site of gastric ulcer moves from pylorus to fundus. A marked increase in acid-and-pepsin secretion into the gastric cavity was observed in patients suffering from chronic duodenal ulcer showing the lowest level of MJPR (3.40 +/- 0.50). | [
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PMID:27413 | In vitro studies on mechanism for concentrating function of gallbladder. | The mechanism for concentrating function of gallbladder was studied in vitro, turning and attention to changes in the gallbladder bile composition, especially a marked increase of Na+ and a marked decrease of Cl- in the concentrating process of hepatic bile to gallbladder one. The following results were obtained by the present investigations: 1) bile acids present in bile regulate the rate of Na+ and Cl- transport: 2) the epithelial membrane of the gallbladder can be regarded as being a simple semi-permeable membrane. | In vitro studies on mechanism for concentrating function of gallbladder. The mechanism for concentrating function of gallbladder was studied in vitro, turning and attention to changes in the gallbladder bile composition, especially a marked increase of Na+ and a marked decrease of Cl- in the concentrating process of hepatic bile to gallbladder one. The following results were obtained by the present investigations: 1) bile acids present in bile regulate the rate of Na+ and Cl- transport: 2) the epithelial membrane of the gallbladder can be regarded as being a simple semi-permeable membrane. | [
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PMID:27414 | Food sensitivity in reflux esophagitis. | We examined 66 patients with pain of possible esophageal origin for sensitivity to intraesophageal infusions of coffee, orange juice, spicy tomato drink, or HCl of varying concentrations as an addendum to their acid infusion (Bernstein) tests. Compared to Berstein-negative subjects, acid-sensitive patients were sensitive to infusion of coffee (P less than 0.01), orange juice (P less than 0.001), and tomato drink (P less than 0.001). Patients were largely insensitive to HCl solutions with a titratable acidity of 1 mEq per liter or less, less than the least acidic food solution tested. However, Berstein-positive patients were still highly sensitive to infusions of coffee, orange juice, and tomato drink adjusted to pH 7 (P less than 0.001). Patients were unable to differentiate symptoms caused by acid or food infusions, and solutions did not differ in the duration of infusion needed either to cause symptoms or to relieve them by saline. We conclude that the pain of esophagitis is nonspecific and can be precipitated by variety of seemingly unrelated substances. | Food sensitivity in reflux esophagitis. We examined 66 patients with pain of possible esophageal origin for sensitivity to intraesophageal infusions of coffee, orange juice, spicy tomato drink, or HCl of varying concentrations as an addendum to their acid infusion (Bernstein) tests. Compared to Berstein-negative subjects, acid-sensitive patients were sensitive to infusion of coffee (P less than 0.01), orange juice (P less than 0.001), and tomato drink (P less than 0.001). Patients were largely insensitive to HCl solutions with a titratable acidity of 1 mEq per liter or less, less than the least acidic food solution tested. However, Berstein-positive patients were still highly sensitive to infusions of coffee, orange juice, and tomato drink adjusted to pH 7 (P less than 0.001). Patients were unable to differentiate symptoms caused by acid or food infusions, and solutions did not differ in the duration of infusion needed either to cause symptoms or to relieve them by saline. We conclude that the pain of esophagitis is nonspecific and can be precipitated by variety of seemingly unrelated substances. | [
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PMID:27416 | Intraluminal pH esophageal motility. | The aim of the present study was to investigate the effect of intraluminal pH on motor activity of the lower esophageal body. Liquid boluses of different pH values (7 to 2) were infused in the esophagus of 14 normal subjects during manometric recording of motor activity. Each test infusion elicited an esophageal motor response, either secondary peristalsis or simultaneous contractions. Secondary peristalsis was elicited by similar volumes of perfusates at pH 7, 6, 5, but significantly less volumes were needed with perfusates pH 4, 3, and 2. Simultaneous motor activity was not affected by intraesophageal pH value in the range investigated. | Intraluminal pH esophageal motility. The aim of the present study was to investigate the effect of intraluminal pH on motor activity of the lower esophageal body. Liquid boluses of different pH values (7 to 2) were infused in the esophagus of 14 normal subjects during manometric recording of motor activity. Each test infusion elicited an esophageal motor response, either secondary peristalsis or simultaneous contractions. Secondary peristalsis was elicited by similar volumes of perfusates at pH 7, 6, 5, but significantly less volumes were needed with perfusates pH 4, 3, and 2. Simultaneous motor activity was not affected by intraesophageal pH value in the range investigated. | [
