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PMID:24510 | Psychological aspects of chronic pain. | Except in emergencies, the physician's obligation is not to relieve pain, but to diagnose and treat pathology. Psychological factors complicate organic pain, and make the diagnosis of "psychogenic" pain virtually impossible on the basis of psychological tests alone. In acute (short-term) pain, anti-anxiety agents are useful, but in chronic pain antidepressants are usually more appropriate. Continuing anxiolytic drugs past the acute stage tends to potentiate depressions. In addition to antidepressants, modalities which help in the management of chronic pain are physical therapy, transcutaneous electrical neurostimulation, use of weak analgesics on a "clock" rather than an "as needed" basis, and behavior modification. Explicit agreement on the features of the doctor--patient relationship is almost always essential for successfully managing these difficult and demanding patients. | Psychological aspects of chronic pain. Except in emergencies, the physician's obligation is not to relieve pain, but to diagnose and treat pathology. Psychological factors complicate organic pain, and make the diagnosis of "psychogenic" pain virtually impossible on the basis of psychological tests alone. In acute (short-term) pain, anti-anxiety agents are useful, but in chronic pain antidepressants are usually more appropriate. Continuing anxiolytic drugs past the acute stage tends to potentiate depressions. In addition to antidepressants, modalities which help in the management of chronic pain are physical therapy, transcutaneous electrical neurostimulation, use of weak analgesics on a "clock" rather than an "as needed" basis, and behavior modification. Explicit agreement on the features of the doctor--patient relationship is almost always essential for successfully managing these difficult and demanding patients. | [
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PMID:24527 | Some clinical aspects of caprine reproduction. | Several clinical reproductive problems in the female dairy goat are considered, including estrus detection, artificial insemination, pregnancy diagnosis, abortions, and periparturient difficulties. Sexual behavior of the buck, onset of puberty, techniques for semen collection and evaluation, the production of teaser animals, and methods of castration are also discussed. Recognition of the intersex goat, which may have a masculine, feminine, or intermediate phenotype, is necessary in herds where naturally polled animals are bred. | Some clinical aspects of caprine reproduction. Several clinical reproductive problems in the female dairy goat are considered, including estrus detection, artificial insemination, pregnancy diagnosis, abortions, and periparturient difficulties. Sexual behavior of the buck, onset of puberty, techniques for semen collection and evaluation, the production of teaser animals, and methods of castration are also discussed. Recognition of the intersex goat, which may have a masculine, feminine, or intermediate phenotype, is necessary in herds where naturally polled animals are bred. | [
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PMID:24528 | The effects of obstetrical medication on the behavior of Israeli newborn infants and some comparisons with Uruguayan and American infants. | The effects of obstetrical medication on neonatal behavior were studied using a sample of Israeli infants from medicated and nonmedicated mothers. Few significant behavioral differences were detected in the first month of life, and there was no difference in Bayley Infant Scale performance at 3 months of age. Comparative analyses using samples of American and Uruguayan infants and additional data from other studies of American newborns led to the conclusion that light levels of obstetrical medication do not appear to have significant effects on neonatal behavior. However, this may well be qualified by some initial population differences, by the measurements commonly used in the studies reviewed, and possibly by critical cutoff points dividing the levels of medication that will and will not affect neonatal behavior. | The effects of obstetrical medication on the behavior of Israeli newborn infants and some comparisons with Uruguayan and American infants. The effects of obstetrical medication on neonatal behavior were studied using a sample of Israeli infants from medicated and nonmedicated mothers. Few significant behavioral differences were detected in the first month of life, and there was no difference in Bayley Infant Scale performance at 3 months of age. Comparative analyses using samples of American and Uruguayan infants and additional data from other studies of American newborns led to the conclusion that light levels of obstetrical medication do not appear to have significant effects on neonatal behavior. However, this may well be qualified by some initial population differences, by the measurements commonly used in the studies reviewed, and possibly by critical cutoff points dividing the levels of medication that will and will not affect neonatal behavior. | [
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PMID:24529 | Implications of disorders of purine metabolism for the kidney and the urinary tract. | The spectrum of kidney and urinary tract disorders related to purines comprises acute hyperuricosuric nephropathy, chronic urate nephropathy and urolithiasis. Two factors in the development of acute hyperuricosuric nephropathy are increased uric acid concentration and low pH in the tubular fluid. Chronic urate nephropathy still possess several problems: incidence (although this seems to be decreasing, presumably owing to effective prevention), the source of interstitial urate, the cause of the interstitial deposition of urate, and the role of urate deposits in the pathogenesis of the interstitial nephropathy. The relation of the experimental nephropathy to the pathogenesis of chronic urate nephropathy in the human is not yet clear but a model is proposed according to which interstitial urate derives from two sources: hyperuricaemic plasma and hyperuricosuric tubular fluid. Urolithiasis related to purines leads to uric acid-urate stones, xanthine stones, 2,8-dihydroxyadenine stones, iatrogenic xanthine and oxipurinol stones, and possibly calcium stones. Pathogenetic factors in uric acid lithiasis are hyperuricosuria (whether due to an inborn enzyme abnormality or of unknown aetiology) and low urinary pH; oliguria is a contributory factor. There remain several open questions about uric acid lithiasis: incidence, the shift of its location from lower to upper urinary tract, the interplay of pathogenetic factors, and the role of compounds which inhibit crystallization. | Implications of disorders of purine metabolism for the kidney and the urinary tract. The spectrum of kidney and urinary tract disorders related to purines comprises acute hyperuricosuric nephropathy, chronic urate nephropathy and urolithiasis. Two factors in the development of acute hyperuricosuric nephropathy are increased uric acid concentration and low pH in the tubular fluid. Chronic urate nephropathy still possess several problems: incidence (although this seems to be decreasing, presumably owing to effective prevention), the source of interstitial urate, the cause of the interstitial deposition of urate, and the role of urate deposits in the pathogenesis of the interstitial nephropathy. The relation of the experimental nephropathy to the pathogenesis of chronic urate nephropathy in the human is not yet clear but a model is proposed according to which interstitial urate derives from two sources: hyperuricaemic plasma and hyperuricosuric tubular fluid. Urolithiasis related to purines leads to uric acid-urate stones, xanthine stones, 2,8-dihydroxyadenine stones, iatrogenic xanthine and oxipurinol stones, and possibly calcium stones. Pathogenetic factors in uric acid lithiasis are hyperuricosuria (whether due to an inborn enzyme abnormality or of unknown aetiology) and low urinary pH; oliguria is a contributory factor. There remain several open questions about uric acid lithiasis: incidence, the shift of its location from lower to upper urinary tract, the interplay of pathogenetic factors, and the role of compounds which inhibit crystallization. | [
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PMID:24530 | Characterization of human adenosine deaminase. | A deficiency of adenosine deaminase in man is associated with one form of severe combined immunodeficiency disease. In an attempt to define the nature of this relationship we have characterized the normal human enzyme and examined the role of this enzyme in monocyte-macrophage activation. The human enzyme was purified 800 000-fold to apparent homogeneity from human erythrocytes with 31% recovery by immunoabsorbent chromatography. The homogeneous protein contains carbohydrate and has a subunit molecular weight of 42 000, estimated by sodium dodecyl sulphate gel electrophoresis. The enzyme was found to exist in either a soluble or a particulate form. The active soluble forms are interconvertible with apparent molecular weights of 36 000 (small), 114 000 (intermediate), and 298 000 (large). However, conversion of the small form into the large form needs a protein with a molecular weight of 200 000 which has no adenosine deaminase activity. | Characterization of human adenosine deaminase. A deficiency of adenosine deaminase in man is associated with one form of severe combined immunodeficiency disease. In an attempt to define the nature of this relationship we have characterized the normal human enzyme and examined the role of this enzyme in monocyte-macrophage activation. The human enzyme was purified 800 000-fold to apparent homogeneity from human erythrocytes with 31% recovery by immunoabsorbent chromatography. The homogeneous protein contains carbohydrate and has a subunit molecular weight of 42 000, estimated by sodium dodecyl sulphate gel electrophoresis. The enzyme was found to exist in either a soluble or a particulate form. The active soluble forms are interconvertible with apparent molecular weights of 36 000 (small), 114 000 (intermediate), and 298 000 (large). However, conversion of the small form into the large form needs a protein with a molecular weight of 200 000 which has no adenosine deaminase activity. | [
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PMID:24531 | The purinergic nerve hypothesis. | Experiments over the past decade have revealed a third component in the autonomic nervous system which is neither adrenergic nor cholinergic. These nerves are strongly represented in the gastrointestinal tract of a wide range of vertebrate species and have also been identified in lung, trachea, retractor penis, bladder, oesophagus, eye, seminal vesicle and in some parts of the cardiovascular system and brain. Evidence has been presented that the principal active substance released by these nerves in the gut is a purine nucleotide, probably ATP, and they have therefore been termed 'purinergic'. The evidence includes: (1) synthesis and storage of ATP in nerves; (2) release of ATP from the nerves when they are stimulated; (3) mimicry by exogenously applied ATP of the action of nerve-released transmitter; (4) the presence of Mg2+-activated ATPase, 5'-nucleotidase and adenosine deaminase, enzymes which inactivate ATP; (5) the similar blocking and potentiating effects produced by drugs on the responses to exogenously applied ATP and nerve stimulation. A tentative model for the synthesis, storage, release and inactivation of ATP during purinergic nerve transmission is proposed. Some properties of purinergic receptors are described. | The purinergic nerve hypothesis. Experiments over the past decade have revealed a third component in the autonomic nervous system which is neither adrenergic nor cholinergic. These nerves are strongly represented in the gastrointestinal tract of a wide range of vertebrate species and have also been identified in lung, trachea, retractor penis, bladder, oesophagus, eye, seminal vesicle and in some parts of the cardiovascular system and brain. Evidence has been presented that the principal active substance released by these nerves in the gut is a purine nucleotide, probably ATP, and they have therefore been termed 'purinergic'. The evidence includes: (1) synthesis and storage of ATP in nerves; (2) release of ATP from the nerves when they are stimulated; (3) mimicry by exogenously applied ATP of the action of nerve-released transmitter; (4) the presence of Mg2+-activated ATPase, 5'-nucleotidase and adenosine deaminase, enzymes which inactivate ATP; (5) the similar blocking and potentiating effects produced by drugs on the responses to exogenously applied ATP and nerve stimulation. A tentative model for the synthesis, storage, release and inactivation of ATP during purinergic nerve transmission is proposed. Some properties of purinergic receptors are described. | [
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PMID:24535 | Functional maturation of the epididymis in rabbit and rhesus monkey. | The changes in the weight, histology and biochemical composition of the epididymis (caput, corpus and cauda segments) in prepuberal rabbit and rhesus monkey in response to testosterone treatment were investigated. The increase in the weight of the organ was accompanied by increased levels of RNA and DNA. Androgen therapy caused an increase in the concentration of sialic acid, phospholipids and glycerylphosphorylcholine and activities of alkaline phosphatase, acid phosphatase and hyaluronidase. The cauda region of the epididymis exhibited relatively higher levels of sialic acid and glycerylphosphorylcholine. These findings are discussed in relation to the functional maturation of the organ and the role of androgen in this process. | Functional maturation of the epididymis in rabbit and rhesus monkey. The changes in the weight, histology and biochemical composition of the epididymis (caput, corpus and cauda segments) in prepuberal rabbit and rhesus monkey in response to testosterone treatment were investigated. The increase in the weight of the organ was accompanied by increased levels of RNA and DNA. Androgen therapy caused an increase in the concentration of sialic acid, phospholipids and glycerylphosphorylcholine and activities of alkaline phosphatase, acid phosphatase and hyaluronidase. The cauda region of the epididymis exhibited relatively higher levels of sialic acid and glycerylphosphorylcholine. These findings are discussed in relation to the functional maturation of the organ and the role of androgen in this process. | [
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PMID:24536 | Further evidence of teratogenic effects apparently produced by neurotransmitters during brain differentiation. | 1. Adult male rats treated neonatally with the monoamine oxidase inhibitor pargyline showed significantly increased nuclear volumes of the nerve cells in the medial and central amygdaloid nuclei and significantly decreased concentrations of noradrenaline and dopamine in the hypothalamus as compared to control males. 2. Adult male rats treated neonatally with the monoamine depletor reserpine also displayed increased nuclear volumes of the nerve cells in the medial and central amygdaloid nuclei in comparison to the control males but significantly decreased nuclear volumes in comparison to the pargylinized males. Furthermore, the number of synapses in the stratum radiatum (CA 1-region) of the hippocampus was significantly decreased in the reserpinized males in comparison to the control males. 3. Adult male rats treated neonatally with the acetylcholine esterase inhibitor pyridostigmine showed a slight decrease of the noradrenaline concentration in the hypothalamus. These findings suggest that changes of neurotransmitter concentrations and/or turnover rates apparently produced by psychotrophic drugs during brain differentiation are able to exert teratogenic effects. | Further evidence of teratogenic effects apparently produced by neurotransmitters during brain differentiation. 1. Adult male rats treated neonatally with the monoamine oxidase inhibitor pargyline showed significantly increased nuclear volumes of the nerve cells in the medial and central amygdaloid nuclei and significantly decreased concentrations of noradrenaline and dopamine in the hypothalamus as compared to control males. 2. Adult male rats treated neonatally with the monoamine depletor reserpine also displayed increased nuclear volumes of the nerve cells in the medial and central amygdaloid nuclei in comparison to the control males but significantly decreased nuclear volumes in comparison to the pargylinized males. Furthermore, the number of synapses in the stratum radiatum (CA 1-region) of the hippocampus was significantly decreased in the reserpinized males in comparison to the control males. 3. Adult male rats treated neonatally with the acetylcholine esterase inhibitor pyridostigmine showed a slight decrease of the noradrenaline concentration in the hypothalamus. These findings suggest that changes of neurotransmitter concentrations and/or turnover rates apparently produced by psychotrophic drugs during brain differentiation are able to exert teratogenic effects. | [
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PMID:24537 | Different effects of amygdaloid lesions on compensatory ovarian hypertrophy in cyclic and prepubertal female rats. | Cyclic and 28-day-old immature female rats were hemiovariectomized and partly bilaterally lesioned in the cortical amygdaloid nucleus (CAN). The compensatory ovarian hypertrophy (COH) recorded in the adult females on the 8th day after hemicastration was completely prevented by the amygdaloid lesions. In contrast, damage to the CAN did not alter the ovarian weight in prepubertal females, although COH was also induced in these animals by unilateral ovariectomy. | Different effects of amygdaloid lesions on compensatory ovarian hypertrophy in cyclic and prepubertal female rats. Cyclic and 28-day-old immature female rats were hemiovariectomized and partly bilaterally lesioned in the cortical amygdaloid nucleus (CAN). The compensatory ovarian hypertrophy (COH) recorded in the adult females on the 8th day after hemicastration was completely prevented by the amygdaloid lesions. In contrast, damage to the CAN did not alter the ovarian weight in prepubertal females, although COH was also induced in these animals by unilateral ovariectomy. | [
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PMID:24538 | Investigation of the mechanism of the methylmalonyl-CoA mutase reaction with the substrate analogue: ethylmalonyl-CoA. | 1. Ethylmalonyl-CoA was found to be a substrate for methylmalonyl-CoA mutase from Propionibacterium shermanii, the product being mainly (2R)-methylsuccinyl-CoA along with some (2S)-diastereoisomer. 2. The relevant 1H-nuclear magnetic resonance signals of methylsuccinic acid and of its dimethyl ester were assigned to the diastereotopic methylene hydrogens using sterospecifically dideuterated specimens of known configuration. 3. [2(-2)H1]Ethylmalonyl-CoA was converted by methylmalonyl-CoA mutase in 2H2O mainly to (2R, 3S)-[3(-2)H1]methylsuccinyl-CoA. No dideuterated product was observed. 4. Starting from (1R)-[1(-2)H1]-ethathanol, (1S)-[1(-2)H1]ethanol and [2H6] ethanol the following deuterated specimens of ethylmalonic acid were synthesised and characterised: (3S)-[3(-2)H1], (3R)-[3(-2)H1] and [3(-2)H2, 4(-2)H3], respectively. 5. Conversion of (3S)-[3(-2)H1]-ethylmalonyl-CoA (70% 2H1 and 2% 2H2 species) on the mutase in water afforded mainly (2R)-[2(-2)H1]methylsuccinyl-CoA along with some (2S)-diastereoisomer. No deuterium loss was observed. 6. Methylmalonyl-CoA mutase converted (3R)-[3(-2)H1]ethylmalonyl-CoA (81% 2H1 and 2% 2H2 species) to the following methylsuccinyl-CoA species: 33% [3(-2)H1], the deuterium being in the threo position with respect to the methyl group; 21% [2(-2)H1]; 46% unlabelled. The ratio of the species with (2R) and (2S) configuration was about 60:40. 7. Reaction of [3(-2)H2, 4(-2)H3]ethylmalonyl-CoA (94.5% [2H5] species) with the mutase gave the following labelled methylsuccinyl-CoA species:53.4% [methyl-2H3, 2(-2)H1, 3(-2)H1], the 3-deuterium being in the threo position with respect to the methyl group; 37.6% [methyl-2H3, 2(-2)H1]; 5% [methyl(-2)H3, 2(-2)H1, 2(-2)H1, 3(-2)H1] the 3-deuterium being in erythro position with respect to the methyl group; 4% [methyl(-2)H3, 3(-2)H1]. The ratio of the species with (2R) and (2S) configuration was about 70:30. 8. Implications of these findings for the mechanism of the rearrangements catalysed by coenzyme B12 are discussed. | Investigation of the mechanism of the methylmalonyl-CoA mutase reaction with the substrate analogue: ethylmalonyl-CoA. 1. Ethylmalonyl-CoA was found to be a substrate for methylmalonyl-CoA mutase from Propionibacterium shermanii, the product being mainly (2R)-methylsuccinyl-CoA along with some (2S)-diastereoisomer. 2. The relevant 1H-nuclear magnetic resonance signals of methylsuccinic acid and of its dimethyl ester were assigned to the diastereotopic methylene hydrogens using sterospecifically dideuterated specimens of known configuration. 3. [2(-2)H1]Ethylmalonyl-CoA was converted by methylmalonyl-CoA mutase in 2H2O mainly to (2R, 3S)-[3(-2)H1]methylsuccinyl-CoA. No dideuterated product was observed. 4. Starting from (1R)-[1(-2)H1]-ethathanol, (1S)-[1(-2)H1]ethanol and [2H6] ethanol the following deuterated specimens of ethylmalonic acid were synthesised and characterised: (3S)-[3(-2)H1], (3R)-[3(-2)H1] and [3(-2)H2, 4(-2)H3], respectively. 5. Conversion of (3S)-[3(-2)H1]-ethylmalonyl-CoA (70% 2H1 and 2% 2H2 species) on the mutase in water afforded mainly (2R)-[2(-2)H1]methylsuccinyl-CoA along with some (2S)-diastereoisomer. No deuterium loss was observed. 6. Methylmalonyl-CoA mutase converted (3R)-[3(-2)H1]ethylmalonyl-CoA (81% 2H1 and 2% 2H2 species) to the following methylsuccinyl-CoA species: 33% [3(-2)H1], the deuterium being in the threo position with respect to the methyl group; 21% [2(-2)H1]; 46% unlabelled. The ratio of the species with (2R) and (2S) configuration was about 60:40. 7. Reaction of [3(-2)H2, 4(-2)H3]ethylmalonyl-CoA (94.5% [2H5] species) with the mutase gave the following labelled methylsuccinyl-CoA species:53.4% [methyl-2H3, 2(-2)H1, 3(-2)H1], the 3-deuterium being in the threo position with respect to the methyl group; 37.6% [methyl-2H3, 2(-2)H1]; 5% [methyl(-2)H3, 2(-2)H1, 2(-2)H1, 3(-2)H1] the 3-deuterium being in erythro position with respect to the methyl group; 4% [methyl(-2)H3, 3(-2)H1]. The ratio of the species with (2R) and (2S) configuration was about 70:30. 8. Implications of these findings for the mechanism of the rearrangements catalysed by coenzyme B12 are discussed. | [
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PMID:24539 | The value of bilateral biopsy in unilateral cryptorchidism. | Here are the results of bilateral biopsies on 52 cases of unilateral cryptorchidia. In 12 patients the scrotal testicle showed equal lesions or more important lesions in the undescended testicle. It points out the importance of systematizing the histological testicular study, keeping in mind: (1) spermatogenic index; (2) variation in spermatogonias; (3) tubular diameter, and (4) interstitial tissue. Stress is made on the innocuousness of the bilateral testicular biopsy and its indispensability since it is the only way to recognize a dysgenetic cryptorchidia and evaluate the future fertility of the patient. | The value of bilateral biopsy in unilateral cryptorchidism. Here are the results of bilateral biopsies on 52 cases of unilateral cryptorchidia. In 12 patients the scrotal testicle showed equal lesions or more important lesions in the undescended testicle. It points out the importance of systematizing the histological testicular study, keeping in mind: (1) spermatogenic index; (2) variation in spermatogonias; (3) tubular diameter, and (4) interstitial tissue. Stress is made on the innocuousness of the bilateral testicular biopsy and its indispensability since it is the only way to recognize a dysgenetic cryptorchidia and evaluate the future fertility of the patient. | [