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PMID:27418 | [Riboflavin biosynthesis operon of Bacillus subtilis. XIII. Genetic and biochemical study of mutants with regard to intermediate stages of biosynthesis]. | New riboflavin dependent mutants of Bacillus subtilis accumulating different pteridines were studied. The data obtained show that the formation of ribityl side chain proceeds in a few steps at least on a part of riboflavin precursors. The oxidation of connected ribosyl into ribulose with subsequent restoration of it into ribityl proceeds at first. The corresponding genes are located on terminal part of riboflavin operon, as show the results of two-factor transformational crosses with different donors and recipients. | [Riboflavin biosynthesis operon of Bacillus subtilis. XIII. Genetic and biochemical study of mutants with regard to intermediate stages of biosynthesis]. New riboflavin dependent mutants of Bacillus subtilis accumulating different pteridines were studied. The data obtained show that the formation of ribityl side chain proceeds in a few steps at least on a part of riboflavin precursors. The oxidation of connected ribosyl into ribulose with subsequent restoration of it into ribityl proceeds at first. The corresponding genes are located on terminal part of riboflavin operon, as show the results of two-factor transformational crosses with different donors and recipients. | [
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PMID:27425 | Experimental and clinical investigations on stem cell take and colony formation. | First, lymphocyte transplantation into total body-irradiated rats is discussed. The effect on spleen colony formation caused by the transplantation of untreated lymphocytes, as well as of lymphocytes previously incubated with PHA, or with PHA plus L-asparaginase, or with lymphokines, was studied. Then the effect of the urinary colony-stimulating factor in vitro, and the in vitro feeder-layer activity of leucocytes on colony formation of human and mice bone marrow cells in haematological diseases is dealth with. The injection of rat lymphocytes previously incubated for 24 hours with PHA resulted in a higher number and a larger size of colonies in the spleen of the recipient rats. Lymphocytes preincubated with lymphokines gave rise to the formation of spleen colonies which were larger than those developing after the injection of untreated lymphocytes. When the lymphocytes were previously incubated with PHA plus L-asparaginase, PHA failed to stimulate colony formation in the spleen. The phenomenon is explained by assuming that PHA, as an aspecific stimulator of cell division, initiated the division of CFUs, thus the CFUs content of the preincubated samples increased, resulting in an increase in the number of colonies formed after the transplantation of lymphocytes pretreated with PHA. Another possible explanation is that CFUs division, or their spleen take is enhanced by the immunocompetent lymphocytes activated by PHA, either directly or via soluble mediators produced or released by immunocompetent lymphocytes such as lymphokines. The study of colony-forming cells and colony-stimulating activity in primary myelofibrosis (PM) showed an increase in the number of circulating CFUc in this condition, and an abnormal density of these cells reaching a peak below 1.062. The lowering of CSA in the first two peripheral blood gradient fractions agrees with the observation in the same fractions of a high percentage of CFUc at the expense of the CSC population. Thus, double cell population seems to exist in PM. One is greatly abnormal with a low specific density and high plating efficiency, whereas the other population is almost normal, showing a higher specific density and a lower plating efficiency. | Experimental and clinical investigations on stem cell take and colony formation. First, lymphocyte transplantation into total body-irradiated rats is discussed. The effect on spleen colony formation caused by the transplantation of untreated lymphocytes, as well as of lymphocytes previously incubated with PHA, or with PHA plus L-asparaginase, or with lymphokines, was studied. Then the effect of the urinary colony-stimulating factor in vitro, and the in vitro feeder-layer activity of leucocytes on colony formation of human and mice bone marrow cells in haematological diseases is dealth with. The injection of rat lymphocytes previously incubated for 24 hours with PHA resulted in a higher number and a larger size of colonies in the spleen of the recipient rats. Lymphocytes preincubated with lymphokines gave rise to the formation of spleen colonies which were larger than those developing after the injection of untreated lymphocytes. When the lymphocytes were previously incubated with PHA plus L-asparaginase, PHA failed to stimulate colony formation in the spleen. The phenomenon is explained by assuming that PHA, as an aspecific stimulator of cell division, initiated the division of CFUs, thus the CFUs content of the preincubated samples increased, resulting in an increase in the number of colonies formed after the transplantation of lymphocytes pretreated with PHA. Another possible explanation is that CFUs division, or their spleen take is enhanced by the immunocompetent lymphocytes activated by PHA, either directly or via soluble mediators produced or released by immunocompetent lymphocytes such as lymphokines. The study of colony-forming cells and colony-stimulating activity in primary myelofibrosis (PM) showed an increase in the number of circulating CFUc in this condition, and an abnormal density of these cells reaching a peak below 1.062. The lowering of CSA in the first two peripheral blood gradient fractions agrees with the observation in the same fractions of a high percentage of CFUc at the expense of the CSC population. Thus, double cell population seems to exist in PM. One is greatly abnormal with a low specific density and high plating efficiency, whereas the other population is almost normal, showing a higher specific density and a lower plating efficiency. | [
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PMID:27426 | Effect of lanthanum ion on platelet electrophoretic mobility. | The effect of trivalent La ions on the elctrophoretic mobility of washed human platelets has been studied. The mobility of the human platelet in 0.145 M sodium chloride at pH 7.2 was -0.84 +/- 0.06 micron per sec per volt per cm with an isoelectric point approximately at pH 3.4 La3+ at a concentration corresponding to the physiological level of Ca2+ changed the sign of the electrophoretic mobility of the platelets at pH 5.5 and pH 4.0, but had only a faint or practically no effect at pH 3.0 and pH 2.0. It is assumed that La 3+ exerts its effect when bound to the phosphate groups on the surface. | Effect of lanthanum ion on platelet electrophoretic mobility. The effect of trivalent La ions on the elctrophoretic mobility of washed human platelets has been studied. The mobility of the human platelet in 0.145 M sodium chloride at pH 7.2 was -0.84 +/- 0.06 micron per sec per volt per cm with an isoelectric point approximately at pH 3.4 La3+ at a concentration corresponding to the physiological level of Ca2+ changed the sign of the electrophoretic mobility of the platelets at pH 5.5 and pH 4.0, but had only a faint or practically no effect at pH 3.0 and pH 2.0. It is assumed that La 3+ exerts its effect when bound to the phosphate groups on the surface. | [
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PMID:27427 | Antihaemophilic factor (factor VIII) from human granulocytes. | Factor VIII has been isolated and purified from human granulocytes using chromatography on Sephadex G-200 AND DEAE-Sephadex A-50. The final preparation was purified 60-fold. Some properties of this partly purified factor VIII were compared with the human plasma AHF preparation. Optimum pH activity and the highest activity after incubation at different pH values was found in buffer at pH 7.0. The preparation was thermolabile and had a molecular weight of about 214,000. Absorption curves indicated that it is a protein exhibiting four fractions in polyacrylamide gel disk electrophoresis. AHF preparations from plasma and granulocytes were almost identical in the characteristics tested. | Antihaemophilic factor (factor VIII) from human granulocytes. Factor VIII has been isolated and purified from human granulocytes using chromatography on Sephadex G-200 AND DEAE-Sephadex A-50. The final preparation was purified 60-fold. Some properties of this partly purified factor VIII were compared with the human plasma AHF preparation. Optimum pH activity and the highest activity after incubation at different pH values was found in buffer at pH 7.0. The preparation was thermolabile and had a molecular weight of about 214,000. Absorption curves indicated that it is a protein exhibiting four fractions in polyacrylamide gel disk electrophoresis. AHF preparations from plasma and granulocytes were almost identical in the characteristics tested. | [