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PMID:24540 | Studies on beta-adrenoceptors mediating changes in mechanical events and adenosine 3',5'-monophosphate levels. Rat atria. | Increasing concentrations of the functional antagonist carbachol resulted in a shift to the right of the dose-response curves for (--)-isoproterenol-induced positive chronotropic and inotropic responses and a reduction of the maximum degree of response that could be produced relative to that produced by theophylline. Carbachol only reduced the maximum responses to the partial agonist (--)-soterenol. The ED50 value for isoproterenol-induced increases in adenosine 3',5'-monophosphate (cyclic AMP) levels was more than 10-fold larger than those for production of mechanical events and soterenol did not produce a measurable increase in cyclic AMP above baseline values. Soterenol was analyzed as a competitive antagonist of isoproterenol-induced responses and the apparent dissociation constants (KA) for soterenol calculated to be 1.21 X 10(-7) M for receptors mediating positive chronotropic responses, 3.56 X 10(-7) M for receptors mediating positive inotropic responses, and 2.96 X 10(-7) M for receptors mediating increases in cyclic AMP levels. By two additional theoretical models of drug-receptor interactions, the KA values for soterenol were between 1.5 and 4.5 X 10(-7) M. These KA values were essentially the same as the ED50 values for soterenol-induced mechanical effects. By one of the models, the KA for isoproterenol was calculated to be about 2.14 X 10(-8) M, a value close to the ED50 value for isoproterenol-induced increases in cyclic AMP (5.45 X 10(-8) M). These results suggest that the beta-adrenoceptors mediating the three responses in rat atria are of the same type and that differences in concentrations required to produce mechanical vs. cyclic AMP changes results with agonists possessing high efficacy for which there is a receptor reserve. | Studies on beta-adrenoceptors mediating changes in mechanical events and adenosine 3',5'-monophosphate levels. Rat atria. Increasing concentrations of the functional antagonist carbachol resulted in a shift to the right of the dose-response curves for (--)-isoproterenol-induced positive chronotropic and inotropic responses and a reduction of the maximum degree of response that could be produced relative to that produced by theophylline. Carbachol only reduced the maximum responses to the partial agonist (--)-soterenol. The ED50 value for isoproterenol-induced increases in adenosine 3',5'-monophosphate (cyclic AMP) levels was more than 10-fold larger than those for production of mechanical events and soterenol did not produce a measurable increase in cyclic AMP above baseline values. Soterenol was analyzed as a competitive antagonist of isoproterenol-induced responses and the apparent dissociation constants (KA) for soterenol calculated to be 1.21 X 10(-7) M for receptors mediating positive chronotropic responses, 3.56 X 10(-7) M for receptors mediating positive inotropic responses, and 2.96 X 10(-7) M for receptors mediating increases in cyclic AMP levels. By two additional theoretical models of drug-receptor interactions, the KA values for soterenol were between 1.5 and 4.5 X 10(-7) M. These KA values were essentially the same as the ED50 values for soterenol-induced mechanical effects. By one of the models, the KA for isoproterenol was calculated to be about 2.14 X 10(-8) M, a value close to the ED50 value for isoproterenol-induced increases in cyclic AMP (5.45 X 10(-8) M). These results suggest that the beta-adrenoceptors mediating the three responses in rat atria are of the same type and that differences in concentrations required to produce mechanical vs. cyclic AMP changes results with agonists possessing high efficacy for which there is a receptor reserve. | [
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PMID:24541 | Cardiovascular responses to intraventricular adrenaline in spontaneous hypertensive rats. | The effects of intracerebroventricular (i.c.v.) injections of adrenaline on the blood pressure and heart rate of spontaneous hypertensive (SH) rats have been investigated. Adrenaline induced dose-related falls in blood pressure and heart rate in both conscious and urethane anaesthetised rats. In conscious rats, the hypotension and metoprolol, but were unaffected by pretreatment with phentolamine, piperoxan, fluphenazine or methysergide. However, in urethane-anaesthetised rats, the hypotension and bradycardia induced by i.c.v. adrenaline was not significantly affected by i.c.v. pretreatment with propranolol or oxprenolol, while metoprolol significantly antagonised only the bradycardia. Piperoxan, fluphenazine and methysergide were also without effect. Pretreatment with mecamylamine (i.p.) abolished the cardiovascular depressor effects of i.c.v. adrenaline in both conscious and urethane anaesthetised SH rats. It is concluded that the cardiovascular depressor effects of i.c.v. adrenaline are mediated by central adrenoceptors in SH rats and that, in conscious rats, these depressor effects may be mediated by central beta-adrenoceptors rather than alpha-adrenoceptors. | Cardiovascular responses to intraventricular adrenaline in spontaneous hypertensive rats. The effects of intracerebroventricular (i.c.v.) injections of adrenaline on the blood pressure and heart rate of spontaneous hypertensive (SH) rats have been investigated. Adrenaline induced dose-related falls in blood pressure and heart rate in both conscious and urethane anaesthetised rats. In conscious rats, the hypotension and metoprolol, but were unaffected by pretreatment with phentolamine, piperoxan, fluphenazine or methysergide. However, in urethane-anaesthetised rats, the hypotension and bradycardia induced by i.c.v. adrenaline was not significantly affected by i.c.v. pretreatment with propranolol or oxprenolol, while metoprolol significantly antagonised only the bradycardia. Piperoxan, fluphenazine and methysergide were also without effect. Pretreatment with mecamylamine (i.p.) abolished the cardiovascular depressor effects of i.c.v. adrenaline in both conscious and urethane anaesthetised SH rats. It is concluded that the cardiovascular depressor effects of i.c.v. adrenaline are mediated by central adrenoceptors in SH rats and that, in conscious rats, these depressor effects may be mediated by central beta-adrenoceptors rather than alpha-adrenoceptors. | [
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PMID:24542 | Beta-adrenoceptors modulate noradrenaline release from axonal sprouts in cultured rat superior cervical ganglia. | Superior cervical ganglia of rats grown in organ culture were used to study the effect of beta-receptor stimulants and antagonists on 3H-noradrenaline release in response to stimulation by KC1 (75 mM). (--)-Isoprenaline 1X 10(-9)--1 X 10(-7) M) increased 20--25% the release of 3H-noradrenaline from cultured ganglia exposed to KC1. Isoprenaline did not modify either the spontaneous (non-calcium dependent) release of 3H-noradrenaline from cultured ganglia, or the KC1-stimulated release from fresh ganglia. The effect of (--)-isoprenaline was blocked by (--)-propranolol 5 X 10(-9) -- 1 X 10(-8) M and by butoxamine 10(-6) M, but not by (+)-propranolol (1 -- 5 X 10(-8) M), practolol (1 X 10(-8) -- 1 X 10(-6) M), or sotalol (1 X 10(-7) -- 1 X 10(-6) M). Isoprenaline induced augmentation of 3H-noradrenaline release and its antagonism by (--)-propranolol still occurred in the presence of DMI. It is suggested that presynaptic beta-receptors in sympathetic nerve terminals may be involved in a positive feedback of noradrenaline release. | Beta-adrenoceptors modulate noradrenaline release from axonal sprouts in cultured rat superior cervical ganglia. Superior cervical ganglia of rats grown in organ culture were used to study the effect of beta-receptor stimulants and antagonists on 3H-noradrenaline release in response to stimulation by KC1 (75 mM). (--)-Isoprenaline 1X 10(-9)--1 X 10(-7) M) increased 20--25% the release of 3H-noradrenaline from cultured ganglia exposed to KC1. Isoprenaline did not modify either the spontaneous (non-calcium dependent) release of 3H-noradrenaline from cultured ganglia, or the KC1-stimulated release from fresh ganglia. The effect of (--)-isoprenaline was blocked by (--)-propranolol 5 X 10(-9) -- 1 X 10(-8) M and by butoxamine 10(-6) M, but not by (+)-propranolol (1 -- 5 X 10(-8) M), practolol (1 X 10(-8) -- 1 X 10(-6) M), or sotalol (1 X 10(-7) -- 1 X 10(-6) M). Isoprenaline induced augmentation of 3H-noradrenaline release and its antagonism by (--)-propranolol still occurred in the presence of DMI. It is suggested that presynaptic beta-receptors in sympathetic nerve terminals may be involved in a positive feedback of noradrenaline release. | [
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PMID:24543 | Evidence that the efficacy (intrinsic activity) of fenoterol is higher than that of salbutamol on beta-adrenoceptors in guinea-pig trachea. | The functional antagonism of carbachol by fenoterol and salbutamol (beta-adrenoceptor agonists) has been used to demonstrate that the efficacy (intrinsic activity) of fenoterol was about twice that of salbutamol on guinea-pig trachea. The mean maximum shifts of the carbachol concentration--response lines by fenoterol and salbutamol were (log units) 1.07 +/- 0.07 (n = 5) and 0.64 +/- 0.07 (n = 5) respectively. This difference in their efficacies could be demonstrated as differences in maximum relaxation on tracheal preparations contracted with carbachol, although this was dependent on the concentration of carbachol used. beta-Adrenoceptor blocking properties of salbutamol (1 mM) but not fenoterol (1 mM) could be demonstrated on trachea in that salbutamol, but not fenoterol, antagonised the shift in the carbachol concentration--response line produced by isoprenaline. The implications of these findings in relation to the use of fenoterol and salbutamol as bronchodilators is discussed. | Evidence that the efficacy (intrinsic activity) of fenoterol is higher than that of salbutamol on beta-adrenoceptors in guinea-pig trachea. The functional antagonism of carbachol by fenoterol and salbutamol (beta-adrenoceptor agonists) has been used to demonstrate that the efficacy (intrinsic activity) of fenoterol was about twice that of salbutamol on guinea-pig trachea. The mean maximum shifts of the carbachol concentration--response lines by fenoterol and salbutamol were (log units) 1.07 +/- 0.07 (n = 5) and 0.64 +/- 0.07 (n = 5) respectively. This difference in their efficacies could be demonstrated as differences in maximum relaxation on tracheal preparations contracted with carbachol, although this was dependent on the concentration of carbachol used. beta-Adrenoceptor blocking properties of salbutamol (1 mM) but not fenoterol (1 mM) could be demonstrated on trachea in that salbutamol, but not fenoterol, antagonised the shift in the carbachol concentration--response line produced by isoprenaline. The implications of these findings in relation to the use of fenoterol and salbutamol as bronchodilators is discussed. | [
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PMID:24544 | Antagonism by mianserin and classical alpha-adrenoceptor blocking drugs of some cardiovascular and behavioral effects of clonidine. | Antagonism of pressor responses to sympathetic outflow stimulation and alpha-adrenoceptor agonists in pithed spontaneously hypertensive rats was used to estimate postsynaptic alpha-adrenoceptor blocking activity of mianserin, phentolamine, phenoxybenzamine, piperoxan and yohimbine. Estimation of presynaptic alpha-adrenoceptor blocking activity of these drugs was obtained by studying their ability to antagonize clonidine-induced suppression of positive chronotropic responses to sympathetic outflow stimulation. In this manner, evidence was obtained that mianserin causes selective presynaptic alpha-adrenoceptor blockade. Mianserin, piperoxan and yohimbine antagonized clonidine-induced avoidance blockade or hypotension in spontaneously hypertensive rats, but methysergide, phenoxybenzamine and phentolamine were ineffective. These results suggest that mianserin may antagonize the central effects of clonidine by blockade of noradrenergic presynaptic or autoreceptors and possibly explain the antidepressant effect of mianserin as due to indirect activation of central noradrenergic neurons. | Antagonism by mianserin and classical alpha-adrenoceptor blocking drugs of some cardiovascular and behavioral effects of clonidine. Antagonism of pressor responses to sympathetic outflow stimulation and alpha-adrenoceptor agonists in pithed spontaneously hypertensive rats was used to estimate postsynaptic alpha-adrenoceptor blocking activity of mianserin, phentolamine, phenoxybenzamine, piperoxan and yohimbine. Estimation of presynaptic alpha-adrenoceptor blocking activity of these drugs was obtained by studying their ability to antagonize clonidine-induced suppression of positive chronotropic responses to sympathetic outflow stimulation. In this manner, evidence was obtained that mianserin causes selective presynaptic alpha-adrenoceptor blockade. Mianserin, piperoxan and yohimbine antagonized clonidine-induced avoidance blockade or hypotension in spontaneously hypertensive rats, but methysergide, phenoxybenzamine and phentolamine were ineffective. These results suggest that mianserin may antagonize the central effects of clonidine by blockade of noradrenergic presynaptic or autoreceptors and possibly explain the antidepressant effect of mianserin as due to indirect activation of central noradrenergic neurons. | [
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PMID:24562 | [A double adsorption technique for the determination of corticosteroid-binding globulin in human serum (author's transl)]. | Despite the widespread use of many different estrogenic compounds in hormonal contraception and estrogen replacement, in the human a simple test for comparing the potencies of the compounds is lacking. Many of the estrogenic compounds are difficult to measure, and animal studies may not relate directly to those done on humans in terms of potency. Corticosteroid-binding globulin (CBG) in human serum has been known since 1958 to increase in response to estrogen administration, and the degree of its elevation has been shown to be closely dose-related and reasonably sensitive. Therefore, CBG represents a biological parameter that provides for the specific quantal analysis of estrogenicity in the human. In this paper a simple and reliable method for the determination of the cortisol-binding capacity of CBG (CBG-BC) in human serum is described. The procedure is based on the amount of cortisol required to saturate the CBG binding sites. All endogenous steroids are initially removed from the serum by adsorption on dextran-coated charcoal (DCC). A mixture of a known amount of unlabeled and labeled cortisol is added to the sample in order to saturate the "stripped" binding sites of CBG. The unbound cortisol is removed by DCC at 4 degrees C, and the supernatant represents the total cortisol-binding capacity of the serum sample. To distinguish between cortisol-binding to CBG and to albumin, "stripped" serum is heated for 30 min at 60 degrees C, the temperature which does not influence cortisol binding by albumin but which does inactivate CBG. The samples are subsequently equilibrated with a mixture of labeled and unlabeled cortisol. The free cortisol is removed by DCC and the bound cortisol remaining in the supernatant is designated as the albumin-bound cortisol. Thus, CBG-BC is assayed by a double adsorption technique as differential binding to native and heat-inactivated serum. The cortisol determined by this method corresponds to CBG-bound cortisol, and CBG-BC is expressed as micrograms of cortisol bound per 100 ml serum. The precision and accuracy of the method were found to be good. The values of CBG-BC obtained by this method were 14.9 +/- 1.9 (SD) microgram/100ml in adult men and 16.0 +/- 1.8 microgram/100ml in adult women. Daily serum samples in five women revealed no significant variations in CBG-BC throughout the normal menstrual cycle despite fluctuating levels of serum estradiol. CBG-BC was also found not to vary significantly with age. In pregnancy, however CBG-BC increased significantly. | [A double adsorption technique for the determination of corticosteroid-binding globulin in human serum (author's transl)]. Despite the widespread use of many different estrogenic compounds in hormonal contraception and estrogen replacement, in the human a simple test for comparing the potencies of the compounds is lacking. Many of the estrogenic compounds are difficult to measure, and animal studies may not relate directly to those done on humans in terms of potency. Corticosteroid-binding globulin (CBG) in human serum has been known since 1958 to increase in response to estrogen administration, and the degree of its elevation has been shown to be closely dose-related and reasonably sensitive. Therefore, CBG represents a biological parameter that provides for the specific quantal analysis of estrogenicity in the human. In this paper a simple and reliable method for the determination of the cortisol-binding capacity of CBG (CBG-BC) in human serum is described. The procedure is based on the amount of cortisol required to saturate the CBG binding sites. All endogenous steroids are initially removed from the serum by adsorption on dextran-coated charcoal (DCC). A mixture of a known amount of unlabeled and labeled cortisol is added to the sample in order to saturate the "stripped" binding sites of CBG. The unbound cortisol is removed by DCC at 4 degrees C, and the supernatant represents the total cortisol-binding capacity of the serum sample. To distinguish between cortisol-binding to CBG and to albumin, "stripped" serum is heated for 30 min at 60 degrees C, the temperature which does not influence cortisol binding by albumin but which does inactivate CBG. The samples are subsequently equilibrated with a mixture of labeled and unlabeled cortisol. The free cortisol is removed by DCC and the bound cortisol remaining in the supernatant is designated as the albumin-bound cortisol. Thus, CBG-BC is assayed by a double adsorption technique as differential binding to native and heat-inactivated serum. The cortisol determined by this method corresponds to CBG-bound cortisol, and CBG-BC is expressed as micrograms of cortisol bound per 100 ml serum. The precision and accuracy of the method were found to be good. The values of CBG-BC obtained by this method were 14.9 +/- 1.9 (SD) microgram/100ml in adult men and 16.0 +/- 1.8 microgram/100ml in adult women. Daily serum samples in five women revealed no significant variations in CBG-BC throughout the normal menstrual cycle despite fluctuating levels of serum estradiol. CBG-BC was also found not to vary significantly with age. In pregnancy, however CBG-BC increased significantly. | [
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PMID:24563 | The protective effect of ketotifen in bronchial asthma. | Thirty-six patients with confirmed allergic bronchial asthma were divided into 3 parallel groups and treated with ketotifen 1 mg b.i.d. (Group I), ketotifen 2 mg b.i.d. (Group II), and clemastine 1 mg b.i.d. (Group III) respectively for 6 months. Nine out of 12 patients in Group I and 10 out of 12 in Group II experienced a statistically significant improvement in dyspnoea and in pulmonary function (Tiffeneau test). Likewise there was a marked reduction in the number, severity and average duration (minutes per week) of asthmatic attacks. By the 12th week, 8 out of 12 patients in the clemastine group had dropped out because of inefficacy. Ketotifen was very well tolerated by all the patients. In both ketotifen groups pathological eosinophil counts returned to normal during treatment. It can be concluded that ketotifen is effective in the long-term prophylaxis of bronchial asthma. | The protective effect of ketotifen in bronchial asthma. Thirty-six patients with confirmed allergic bronchial asthma were divided into 3 parallel groups and treated with ketotifen 1 mg b.i.d. (Group I), ketotifen 2 mg b.i.d. (Group II), and clemastine 1 mg b.i.d. (Group III) respectively for 6 months. Nine out of 12 patients in Group I and 10 out of 12 in Group II experienced a statistically significant improvement in dyspnoea and in pulmonary function (Tiffeneau test). Likewise there was a marked reduction in the number, severity and average duration (minutes per week) of asthmatic attacks. By the 12th week, 8 out of 12 patients in the clemastine group had dropped out because of inefficacy. Ketotifen was very well tolerated by all the patients. In both ketotifen groups pathological eosinophil counts returned to normal during treatment. It can be concluded that ketotifen is effective in the long-term prophylaxis of bronchial asthma. | [
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PMID:24570 | Effect of lactulose on ammonia production in a fecal incubation system. | An in vitro fecal incubation system was used to demonstrate how lactulose influences ammonia metabolism in the colon. Lactulose and other fermentable substrates (glucose, mannitol, and sorbitol), pH and organic acid were varied independently so that their different effects could be determined. Fermentable substrate caused a fall in ammonia concentration during the period of fermentation. Acidification to pH 5.0 or less, with hydrochloric acid or a lactic-acetic acid mixture, significantly reduced ammonia generation, but unlike fermentable substrates, did not lower the existing ammonia concentration. The lactic-acetic acid mixture did not reduce ammonia generation significantly below that found with acidification by hydrochloric acid. The effect of lactulose in reducing ammonia concentration is attributed to its role as a bacterial substrate in either increasing bacterial assimilation of ammonia or reducing deamination of nitrogenous compounds. The effect of low pH in reducing generation of ammonia appears to be part of a general reduction in bacterial metabolism. | Effect of lactulose on ammonia production in a fecal incubation system. An in vitro fecal incubation system was used to demonstrate how lactulose influences ammonia metabolism in the colon. Lactulose and other fermentable substrates (glucose, mannitol, and sorbitol), pH and organic acid were varied independently so that their different effects could be determined. Fermentable substrate caused a fall in ammonia concentration during the period of fermentation. Acidification to pH 5.0 or less, with hydrochloric acid or a lactic-acetic acid mixture, significantly reduced ammonia generation, but unlike fermentable substrates, did not lower the existing ammonia concentration. The lactic-acetic acid mixture did not reduce ammonia generation significantly below that found with acidification by hydrochloric acid. The effect of lactulose in reducing ammonia concentration is attributed to its role as a bacterial substrate in either increasing bacterial assimilation of ammonia or reducing deamination of nitrogenous compounds. The effect of low pH in reducing generation of ammonia appears to be part of a general reduction in bacterial metabolism. | [
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PMID:24576 | How thyroid disease presents in the elderly. | Some of the symptoms and signs of hypothyroidism and hyperthyroidism in elderly patients may be mistakenly attributed to "old age." Weight loss, muscle weakness, tremor, angina, congestive heart failure--all signs of hyperthyroidism--are also concomitants of aging. Fatigue, sluggishness, withdrawal behavior, senile atrophic skin changes--all signs of hypothroidism--are also a part of the normal aging process. Although screening elderly people for thyroid disease is economically unsound, the physician should maintain a high index of suspicion of its presence. Laboratory tests must be interpreted with extra care. Values of 131I uptake, serum T4 and T3, thyroid-stimulating hormone, and thyrotropin-releasing hormone are all helpful in diagnosis. Thyroid disease is easily treated in elderly patients, and results often are dramatic. Propranolol is effective in thyrotoxic patients when symptoms require prompt relief. The definitive treatment, however, is 131I; antithyroid drugs are difficult to manage. Hypothyroidism is easily treated with T4. | How thyroid disease presents in the elderly. Some of the symptoms and signs of hypothyroidism and hyperthyroidism in elderly patients may be mistakenly attributed to "old age." Weight loss, muscle weakness, tremor, angina, congestive heart failure--all signs of hyperthyroidism--are also concomitants of aging. Fatigue, sluggishness, withdrawal behavior, senile atrophic skin changes--all signs of hypothroidism--are also a part of the normal aging process. Although screening elderly people for thyroid disease is economically unsound, the physician should maintain a high index of suspicion of its presence. Laboratory tests must be interpreted with extra care. Values of 131I uptake, serum T4 and T3, thyroid-stimulating hormone, and thyrotropin-releasing hormone are all helpful in diagnosis. Thyroid disease is easily treated in elderly patients, and results often are dramatic. Propranolol is effective in thyrotoxic patients when symptoms require prompt relief. The definitive treatment, however, is 131I; antithyroid drugs are difficult to manage. Hypothyroidism is easily treated with T4. | [