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PMID:27428 | Some properties of an endo-1,4-beta-D-xylanase from the ligniperdous fungus Trametes hirsuta. | Some properties of purified endo-1,4-beta-D-xylanase (1,4-beta-D-xylan xylanohydrolase, EC 3.2.1.8) from the ligniperdous fungus Trametes hirsuta were investigated. The enzyme was stable between pH 4.0 and 8.0 were optimum activity at pH 5.0--5.5. The temperature optimum was 50 degrees C and the enzyme was stable for up to 30 min at 45 degrees C; however, it was denatured at higher temperatures. The Km for 4-O-methylglucurono-D-xylan was 6.36.10(-3) equivalents of D-xylose per litre, the activation energy was 28 kJ mol-1. The molecular weight determined by means of gel chromatography was 22000--24000. The enzyme was activated by Ca2+ and inhibited by Ag+ and Hg2+. On the basis of the effect of 2-hydroxy-5-nitrobenzyl bromide. N-bromosuccinimide and N-acetylimidazole it may be assumed that trytophan and possibly tyrosine residues influence the enzyme catalysis. | Some properties of an endo-1,4-beta-D-xylanase from the ligniperdous fungus Trametes hirsuta. Some properties of purified endo-1,4-beta-D-xylanase (1,4-beta-D-xylan xylanohydrolase, EC 3.2.1.8) from the ligniperdous fungus Trametes hirsuta were investigated. The enzyme was stable between pH 4.0 and 8.0 were optimum activity at pH 5.0--5.5. The temperature optimum was 50 degrees C and the enzyme was stable for up to 30 min at 45 degrees C; however, it was denatured at higher temperatures. The Km for 4-O-methylglucurono-D-xylan was 6.36.10(-3) equivalents of D-xylose per litre, the activation energy was 28 kJ mol-1. The molecular weight determined by means of gel chromatography was 22000--24000. The enzyme was activated by Ca2+ and inhibited by Ag+ and Hg2+. On the basis of the effect of 2-hydroxy-5-nitrobenzyl bromide. N-bromosuccinimide and N-acetylimidazole it may be assumed that trytophan and possibly tyrosine residues influence the enzyme catalysis. | [
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PMID:27430 | [Psychopharmacologic drugs in the rehabilitation after myocardial infarct]. | The specific use of psychotropic agents in myocardial infarction has become possible due to interdisciplinary cooperation. Especially the differentiated therapeutic application of benzodiazepine derivatives will take into better account the different psychopathologic states in the various phases. The medicamentous facilities are extended by using the central and peripheral effects of the beta-blockers. After a myocardial infarction there are various somatic and mental symptoms. This requires precise medical and psychological management with reference to a diagnostic and combined-therapeutic schedule of treatment. An intensive co-operation with all medical departments is a conditio sine qua non. | [Psychopharmacologic drugs in the rehabilitation after myocardial infarct]. The specific use of psychotropic agents in myocardial infarction has become possible due to interdisciplinary cooperation. Especially the differentiated therapeutic application of benzodiazepine derivatives will take into better account the different psychopathologic states in the various phases. The medicamentous facilities are extended by using the central and peripheral effects of the beta-blockers. After a myocardial infarction there are various somatic and mental symptoms. This requires precise medical and psychological management with reference to a diagnostic and combined-therapeutic schedule of treatment. An intensive co-operation with all medical departments is a conditio sine qua non. | [
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PMID:27437 | [Insect bite allergy. Acute therapy and hyposensitization by means of insect venom]. | Systemic reactions after an insect bite necessitate an allergologic investigation. In addition to taking the patient's history a skin test and an estimation of allergenspecific IgE should be performed. If insect bite allergy is confirmed immediate pharmacological treatment is necessary and hyposensitization therapy is recommended. Purified venom extracts are available. | [Insect bite allergy. Acute therapy and hyposensitization by means of insect venom]. Systemic reactions after an insect bite necessitate an allergologic investigation. In addition to taking the patient's history a skin test and an estimation of allergenspecific IgE should be performed. If insect bite allergy is confirmed immediate pharmacological treatment is necessary and hyposensitization therapy is recommended. Purified venom extracts are available. | [