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PMID:24578 | [Cardiomyopathies]. | A) Definition and classification. Cardiomyopathy is defined as a dysfunction of cardiac muscle of unknown origin and classified according to genetic, morphological and functional criteria as follows: 1. cardiomyopathy of autosomal dominant inheritance with asymmetric septal hypertrophy (ASH) a) obstructive b) non obstructive 2. cardiomyopathy of autosomal dominant inheritance without ASH 3. cardiomyopathy of autosomal recessive inheritance 4. sporadic cardiomyopathy. It is assumed that at least the cardiomyopathies listed here are heterogeneous groups that have to be subdivided as soon as further discriminating findings--structural or enzymatic--are available. B) Diagnosis, prognosis, treatment. The diagnostic methods are evaluated, the prognosis and the treatment of the cardiomyopathies are reviewed. | [Cardiomyopathies]. A) Definition and classification. Cardiomyopathy is defined as a dysfunction of cardiac muscle of unknown origin and classified according to genetic, morphological and functional criteria as follows: 1. cardiomyopathy of autosomal dominant inheritance with asymmetric septal hypertrophy (ASH) a) obstructive b) non obstructive 2. cardiomyopathy of autosomal dominant inheritance without ASH 3. cardiomyopathy of autosomal recessive inheritance 4. sporadic cardiomyopathy. It is assumed that at least the cardiomyopathies listed here are heterogeneous groups that have to be subdivided as soon as further discriminating findings--structural or enzymatic--are available. B) Diagnosis, prognosis, treatment. The diagnostic methods are evaluated, the prognosis and the treatment of the cardiomyopathies are reviewed. | [
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PMID:24577 | Forensic considerations on the comparison of "serum gamma-glutamyltranspeptidase" ("gamma-GT) activity in experimental acute alcoholic intoxication and in alcoholic car drivers who caused road accidents. | The authors studied blood alcohol levels and serum gamma-GT activity in 18 drivers who caused car accidents while intoxicated, and in 20 young volunteer subjects without any hepatic damage, in which an acute alcoholic intoxication was present. In most of the drivers the serum gamma-GT activity was significantly higher than in the volunteers, which strongly suggests a chronic alcoholic intoxication. In the 20 volunteers, who were occasional drinkers, the serum gamma-GT activity did not show pathological values even in a state of heavy intoxication. According to the authors, the serum gamma-GT determination could be made obligatory by law, in order to differentiate between acute or chronic alcoholic intoxication. Should an intoxicated driver be found guilty of an offense, this fact could aggravate the circumstances. Furthermore serum gamma-GT activity determination could be used as an evaluating parameter in granting driving licences. | Forensic considerations on the comparison of "serum gamma-glutamyltranspeptidase" ("gamma-GT) activity in experimental acute alcoholic intoxication and in alcoholic car drivers who caused road accidents. The authors studied blood alcohol levels and serum gamma-GT activity in 18 drivers who caused car accidents while intoxicated, and in 20 young volunteer subjects without any hepatic damage, in which an acute alcoholic intoxication was present. In most of the drivers the serum gamma-GT activity was significantly higher than in the volunteers, which strongly suggests a chronic alcoholic intoxication. In the 20 volunteers, who were occasional drinkers, the serum gamma-GT activity did not show pathological values even in a state of heavy intoxication. According to the authors, the serum gamma-GT determination could be made obligatory by law, in order to differentiate between acute or chronic alcoholic intoxication. Should an intoxicated driver be found guilty of an offense, this fact could aggravate the circumstances. Furthermore serum gamma-GT activity determination could be used as an evaluating parameter in granting driving licences. | [
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PMID:24583 | Modification of the immunologic properties of the cell surface. | Living CF1 mouse-transplantable spontaneous mammary adenocarcinoma cells were modified with glutaraldehyde, formalin, 2,4,6-trinitrophenylate, Vibrio cholerae neurominidase, iodoacetate, heparin, histamine and adenosine 3'5'-monophosphate (cAMP), then used to immunize syngeneic CF1 mice. Animals immunized with the fixed (formalinized or glutaraldehyde fixed) neuraminidase-treated cells or the membrane of these cells, rejected two challenging doses of 10(8) viable unmodified adenocarcinoma cells. Animals immunized with adenocarcinoma cells treated with neuraminidase (170 IU/5x10(5) cells) or with the spontaneous adenocarcinoma-cell surface glycoprotein, rejected the first challenging dose but developed tumors and died on the second challenge with the viable untreated adenocarcinoma cells. Animals immunized with the adenocarcinoma cells pretreated with trinitrophenylate, glutaraldehyde or formalin, developed temporary resistance to the spontaneous mammary adenocarcinoma. Adenocarcinoma cells pretreated with NaF, iodoacetate, heparin, EDTA, Colchicine or histamine showed reduced oncogenicity and stronger resistance in mice to the development of a mammary tumor than to a smaller number (10(3) AdCa cells) of untreated viable adenocarcinoma cells. Cells treated with adenosine 3'5'-monophosphate accelerated tumor development. | Modification of the immunologic properties of the cell surface. Living CF1 mouse-transplantable spontaneous mammary adenocarcinoma cells were modified with glutaraldehyde, formalin, 2,4,6-trinitrophenylate, Vibrio cholerae neurominidase, iodoacetate, heparin, histamine and adenosine 3'5'-monophosphate (cAMP), then used to immunize syngeneic CF1 mice. Animals immunized with the fixed (formalinized or glutaraldehyde fixed) neuraminidase-treated cells or the membrane of these cells, rejected two challenging doses of 10(8) viable unmodified adenocarcinoma cells. Animals immunized with adenocarcinoma cells treated with neuraminidase (170 IU/5x10(5) cells) or with the spontaneous adenocarcinoma-cell surface glycoprotein, rejected the first challenging dose but developed tumors and died on the second challenge with the viable untreated adenocarcinoma cells. Animals immunized with the adenocarcinoma cells pretreated with trinitrophenylate, glutaraldehyde or formalin, developed temporary resistance to the spontaneous mammary adenocarcinoma. Adenocarcinoma cells pretreated with NaF, iodoacetate, heparin, EDTA, Colchicine or histamine showed reduced oncogenicity and stronger resistance in mice to the development of a mammary tumor than to a smaller number (10(3) AdCa cells) of untreated viable adenocarcinoma cells. Cells treated with adenosine 3'5'-monophosphate accelerated tumor development. | [
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PMID:24584 | The persistent PHA-responsive population in the mouse thymus. II. Recirculatory characteristics and immunological properties. | Experiments were performed to ascertain whether the persistent PHA-responsive cell population found in mouse thymus grafts was capable both of localizing to peripheral lymphoid organs when deliberately injected into suitable recipient mice, and of responding to antigenic stimulation. It was shown that the persistent population could not be found in large numbers in recipient spleens or lymph nodes at late time after transfer. These cells could, however, be found in large numbers in the peripheral blood of recipients. Thus they appeared to be able to circulate in the blood but not be able to extravascularize in lymphoid organs. Since these cells did not localize to spleen or lymph nodes, it was not surprising to find that they did not respond to the skin sensitizing agent oxazolone in draining lymph nodes or spleen, or to histocompatibility antigens (graft-vs-host). When tested against allogeneic cells in vitro (mixed lymphocyte reaction), it was shown that the persistent population was over nine times less reactive than in its reactivity to PHA, when compared to the non-persistent population. The functional significance of these thymocyte populations is discussed. | The persistent PHA-responsive population in the mouse thymus. II. Recirculatory characteristics and immunological properties. Experiments were performed to ascertain whether the persistent PHA-responsive cell population found in mouse thymus grafts was capable both of localizing to peripheral lymphoid organs when deliberately injected into suitable recipient mice, and of responding to antigenic stimulation. It was shown that the persistent population could not be found in large numbers in recipient spleens or lymph nodes at late time after transfer. These cells could, however, be found in large numbers in the peripheral blood of recipients. Thus they appeared to be able to circulate in the blood but not be able to extravascularize in lymphoid organs. Since these cells did not localize to spleen or lymph nodes, it was not surprising to find that they did not respond to the skin sensitizing agent oxazolone in draining lymph nodes or spleen, or to histocompatibility antigens (graft-vs-host). When tested against allogeneic cells in vitro (mixed lymphocyte reaction), it was shown that the persistent population was over nine times less reactive than in its reactivity to PHA, when compared to the non-persistent population. The functional significance of these thymocyte populations is discussed. | [
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PMID:24585 | Ther persistent PHA-responsive population in the mouse thymus. II. Recirculatory characteristics and immunological properties. | Experiments were performed to ascertain whether the persistent PHA-responsive cell population found in mouse thymus grafts was capable both of localizing to peripheral lymphoid organs when deliverately injected into suitable recipient mice, and of resonding to antigenic stimulation. It was shown that the persistent population could not be found in large numbers in recipient spleens or lymph nodes at late time after transfer. These cells could, however, be found in large numbers in the peripheral blood of recipients. Thus they appeared to be able to circulate in the blood but not able to extravascularize in lymphoid organs. Since these cells did not localize to spleen or lymph nodes, it was not surprising to find that they did not response to the skin sensitizing agent oxazolone in draining lymph nodes or spleen, or to histocompatibility antigens (graft-vs-host). When tested against allogeneic cells in vitro (mixed lymphocyte reaction), it was shown that the persistent population was over nine times less reactive than in its reactivity to PHA, when compared to the non-persistent population. The functional significance of these thymocyte populations is discussed. | Ther persistent PHA-responsive population in the mouse thymus. II. Recirculatory characteristics and immunological properties. Experiments were performed to ascertain whether the persistent PHA-responsive cell population found in mouse thymus grafts was capable both of localizing to peripheral lymphoid organs when deliverately injected into suitable recipient mice, and of resonding to antigenic stimulation. It was shown that the persistent population could not be found in large numbers in recipient spleens or lymph nodes at late time after transfer. These cells could, however, be found in large numbers in the peripheral blood of recipients. Thus they appeared to be able to circulate in the blood but not able to extravascularize in lymphoid organs. Since these cells did not localize to spleen or lymph nodes, it was not surprising to find that they did not response to the skin sensitizing agent oxazolone in draining lymph nodes or spleen, or to histocompatibility antigens (graft-vs-host). When tested against allogeneic cells in vitro (mixed lymphocyte reaction), it was shown that the persistent population was over nine times less reactive than in its reactivity to PHA, when compared to the non-persistent population. The functional significance of these thymocyte populations is discussed. | [
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PMID:24586 | Cell transplantation into immunodeficient chicken embryos. Reconstituting capacity of cells from the yolk sac at different stages of development and from the liver, thymus, bursa of Fabricius, spleen and bone marrow of 15-day embryos. | Cyclophosphamide-treated 18-day-old chicken embryos were transplanted with histocompatible cells from the yolk sac at different stages of development and from the liver, thymus, bursa of Fabricius, spleen and bone marrow of 15-day-old-embryos. At the age of 36 days, the cell recipients were studied to determine the reconstitution capacity of the transplanted cells. The parameters used include the survival pattern, gain of body weight, antibody-forming capacity, response of peripheral blood lymphocytes to Con A, weight and microscopic morphology of the bursa of Fabricius, and weight of spleen and thymus. By all the criteria employed, only bursa cells were capable of a functional and morphological reconstitution of the recipient's humoral immune system. These data indicate that the role of the yolk sac as the first generator of prebursal stem cells remains questionable. In addition, these findings confirm the previous observations that, as a differentiation site of the B-cell lineage, the bursa of Fabricius precedes the bone marrow during ontogenetic development. | Cell transplantation into immunodeficient chicken embryos. Reconstituting capacity of cells from the yolk sac at different stages of development and from the liver, thymus, bursa of Fabricius, spleen and bone marrow of 15-day embryos. Cyclophosphamide-treated 18-day-old chicken embryos were transplanted with histocompatible cells from the yolk sac at different stages of development and from the liver, thymus, bursa of Fabricius, spleen and bone marrow of 15-day-old-embryos. At the age of 36 days, the cell recipients were studied to determine the reconstitution capacity of the transplanted cells. The parameters used include the survival pattern, gain of body weight, antibody-forming capacity, response of peripheral blood lymphocytes to Con A, weight and microscopic morphology of the bursa of Fabricius, and weight of spleen and thymus. By all the criteria employed, only bursa cells were capable of a functional and morphological reconstitution of the recipient's humoral immune system. These data indicate that the role of the yolk sac as the first generator of prebursal stem cells remains questionable. In addition, these findings confirm the previous observations that, as a differentiation site of the B-cell lineage, the bursa of Fabricius precedes the bone marrow during ontogenetic development. | [
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PMID:24590 | Effects of fluoride on carbohydrate metabolism by washed cells of Streptococcus mutans grown at various pH values in a chemostat. | Streptococcus mutans Ingbritt was grown anaerobically in a chemostat, at a rate (mean generation time, 13 h) similar to that in dental plaque, in a complex medium with excess glucose and at pH values of 6.5, 6.0, and 5.5. The yield of cells was constant at pH 6.5 and 6.0 (2.00 mg/ml) but fell to 1.25 at pH 5.5; Y(glucose) was relatively constant under all conditions. Lactic acid was the major end product. Amino acid analysis of the culture supernatants indicated that growth was probably limited by the availability of cysteine. Cells were harvested and monitored for their capacity to produce acid from endogenous polysaccharide and exogenous sugars in the presence and absence of NaF, as well as for their glucose phosphoenolpyruvate (PEP)-phosphotransferase activity. Surprisingly, cells grown at pH 5.5 possessed two to three times more glycolytic activity, as measured by the rate of acid production, than cells grown at pH 6.5 and 6.0 when incubated in a washed suspension at constant pH with a sugar source. Furthermore, the cells grown at pH 5.5 were about twice as resistant to the effect of NaF in reducing the rate of acid production in this system. Fluoride inhibition could be reversed by increasing the pH of the system. Cells grown at all three pH values showed significant acid production from endogenous reserves, despite the fact that the glucoamylase-specific glycogen content of the cells dropped from 33% of the total carbohydrate during pH 6.5 growth to only 3% after growth at pH 6.0 and 6.5. Incubation of washed cells for 18 h in phosphate buffer resulted in the loss of 62% of the total carbohydrate, indicating that nonglycogen cellular polysaccharide was metabolized. A comparison of the fluoride effect on endogenous and exogenous metabolism under pH fall conditions showed that, with pH 6.5- and 6.0-grown cells, the inhibitor was more effective in the presence of an exogenous carbon source than in its absence. This effect was not seen with pH 5.5-grown cells. The decreased sensitivity of the pH 5.5-grown cells to fluoride was probably associated with the decreased glucose PEP-phosphotransferase activity (11%) in these cells compared with the activity of those grown at pH 6.5. This evidence supports the hypothesis that S. mutans possesses at least two glucose transport systems, one of which is relatively fluoride insensitive. | Effects of fluoride on carbohydrate metabolism by washed cells of Streptococcus mutans grown at various pH values in a chemostat. Streptococcus mutans Ingbritt was grown anaerobically in a chemostat, at a rate (mean generation time, 13 h) similar to that in dental plaque, in a complex medium with excess glucose and at pH values of 6.5, 6.0, and 5.5. The yield of cells was constant at pH 6.5 and 6.0 (2.00 mg/ml) but fell to 1.25 at pH 5.5; Y(glucose) was relatively constant under all conditions. Lactic acid was the major end product. Amino acid analysis of the culture supernatants indicated that growth was probably limited by the availability of cysteine. Cells were harvested and monitored for their capacity to produce acid from endogenous polysaccharide and exogenous sugars in the presence and absence of NaF, as well as for their glucose phosphoenolpyruvate (PEP)-phosphotransferase activity. Surprisingly, cells grown at pH 5.5 possessed two to three times more glycolytic activity, as measured by the rate of acid production, than cells grown at pH 6.5 and 6.0 when incubated in a washed suspension at constant pH with a sugar source. Furthermore, the cells grown at pH 5.5 were about twice as resistant to the effect of NaF in reducing the rate of acid production in this system. Fluoride inhibition could be reversed by increasing the pH of the system. Cells grown at all three pH values showed significant acid production from endogenous reserves, despite the fact that the glucoamylase-specific glycogen content of the cells dropped from 33% of the total carbohydrate during pH 6.5 growth to only 3% after growth at pH 6.0 and 6.5. Incubation of washed cells for 18 h in phosphate buffer resulted in the loss of 62% of the total carbohydrate, indicating that nonglycogen cellular polysaccharide was metabolized. A comparison of the fluoride effect on endogenous and exogenous metabolism under pH fall conditions showed that, with pH 6.5- and 6.0-grown cells, the inhibitor was more effective in the presence of an exogenous carbon source than in its absence. This effect was not seen with pH 5.5-grown cells. The decreased sensitivity of the pH 5.5-grown cells to fluoride was probably associated with the decreased glucose PEP-phosphotransferase activity (11%) in these cells compared with the activity of those grown at pH 6.5. This evidence supports the hypothesis that S. mutans possesses at least two glucose transport systems, one of which is relatively fluoride insensitive. | [
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PMID:24591 | Observation of beta-hemolysis among three strains of Streptococcus mutans. | Streptococcus mutans is normally alpha- or gamma-hemolytic on blood agar plates. However, three recently isolated S. mutans strains were observed to elicit beta-hemolysis. The production and nature of a hemolytic substance were studied. | Observation of beta-hemolysis among three strains of Streptococcus mutans. Streptococcus mutans is normally alpha- or gamma-hemolytic on blood agar plates. However, three recently isolated S. mutans strains were observed to elicit beta-hemolysis. The production and nature of a hemolytic substance were studied. | [
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PMID:24592 | Splenic foci of cytotoxic lymphocytes in a graft-versus-host reaction. | It has been established that cytotoxic lymphocytes are generated in discrete foci in the spleens of mice undergoing a graft-versus-host reaction. The foci segregated randomly in such spleens. 2.8 X 10(4) lymph node cells, 2.3 X 10(5) spleen cells and 2.7 X 10(6) thymus cells from CBA mice generated a mean of one focus in lethally irradiated (CBA X DBA)F1 recipient mice. Cytotoxic foci were detected when cytotoxicity was measured by two different cytotoxicity assays, the release of [51Cr]-sodium chromate from [51Cr]-labelled target cells and colony inhibition. The seeding efficiency of cytotoxic focus-forming cells has been estimated at 19%. | Splenic foci of cytotoxic lymphocytes in a graft-versus-host reaction. It has been established that cytotoxic lymphocytes are generated in discrete foci in the spleens of mice undergoing a graft-versus-host reaction. The foci segregated randomly in such spleens. 2.8 X 10(4) lymph node cells, 2.3 X 10(5) spleen cells and 2.7 X 10(6) thymus cells from CBA mice generated a mean of one focus in lethally irradiated (CBA X DBA)F1 recipient mice. Cytotoxic foci were detected when cytotoxicity was measured by two different cytotoxicity assays, the release of [51Cr]-sodium chromate from [51Cr]-labelled target cells and colony inhibition. The seeding efficiency of cytotoxic focus-forming cells has been estimated at 19%. | [
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PMID:24595 | Decrease of peripheral resistance after acute intravenous application of a new beta-receptor blocking agents, bufuralol HCl. | The hemodynamic effects of a new beta-receptor blocking agent, bufuraolo HCl, were compared with those of pindolol. Contrary to pindolol, bufuralol HCl induces a decrease of the peripheral resistance immediately. Under exercise conditions, the peripheral resistance increases under pindolol whereas it remains constant under bufuralol HCl in spite of reduced cardiac output. As an explanation, an effect of bufuralol HCl on peripheral resistance is discussed which might be independent of the beta-receptor blocking effects in the periphery. | Decrease of peripheral resistance after acute intravenous application of a new beta-receptor blocking agents, bufuralol HCl. The hemodynamic effects of a new beta-receptor blocking agent, bufuraolo HCl, were compared with those of pindolol. Contrary to pindolol, bufuralol HCl induces a decrease of the peripheral resistance immediately. Under exercise conditions, the peripheral resistance increases under pindolol whereas it remains constant under bufuralol HCl in spite of reduced cardiac output. As an explanation, an effect of bufuralol HCl on peripheral resistance is discussed which might be independent of the beta-receptor blocking effects in the periphery. | [
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PMID:24596 | Cefazolin in the treatment of pneumonia. | Cefazolin is a semi-synthetic derivative of cephalosporin C that has a lower cross-immunogenicity with penicillins than do the other cephalosporins. This agent was evaluated as an alternative to penicillin in the therapy of patients with pneumococcal pneumonia. Thirty patient were treated with cefazolin, most receiving 125 or 250 mg IM every 12 hours for 5-10 days. Satisfactory clinical responses were obtained in 29 of these 30 patients, and none complained of pain following IM injections. Three patients developed eosinophilia while receiving cefazolin, and one of these also had a maculopapular eruption that may have been an allergic reaction to cefazolin. Serum levels of cefazolin were measured at 1, 6, and 12 hours after administration. Susceptibilities of 100 isolates of Streptococcus pneumoniae, including these patients' organisms, were determined by broth dilution. Both cefazolin and cephalothin were bactericidal for all 100 isolates at concentrations of 2 microgram/ml or less. Cefazolin appears to be an entirely adequate alternative to penicillin for the therapy of pneumococcalpneumonia. This agent is effective in low dosages, and adequate serum levels are maintained for long periods of time, permitting twice-daily administration. | Cefazolin in the treatment of pneumonia. Cefazolin is a semi-synthetic derivative of cephalosporin C that has a lower cross-immunogenicity with penicillins than do the other cephalosporins. This agent was evaluated as an alternative to penicillin in the therapy of patients with pneumococcal pneumonia. Thirty patient were treated with cefazolin, most receiving 125 or 250 mg IM every 12 hours for 5-10 days. Satisfactory clinical responses were obtained in 29 of these 30 patients, and none complained of pain following IM injections. Three patients developed eosinophilia while receiving cefazolin, and one of these also had a maculopapular eruption that may have been an allergic reaction to cefazolin. Serum levels of cefazolin were measured at 1, 6, and 12 hours after administration. Susceptibilities of 100 isolates of Streptococcus pneumoniae, including these patients' organisms, were determined by broth dilution. Both cefazolin and cephalothin were bactericidal for all 100 isolates at concentrations of 2 microgram/ml or less. Cefazolin appears to be an entirely adequate alternative to penicillin for the therapy of pneumococcalpneumonia. This agent is effective in low dosages, and adequate serum levels are maintained for long periods of time, permitting twice-daily administration. | [