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PMID:27438 | [Acid-base equilibrium in the cerebrospinal fluid in diseases of the central nervous parenchyma]. | A number of case studies reveal and dynamic changes of the acid-base balance in the blood and cerebrospinal fluid, such as the increasing decompensation of the metabolic lactate acidosis in the cerebrospinal fluid in lethal cases or the gradual regression in patients who were cured. Individual metabolic cerebrospinal parameters are correlated with the disturbance of consciousness. The cerebrospinal fluid parameters of patients with clouded consiciousness constitute prognostically critical values: lactate 3.6 mmol/l, pyruvate 0.19 mmol/l, pH 7.249 and bicarbonate 18.1 mmol/l. Affections of the central nervous parenchyma are better reflected in the cerebrospinal fluid than in the blood. The determination of the CSF/blood relationship of individual parameters shows a significant increase of the lactate quotient as well as a decrease of the pH quotient and of bicarbonate quotient. Respiratory parameters do not show any statistically significant changes. Statistically significant correlations show that, in the case of primary central nervous diseases, the increased hydrogen ion supply in the cerebrospinal fluid is quickly intercepted by the bicarbonate buffer and that, with a corresponding pH decrease, there is a rapid decompensation of the acid-base balance. | [Acid-base equilibrium in the cerebrospinal fluid in diseases of the central nervous parenchyma]. A number of case studies reveal and dynamic changes of the acid-base balance in the blood and cerebrospinal fluid, such as the increasing decompensation of the metabolic lactate acidosis in the cerebrospinal fluid in lethal cases or the gradual regression in patients who were cured. Individual metabolic cerebrospinal parameters are correlated with the disturbance of consciousness. The cerebrospinal fluid parameters of patients with clouded consiciousness constitute prognostically critical values: lactate 3.6 mmol/l, pyruvate 0.19 mmol/l, pH 7.249 and bicarbonate 18.1 mmol/l. Affections of the central nervous parenchyma are better reflected in the cerebrospinal fluid than in the blood. The determination of the CSF/blood relationship of individual parameters shows a significant increase of the lactate quotient as well as a decrease of the pH quotient and of bicarbonate quotient. Respiratory parameters do not show any statistically significant changes. Statistically significant correlations show that, in the case of primary central nervous diseases, the increased hydrogen ion supply in the cerebrospinal fluid is quickly intercepted by the bicarbonate buffer and that, with a corresponding pH decrease, there is a rapid decompensation of the acid-base balance. | [
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PMID:27440 | Unilateral cryptorchidism with compensatory hypertrophy of descended testicle in prepubertal boys. | 5 prepubertal boys with unilateral cryptorchidism and compensatory hypertrophy of the descended testicle, 22 prepubertal boys with unilateral cryptorchidism and without CTH, and 14 prepubertal normal boys were submitted to LH-RH and to HCG tests in order to study the hormonal behaviour in CTH phenomenon before puberty. High but normal peaks of plasma LH and FSH were observed after LH-RH in CTH boys who showed a significant increase of testosterone after HCG stimulation. On the contrary the LH response to LH-RH and the testosterone response to HCG of the boys with unilateral cryptorchidism and without CTH were, as expected, significantly lower than in the control ones. | Unilateral cryptorchidism with compensatory hypertrophy of descended testicle in prepubertal boys. 5 prepubertal boys with unilateral cryptorchidism and compensatory hypertrophy of the descended testicle, 22 prepubertal boys with unilateral cryptorchidism and without CTH, and 14 prepubertal normal boys were submitted to LH-RH and to HCG tests in order to study the hormonal behaviour in CTH phenomenon before puberty. High but normal peaks of plasma LH and FSH were observed after LH-RH in CTH boys who showed a significant increase of testosterone after HCG stimulation. On the contrary the LH response to LH-RH and the testosterone response to HCG of the boys with unilateral cryptorchidism and without CTH were, as expected, significantly lower than in the control ones. | [