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PMID:24597 | Comparative potency of intravenous penbutolol and propranolol in man. | A study on the comparative potency of intravenously administered penbutolol, a new non-selective beta adrenoceptor antagonist, on the resting and post-exercise heart rate and rate-pressure product was carried out in 5 normal human subjects. The effects produced by 0.1, 0.2 and 0.3 mg of penbutolol and of 0.025 mg/kg (mean dose 1.28 mg) of propranolol was recorded. The dose of penbutolol producing an effect equivalent to that of propranolol was calculating from the log dose-response curve of penbutolol. In these tests, penbutolol was 7.90 and 7.66 times more potent than propranolol on a weight to weight basis. Penbutolol was well tolerated and no side-effects were observed. | Comparative potency of intravenous penbutolol and propranolol in man. A study on the comparative potency of intravenously administered penbutolol, a new non-selective beta adrenoceptor antagonist, on the resting and post-exercise heart rate and rate-pressure product was carried out in 5 normal human subjects. The effects produced by 0.1, 0.2 and 0.3 mg of penbutolol and of 0.025 mg/kg (mean dose 1.28 mg) of propranolol was recorded. The dose of penbutolol producing an effect equivalent to that of propranolol was calculating from the log dose-response curve of penbutolol. In these tests, penbutolol was 7.90 and 7.66 times more potent than propranolol on a weight to weight basis. Penbutolol was well tolerated and no side-effects were observed. | [
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PMID:24598 | Scanning electron microscopy of human female reproductive tract and amniotic fluid cells. | Scanning electron microscopy was used to examine surface ultrastructural characteristics of cells of the epithelium of female reproductive tract, cervical mucus, and amniotic fluid cells. The female epithelium undergoes hormone-dependent cyclical morphological alterations in cell shape, apical microvilli, ciliation, and secretory patterns. The frequency distribution of ciliated cells varies in different parts of the female reproductive tract and different segments of the same organ. In the endocervix, a few ciliated cells are randomly distributed. The density of ciliated cells is variable in several areas of the endometrium. The sparse distribution of ciliated cells in the central portion of the uterine wall may indicate a possible implantation site. According to surface ultrastructure of cells, three segments are noted in the uterotubal junction: cornual endometrium, interstitial tubal epithelium, and transitional area. The vaginal epithelium is made of flat polygonal cells with interdigitating borders and a fine interlacing network of microridges. After the menopause, the microridge pattern and distinct cell borders are lost or severely modified. Two types of fibrous structures can be found in cervical mucus: a) microfibrils that vary in diameter from 500 to 1,500 A and from bundles or networks, and b) long, thick fibers that vary in diameter from 0.5 to 5 micrometer run parallel to each other, and appear to be made up of microfibrils. Amniotic fluid cells, derived from amniotic epithelium and fetal ectoderm, consist of non-nucleated and nucleated cells with extensive microplicae. | Scanning electron microscopy of human female reproductive tract and amniotic fluid cells. Scanning electron microscopy was used to examine surface ultrastructural characteristics of cells of the epithelium of female reproductive tract, cervical mucus, and amniotic fluid cells. The female epithelium undergoes hormone-dependent cyclical morphological alterations in cell shape, apical microvilli, ciliation, and secretory patterns. The frequency distribution of ciliated cells varies in different parts of the female reproductive tract and different segments of the same organ. In the endocervix, a few ciliated cells are randomly distributed. The density of ciliated cells is variable in several areas of the endometrium. The sparse distribution of ciliated cells in the central portion of the uterine wall may indicate a possible implantation site. According to surface ultrastructure of cells, three segments are noted in the uterotubal junction: cornual endometrium, interstitial tubal epithelium, and transitional area. The vaginal epithelium is made of flat polygonal cells with interdigitating borders and a fine interlacing network of microridges. After the menopause, the microridge pattern and distinct cell borders are lost or severely modified. Two types of fibrous structures can be found in cervical mucus: a) microfibrils that vary in diameter from 500 to 1,500 A and from bundles or networks, and b) long, thick fibers that vary in diameter from 0.5 to 5 micrometer run parallel to each other, and appear to be made up of microfibrils. Amniotic fluid cells, derived from amniotic epithelium and fetal ectoderm, consist of non-nucleated and nucleated cells with extensive microplicae. | [
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PMID:24599 | Variation in human semen viscoelastic properties with respect to time post ejaculation and frequency of ejaculation. | The present study concerns the rheological characterization of the coagulation-liquefaction process of human semen. Results obtained using a multiple point capillary viscometer reveal marked variations in the elasticity and viscosity of an individual's semen with time immediately following ejaculation and frequency of ejaculation. Similarity among all cases examined for each material property is revealed by relating times post ejaculation to semen liquefaction time, thereby coupling liquefaction time with specific material property values. Further, the final state of liquefied semen is found to be characterized by Newtonian behavior (mean absolute viscosity = 3.37 centipoise). The semen's liquefaction time and ejaculate volume are determined to be functions of the frequency of ejaculation. Steady state ejaculate volume is found to decrease linearly with increasing ejaculation frequency, thereby providing a measure of glandular secretory rate. When collectively considered, these findings provide possible means for monitoring an individual's glandular behavior over an extended period of time and comparing such behavior to established standards. | Variation in human semen viscoelastic properties with respect to time post ejaculation and frequency of ejaculation. The present study concerns the rheological characterization of the coagulation-liquefaction process of human semen. Results obtained using a multiple point capillary viscometer reveal marked variations in the elasticity and viscosity of an individual's semen with time immediately following ejaculation and frequency of ejaculation. Similarity among all cases examined for each material property is revealed by relating times post ejaculation to semen liquefaction time, thereby coupling liquefaction time with specific material property values. Further, the final state of liquefied semen is found to be characterized by Newtonian behavior (mean absolute viscosity = 3.37 centipoise). The semen's liquefaction time and ejaculate volume are determined to be functions of the frequency of ejaculation. Steady state ejaculate volume is found to decrease linearly with increasing ejaculation frequency, thereby providing a measure of glandular secretory rate. When collectively considered, these findings provide possible means for monitoring an individual's glandular behavior over an extended period of time and comparing such behavior to established standards. | [
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PMID:24600 | Ovulation induction with human menopausal gonadotrophins: an evaluation of a variable daily dosage regimen. | Human menopausal gonadotrophins (HMG) were used together with human chorionic gonadotrophin (HCG) in 19 women and 39 treatment cycles in an attempt to induce ovulation. Daily 24 hours urinary total estrogen excretion rates were measured and HMG daily dosage was varied according to levels obtained. HCG injections were timed to coincide with an estimated urinary total estrogen excretion rate of 100-150 g per 24 hours. Thirty-one ovulatory cycles occurred (79%) and there were nine pregnancies (23%) of which five were multiple. Eleven cycles were complicated by hyperstimulation (25.6%) of which six were severe. The variable HMG dosage regimen was found to offer no advantages when compared with our standard daily dosage regimen. A rapid rise of estrogen excretion occurred in over 80% of hyperstimulation cycles, including all severe ones, and it was found that this rise could occur after the last dosage of HMG had been given. Because of this, it is proposed that HCG injections be delayed until 48 hours after the last injection of HMG. The finding of a value for the last available 24 hour urinary total estrogen excretion of less than 150 microgram can be taken as an indicator that hyperstimulation is unlikely to occur, and that HCG can safely be given. No indication was found that such a procedure would diminish ovulation or pregnancy rates. | Ovulation induction with human menopausal gonadotrophins: an evaluation of a variable daily dosage regimen. Human menopausal gonadotrophins (HMG) were used together with human chorionic gonadotrophin (HCG) in 19 women and 39 treatment cycles in an attempt to induce ovulation. Daily 24 hours urinary total estrogen excretion rates were measured and HMG daily dosage was varied according to levels obtained. HCG injections were timed to coincide with an estimated urinary total estrogen excretion rate of 100-150 g per 24 hours. Thirty-one ovulatory cycles occurred (79%) and there were nine pregnancies (23%) of which five were multiple. Eleven cycles were complicated by hyperstimulation (25.6%) of which six were severe. The variable HMG dosage regimen was found to offer no advantages when compared with our standard daily dosage regimen. A rapid rise of estrogen excretion occurred in over 80% of hyperstimulation cycles, including all severe ones, and it was found that this rise could occur after the last dosage of HMG had been given. Because of this, it is proposed that HCG injections be delayed until 48 hours after the last injection of HMG. The finding of a value for the last available 24 hour urinary total estrogen excretion of less than 150 microgram can be taken as an indicator that hyperstimulation is unlikely to occur, and that HCG can safely be given. No indication was found that such a procedure would diminish ovulation or pregnancy rates. | [
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PMID:24601 | Ovulation timing by a radioreceptor assay for human luteinizing hormone. | Since predetermination of ovulation would be helpful in treatment of sterility, a quick, sensitive, and specific radioreceptor assay (RRA) for measurement of actual LH concentrations in human serum has been developed. Using partially purified membrane receptors from bovine testes, our assay system enabled precise measurement of LH within 4 hours. The detection limit of the present method is 0.78 ng LER 960/ml serum. The method was used to detect ovulation during four observation cycles each in eleven women who were undergoing treatment for infertility, such as recommended intercourse or artificial insemination because of reduced fertility of their husbands. In all women ovulations could be predicted by LH surge and were verified by laparoscopy within 36 hours. Insemination was carried out at the time of the characteristic increase of LH values around mid-cycle. Pregnancy occurred in three women during the observation period. | Ovulation timing by a radioreceptor assay for human luteinizing hormone. Since predetermination of ovulation would be helpful in treatment of sterility, a quick, sensitive, and specific radioreceptor assay (RRA) for measurement of actual LH concentrations in human serum has been developed. Using partially purified membrane receptors from bovine testes, our assay system enabled precise measurement of LH within 4 hours. The detection limit of the present method is 0.78 ng LER 960/ml serum. The method was used to detect ovulation during four observation cycles each in eleven women who were undergoing treatment for infertility, such as recommended intercourse or artificial insemination because of reduced fertility of their husbands. In all women ovulations could be predicted by LH surge and were verified by laparoscopy within 36 hours. Insemination was carried out at the time of the characteristic increase of LH values around mid-cycle. Pregnancy occurred in three women during the observation period. | [
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PMID:24602 | Endocoagulation: a new and completely safe medical current for sterilization. | Tubal sterilization via pelviscopy with low current represents a unique method: the risk of high-frequency is thereby eliminated, and the application of the Crocodile-Forceps is relatively simple. The mouth of the Crocodile-Forceps grasps the tube under pelviscopic control. Tubal coagulation succeeds under electronic control at 100 degrees C and the area of coagulation measures 6-8 mm in length. In case additional separation of the tube is desired it may be performed bloodlessly with a scissor-like movement. The Crocodile-Clamp is electronically monitored by the "Tube-Coagulator" or the universal endoscopic "Endo-Coagulator." Since 1973 964 tubal coagulations by means of cutting have been performed without postoperative hemorrhage or other complications (pregnancy rate 0). | Endocoagulation: a new and completely safe medical current for sterilization. Tubal sterilization via pelviscopy with low current represents a unique method: the risk of high-frequency is thereby eliminated, and the application of the Crocodile-Forceps is relatively simple. The mouth of the Crocodile-Forceps grasps the tube under pelviscopic control. Tubal coagulation succeeds under electronic control at 100 degrees C and the area of coagulation measures 6-8 mm in length. In case additional separation of the tube is desired it may be performed bloodlessly with a scissor-like movement. The Crocodile-Clamp is electronically monitored by the "Tube-Coagulator" or the universal endoscopic "Endo-Coagulator." Since 1973 964 tubal coagulations by means of cutting have been performed without postoperative hemorrhage or other complications (pregnancy rate 0). | [
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PMID:24604 | In vivo cholinergic stimulation of the rabbit oviductal musculature. | The in vivo activity of the rabbit oviductal musculature was evaluated in terms of constant rate perfusion resistance. Efforts to compare adrenergic and cholinergic stimulation showed that both pilocarpine and bethanechol chloride increased muscular activity 17-24% of that induced by epinephrine. The cholinergic stimulatory effect was reduced by adrenergic alpha blockade and completely abolished by cholinergic blockade or norepinephrine depletion. The data suggest that cholinergic stimulation of the tubal musculature is mediated, in part, via postganglionic adrenergic nerve fibers. It would appear that the oviductal musculature is not a primary target tissue for systemically administered autonomic agonists or antagonists. | In vivo cholinergic stimulation of the rabbit oviductal musculature. The in vivo activity of the rabbit oviductal musculature was evaluated in terms of constant rate perfusion resistance. Efforts to compare adrenergic and cholinergic stimulation showed that both pilocarpine and bethanechol chloride increased muscular activity 17-24% of that induced by epinephrine. The cholinergic stimulatory effect was reduced by adrenergic alpha blockade and completely abolished by cholinergic blockade or norepinephrine depletion. The data suggest that cholinergic stimulation of the tubal musculature is mediated, in part, via postganglionic adrenergic nerve fibers. It would appear that the oviductal musculature is not a primary target tissue for systemically administered autonomic agonists or antagonists. | [
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PMID:24605 | Seminal proteins in patients with agglomerations of spermatozoa. | Seminal proteins (IgA, IgG, albumin) were measured in normozoospermic patients and in patients with normozoospermia associated with agglomerations of spermatozoa. The type of agglomeration was not taken into account. A slight yet significant increase in seminal IgA was detected in patients with agglomerations whereas the other proteins remained unchanged. By testing seminal plasma against C1q by double diffusion it was shown that many patients of both groups had nonimmune C1q precipitins. The significance of such precipitins in semen is not known. | Seminal proteins in patients with agglomerations of spermatozoa. Seminal proteins (IgA, IgG, albumin) were measured in normozoospermic patients and in patients with normozoospermia associated with agglomerations of spermatozoa. The type of agglomeration was not taken into account. A slight yet significant increase in seminal IgA was detected in patients with agglomerations whereas the other proteins remained unchanged. By testing seminal plasma against C1q by double diffusion it was shown that many patients of both groups had nonimmune C1q precipitins. The significance of such precipitins in semen is not known. | [
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PMID:24606 | Decrease of sperm antibody titer in males, and conception after treatment of chronic prostatitis. | Although autoimmunization to spermatozoa is a cause of male infertility, the cause of antibody formation is unknown in most cases. It has been shown that the titer is usually unchanged for as much as 16 years in the same individual. Trials to reduce the titer with varying methods have not been successful. A new possibility of treatment was indicated by the finding of a higher incidence of prostatitis in men with sperm antibodies than in a control group. Following treatment of prostatitis we observed a reduction of the antibody titer in eight cases. In five cases the cervical mucus penetrating capacity of the spermatozoa improved, and conception occurred. | Decrease of sperm antibody titer in males, and conception after treatment of chronic prostatitis. Although autoimmunization to spermatozoa is a cause of male infertility, the cause of antibody formation is unknown in most cases. It has been shown that the titer is usually unchanged for as much as 16 years in the same individual. Trials to reduce the titer with varying methods have not been successful. A new possibility of treatment was indicated by the finding of a higher incidence of prostatitis in men with sperm antibodies than in a control group. Following treatment of prostatitis we observed a reduction of the antibody titer in eight cases. In five cases the cervical mucus penetrating capacity of the spermatozoa improved, and conception occurred. | [
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PMID:24607 | pH controlled diazo coupling of aldolase. Selective formation of diazothioether chromophores and retention of enzyme activity. | pH Conditions have been found which achieve selective reaction of diazotized p-amino benzoate with cysteine residues of rabbit muscle aldolase. The difference in reactivity of the two sulphydryl groups involved, (Cys--237 and Cys--287) permits one to form either four or eight diazothioethers on the tetrameric enzyme and obtain a homogeneous protein. In both cases the enzyme became slightly more active in the fructose-1, 6-bisphosphate cleavage, the KM value being retained. The results have been discussed with regard to chemically modifying an enzyme to change its physical, chemical and immunological properties, whilst leaving the catalytical activity unmodified. | pH controlled diazo coupling of aldolase. Selective formation of diazothioether chromophores and retention of enzyme activity. pH Conditions have been found which achieve selective reaction of diazotized p-amino benzoate with cysteine residues of rabbit muscle aldolase. The difference in reactivity of the two sulphydryl groups involved, (Cys--237 and Cys--287) permits one to form either four or eight diazothioethers on the tetrameric enzyme and obtain a homogeneous protein. In both cases the enzyme became slightly more active in the fructose-1, 6-bisphosphate cleavage, the KM value being retained. The results have been discussed with regard to chemically modifying an enzyme to change its physical, chemical and immunological properties, whilst leaving the catalytical activity unmodified. | [
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PMID:24610 | The measurement of histochemically-stained NADP-dependent isocitrate dehydrogenase in the sebaceous glands of hairless hamster. | The effect of substrate, co-factor and Nitroblue Tetrazolium concentration on the production of formazan by the action of NADP-dependent isocitrate dehydrogenase was studied in the sebaceous glands of the hairless hamster. The measurement (in average optical density units per unit area) of formazan in histochemically stained skin sections was carried out by television scanning microdensitometry (Quantimet 720D). Having established optimal conditions for this enzyme, a second study was carried out to determine the effect of different power objectives and wide-band wavelength light instead of white light on the average optical density per unit area, recorded by the instrument, of the formazan produced in a defined number of sebaceous glands in a single skin section. It was found that there was no difference in the average optical density per unit area recorded by the instrument at different power objectives and a peak value of average optical density per unit area could be obtained using a K4 or a K5 Balzer filter (550-650 nm). | The measurement of histochemically-stained NADP-dependent isocitrate dehydrogenase in the sebaceous glands of hairless hamster. The effect of substrate, co-factor and Nitroblue Tetrazolium concentration on the production of formazan by the action of NADP-dependent isocitrate dehydrogenase was studied in the sebaceous glands of the hairless hamster. The measurement (in average optical density units per unit area) of formazan in histochemically stained skin sections was carried out by television scanning microdensitometry (Quantimet 720D). Having established optimal conditions for this enzyme, a second study was carried out to determine the effect of different power objectives and wide-band wavelength light instead of white light on the average optical density per unit area, recorded by the instrument, of the formazan produced in a defined number of sebaceous glands in a single skin section. It was found that there was no difference in the average optical density per unit area recorded by the instrument at different power objectives and a peak value of average optical density per unit area could be obtained using a K4 or a K5 Balzer filter (550-650 nm). | [
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PMID:24609 | A procedure for the assay of DNA damage in mammalian cells by alkaline elution and microfluorometric DNA determination. | The microfluorimetric assay of Kissane and Robins (J. Biol. Chem. 233, 184-188, 1958) has been modified to monitor DNA content in alkaline elution fractions. The elution profiles of DNA from treated and control cultured cells demonstrated a good correlation whether DNA in the elution fractions was determined by the fluorometric method or by measuring its radioactivity. The DNA recovery was of about 80% and the measured material was DNase digestible. | A procedure for the assay of DNA damage in mammalian cells by alkaline elution and microfluorometric DNA determination. The microfluorimetric assay of Kissane and Robins (J. Biol. Chem. 233, 184-188, 1958) has been modified to monitor DNA content in alkaline elution fractions. The elution profiles of DNA from treated and control cultured cells demonstrated a good correlation whether DNA in the elution fractions was determined by the fluorometric method or by measuring its radioactivity. The DNA recovery was of about 80% and the measured material was DNase digestible. | [