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PMID:27442 | Carcinoma in situ of the testis: frequency and relationship to invasive germ cell tumours in infertile men. | A light microscopical study on a total of 812 consecutive testicular biopsies from 555 infertile men revealed intratubular changes in germ cells compatible with a carcinoma in situ pattern in six oligospermic patients (I.I%); the changes were found in both testes in two of these men. Four of the six patients developed an invasive germ cell tumour within follow-up period of 1.3 to 4.5 years. The results confirm the malignant nature of these intratubular atypical germ cells. It is concluded that testicular biopsy may be useful for early detection and cure of germ cell carcinoma in patients at risk, i.e. patients with cryptorchidism, infertile men or patients with previous cancer of one testis. | Carcinoma in situ of the testis: frequency and relationship to invasive germ cell tumours in infertile men. A light microscopical study on a total of 812 consecutive testicular biopsies from 555 infertile men revealed intratubular changes in germ cells compatible with a carcinoma in situ pattern in six oligospermic patients (I.I%); the changes were found in both testes in two of these men. Four of the six patients developed an invasive germ cell tumour within follow-up period of 1.3 to 4.5 years. The results confirm the malignant nature of these intratubular atypical germ cells. It is concluded that testicular biopsy may be useful for early detection and cure of germ cell carcinoma in patients at risk, i.e. patients with cryptorchidism, infertile men or patients with previous cancer of one testis. | [
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PMID:27443 | G6PD Vientiane: a new glucose-6-phosphate dehydrogenase variant with increased stability. | A new G6PD variant, called G6PD Vientiane, has been discovered in a patient from Laos. The characteristics of this variant are: mild enzyme deficiency (about 50% of the normal activity) in the granulocytes and the red cells, with normal G6PD-related antigen concentration; increased stability; normal Km glucose 6-phosphate and NADP+; increased inhibition constant by NADPH; decreased inhibition by ATP; slightly increased utilization of the substrate analogue; abnormal pH curve, with maximum activity at pH 9.5; slightly reduced starch gel electrophoretic migration. The implications of the molecular stability of a deficient mutant variant are discussed. | G6PD Vientiane: a new glucose-6-phosphate dehydrogenase variant with increased stability. A new G6PD variant, called G6PD Vientiane, has been discovered in a patient from Laos. The characteristics of this variant are: mild enzyme deficiency (about 50% of the normal activity) in the granulocytes and the red cells, with normal G6PD-related antigen concentration; increased stability; normal Km glucose 6-phosphate and NADP+; increased inhibition constant by NADPH; decreased inhibition by ATP; slightly increased utilization of the substrate analogue; abnormal pH curve, with maximum activity at pH 9.5; slightly reduced starch gel electrophoretic migration. The implications of the molecular stability of a deficient mutant variant are discussed. | [
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PMID:27444 | Growth and embryo formation in wild-carrot suspension cultures with ammonium ion as a sole nitrogen source. | Wild-carrot (Daucus carota L.) suspension cultures grew and produced embryos on ammonium ion as a sole source of nitrogen in the absence of any exogenous Kreb's cycle acid when the pH of the medium was controlled by continuous titration with with KOH or KHCO3. | Growth and embryo formation in wild-carrot suspension cultures with ammonium ion as a sole nitrogen source. Wild-carrot (Daucus carota L.) suspension cultures grew and produced embryos on ammonium ion as a sole source of nitrogen in the absence of any exogenous Kreb's cycle acid when the pH of the medium was controlled by continuous titration with with KOH or KHCO3. | [