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PMID:24613 | Binding of aminoglycoside antibiotics to acidic mucopolysaccharides. | Binding of aminoglycoside antibiotics to acidic mucopolysaccharides has been studied by means of physicochemical methods. Reactivity was affected markedly by the ionic environment , e.g. pH and ionic strength of the medium, the concentrations and the molar ratios of the constituents. The ionic character of binding was further confirmed by gel chromatography. The reduction of metachromasis by an aminoglycoside was also observed. Their affinity is correlated with localization of the aminoglycosides in vivo. According to reactivity, the following descending order of affinity was obtained for each family: neomycin, gentamicin, sagamicin, kanamycin and streptomycin; heparin, chondroitin sulfate and hyaluronic acid. This sequence of aminoglycosides corresponds to the extent of oto-, nephro- and neuro-(acute)toxicity, suggesting that their affinity for acidic mucopolysaccharides contribute to their tissue toxicity. | Binding of aminoglycoside antibiotics to acidic mucopolysaccharides. Binding of aminoglycoside antibiotics to acidic mucopolysaccharides has been studied by means of physicochemical methods. Reactivity was affected markedly by the ionic environment , e.g. pH and ionic strength of the medium, the concentrations and the molar ratios of the constituents. The ionic character of binding was further confirmed by gel chromatography. The reduction of metachromasis by an aminoglycoside was also observed. Their affinity is correlated with localization of the aminoglycosides in vivo. According to reactivity, the following descending order of affinity was obtained for each family: neomycin, gentamicin, sagamicin, kanamycin and streptomycin; heparin, chondroitin sulfate and hyaluronic acid. This sequence of aminoglycosides corresponds to the extent of oto-, nephro- and neuro-(acute)toxicity, suggesting that their affinity for acidic mucopolysaccharides contribute to their tissue toxicity. | [
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PMID:24614 | Fluphenazine decanoate vs oral antipsychotics: a comparison of their effectiveness in the treatment of schizophrenia as measured by a reduction in hospital readmissions. | A significant number of chronic psychotic patients are readmitted to state mental hospitals each year due in large part to their inability or unwillingness to continue taking antipsychotic medication on a voluntary basis. This paper describes an ex post facto study comparing the effectiveness of fluphenazine decanoate, a long acting antipsychotic medication, with a variety of oral antipsychotics in reducing the number of readmissions of two groups of chronic psychotic patients to a state operated mental health facility. The results demonstrated that a group of 61 patients receiving fluphenazine decanoate had signigicantly fewer readmissions during the one year study period than did a comparison group on oral antipsychotics. The findings indicate that long acting antipsychotic medication, as part of an intensive outpatient treatment program, strongly militates against the tendency of chronic psychotic patients to discontinue treatment. | Fluphenazine decanoate vs oral antipsychotics: a comparison of their effectiveness in the treatment of schizophrenia as measured by a reduction in hospital readmissions. A significant number of chronic psychotic patients are readmitted to state mental hospitals each year due in large part to their inability or unwillingness to continue taking antipsychotic medication on a voluntary basis. This paper describes an ex post facto study comparing the effectiveness of fluphenazine decanoate, a long acting antipsychotic medication, with a variety of oral antipsychotics in reducing the number of readmissions of two groups of chronic psychotic patients to a state operated mental health facility. The results demonstrated that a group of 61 patients receiving fluphenazine decanoate had signigicantly fewer readmissions during the one year study period than did a comparison group on oral antipsychotics. The findings indicate that long acting antipsychotic medication, as part of an intensive outpatient treatment program, strongly militates against the tendency of chronic psychotic patients to discontinue treatment. | [
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PMID:24615 | Heal illness syndrome and lithium intoxication. | This paper reviews heat illness syndrome and lithium intoxication and shows striking similarities in symptoms, predisposing factors, and physiology of both conditions. It is speculated that there is a common underlying mechanism present to account for this. | Heal illness syndrome and lithium intoxication. This paper reviews heat illness syndrome and lithium intoxication and shows striking similarities in symptoms, predisposing factors, and physiology of both conditions. It is speculated that there is a common underlying mechanism present to account for this. | [
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PMID:24616 | New approach to the rehabilitation of the hard core drug addict (heroin methadone addicts) a pilot community study. | The limitation of the methadone maintenance program has pressed for a re-evaluation of our understanding of the underlying causes of addiction. Apparently, one of theunderlying causes or end result produced by the drug itself is that of depression. It explains why the addicts try to maintain to the maximum, the euphoric state and are unwilling to rehabilitate. A pilot study was conducted for the treatment of ex-methadone addicts with large dosages of antidepressants and anxiolytics. Out of 117 ex-methadone and soft drug addicts treated with antidepressants and anxiolytics, 46% ex-methadone and 46.3% soft drug abusers failed to become abstinent. The failures were basically related to the management of treatment. The study suggests a possibility for detoxification from methadone and control of relapse by antidepressants and anxiolytics. | New approach to the rehabilitation of the hard core drug addict (heroin methadone addicts) a pilot community study. The limitation of the methadone maintenance program has pressed for a re-evaluation of our understanding of the underlying causes of addiction. Apparently, one of theunderlying causes or end result produced by the drug itself is that of depression. It explains why the addicts try to maintain to the maximum, the euphoric state and are unwilling to rehabilitate. A pilot study was conducted for the treatment of ex-methadone addicts with large dosages of antidepressants and anxiolytics. Out of 117 ex-methadone and soft drug addicts treated with antidepressants and anxiolytics, 46% ex-methadone and 46.3% soft drug abusers failed to become abstinent. The failures were basically related to the management of treatment. The study suggests a possibility for detoxification from methadone and control of relapse by antidepressants and anxiolytics. | [
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PMID:24617 | Metropolitan jail psychiatric clinic: a year's experience. | The behavioral, demographic and psychiatric characteristics of 524 inmates examined over a one year period in a metropolitan jail psychiatric treatment clinic are described. The psychiatric problems were sufficiently acute that over one-third of all patients were examined and treated within 24 hours of booking. Two psychiatric syndrome and intervention profiles emerged. The most commonly seen were misdemeanants, with psychotic illnesses who required long term antipsychotic medications and close contact with community treatment resources. Situational reactions with suicidal ideation in patients with character disorder diagnoses were seen less frequently. They were treated with short term anti-anxiety medications and environmental manipulation. | Metropolitan jail psychiatric clinic: a year's experience. The behavioral, demographic and psychiatric characteristics of 524 inmates examined over a one year period in a metropolitan jail psychiatric treatment clinic are described. The psychiatric problems were sufficiently acute that over one-third of all patients were examined and treated within 24 hours of booking. Two psychiatric syndrome and intervention profiles emerged. The most commonly seen were misdemeanants, with psychotic illnesses who required long term antipsychotic medications and close contact with community treatment resources. Situational reactions with suicidal ideation in patients with character disorder diagnoses were seen less frequently. They were treated with short term anti-anxiety medications and environmental manipulation. | [
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PMID:24618 | The use of benzodiazepines in prison populations. | Experience with the use of tranquilizing drugs in the control of anxiety and in attempts at behavior modification, prompted a controlled study in the use of the benzodiazepines at the Utah state prison. It appeared that the benefits derived from the administration of these drugs in prisoner control were nearly outweighed by the frequent appearance of paradoxical rage reactions and increase in hostility and aggressive tendencies in these individuals. A double blind, cohort, prospective, randomized study was set up comparing Valium and Serax in their antianxiety qualities and their tendency to produce increased aggression and paradoxical rage reactions. Guidelines are proposed for the use of benzodiazepines and Serax is suggested as a superior drug to Valium for this purpose if indeed a tranquilizer of this type is indicated. | The use of benzodiazepines in prison populations. Experience with the use of tranquilizing drugs in the control of anxiety and in attempts at behavior modification, prompted a controlled study in the use of the benzodiazepines at the Utah state prison. It appeared that the benefits derived from the administration of these drugs in prisoner control were nearly outweighed by the frequent appearance of paradoxical rage reactions and increase in hostility and aggressive tendencies in these individuals. A double blind, cohort, prospective, randomized study was set up comparing Valium and Serax in their antianxiety qualities and their tendency to produce increased aggression and paradoxical rage reactions. Guidelines are proposed for the use of benzodiazepines and Serax is suggested as a superior drug to Valium for this purpose if indeed a tranquilizer of this type is indicated. | [
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PMID:24619 | A pharmacological and theoretical comparison of high and low potency neuroleptics. | Side effects of high and low potency neuroleptics are contrasted. Low potency drugs are more likely to cause central, autonomic and peripheral side effects of all types with the exception of extrapyramidal symptoms. The tendency to produce tardive dyskinesia cannot be compared as relevant data do not currently exist. Greater affinity for the dopamine receptor and twenty to seventy-fold greater milligram and molar potency are advanced as arguments for greater specificity of high potency drugs in the treatment of psychoses. It is proposed that the high potency neuroleptics might be considered the drugs of choice in the treatment of schizophrenia and schizophreniform psychoses. | A pharmacological and theoretical comparison of high and low potency neuroleptics. Side effects of high and low potency neuroleptics are contrasted. Low potency drugs are more likely to cause central, autonomic and peripheral side effects of all types with the exception of extrapyramidal symptoms. The tendency to produce tardive dyskinesia cannot be compared as relevant data do not currently exist. Greater affinity for the dopamine receptor and twenty to seventy-fold greater milligram and molar potency are advanced as arguments for greater specificity of high potency drugs in the treatment of psychoses. It is proposed that the high potency neuroleptics might be considered the drugs of choice in the treatment of schizophrenia and schizophreniform psychoses. | [
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PMID:24622 | Immunochemical study on the pathway of electron flow in reduced nicotinamide adenine dinucleotide-dependent microsomal lipid peroxidation. | NADH could support the lipid peroxidation of rat liver microsomes in the presence of ferric ions chelated by ADP(ADP-Fe). The reaction had a broad pH optimum (pH 5.8--7.4) and was more active in the acidic pH range. Antibodies to NADH-cytochrome b5 reductase [EC 1.6.2.2] and cytochrome b5 inhibited NADH-dependent lipid peroxidation in the presence of ADP-Fe, whereas the antibody against NADPH-cytochrome c reductase [EC 1.6.2.4] showed no inhibition. These oberservations suggest that the electron from NADH was supplied to the lipid peroxidation reaction via NADH-cytochrome b5 reductase and cytochrome b5. On the other hand, NADPH-supported lipid peroxidation was strongly inhibited by the antibody against NADPH-cytochrome c reductase, confirming the participation of this this flavoprotein in the NADPH-dependent reaction. In the presence of both ADP-Fe and ferric ions chelated by EDTA(EDTA-Fe), NADH-dependent lipid peroxidation was highly stimulated up to the level of the NADPH-dependent reaction. In this case, the antibody against cytochrome b5 could not inhibit the reaction, while the antibody against NADH-cytochrome b5 reductase did inhibit it, suggesting the direct transfer of electrons from NADH-cytochrome b5 reductase to EDTA-Fe complex. | Immunochemical study on the pathway of electron flow in reduced nicotinamide adenine dinucleotide-dependent microsomal lipid peroxidation. NADH could support the lipid peroxidation of rat liver microsomes in the presence of ferric ions chelated by ADP(ADP-Fe). The reaction had a broad pH optimum (pH 5.8--7.4) and was more active in the acidic pH range. Antibodies to NADH-cytochrome b5 reductase [EC 1.6.2.2] and cytochrome b5 inhibited NADH-dependent lipid peroxidation in the presence of ADP-Fe, whereas the antibody against NADPH-cytochrome c reductase [EC 1.6.2.4] showed no inhibition. These oberservations suggest that the electron from NADH was supplied to the lipid peroxidation reaction via NADH-cytochrome b5 reductase and cytochrome b5. On the other hand, NADPH-supported lipid peroxidation was strongly inhibited by the antibody against NADPH-cytochrome c reductase, confirming the participation of this this flavoprotein in the NADPH-dependent reaction. In the presence of both ADP-Fe and ferric ions chelated by EDTA(EDTA-Fe), NADH-dependent lipid peroxidation was highly stimulated up to the level of the NADPH-dependent reaction. In this case, the antibody against cytochrome b5 could not inhibit the reaction, while the antibody against NADH-cytochrome b5 reductase did inhibit it, suggesting the direct transfer of electrons from NADH-cytochrome b5 reductase to EDTA-Fe complex. | [
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PMID:24623 | The structure and function of acid proteases. VIII. Purification and characterization of cathepsins D from Japanese monkey lung. | Two kinds of cathepsin D were found in Japanese monkey lung and were named cathepsins D-I and D-II. Cathepsin D-I was partially purified by ammonium sulfate fractionation and DEAE-cellulose column chromatography. It had properties common to other ordinary cathepsins D in terms of the elution position from a DEAE-cellulose column at pH 8.0, the pH-dependence of activity toward acid-denatured hemoglobin, and the molecular weight of 35,000 as determined by Sephadex G-100 gel filtration. On the other hand, cathepsin D-II was purified about 1,000-fold by a combination of ammonium sulfate fractionation and column chromatographies on DEAE-cellulose and Sephadex G-100. It was a very acidic protein as judged from its elution position from a DEAE-cellulose column at pH 8.0, and the high mobility toward the anode on disc gel electrophoresis at pH 8.6. Its molecular weight was determined to be 35,000 by Sephadex G-100 gel filtration and 39,000 by SDS-polyacrylamide gel electrophoresis. It was optimally active at pH 2.8 against acid-denatured hemoglobin as a substrate, showing 80% of the optimal activity at pH 1.0, and almost no activity above pH 4.0. This pH-profile of activity was similar to that of monkey pepsin C (gastricsin). It did not hydrolyze N-acetyl-L-phenylalanyl-3,5-diiodo-L-tyrosine, a synthetic substrate for pepsin, but was inhibited by a series of pepsin inhibitors such as pepstatin, 1,2-epoxy-3-(p-nitrophenoxy)propane, p-bromophenacyl bromide, and diazoacetyl-DL-norleucine methyl ester, although the diazo reagent was a rather weak inhibitor of the enzyme. The amino acid composition of cathepsin D-II was found to be fairly different from those of other cathepsins D. However, it showed a striking resemblance to that of Japanese monkey pepsinogen C, suggesting some evolutionary relationship between them. | The structure and function of acid proteases. VIII. Purification and characterization of cathepsins D from Japanese monkey lung. Two kinds of cathepsin D were found in Japanese monkey lung and were named cathepsins D-I and D-II. Cathepsin D-I was partially purified by ammonium sulfate fractionation and DEAE-cellulose column chromatography. It had properties common to other ordinary cathepsins D in terms of the elution position from a DEAE-cellulose column at pH 8.0, the pH-dependence of activity toward acid-denatured hemoglobin, and the molecular weight of 35,000 as determined by Sephadex G-100 gel filtration. On the other hand, cathepsin D-II was purified about 1,000-fold by a combination of ammonium sulfate fractionation and column chromatographies on DEAE-cellulose and Sephadex G-100. It was a very acidic protein as judged from its elution position from a DEAE-cellulose column at pH 8.0, and the high mobility toward the anode on disc gel electrophoresis at pH 8.6. Its molecular weight was determined to be 35,000 by Sephadex G-100 gel filtration and 39,000 by SDS-polyacrylamide gel electrophoresis. It was optimally active at pH 2.8 against acid-denatured hemoglobin as a substrate, showing 80% of the optimal activity at pH 1.0, and almost no activity above pH 4.0. This pH-profile of activity was similar to that of monkey pepsin C (gastricsin). It did not hydrolyze N-acetyl-L-phenylalanyl-3,5-diiodo-L-tyrosine, a synthetic substrate for pepsin, but was inhibited by a series of pepsin inhibitors such as pepstatin, 1,2-epoxy-3-(p-nitrophenoxy)propane, p-bromophenacyl bromide, and diazoacetyl-DL-norleucine methyl ester, although the diazo reagent was a rather weak inhibitor of the enzyme. The amino acid composition of cathepsin D-II was found to be fairly different from those of other cathepsins D. However, it showed a striking resemblance to that of Japanese monkey pepsinogen C, suggesting some evolutionary relationship between them. | [
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PMID:24626 | Successive purification of several enzymes having affinities for phosphoric groups of substrates by affinity chromatography on P-cellulose. | Glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, glutathione reductase and pyruvate kinase of Candida utilis and baker's yeast, when in anionic form, were adsorbed on a cation exchanger, P-cellulose, due to affinities similar to those for the phosphoric groups of their respective substrates; thus, glucose-6-phosphate dehydrogenase was readily eluted by either NADP+ or NADPH, glutathione reductase by NADPH, 6-phosphogluconate dehydrogenase by 6-phosphogluconate, and pyruvate kinase by either ATP or ADP. This type of chromatography may be called "affinity-adsorption-elution chromatography"; the main principle is different from that of so-called affinity-elution chromatography. Based on these findings, a large-scale procedure suitable for successive purification of several enzymes having affinities for the phosphoric groups of their substrates was devised. As an example, glucose-6-phosphate dehydrogenase was highly purified from baker's yeast and crystallized. | Successive purification of several enzymes having affinities for phosphoric groups of substrates by affinity chromatography on P-cellulose. Glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, glutathione reductase and pyruvate kinase of Candida utilis and baker's yeast, when in anionic form, were adsorbed on a cation exchanger, P-cellulose, due to affinities similar to those for the phosphoric groups of their respective substrates; thus, glucose-6-phosphate dehydrogenase was readily eluted by either NADP+ or NADPH, glutathione reductase by NADPH, 6-phosphogluconate dehydrogenase by 6-phosphogluconate, and pyruvate kinase by either ATP or ADP. This type of chromatography may be called "affinity-adsorption-elution chromatography"; the main principle is different from that of so-called affinity-elution chromatography. Based on these findings, a large-scale procedure suitable for successive purification of several enzymes having affinities for the phosphoric groups of their substrates was devised. As an example, glucose-6-phosphate dehydrogenase was highly purified from baker's yeast and crystallized. | [
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PMID:24627 | Isolation and some properties of a deoxyribonuclease from Turbo cornutus. | 1. Four DNases were found in the dried liver extract of a top shell, Turbo cornutus. The major one was purified 120-fold by phosphocellulose column chromatography, sulfoethylcellulose column chromatography and gel-filtration on Sephadex G-150. The yield was 2.7%. 2. The enzyme activity was not affected by Mg2+ (10(-3)--10(-2)M), EDTA (10(-3)--10(-2)M), or NaCl (10(-1)M). It showed a pH optimum of 4.7--4.8. Ionic strength was found to be critical for the maximal activity. The isoelectric point was 8.5--9.0. On heating at 50 degrees C C for 5 min the enzymic activity fell to half the initial value. 3. The enzyme preparation degraded native as well as heat-denatured DNA, but not RNA. It degraded heat-denatured DNA endonucleolytically to give oligonucleotides with 3'-phosphates. 4. The 3'-phosphate and 5'-hydroxy termini of oligonucleotides were investigated. At both the 3'- and 5'-terminal positions, purine nucleotides were predominant. | Isolation and some properties of a deoxyribonuclease from Turbo cornutus. 1. Four DNases were found in the dried liver extract of a top shell, Turbo cornutus. The major one was purified 120-fold by phosphocellulose column chromatography, sulfoethylcellulose column chromatography and gel-filtration on Sephadex G-150. The yield was 2.7%. 2. The enzyme activity was not affected by Mg2+ (10(-3)--10(-2)M), EDTA (10(-3)--10(-2)M), or NaCl (10(-1)M). It showed a pH optimum of 4.7--4.8. Ionic strength was found to be critical for the maximal activity. The isoelectric point was 8.5--9.0. On heating at 50 degrees C C for 5 min the enzymic activity fell to half the initial value. 3. The enzyme preparation degraded native as well as heat-denatured DNA, but not RNA. It degraded heat-denatured DNA endonucleolytically to give oligonucleotides with 3'-phosphates. 4. The 3'-phosphate and 5'-hydroxy termini of oligonucleotides were investigated. At both the 3'- and 5'-terminal positions, purine nucleotides were predominant. | [
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PMID:24628 | Aliesterase activity in normal and postheparin human blood sera. | An enzyme with characteristics typical of aliesterase has been found in human blood serum using a gas solid chromatographic assay technique. This conflicts with the findings of several authors that aliesterase is absent in the human blood. Another aliesterase is released into the blood stream after intravenous administration of heparin. Partial purification of the aliesterase in normal (preheparin) and postheparin sera was effected by column chromatography using CM- and DEAE-Sephadex. The preheparin aliesterase and postheparin aliesterase have different pH optima of 7.0 and 8.5 respectively. The preheparin aliesterase activity was very sensitive to sodium fluoride and insensitive to a negatively charged detergent, sodium lauryl sulfate, unlike the postheparin esterase which was highly sensitive to sodium lauryl sulfate and comparatively less sensitive to sodium fluoride. | Aliesterase activity in normal and postheparin human blood sera. An enzyme with characteristics typical of aliesterase has been found in human blood serum using a gas solid chromatographic assay technique. This conflicts with the findings of several authors that aliesterase is absent in the human blood. Another aliesterase is released into the blood stream after intravenous administration of heparin. Partial purification of the aliesterase in normal (preheparin) and postheparin sera was effected by column chromatography using CM- and DEAE-Sephadex. The preheparin aliesterase and postheparin aliesterase have different pH optima of 7.0 and 8.5 respectively. The preheparin aliesterase activity was very sensitive to sodium fluoride and insensitive to a negatively charged detergent, sodium lauryl sulfate, unlike the postheparin esterase which was highly sensitive to sodium lauryl sulfate and comparatively less sensitive to sodium fluoride. | [
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PMID:24629 | Phosphoglycolate phosphatase. Effect of cation and pH on activity. | P-glycolate phosphatase requires divalent cations for activity. Activity-pH curves identified 2 active site residues with pK values at pH 5.7 and pH 9.1 in the presence of magnesium and at pH 5.7 and pH 7.5 in the presence of manganese or cobalt. Saturation velocity kinetics enabled the identification of two distinct divalent cation binding sites. The first, nonspecific site has a K0.5 of 2 to 7 x 10(-5) M, depending on the cation and the pH. The second site, which is specific for magnesium, binds this cation in a negatively cooperative fashion. The affinity at pH 8.1 varies approximately 100-fold from the first magnesium bound to the fourth. The negative cooperativity is greatest at high pH. Because the pH range of activity is very broad, both the phosphate monoanion and dianion of P-glycolate must be bound as the substrate. The concentration of these two species at the apparent Km is independent of magnesium concentration. The P-glycolate.magnesium complex is kinetically inactive. | Phosphoglycolate phosphatase. Effect of cation and pH on activity. P-glycolate phosphatase requires divalent cations for activity. Activity-pH curves identified 2 active site residues with pK values at pH 5.7 and pH 9.1 in the presence of magnesium and at pH 5.7 and pH 7.5 in the presence of manganese or cobalt. Saturation velocity kinetics enabled the identification of two distinct divalent cation binding sites. The first, nonspecific site has a K0.5 of 2 to 7 x 10(-5) M, depending on the cation and the pH. The second site, which is specific for magnesium, binds this cation in a negatively cooperative fashion. The affinity at pH 8.1 varies approximately 100-fold from the first magnesium bound to the fourth. The negative cooperativity is greatest at high pH. Because the pH range of activity is very broad, both the phosphate monoanion and dianion of P-glycolate must be bound as the substrate. The concentration of these two species at the apparent Km is independent of magnesium concentration. The P-glycolate.magnesium complex is kinetically inactive. | [
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PMID:24630 | gamma-Glutamyl cyclotransferase from rat kidney. Sulfhydryl groups and isolation of a stable form of the enzyme. | gamma-Glutamyl cyclotransferase, highly purified from rat kidney, contains several readily accessible sulfhydryl groups whose modification appears to be associated with the appearance of multiple enzyme forms as determined by isoelectric focusing and ion exchange chromatography. The enzyme was obtained in a 1000-fold purified and apparently homogeneous form by a procedures involving treatment with dithiothreitol followed by chromatography on thiol-Sepharose. The enzyme was also isolated in a highly active, apparently homogeneous, and stable form after reduction and treatment with iodoacetamide. The amino acid compositions and other properties of the two forms of the enzyme were very similar. Studies on the activity of the enzyme toward a variety of gamma-glutamyl amino acids and di-gamma-glutamyl amino acids showed that the enzyme is much more active toward certain di-gamma-glutamyl amino acids than toward the corresponding gamma-glutamyl amino acids; thus, the preferred substrates have the general structure gamma-Glu-gamma-Glu-NH-R in which the nature of the R moiety has relatively little effect on activity. | gamma-Glutamyl cyclotransferase from rat kidney. Sulfhydryl groups and isolation of a stable form of the enzyme. gamma-Glutamyl cyclotransferase, highly purified from rat kidney, contains several readily accessible sulfhydryl groups whose modification appears to be associated with the appearance of multiple enzyme forms as determined by isoelectric focusing and ion exchange chromatography. The enzyme was obtained in a 1000-fold purified and apparently homogeneous form by a procedures involving treatment with dithiothreitol followed by chromatography on thiol-Sepharose. The enzyme was also isolated in a highly active, apparently homogeneous, and stable form after reduction and treatment with iodoacetamide. The amino acid compositions and other properties of the two forms of the enzyme were very similar. Studies on the activity of the enzyme toward a variety of gamma-glutamyl amino acids and di-gamma-glutamyl amino acids showed that the enzyme is much more active toward certain di-gamma-glutamyl amino acids than toward the corresponding gamma-glutamyl amino acids; thus, the preferred substrates have the general structure gamma-Glu-gamma-Glu-NH-R in which the nature of the R moiety has relatively little effect on activity. | [
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PMID:24631 | Sea urchin sperm guanylate cyclase antibody. Cross-reactivity various rat tissue guanylate cyclases. | After the repeated injection of sea urchin sperm guanylate cyclase into rabbits, antibodies to the enzyme were formed. These antibodies inhibited the particulate or the Triton-dispersed forms of the sperm enzyme by greater than 97%. The sperm adenylate cyclase, cyclic GMP phosphodiesterase, adenosine triphosphatase, guanosine triphosphatase, and 5'-nucleotidase enzymes were not affected by the antiserum. The antiserum inhibited the Triton-dispersed guanylate cyclase from rat heart, liver, lung, spleen, and kidney but did not inhibit the soluble form of the enzyme from any of these tissues. The inhibition of the Triton-dispersed enzyme in these tissues was partial, however, ranging from 30% (liver) to 70% (heart). These results provide evidence that adenylate cyclase is antigenically different from guanylate cyclase, and that the soluble form of guanylate cyclase is antigenically different from a particulate form of the enzyme in various rat tissues. | Sea urchin sperm guanylate cyclase antibody. Cross-reactivity various rat tissue guanylate cyclases. After the repeated injection of sea urchin sperm guanylate cyclase into rabbits, antibodies to the enzyme were formed. These antibodies inhibited the particulate or the Triton-dispersed forms of the sperm enzyme by greater than 97%. The sperm adenylate cyclase, cyclic GMP phosphodiesterase, adenosine triphosphatase, guanosine triphosphatase, and 5'-nucleotidase enzymes were not affected by the antiserum. The antiserum inhibited the Triton-dispersed guanylate cyclase from rat heart, liver, lung, spleen, and kidney but did not inhibit the soluble form of the enzyme from any of these tissues. The inhibition of the Triton-dispersed enzyme in these tissues was partial, however, ranging from 30% (liver) to 70% (heart). These results provide evidence that adenylate cyclase is antigenically different from guanylate cyclase, and that the soluble form of guanylate cyclase is antigenically different from a particulate form of the enzyme in various rat tissues. | [
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PMID:24632 | Purification and properties of mosquito DNA polymerase. | For the first time, DNA polymerase in a postembryonic insect has been purified and characterized. This enzyme from mosquito larvae was purified 1700-fold and was free of deoxyribonuclease and protease activities, which hindered previous investigations of insect polymerases. The enzyme had a molecular weight of 132,000 by gen filtration and aggregated to higher molecular weights when concentrated. With an activated DNA template, the pH optimum was 7.2 in phosphate buffer, and the Mg2+ concentration optimum was 5 to 10 mM. Polymerase activity was inhibited by the antisulfhydryl reagents, N-ethylmaleimide and p-mercuribenzoate, and by KCl. These properties indicate that the mosquito enzyme resembles mammalian alpha-polymerase but differs in its lack of inhibition to low ethanol concentrations. There was no evidence of a beta-polymerase form in the mosquito. | Purification and properties of mosquito DNA polymerase. For the first time, DNA polymerase in a postembryonic insect has been purified and characterized. This enzyme from mosquito larvae was purified 1700-fold and was free of deoxyribonuclease and protease activities, which hindered previous investigations of insect polymerases. The enzyme had a molecular weight of 132,000 by gen filtration and aggregated to higher molecular weights when concentrated. With an activated DNA template, the pH optimum was 7.2 in phosphate buffer, and the Mg2+ concentration optimum was 5 to 10 mM. Polymerase activity was inhibited by the antisulfhydryl reagents, N-ethylmaleimide and p-mercuribenzoate, and by KCl. These properties indicate that the mosquito enzyme resembles mammalian alpha-polymerase but differs in its lack of inhibition to low ethanol concentrations. There was no evidence of a beta-polymerase form in the mosquito. | [
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PMID:24635 | Purification and properties of an NADPH-dependent 21-oxo-20-hydroxysteroid reductase (17beta-aldol reductase) from sheep liver. Isolation of the 20beta-glycol product. | This investigation was undertaken to test the hypothesis that steroidal 20-hydroxy-21-aldehydes are intermediates in an alternative pathway of corticosteroid metabolism leading to steroidal 20,21-diols. A NADPH-dependent 21-oxo-20-hydroxysteroid reductase which catalyzed the reduction of 11beta,17,20beta-trihydroxy-3-keto-4-pregnen-21-al (isocortisol) to 11beta,17,20beta,21-tetrahydroxy-4-pregnen-3-one (Reichstein's compound E) was prepared from sheep liver. Other steroidal 17-aldols were also good substrates. Some steroidal 17-oxoaldehydes, D-, and L-glyceraldehyde were reduced, but less effectively than the steroidal aldols. Steroidal ketols and 21-oic acids were not substrates. The enzyme contains--SH groups, has a pH optimum of 6.9 to 7.5 and a molecular weight of about 28,000. Reversibility of the enzymic reaction could not be demonstrated. The reduction product obtained from isocortisol was isolated and characterized. Other 17-glycols were derived from their respective steroid aldols. Reductase activity was also present in hamster and rat liver. From a comparison of 21-oxo-20-hydroxysteroid reductase and 21-hydroxysteroid dehydrogenase with respect to pH optima, substrate specificity, stability to heat, and kinetic constants, we conclude that the two enzymes are distinct. | Purification and properties of an NADPH-dependent 21-oxo-20-hydroxysteroid reductase (17beta-aldol reductase) from sheep liver. Isolation of the 20beta-glycol product. This investigation was undertaken to test the hypothesis that steroidal 20-hydroxy-21-aldehydes are intermediates in an alternative pathway of corticosteroid metabolism leading to steroidal 20,21-diols. A NADPH-dependent 21-oxo-20-hydroxysteroid reductase which catalyzed the reduction of 11beta,17,20beta-trihydroxy-3-keto-4-pregnen-21-al (isocortisol) to 11beta,17,20beta,21-tetrahydroxy-4-pregnen-3-one (Reichstein's compound E) was prepared from sheep liver. Other steroidal 17-aldols were also good substrates. Some steroidal 17-oxoaldehydes, D-, and L-glyceraldehyde were reduced, but less effectively than the steroidal aldols. Steroidal ketols and 21-oic acids were not substrates. The enzyme contains--SH groups, has a pH optimum of 6.9 to 7.5 and a molecular weight of about 28,000. Reversibility of the enzymic reaction could not be demonstrated. The reduction product obtained from isocortisol was isolated and characterized. Other 17-glycols were derived from their respective steroid aldols. Reductase activity was also present in hamster and rat liver. From a comparison of 21-oxo-20-hydroxysteroid reductase and 21-hydroxysteroid dehydrogenase with respect to pH optima, substrate specificity, stability to heat, and kinetic constants, we conclude that the two enzymes are distinct. | [
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PMID:24637 | Differential inhibition of glutamine and gamma-glutamylcysteine synthetases by alpha-alkyl analogs of methionine sulfoximine that induce convulsions. | The alpha-methyl and alpha-ethyl analogs of methionine sulfoximine, like methionine sulfoximine, induce convulsions in mice and inhibit glutamine synthetase irreversibly; alpha-ethylmethionine sulfoximine is approximately 50% as inhibitory as methionine sulfoximine and alpha-methylmethionine sulfoximine. However, whereas alpha-methylmethionine sulfoximine and methionine sulfoximine inhibit gamma-glutamylcysteine synthetase markedly, alpha-ethylmethionine sulfoximine does not, nor does administration of the alpha-ethyl analog produce the decrease in tissue glutathione levels found after giving methionine sulfoximine or its alpha-methyl analog. The findings strongly indicate that methionine sulfoximine-induced convulsions are closely associated with inhibition of glutamine synthetase rather than with inhibition of gamma-glutamylcysteine synthetase. The alpha-alkyl methionine sulfoximine analogs cannot be catabolized via the corresponding alpha-keto or alpha-imino acids, and, like other alpha-substituted amino acids, are probably not metabolized to a significant extent in vivo; this suggests that the amino acid sulfoximine molecules themselves, rather than their metabolites, are directly involved in the induction of convulsions. Possible explanations for the reported lack of correlation between the occurrence of convulsions and the levels of glutamine synthetase activity (and its substrates and product) are considered. The findings suggest that studies on the mechanism of induction of convulsions may be extended significantly and refined in biochemical terms by the use of other structurally modified convulsant molecules. | Differential inhibition of glutamine and gamma-glutamylcysteine synthetases by alpha-alkyl analogs of methionine sulfoximine that induce convulsions. The alpha-methyl and alpha-ethyl analogs of methionine sulfoximine, like methionine sulfoximine, induce convulsions in mice and inhibit glutamine synthetase irreversibly; alpha-ethylmethionine sulfoximine is approximately 50% as inhibitory as methionine sulfoximine and alpha-methylmethionine sulfoximine. However, whereas alpha-methylmethionine sulfoximine and methionine sulfoximine inhibit gamma-glutamylcysteine synthetase markedly, alpha-ethylmethionine sulfoximine does not, nor does administration of the alpha-ethyl analog produce the decrease in tissue glutathione levels found after giving methionine sulfoximine or its alpha-methyl analog. The findings strongly indicate that methionine sulfoximine-induced convulsions are closely associated with inhibition of glutamine synthetase rather than with inhibition of gamma-glutamylcysteine synthetase. The alpha-alkyl methionine sulfoximine analogs cannot be catabolized via the corresponding alpha-keto or alpha-imino acids, and, like other alpha-substituted amino acids, are probably not metabolized to a significant extent in vivo; this suggests that the amino acid sulfoximine molecules themselves, rather than their metabolites, are directly involved in the induction of convulsions. Possible explanations for the reported lack of correlation between the occurrence of convulsions and the levels of glutamine synthetase activity (and its substrates and product) are considered. The findings suggest that studies on the mechanism of induction of convulsions may be extended significantly and refined in biochemical terms by the use of other structurally modified convulsant molecules. | [
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PMID:24638 | Stabilization of the relaxed state of aspartate transcarbamoylase by modification with a bifunctional reagent. | Native aspartate transcarbamoylase from Escherichia coli was modified with the bifunctional reagent tartaryl diazide in the presence of the substrate carbamoyl phosphate and the substrate analog succinate. The product had the same sedimentation coefficient as the native enzyme but showed a marked increase in affinity for the substrate aspartate with a hyperbolic saturation curve. The Michaelis constant for aspartate (7.4 mM) is similar to that estimated for the relaxed state of the enzyme. The high substrate affinity was not produced if modification was conducted in the absence of substrate analogs or with a monofunctional reagent. The modified enzyme was also desensitized towards the allosteric effectors ATP and CTP. It appears to represent a stabilized relaxed state whose conversion to the taut state is presumably prevented by cross-linking. | Stabilization of the relaxed state of aspartate transcarbamoylase by modification with a bifunctional reagent. Native aspartate transcarbamoylase from Escherichia coli was modified with the bifunctional reagent tartaryl diazide in the presence of the substrate carbamoyl phosphate and the substrate analog succinate. The product had the same sedimentation coefficient as the native enzyme but showed a marked increase in affinity for the substrate aspartate with a hyperbolic saturation curve. The Michaelis constant for aspartate (7.4 mM) is similar to that estimated for the relaxed state of the enzyme. The high substrate affinity was not produced if modification was conducted in the absence of substrate analogs or with a monofunctional reagent. The modified enzyme was also desensitized towards the allosteric effectors ATP and CTP. It appears to represent a stabilized relaxed state whose conversion to the taut state is presumably prevented by cross-linking. | [
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PMID:24639 | Inactivation of human gamma-glutamylcysteine synthetase by cystamine. Demonstration and quantification of enzyme-ligand complexes. | Human erythrocyte gamma-glutamylcysteine synthetase is inactivated by the disulfide cystamine (2,2'-dithiobis-(ethylamine)) at pH 8.2 with a rate constant of 1020 min-1 mM-1. Magnesium ion and various combinations of substrates and products confer differing degrees of protection against cystamine inactivation, thus allowing the detection and quantification of certain enzyme-ligand interactions. By measuring inactivation rates as a function of ligand concentrations in incomplete reaction mixtures, we have obtained evidence for the following complexes: enzyme . Mg2+; enzyme . Mg2+ . MgATP2-; enzyme . Mg2+ . L-glutamate; enzyme . Mg2+ . MgATP2- . L-glutamate; enzyme . Mg2+ . L-gamma-glutamyl-L-alpha-aminobutyrate. The data also imply the existence of enzyme . (Mg2+)2 . MgATP2- . L-glutamate and several enzyme forms resulting from the weak binding to L-alpha-aminobutyrate. The methods used permit the calculation of cystamine inactivation rates for most of these enzyme forms and also give values for the equilibrium constants describing their formation. | Inactivation of human gamma-glutamylcysteine synthetase by cystamine. Demonstration and quantification of enzyme-ligand complexes. Human erythrocyte gamma-glutamylcysteine synthetase is inactivated by the disulfide cystamine (2,2'-dithiobis-(ethylamine)) at pH 8.2 with a rate constant of 1020 min-1 mM-1. Magnesium ion and various combinations of substrates and products confer differing degrees of protection against cystamine inactivation, thus allowing the detection and quantification of certain enzyme-ligand interactions. By measuring inactivation rates as a function of ligand concentrations in incomplete reaction mixtures, we have obtained evidence for the following complexes: enzyme . Mg2+; enzyme . Mg2+ . MgATP2-; enzyme . Mg2+ . L-glutamate; enzyme . Mg2+ . MgATP2- . L-glutamate; enzyme . Mg2+ . L-gamma-glutamyl-L-alpha-aminobutyrate. The data also imply the existence of enzyme . (Mg2+)2 . MgATP2- . L-glutamate and several enzyme forms resulting from the weak binding to L-alpha-aminobutyrate. The methods used permit the calculation of cystamine inactivation rates for most of these enzyme forms and also give values for the equilibrium constants describing their formation. | [
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PMID:24642 | Characterization of microtubule protein oligomers by analytical ultracentrifugation. | Samples of microtubule protein prepared by repeated cycles of assembly-disassembly were examined at low temperatures by sedimentation velocity ultracentrifugation. Sedimenting boundaries corresponding to the 6 S tubulin subunit as well as to two oligomeric species with s020,w values of 18.6 S and 30.6 S were observed. The 30 S to 6 S mass ratio varied with total protein concentration, suggesting that a concentration-dependent equilibrium exists between 6 S and oligomeric species of tubulin. A study of the effects of pH on the mass distribution among the species demonstrated that the 6 S species was favored at low pH values (5.8 to 6.5), the 30 S oligomer was favored at moderate pH values (6.5 to 7.4), and the 18 S oligomer was formed in increasing proportions at higher pH values (7.4 to 8.2). Incubation of purified microtubule protein solutions with increasing concentrations of NaCl initially favored conversion of the 30 S oligomer to the 18 S species with further increases in salt concentration resulting in the dissolution of both the 30 S and 18 S oligomers. The depolymerizing effects of high salt concentrations were substantially reversible, providing further evidence for 6 S-oligomer equilibria. The manipulation of the solution variables of pH and ionic strength in a systematic fashion led to the construction of a "phase diagram" for the microtubule protein species which provided a relatively complete description of the mass distribution among the 6 S, 18 S, and 30 S species over a range of physiological and near-physiological solution conditions. | Characterization of microtubule protein oligomers by analytical ultracentrifugation. Samples of microtubule protein prepared by repeated cycles of assembly-disassembly were examined at low temperatures by sedimentation velocity ultracentrifugation. Sedimenting boundaries corresponding to the 6 S tubulin subunit as well as to two oligomeric species with s020,w values of 18.6 S and 30.6 S were observed. The 30 S to 6 S mass ratio varied with total protein concentration, suggesting that a concentration-dependent equilibrium exists between 6 S and oligomeric species of tubulin. A study of the effects of pH on the mass distribution among the species demonstrated that the 6 S species was favored at low pH values (5.8 to 6.5), the 30 S oligomer was favored at moderate pH values (6.5 to 7.4), and the 18 S oligomer was formed in increasing proportions at higher pH values (7.4 to 8.2). Incubation of purified microtubule protein solutions with increasing concentrations of NaCl initially favored conversion of the 30 S oligomer to the 18 S species with further increases in salt concentration resulting in the dissolution of both the 30 S and 18 S oligomers. The depolymerizing effects of high salt concentrations were substantially reversible, providing further evidence for 6 S-oligomer equilibria. The manipulation of the solution variables of pH and ionic strength in a systematic fashion led to the construction of a "phase diagram" for the microtubule protein species which provided a relatively complete description of the mass distribution among the 6 S, 18 S, and 30 S species over a range of physiological and near-physiological solution conditions. | [