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PMID:27445 | Initiation and characterization of a diploid cell line from larval tissues of Aedes dorsalis (Meigen). | Mosquito cell cultures were initiated from the minced tissues of newly hatched Aedes dorsalis (Meigen) larvae. Continuous cell division occurred only after an adaptive period of approximately 6 months. Optimal growth of the cells required a relatively low pH of 6.5. Karyological studies showed that the cells have remained diploid (2n = 6) for 60 serial passages and that the cultures are free of contaminating cells. The cultures also were shown to be free of bacteria (including Mycoplasma), fungi and virions. Subpopulations (strains) of the original parental cultures have been selected and characterized on the basis of morphology, karyology, growth rate and monolayer formation. | Initiation and characterization of a diploid cell line from larval tissues of Aedes dorsalis (Meigen). Mosquito cell cultures were initiated from the minced tissues of newly hatched Aedes dorsalis (Meigen) larvae. Continuous cell division occurred only after an adaptive period of approximately 6 months. Optimal growth of the cells required a relatively low pH of 6.5. Karyological studies showed that the cells have remained diploid (2n = 6) for 60 serial passages and that the cultures are free of contaminating cells. The cultures also were shown to be free of bacteria (including Mycoplasma), fungi and virions. Subpopulations (strains) of the original parental cultures have been selected and characterized on the basis of morphology, karyology, growth rate and monolayer formation. | [
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PMID:27446 | gamma-Glutamyltransferase in human diploid fibroblasts and other mammalian cells. | gamma-Glutamyltransferase was determined in WI-38 human diploid fibroblasts and compared to enzyme levels determined in several other mammalian cell lines including: fibroblast-like cells from human skin, tibia and foreskin; epithelial-like cells from human, bovine and monkey kidney; and transformed cells (Chinese hamster ovary, HeLa S3 and SV-40 transformed WI-38). Transformed cells had the lowest activity found followed in increasing order by fibroblasts, human and bovine epithelial cells and monkey kidney epithelial cells. The enzyme isolated from the plasma membrane of WI-38 cells, like the enzyme from kidney and brain, was found to be irreversibly inhibited by iodoacetamide, reversibly by serine-borate, and had a strong specificity for certain amino acids. The possibility exists that gamma-glutamyltransferase could be involved in transport of amino acids into cells in culture; and glutamine, used in media, is an excellent substrate for the enzyme. | gamma-Glutamyltransferase in human diploid fibroblasts and other mammalian cells. gamma-Glutamyltransferase was determined in WI-38 human diploid fibroblasts and compared to enzyme levels determined in several other mammalian cell lines including: fibroblast-like cells from human skin, tibia and foreskin; epithelial-like cells from human, bovine and monkey kidney; and transformed cells (Chinese hamster ovary, HeLa S3 and SV-40 transformed WI-38). Transformed cells had the lowest activity found followed in increasing order by fibroblasts, human and bovine epithelial cells and monkey kidney epithelial cells. The enzyme isolated from the plasma membrane of WI-38 cells, like the enzyme from kidney and brain, was found to be irreversibly inhibited by iodoacetamide, reversibly by serine-borate, and had a strong specificity for certain amino acids. The possibility exists that gamma-glutamyltransferase could be involved in transport of amino acids into cells in culture; and glutamine, used in media, is an excellent substrate for the enzyme. | [
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PMID:27454 | Filamentous capsulated streptococci from the human respiratory tract: chemical and immunochemical characterization of a glycoprotein capsular antigen of provisional binary capsular type 87. | A filamentous alpha-hemolytic streptococcus of provisional capsular type 87 isolated from the human respiratory tract has been shown to be binary capsulated. One of the capsular antigens appears to be a glycoprotein; the other appears to be a polysaccharide. Transformation reactions with deoxyribonucleic acid from streptococcus type 87 and a number of noncapsulated pneumococci yielded transformed pneumococci with either a glycoprotein capsule or a polysaccharide capsule, but not with both. Capsular precipitin (quellung) reactions were observed when streptococcus type 87 was treated with homologous antiserum or with antisera to either of the two distinct capsular transformants. Each of the transformed pneumococci gave a quellung reaction with its homologous antiserum or with antiserum to streptococcus type 87, but