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PMID:24643 | Relation of protein synthesis and transglutaminase activity to formation of the cross-linked envelope during terminal differentiation of the cultured human epidermal keratinocyte. | When serially cultivated human epidermal keratinocytes are placed in suspension culture they stop growing and form, beneath the plasma membrane, an insoluble envelope consisting of protein cross-linked by epsilon- (gamma-glutamyl)lysine. The formation of envelopes in suspended cells is preceded by a sharp decline in the rate of protein synthesis, and most envelopes appear only after the average rate of protein synthesis has fallen to a very low level. If protein synthesis is reduced over 98 percent with cycloheximide or emetine at the time that surface-grown cells are placed in suspension culture, cross-linked envelopes form in most of the cells. This shows that the precursor of the envelope and the cross-linking enzyme are already in the cytoplasm in most cells of growing surface cultures. The process of envelope formation by suspension cultures is actually accelerated by the inhibitors of protein synthesis; an increased number of cells with cross-linked envelopes is observable within 4-6 h after the addition of cycloheximide. The inhibitor also induces a large fraction of the cells of surface cultures to form enveloped within a few days. These findings suggest that arrest of protein synthesis leads to activation of the cross-linking process. Agents known to inhibit transglutaminase-mediated protein cross-linking-putrescine, iodoacetamide, and ethylene glycol-bis(beta-aminoethyl ether)N,N,N',N'-tetraacetate (EGTA)- also prevent envelope formation. Though the activity of the cross-linking transglutaminase depends on the presence of cellular Ca++, we have not been able to activate the cross-linking process by high external Ca++ concentration or ionophores. | Relation of protein synthesis and transglutaminase activity to formation of the cross-linked envelope during terminal differentiation of the cultured human epidermal keratinocyte. When serially cultivated human epidermal keratinocytes are placed in suspension culture they stop growing and form, beneath the plasma membrane, an insoluble envelope consisting of protein cross-linked by epsilon- (gamma-glutamyl)lysine. The formation of envelopes in suspended cells is preceded by a sharp decline in the rate of protein synthesis, and most envelopes appear only after the average rate of protein synthesis has fallen to a very low level. If protein synthesis is reduced over 98 percent with cycloheximide or emetine at the time that surface-grown cells are placed in suspension culture, cross-linked envelopes form in most of the cells. This shows that the precursor of the envelope and the cross-linking enzyme are already in the cytoplasm in most cells of growing surface cultures. The process of envelope formation by suspension cultures is actually accelerated by the inhibitors of protein synthesis; an increased number of cells with cross-linked envelopes is observable within 4-6 h after the addition of cycloheximide. The inhibitor also induces a large fraction of the cells of surface cultures to form enveloped within a few days. These findings suggest that arrest of protein synthesis leads to activation of the cross-linking process. Agents known to inhibit transglutaminase-mediated protein cross-linking-putrescine, iodoacetamide, and ethylene glycol-bis(beta-aminoethyl ether)N,N,N',N'-tetraacetate (EGTA)- also prevent envelope formation. Though the activity of the cross-linking transglutaminase depends on the presence of cellular Ca++, we have not been able to activate the cross-linking process by high external Ca++ concentration or ionophores. | [
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PMID:24644 | Serum standards for the bioassay of aminoglycosides in cerebrospinal fluid. | Four diluents were compared as reference standards for the assay of gentamicin in cerebrospinal fluid (CSF): human CSF, human serum, distilled water, and 150 mmol NaCl/4.5 mmol CaCl2. Standards prepared in pooled human serum were the best alternative to CSF for the assay of gentamicin and were also useful for the assay of tobramycin, netilmicin, amikacin, and sisomicin. The pH (6.0-9.8) of CSF did not alter the results of the assay. | Serum standards for the bioassay of aminoglycosides in cerebrospinal fluid. Four diluents were compared as reference standards for the assay of gentamicin in cerebrospinal fluid (CSF): human CSF, human serum, distilled water, and 150 mmol NaCl/4.5 mmol CaCl2. Standards prepared in pooled human serum were the best alternative to CSF for the assay of gentamicin and were also useful for the assay of tobramycin, netilmicin, amikacin, and sisomicin. The pH (6.0-9.8) of CSF did not alter the results of the assay. | [
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PMID:24645 | The effects of one-day treatment of anxiety with high doses of halazepam. | The main purposes of this study were to examine the effects of one-day treatment of anxiety and to test whether significant relief of anxiety can be achieved in a short time period by frequency individual adjustment of doses. A double-blind crossover study of halazepam and placebo was carried out with 22 patients, 20 completing the study. Each treatment lasted for one day. Oral medication was administered starting with a dose of 40 mg and the dose adjusted every 2 hours. The total daily dose did not exceed 600 mg. Several rating and self-rating scales were used. All observer rating scales and several self-rating scales discriminated at a significant level. The design appeart to be suitable for the rapid screening for anti-anxiety properties of new drugs. | The effects of one-day treatment of anxiety with high doses of halazepam. The main purposes of this study were to examine the effects of one-day treatment of anxiety and to test whether significant relief of anxiety can be achieved in a short time period by frequency individual adjustment of doses. A double-blind crossover study of halazepam and placebo was carried out with 22 patients, 20 completing the study. Each treatment lasted for one day. Oral medication was administered starting with a dose of 40 mg and the dose adjusted every 2 hours. The total daily dose did not exceed 600 mg. Several rating and self-rating scales were used. All observer rating scales and several self-rating scales discriminated at a significant level. The design appeart to be suitable for the rapid screening for anti-anxiety properties of new drugs. | [
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PMID:24646 | A study in the management of anxiety with lorazepam. | A four-week double-blind comparison between lorazepam and placebo involving 60 patients with anxiety revealed that lorazepam in a mean dose of approximately 3 mg on a b.i.d. regimen is significantly and clinically more effective than placebo for almost all items of the Global, Hamilton, and 35-Item scales at nearly all evaluation periods. Moreover, lorazapem was associated with a 50% greater improvement rate than was placebo. Except for only one patient who was withdrawn from the study because of severe sedation, lorazepam was well tolerated and did not interact adversely with concomitant nonpsychoactive medication. | A study in the management of anxiety with lorazepam. A four-week double-blind comparison between lorazepam and placebo involving 60 patients with anxiety revealed that lorazepam in a mean dose of approximately 3 mg on a b.i.d. regimen is significantly and clinically more effective than placebo for almost all items of the Global, Hamilton, and 35-Item scales at nearly all evaluation periods. Moreover, lorazapem was associated with a 50% greater improvement rate than was placebo. Except for only one patient who was withdrawn from the study because of severe sedation, lorazepam was well tolerated and did not interact adversely with concomitant nonpsychoactive medication. | [
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PMID:24647 | Acid base status during treatment of chronic uremia with diafiltration. | Treatment of chronic uremia by hemodiafiltration requires replacement of the filtrate. Using Ringer's solution alone, there is a depression of pH because of bicarbonate loss. To bring the acid base status back to normal, sodium lactate in increasing concentrations (283 mg% = 32 mM/1, 361 mg% = 40 mM/1; 462 mg% = 51 mM/1; 508 mg% = 57 mM/1) was added to the replacement fluid. The optimal concentration is 450 mg% (=50 mM/1) sodium lactate, provided the following conditions are fulfilled: (a) substitution after the filter; (b) mixing ratio of blood and substitution fluid 1:2. Using 12-15 liters of substitution fluid during a 5 hr treatment, the added lactate amounts to 60 g (=0.54 M). With continuous addition of lactate, the serum concentration of lactate is 3.5 times normal and the concentration of serum pyruvate 4 times normal. An excess lactate concentration, according to Huckabee [1,2], was thus not observed. The sieving coefficients were the following: sodium, potassium, urea, lactate, pyruvate, and phosphate 1; chloride greater then 1; calcium and protein less than 1. Serum osmolality fell, on the average, 9 mOsmol/1 during diafiltration. | Acid base status during treatment of chronic uremia with diafiltration. Treatment of chronic uremia by hemodiafiltration requires replacement of the filtrate. Using Ringer's solution alone, there is a depression of pH because of bicarbonate loss. To bring the acid base status back to normal, sodium lactate in increasing concentrations (283 mg% = 32 mM/1, 361 mg% = 40 mM/1; 462 mg% = 51 mM/1; 508 mg% = 57 mM/1) was added to the replacement fluid. The optimal concentration is 450 mg% (=50 mM/1) sodium lactate, provided the following conditions are fulfilled: (a) substitution after the filter; (b) mixing ratio of blood and substitution fluid 1:2. Using 12-15 liters of substitution fluid during a 5 hr treatment, the added lactate amounts to 60 g (=0.54 M). With continuous addition of lactate, the serum concentration of lactate is 3.5 times normal and the concentration of serum pyruvate 4 times normal. An excess lactate concentration, according to Huckabee [1,2], was thus not observed. The sieving coefficients were the following: sodium, potassium, urea, lactate, pyruvate, and phosphate 1; chloride greater then 1; calcium and protein less than 1. Serum osmolality fell, on the average, 9 mOsmol/1 during diafiltration. | [
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PMID:24652 | Kinetics of nitrosation of the herbicide glyphosate. | The herbicide glyphosate was nitrosated by third-order kinetics to N-nitrosoglyphosate. The nitrosation at 25 degrees C was maximum at the reaction pH of 2.5 and had a pH-dependent rate constant of 2.43 M-2 sec-1. An activation energy of 9.5 kcal mole-1 also suggested that glyphosate is nitrosated very readily. Thiocyanate increased the rate 4.6 fold. The possibility of using these results to predict the formation of N-nitrosoglyphosate under normal agricultural practice is discussed. | Kinetics of nitrosation of the herbicide glyphosate. The herbicide glyphosate was nitrosated by third-order kinetics to N-nitrosoglyphosate. The nitrosation at 25 degrees C was maximum at the reaction pH of 2.5 and had a pH-dependent rate constant of 2.43 M-2 sec-1. An activation energy of 9.5 kcal mole-1 also suggested that glyphosate is nitrosated very readily. Thiocyanate increased the rate 4.6 fold. The possibility of using these results to predict the formation of N-nitrosoglyphosate under normal agricultural practice is discussed. | [
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PMID:24664 | Effects of restoring lethally irradiated mice with anti-Thy 1.2-treated bone marrow: graft-vs-host, host-vs-graft, and mitogen reactivity. | In vitro treatment of A/J mouse bone marrow with anti-Thy 1.2 serum and guinea pig complement (GPC) eliminated its ability to induce graft-vs-host (GVH) mortality in lethally irradiated C57BL/6J x A/F1 (BAF1) mice. The anti-Thy 1.2 and GPC treatment of A/J marrow significantly reduced spleen cell activation by phytohemagglutinin (PHA) but not lipopolysaccharide (LPS) stimulation in A/J mice assayed 6 weeks after lethal irradiation and reconstitution with the treated marrow. However, the anti-Thy 1.2 treatment of A/J bone marrow did not impair the ability of the lethally irradiated, reconstituted, syngeneic mice to reject C57BL/6J skin grafts. We conclude that lymphocytes in bone marrow which are susceptible to inactivation by anti-Thy 1.2 mediate allograft reactions and/or that radioresistant cells which persist in the recipient initiate rejection of allogeneic skin grafts. | Effects of restoring lethally irradiated mice with anti-Thy 1.2-treated bone marrow: graft-vs-host, host-vs-graft, and mitogen reactivity. In vitro treatment of A/J mouse bone marrow with anti-Thy 1.2 serum and guinea pig complement (GPC) eliminated its ability to induce graft-vs-host (GVH) mortality in lethally irradiated C57BL/6J x A/F1 (BAF1) mice. The anti-Thy 1.2 and GPC treatment of A/J marrow significantly reduced spleen cell activation by phytohemagglutinin (PHA) but not lipopolysaccharide (LPS) stimulation in A/J mice assayed 6 weeks after lethal irradiation and reconstitution with the treated marrow. However, the anti-Thy 1.2 treatment of A/J bone marrow did not impair the ability of the lethally irradiated, reconstituted, syngeneic mice to reject C57BL/6J skin grafts. We conclude that lymphocytes in bone marrow which are susceptible to inactivation by anti-Thy 1.2 mediate allograft reactions and/or that radioresistant cells which persist in the recipient initiate rejection of allogeneic skin grafts. | [
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PMID:24665 | Generation of low-dose paralysis in the absence of the ability to secrete antibody. | (CBA/N female x BALB/c male)F1 male mice carry an X-linked defect, originating from CBA/N mice, which renders them unable to generate an antibody response to SSS-III. Histocompatible (BALB/c female x CBA/N male) reciprocal F1 male hybrids do not carry the X-linked defect and therefore generate a readily detectable PFC response to SSS-III, which can be adoptively transferred into nonresponding reciprocal F1 male mice. In the present work, we show that this adoptive response could be inhibited in recipient (CBA/N female x BALB/c male)F1 male nonresponding mice in which low dose paralysis had been induced. Evidence is presented which indicates that such suppression is of host rather than donor cell origin. The capacity to develop low-dose paralysis, a phenomenon that is antigen specific and has been attributed to the action of suppressor T cells, indicates that nonresponding (CBA/N female x BALB/c male) F1 males (and presumably the CBA/N progenitor strain) have the ability to recognize this antigen. Furthermore, since these animals fail to make a serum antibody response to SSS-III, the signal that activates suppressor T cells cannot be circulating antibody or antigen-antibody complexes. These findings are most consistent with the view that low-dose paralysis of the response to SSS-III is not dependent on antibody-mediated feedback inhibition; rather, it is an active process mediated by suppressor T cells. | Generation of low-dose paralysis in the absence of the ability to secrete antibody. (CBA/N female x BALB/c male)F1 male mice carry an X-linked defect, originating from CBA/N mice, which renders them unable to generate an antibody response to SSS-III. Histocompatible (BALB/c female x CBA/N male) reciprocal F1 male hybrids do not carry the X-linked defect and therefore generate a readily detectable PFC response to SSS-III, which can be adoptively transferred into nonresponding reciprocal F1 male mice. In the present work, we show that this adoptive response could be inhibited in recipient (CBA/N female x BALB/c male)F1 male nonresponding mice in which low dose paralysis had been induced. Evidence is presented which indicates that such suppression is of host rather than donor cell origin. The capacity to develop low-dose paralysis, a phenomenon that is antigen specific and has been attributed to the action of suppressor T cells, indicates that nonresponding (CBA/N female x BALB/c male) F1 males (and presumably the CBA/N progenitor strain) have the ability to recognize this antigen. Furthermore, since these animals fail to make a serum antibody response to SSS-III, the signal that activates suppressor T cells cannot be circulating antibody or antigen-antibody complexes. These findings are most consistent with the view that low-dose paralysis of the response to SSS-III is not dependent on antibody-mediated feedback inhibition; rather, it is an active process mediated by suppressor T cells. | [
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PMID:24666 | A simplified solid-phase radioimmunoassay for carcinoembryonic antigen. | A simple and rapid solid-phase radioimmunoassay for the measurement of carcinoembryonic antigen (CEA) levels in sera or plasma is described. The procedure involves perchloric acid (PCA) extraction of the samples, followed by a rapid neutralization of excess acid with alkaline buffer solution. The PCA extracts are assayed uing antibody-coated plastic tubes and radio-labeled anti-CEA antibody as a marker. The assay may be completed in one day. The solid-phase direct-binding assay demonstrated a sensitivity of 0.5 ng CEA/ml plasmin and a marked tolerance to variations in pH and ionic strength of the system. A fair correlation between the CEA levels determined by the solid-phase radioimmunoassay and zirconium phosphate gel method was observed. | A simplified solid-phase radioimmunoassay for carcinoembryonic antigen. A simple and rapid solid-phase radioimmunoassay for the measurement of carcinoembryonic antigen (CEA) levels in sera or plasma is described. The procedure involves perchloric acid (PCA) extraction of the samples, followed by a rapid neutralization of excess acid with alkaline buffer solution. The PCA extracts are assayed uing antibody-coated plastic tubes and radio-labeled anti-CEA antibody as a marker. The assay may be completed in one day. The solid-phase direct-binding assay demonstrated a sensitivity of 0.5 ng CEA/ml plasmin and a marked tolerance to variations in pH and ionic strength of the system. A fair correlation between the CEA levels determined by the solid-phase radioimmunoassay and zirconium phosphate gel method was observed. | [
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PMID:24668 | Factors influencing the therapy of experimental Proteus mirabilis meningitis in rabbits. | The therapy of Proteus mirabilis meningitis with gentamicin alone and in combination with chloramphenicol was studied in a rabbit model. Antibiotics were administered for 8 hr. Samples of serum and cerebrospinal fluid (CSF) obtained simultaneously were assayed at 2-hr intervals for antibiotic concentration and counts of bacteria in CSF. The percentage of penetration ([concentration in CSF divided by concentration in serum] x 100%) of gentamicin ranged from 14% to 23%, but very large dosages were required to kill bacteria in the CSF. Although the minimal bactericidal concentration (MBC) of gentamicin was 1 microgram/ml, killing in vivo occurred only when concentrations in CSF were 10--30 times the MBC. The high concentration required for bactericidal activity in vivo may be explained by the reduced pH of infected CSF (mean pH, 6.98; range, 6.69--7.18). The bactericidal action of gentamicin was abolished by the simultaneous administration of chloramphenicol. Titers of bacteria in CSF were reduced 2.60 log10 (mean) with gentamicin therapy vs. 0.92 log10 (mean) with combination (P less than 0.01). | Factors influencing the therapy of experimental Proteus mirabilis meningitis in rabbits. The therapy of Proteus mirabilis meningitis with gentamicin alone and in combination with chloramphenicol was studied in a rabbit model. Antibiotics were administered for 8 hr. Samples of serum and cerebrospinal fluid (CSF) obtained simultaneously were assayed at 2-hr intervals for antibiotic concentration and counts of bacteria in CSF. The percentage of penetration ([concentration in CSF divided by concentration in serum] x 100%) of gentamicin ranged from 14% to 23%, but very large dosages were required to kill bacteria in the CSF. Although the minimal bactericidal concentration (MBC) of gentamicin was 1 microgram/ml, killing in vivo occurred only when concentrations in CSF were 10--30 times the MBC. The high concentration required for bactericidal activity in vivo may be explained by the reduced pH of infected CSF (mean pH, 6.98; range, 6.69--7.18). The bactericidal action of gentamicin was abolished by the simultaneous administration of chloramphenicol. Titers of bacteria in CSF were reduced 2.60 log10 (mean) with gentamicin therapy vs. 0.92 log10 (mean) with combination (P less than 0.01). | [
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PMID:24669 | Changing ecology of acute bacterial empyema: occurrence and mortality at Boston City Hospital during 12 selected years from 1935 to 1972. | The occurrence, etiology, and demography of acute bacterial empyema are presented to reflect the widespread use of sulfonamides, penicillin, and other active antibiotics. In community-acquired (C-A) cases Streptococcus pneumoniae, hemolytic streptococci, and Staphylococcus aureus were the most frequent single organisms identified in initial positive cultures of pleural fluid during 1935. S. pneumoniae declined steadily until 1953 but continued to occur frequently in C-A cases. Hemolytic streptococci became infrequent. S. aureus increased and became the most frequent organism in 1955 and declined to original levels after 1965 while gram-negative rods increased. S. aureus, aerobic gram-negative rods, and enterococci were most frequent in originally mixed infections, hospital-acquired cases, and superinfections. Anaerobes with or without aerobes were mostly seen in C-A cases and did not vary in incidence. Mortality increased with age. Overall mortality was greater during the 10 antibiotic years, associated with a marked decrease in the proportion of patients younger than 50 years and increase in those over 60 years old, and was greater in patients with serious underlying diseases subjected to more complicated surgical procedures. | Changing ecology of acute bacterial empyema: occurrence and mortality at Boston City Hospital during 12 selected years from 1935 to 1972. The occurrence, etiology, and demography of acute bacterial empyema are presented to reflect the widespread use of sulfonamides, penicillin, and other active antibiotics. In community-acquired (C-A) cases Streptococcus pneumoniae, hemolytic streptococci, and Staphylococcus aureus were the most frequent single organisms identified in initial positive cultures of pleural fluid during 1935. S. pneumoniae declined steadily until 1953 but continued to occur frequently in C-A cases. Hemolytic streptococci became infrequent. S. aureus increased and became the most frequent organism in 1955 and declined to original levels after 1965 while gram-negative rods increased. S. aureus, aerobic gram-negative rods, and enterococci were most frequent in originally mixed infections, hospital-acquired cases, and superinfections. Anaerobes with or without aerobes were mostly seen in C-A cases and did not vary in incidence. Mortality increased with age. Overall mortality was greater during the 10 antibiotic years, associated with a marked decrease in the proportion of patients younger than 50 years and increase in those over 60 years old, and was greater in patients with serious underlying diseases subjected to more complicated surgical procedures. | [
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PMID:24670 | Isolation of the etiologic agent of Korean Hemorrhagic fever. | Lung tissues from 73 rodents (Apodemus agrarius coreae) gave specific immunofluorescent reactions when they reacted with sera from patients convalescing from Korean hemorrhagic fever. Similar staaining was observed in the lungs of A. agrarius inoculated with acute-phase sera obtained from two patients with this disease. The unidentified agent was successfully propagated in adult A. agrarius through eight passages representing a cumulative dilution of greater than 10(-17). Experimentally inoculated rodents developed specific fluorescent antigen in the lung, kidney, liver, parotid glands, and bladder. Organs, especially lungs, were positive beginning 10 days and continuing through 69 days after inoculation. The agent could not be cultivated in several types of cell cultures nor in laboratory animals. No fluorescence was observed when infected A. agrarius lung tissues were reacted with antisera to Marburg virus, Ebola virus, and serval arenaviruses. Diagnostic increases in immunofluorescent antibodies occurred in 113 of 116 severe and 11 of 34 milder cases of clinically suspected Korean hemorrhagic fever. Antibodies were present during the first week of symptoms, reached a peak at the end of the second week, and persisted for up to 14 years. Convalescent-phase sera from four persons suffering a similar disease in the Soviet Union were also positive for antibodies. | Isolation of the etiologic agent of Korean Hemorrhagic fever. Lung tissues from 73 rodents (Apodemus agrarius coreae) gave specific immunofluorescent reactions when they reacted with sera from patients convalescing from Korean hemorrhagic fever. Similar staaining was observed in the lungs of A. agrarius inoculated with acute-phase sera obtained from two patients with this disease. The unidentified agent was successfully propagated in adult A. agrarius through eight passages representing a cumulative dilution of greater than 10(-17). Experimentally inoculated rodents developed specific fluorescent antigen in the lung, kidney, liver, parotid glands, and bladder. Organs, especially lungs, were positive beginning 10 days and continuing through 69 days after inoculation. The agent could not be cultivated in several types of cell cultures nor in laboratory animals. No fluorescence was observed when infected A. agrarius lung tissues were reacted with antisera to Marburg virus, Ebola virus, and serval arenaviruses. Diagnostic increases in immunofluorescent antibodies occurred in 113 of 116 severe and 11 of 34 milder cases of clinically suspected Korean hemorrhagic fever. Antibodies were present during the first week of symptoms, reached a peak at the end of the second week, and persisted for up to 14 years. Convalescent-phase sera from four persons suffering a similar disease in the Soviet Union were also positive for antibodies. | [