neither reacted with antiserum to the heterologous transformant. Chemical analysis showed the glycoprotein antigen of streptococcus type 87 to contain, in addition to amino acids, glucose, galactose, glucosamine, and phosphate. The amino acid composition of the glycoprotein capsular antigens from streptococcus type 87 and of those from transformed pneumococci were similar, showing only minor differences. The glycoprotein capsular antigen from streptococcus type 87 gave two closely associated precipitin bands with homologous antiserum or antisera to transformed pneumococci with the glycoprotein capsule. That the two precipitin bands represent two unrelated proteins is precluded largely on the basis of the unlikely probability of 100% cotransformation of the genes coding for both proteins in the pneumococcal transformants that were isolated. Chemical analyses of the various fractions of the glycoprotein indicate that the two precipitin bands may represent a glycoprotein and its corresponding apoprotein. | Filamentous capsulated streptococci from the human respiratory tract: chemical and immunochemical characterization of a glycoprotein capsular antigen of provisional binary capsular type 87. A filamentous alpha-hemolytic streptococcus of provisional capsular type 87 isolated from the human respiratory tract has been shown to be binary capsulated. One of the capsular antigens appears to be a glycoprotein; the other appears to be a polysaccharide. Transformation reactions with deoxyribonucleic acid from streptococcus type 87 and a number of noncapsulated pneumococci yielded transformed pneumococci with either a glycoprotein capsule or a polysaccharide capsule, but not with both. Capsular precipitin (quellung) reactions were observed when streptococcus type 87 was treated with homologous antiserum or with antisera to either of the two distinct capsular transformants. Each of the transformed pneumococci gave a quellung reaction with its homologous antiserum or with antiserum to streptococcus type 87, but neither reacted with antiserum to the heterologous transformant. Chemical analysis showed the glycoprotein antigen of streptococcus type 87 to contain, in addition to amino acids, glucose, galactose, glucosamine, and phosphate. The amino acid composition of the glycoprotein capsular antigens from streptococcus type 87 and of those from transformed pneumococci were similar, showing only minor differences. The glycoprotein capsular antigen from streptococcus type 87 gave two closely associated precipitin bands with homologous antiserum or antisera to transformed pneumococci with the glycoprotein capsule. That the two precipitin bands represent two unrelated proteins is precluded largely on the basis of the unlikely probability of 100% cotransformation of the genes coding for both proteins in the pneumococcal transformants that were isolated. Chemical analyses of the various fractions of the glycoprotein indicate that the two precipitin bands may represent a glycoprotein and its corresponding apoprotein. | [
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PMID:27455 | Filamentous capsulated streptococci from the human respiratory tract: chemical and immunochemical characterization of the polysaccharide capsular antigen of provisional binary capsular type 87. | The polysaccharide capsular antigen of the filamentous binary capsulated streptococcus of provisional type 87 and the polysaccharide capsular antigens of two pneumoccal strains transformed with deoxyribonucleic acid of streptococus type 87 have been purified and analyzed with regard to their component monosaccharides. The purified polysaccharides from the three strains were immunochemically identical. Each was found to contain rhamnose, glucose, galactose, galactosamine, and phosphate. Rhamnose was the immunodominant sugar. | Filamentous capsulated streptococci from the human respiratory tract: chemical and immunochemical characterization of the polysaccharide capsular antigen of provisional binary capsular type 87. The polysaccharide capsular antigen of the filamentous binary capsulated streptococcus of provisional type 87 and the polysaccharide capsular antigens of two pneumoccal strains transformed with deoxyribonucleic acid of streptococus type 87 have been purified and analyzed with regard to their component monosaccharides. The purified polysaccharides from the three strains were immunochemically identical. Each was found to contain rhamnose, glucose, galactose, galactosamine, and phosphate. Rhamnose was the immunodominant sugar. | [
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