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PMID:24674 | Physician's assistants, their physician employers, and the problem of autonomy: consensus or conflict? | Do physician's assistants (PAs) and their physician employers disagree about levels of supervision and autonomy, and does level of physician's assistant autonomy relate in any way to other aspects of practice satisfaction? An indepth study of MD-PA teams in practice reveals that there is greater consensus than conflict concerning the autonomy of the physician's assistant; that the level of physician's assistant autonomy is not related to salary or to physician's assistant employment satisfaction; and that physician-employers who consider their physician's assistants to be more autonomous also tend to feel that the quality of their lives has improved as a result of hiring an assistant. | Physician's assistants, their physician employers, and the problem of autonomy: consensus or conflict? Do physician's assistants (PAs) and their physician employers disagree about levels of supervision and autonomy, and does level of physician's assistant autonomy relate in any way to other aspects of practice satisfaction? An indepth study of MD-PA teams in practice reveals that there is greater consensus than conflict concerning the autonomy of the physician's assistant; that the level of physician's assistant autonomy is not related to salary or to physician's assistant employment satisfaction; and that physician-employers who consider their physician's assistants to be more autonomous also tend to feel that the quality of their lives has improved as a result of hiring an assistant. | [
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PMID:24675 | Nitrogenase synthesis in Klebsiella pneumoniae: comparison of ammonium and oxygen regulation. | Rates of nitrogenase synthesis by Klebsiella pneumoniae were measured by pulse-labelling organisms with a mixture of 14C-labelled amino acids followed by sodium dodecyl sulphate gel electrophoresis and autoradiography. Populations from an NH4+-repressed, SO42--limited chemostat (0.46 mg dry wt ml-1), when released from NH4+ repression, simultaneously synthesized detectable quantities of the three nitrogenase polypeptides 45 min before acetylene-reducing activity was observed. Exposure of populations synthesizing nitrogenase to air or NH4+ (200 microgram N ml-1) repressed synthesis of both component proteins simultaneously, the rate initially decreasing by half in 11 to 12 min; in the presence of NH4+ a second slower phase with an approximate half-life of 30 min was observed. With 5% O2 in N2 the half-lives for the decreases in the rates of synthesis were 30 min for the Fe protein and 33 min for the Mo-Fe protein. Oxygen also repressed nitrogenase in a glutamine synthetase constitutive derivative of K. pneumoniae (strain SK24) which escapes NH4+ repression. Regulation of nitrogenase by O2 may therefore be independent of glutamine synthetase. | Nitrogenase synthesis in Klebsiella pneumoniae: comparison of ammonium and oxygen regulation. Rates of nitrogenase synthesis by Klebsiella pneumoniae were measured by pulse-labelling organisms with a mixture of 14C-labelled amino acids followed by sodium dodecyl sulphate gel electrophoresis and autoradiography. Populations from an NH4+-repressed, SO42--limited chemostat (0.46 mg dry wt ml-1), when released from NH4+ repression, simultaneously synthesized detectable quantities of the three nitrogenase polypeptides 45 min before acetylene-reducing activity was observed. Exposure of populations synthesizing nitrogenase to air or NH4+ (200 microgram N ml-1) repressed synthesis of both component proteins simultaneously, the rate initially decreasing by half in 11 to 12 min; in the presence of NH4+ a second slower phase with an approximate half-life of 30 min was observed. With 5% O2 in N2 the half-lives for the decreases in the rates of synthesis were 30 min for the Fe protein and 33 min for the Mo-Fe protein. Oxygen also repressed nitrogenase in a glutamine synthetase constitutive derivative of K. pneumoniae (strain SK24) which escapes NH4+ repression. Regulation of nitrogenase by O2 may therefore be independent of glutamine synthetase. | [
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PMID:24676 | Factors influencing the susceptibility of Candida albicans to the polyenoic antibiotics nystatin and amphotericin B. | Factors influencing the interaction between Candida albicans and the polyenoic antibiotics nystatin and amphotericin B have been investigated using a K+-specific electrode to measure polyene-mediated efflux of cellular K+. In batch cultures, sensitivity was a function of culture age. Using continuous (chemostat) cultures, the influence of growth-limiting substrate, specific growth rate, growth temperature and growth pH were examined. Carbon-limited cultures showed the highest sensitivity of those substrates tested, and susceptibility increased with growth rate. Within the range 22 to 42 degrees C, growth at lower temperatures resulted in increased sensitivity, whilst a similar trend was observed when the growth pH of cultures was reduced. Further, under the conditions tested, there were considerable variations in free intracellular K+ concentrations. | Factors influencing the susceptibility of Candida albicans to the polyenoic antibiotics nystatin and amphotericin B. Factors influencing the interaction between Candida albicans and the polyenoic antibiotics nystatin and amphotericin B have been investigated using a K+-specific electrode to measure polyene-mediated efflux of cellular K+. In batch cultures, sensitivity was a function of culture age. Using continuous (chemostat) cultures, the influence of growth-limiting substrate, specific growth rate, growth temperature and growth pH were examined. Carbon-limited cultures showed the highest sensitivity of those substrates tested, and susceptibility increased with growth rate. Within the range 22 to 42 degrees C, growth at lower temperatures resulted in increased sensitivity, whilst a similar trend was observed when the growth pH of cultures was reduced. Further, under the conditions tested, there were considerable variations in free intracellular K+ concentrations. | [
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PMID:24677 | Cytoplasmic vacuoles of Rous virus transformed cells are organelles involved in cation uptake. | Cytoplasmic vacuoles induced during transformation of cells by Bryan strain Rous sarcoma virus (RSV-BH) have been studied using the cationic dye, neutral red(NR). Both the rate of uptake and the accumulation of NR are greater in RSV-BH transformed cells than non-transformed cells however, uptake was greater in vacuolated than in non-vacuolated cells, whether or not they were transformed. The NR was incorporated into pre-existing vacuoles in the absence of cytoplasmic staining, suggesting the existence of direct channels from the cell surface to the vacuoles. Other low mol. wt. cationic dyes could also be incorporated into vacuoles, although those with branched structures or cationic weights greater than 330 were excluded. No anionic dyes were incorporated. Infection of cells with a virus mutant, RSV-BH-Ta, induces temperature-dependent vacuolization. After a shift to the vacuole-permissive temperature, vacuoles developed at different rates and with morphological variations with different cations. Vacuoles which had formed in the presence of several cations, (K+, Rb+, tris+, choline+) failed to disappear when cells were incubated at a temperature sufficient to revert vacuoles formed in Na+-containing medium. No short-term effects of Cl-replacements (Br-, I-, or SO2-4) on vacuolization or reversal were observed. The results suggest that these vacuoles are organelles involved in cation uptake. A possible function for these organelles in RSV-BH induced malignancy is discussed. | Cytoplasmic vacuoles of Rous virus transformed cells are organelles involved in cation uptake. Cytoplasmic vacuoles induced during transformation of cells by Bryan strain Rous sarcoma virus (RSV-BH) have been studied using the cationic dye, neutral red(NR). Both the rate of uptake and the accumulation of NR are greater in RSV-BH transformed cells than non-transformed cells however, uptake was greater in vacuolated than in non-vacuolated cells, whether or not they were transformed. The NR was incorporated into pre-existing vacuoles in the absence of cytoplasmic staining, suggesting the existence of direct channels from the cell surface to the vacuoles. Other low mol. wt. cationic dyes could also be incorporated into vacuoles, although those with branched structures or cationic weights greater than 330 were excluded. No anionic dyes were incorporated. Infection of cells with a virus mutant, RSV-BH-Ta, induces temperature-dependent vacuolization. After a shift to the vacuole-permissive temperature, vacuoles developed at different rates and with morphological variations with different cations. Vacuoles which had formed in the presence of several cations, (K+, Rb+, tris+, choline+) failed to disappear when cells were incubated at a temperature sufficient to revert vacuoles formed in Na+-containing medium. No short-term effects of Cl-replacements (Br-, I-, or SO2-4) on vacuolization or reversal were observed. The results suggest that these vacuoles are organelles involved in cation uptake. A possible function for these organelles in RSV-BH induced malignancy is discussed. | [
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PMID:24678 | Growth hormone and prolactin response to bromocriptine in patients with Huntington's chorea. | The growth hormone (hGH) and prolactin (hPRL) responses to oral bromocriptine were studied in two groups of patients with Huntington's chorea and in seven healthy control subjects. The patients included six patients who had previously been treated with phenothiazines and six patients who had not received phenothiazine treatment. All medication was stopped 72 hours before the investigation which involved taking blood samples for up to 210 minutes after taking bromocriptine (2.5 mg). Plasma samples were analysed for hGH and hPRL. There was no significant difference in basal hGH concentrations between the patients and control subjects. The hGH response to bromocriptine varied in the individual patients but the concentrations were significantly lower in the patients compared with the controls between 160 and 210 minutes. The basal concentrations of hPRL were also not different, apart from the findings of elevated hPRL concentrations in three patients previously treated with phenothiazines. The patients and control subjects showed a consistent fall in hPRL concentrations after taking bromocriptine. The lower peak hGH response to bromocriptine found in the patients suggests that there may be an alteration of dopaminergic neurones mediating hGH release. | Growth hormone and prolactin response to bromocriptine in patients with Huntington's chorea. The growth hormone (hGH) and prolactin (hPRL) responses to oral bromocriptine were studied in two groups of patients with Huntington's chorea and in seven healthy control subjects. The patients included six patients who had previously been treated with phenothiazines and six patients who had not received phenothiazine treatment. All medication was stopped 72 hours before the investigation which involved taking blood samples for up to 210 minutes after taking bromocriptine (2.5 mg). Plasma samples were analysed for hGH and hPRL. There was no significant difference in basal hGH concentrations between the patients and control subjects. The hGH response to bromocriptine varied in the individual patients but the concentrations were significantly lower in the patients compared with the controls between 160 and 210 minutes. The basal concentrations of hPRL were also not different, apart from the findings of elevated hPRL concentrations in three patients previously treated with phenothiazines. The patients and control subjects showed a consistent fall in hPRL concentrations after taking bromocriptine. The lower peak hGH response to bromocriptine found in the patients suggests that there may be an alteration of dopaminergic neurones mediating hGH release. | [
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PMID:24680 | Influence of dietary biotin level on growth, metabolism and pathology of rainbow trout. | Supplementation of a dry diet containing herring and soybean meals with D biotin had no effect on the feed comsumption, growth or mortality of rainbow trout in a 28 week experiment. In a second experiment two series of diets were used: practical, based upon herring and soybean meals, and purified, based upon casein and gelatin. The basal levels of biotin in the two series of diets were: 0.51 and 0 mg/kg, respectively. There were four treatment groups in each of the two diet series: the practical diets were either supplemented with 0.5 mg D biotin/kg or 10% of either raw or heated egg white was added to the diet. The purified diets were supplemented with graded levels of D biotin (0, 0.25 0.5, or 1.0 mg/kg). Raw egg white (1%) was added to the biotin unsupplemented diet. Feeding these diets to rainbow trout for 24 weeks showed that both the practical diet containing egg white, and the unsupplemented, purified diet were biotin deficient, all the other diets contained adequate amounts of biotin to support growth and prevent mortality. Neither supplementation of the practical diet nor adding more than 0.25 mg D biotin/kg to the purified diet resulted in any improvement in growth however the biotin levels in the livers were greater for the fish which had received the higher levels of dietary biotin, but these increased levels of biotin did not influence the activity of acetyl CoA carboxylase. The biotin deficient fish were anorexic, and exhibited degeneration of the gills: shortening and thickening of the lamellae and hypertrophy and hyperplasia of the heterogenous epithelial cells of the lamellae. | Influence of dietary biotin level on growth, metabolism and pathology of rainbow trout. Supplementation of a dry diet containing herring and soybean meals with D biotin had no effect on the feed comsumption, growth or mortality of rainbow trout in a 28 week experiment. In a second experiment two series of diets were used: practical, based upon herring and soybean meals, and purified, based upon casein and gelatin. The basal levels of biotin in the two series of diets were: 0.51 and 0 mg/kg, respectively. There were four treatment groups in each of the two diet series: the practical diets were either supplemented with 0.5 mg D biotin/kg or 10% of either raw or heated egg white was added to the diet. The purified diets were supplemented with graded levels of D biotin (0, 0.25 0.5, or 1.0 mg/kg). Raw egg white (1%) was added to the biotin unsupplemented diet. Feeding these diets to rainbow trout for 24 weeks showed that both the practical diet containing egg white, and the unsupplemented, purified diet were biotin deficient, all the other diets contained adequate amounts of biotin to support growth and prevent mortality. Neither supplementation of the practical diet nor adding more than 0.25 mg D biotin/kg to the purified diet resulted in any improvement in growth however the biotin levels in the livers were greater for the fish which had received the higher levels of dietary biotin, but these increased levels of biotin did not influence the activity of acetyl CoA carboxylase. The biotin deficient fish were anorexic, and exhibited degeneration of the gills: shortening and thickening of the lamellae and hypertrophy and hyperplasia of the heterogenous epithelial cells of the lamellae. | [
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PMID:24685 | A membrane model of the human oral mucosa as derived from buccal absorption performance and physicochemical properties of the beta-blocking drugs atenolol and propranolol. | The buccal absorption characteristics and physicochemical properties of the beta-adrenoceptor blocking agents propranolol and atenolol have been investigated to evaluate their permeation properties across biological lipid membranes. The dissociation constants, solubilities of free base, and n-heptane partition coefficients show that propranolol in its unionized form is much more lipophilic than atenolol, both drugs being bases with a similar pKa. Buccal absorption was studied under conditions of varying drug concentration, contact time, and pH, and controlled through the use of a non-absorbable marker. The absorption findings are in general agreement with the pH-partition theory. A new compartmental diffusional model that includes membrane storage and a hypothetical "aqueous pH-buffering surfaces system" allowed a more exhaustive interpretation to be made. A method for the estimation of the intrinsic pH and buffer capacity of the postulated surface system from drug pH-absorption data and partition coefficients alone is described. With human oral mucosa the intrinsic pH was near 6.7, and the buffering capacity of the system (expressed as the ratio deltapH/deltapH eff) about 2.86. The method was validated using published absorption data from the rat small intestine. Absorption of unionized drug through pores is shown to be negligible in the buccal absorption situation. The time course of absorption suggests membrane storage of lipophilic compounds; the in vivo partition coefficient of unionized propranolol relative to the mucous membrane could be calculated for the peusdo-steady state of absorption, i.e. the partition equilibrium between mouth content and membrane, to be approximately 776; this value is of the same order as the in vitro partition coefficient for the erythrocyte/plasma system. The lipid biophase of the buccal membrane is estimated semiquantitatively to be of intermediate polarity (epsilon = 3-4). | A membrane model of the human oral mucosa as derived from buccal absorption performance and physicochemical properties of the beta-blocking drugs atenolol and propranolol. The buccal absorption characteristics and physicochemical properties of the beta-adrenoceptor blocking agents propranolol and atenolol have been investigated to evaluate their permeation properties across biological lipid membranes. The dissociation constants, solubilities of free base, and n-heptane partition coefficients show that propranolol in its unionized form is much more lipophilic than atenolol, both drugs being bases with a similar pKa. Buccal absorption was studied under conditions of varying drug concentration, contact time, and pH, and controlled through the use of a non-absorbable marker. The absorption findings are in general agreement with the pH-partition theory. A new compartmental diffusional model that includes membrane storage and a hypothetical "aqueous pH-buffering surfaces system" allowed a more exhaustive interpretation to be made. A method for the estimation of the intrinsic pH and buffer capacity of the postulated surface system from drug pH-absorption data and partition coefficients alone is described. With human oral mucosa the intrinsic pH was near 6.7, and the buffering capacity of the system (expressed as the ratio deltapH/deltapH eff) about 2.86. The method was validated using published absorption data from the rat small intestine. Absorption of unionized drug through pores is shown to be negligible in the buccal absorption situation. The time course of absorption suggests membrane storage of lipophilic compounds; the in vivo partition coefficient of unionized propranolol relative to the mucous membrane could be calculated for the peusdo-steady state of absorption, i.e. the partition equilibrium between mouth content and membrane, to be approximately 776; this value is of the same order as the in vitro partition coefficient for the erythrocyte/plasma system. The lipid biophase of the buccal membrane is estimated semiquantitatively to be of intermediate polarity (epsilon = 3-4). | [
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PMID:24686 | Biopharmaceutical properties of liquid and tablet antacids: in vivo studies using the intragastric pH-measurement technique. | Three types of liquid and tablet antacids have been studied in vitro and in vivo: aluminium hydroxide, aluminium hydroxide-magnesium carbonate and hydrotalcite. The effects on gastric pH of antacid suspensions and antacid chewing tablets having identical active ingredients have been studied in 36 volunteers, the sequence of both forms of administration being randomized. Gastric acid secretion was continuously stimulated during the experiment by a pentagastrin infusion. Antacid chewing tablets gave inferior results when compared with the same antacid in liquid. Antacid suspensions are therefore preferred in the treatment of acid-peptic disease. | Biopharmaceutical properties of liquid and tablet antacids: in vivo studies using the intragastric pH-measurement technique. Three types of liquid and tablet antacids have been studied in vitro and in vivo: aluminium hydroxide, aluminium hydroxide-magnesium carbonate and hydrotalcite. The effects on gastric pH of antacid suspensions and antacid chewing tablets having identical active ingredients have been studied in 36 volunteers, the sequence of both forms of administration being randomized. Gastric acid secretion was continuously stimulated during the experiment by a pentagastrin infusion. Antacid chewing tablets gave inferior results when compared with the same antacid in liquid. Antacid suspensions are therefore preferred in the treatment of acid-peptic disease. | [
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PMID:24687 | The diffusion of nandrolone through occluded and non-occluded human skin. | Diffusion constants have been measured for nandrolone in human cadaver skin in vitro under conditions which approximate to both the occluded and non-occluded topical therapeutic situation. The results suggest that occlusion affects the diffusion parameters only marginally, if at all, but appears to reduce the ability of the stratum corneum to "irreversibly" bind a fraction of the applied dose. Studies of nandrolone diffusion in freshly excised human epidermis yield similar results to those from measurements on cadaver skin, and it has also been found that epidermal samples excised from non-involved sites of human psoriatic patients behave under these conditions in essentially the same way as normal epidermis. | The diffusion of nandrolone through occluded and non-occluded human skin. Diffusion constants have been measured for nandrolone in human cadaver skin in vitro under conditions which approximate to both the occluded and non-occluded topical therapeutic situation. The results suggest that occlusion affects the diffusion parameters only marginally, if at all, but appears to reduce the ability of the stratum corneum to "irreversibly" bind a fraction of the applied dose. Studies of nandrolone diffusion in freshly excised human epidermis yield similar results to those from measurements on cadaver skin, and it has also been found that epidermal samples excised from non-involved sites of human psoriatic patients behave under these conditions in essentially the same way as normal epidermis. | [
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PMID:24688 | Circling behaviour in the rat following unilateral injections of p-chlorophenylalanine and ethanolamine-O-sulphate into the substantia nigra. | Circling behaviour following the unilateral intranigral injection of the gamma-aminobutyric acid (GABA) transaminase inhibitor ethanolamine-O-sulphate, (EOS), or the tryptophan hydroxylase inhibitor p-chlorophenylalanine (pCPA) was compared, both for spontaneous circling and for circling induced by apomorphine administration. EOS-injected rats exhibited spontaneous contralateral rotation for up to 3 h after such injections. pCPA-injected rats showed only a slight ipsilateral postural asymmetry, both groups of rats elicited ipsilateral rotation upon apomorphine challenge. Increased ipsilateral dopamine release within the corpus striatrum accompanied this response. It is proposed that GABA and 5-HT may be closely related in modifying the nigrostriatal impulse flow. | Circling behaviour in the rat following unilateral injections of p-chlorophenylalanine and ethanolamine-O-sulphate into the substantia nigra. Circling behaviour following the unilateral intranigral injection of the gamma-aminobutyric acid (GABA) transaminase inhibitor ethanolamine-O-sulphate, (EOS), or the tryptophan hydroxylase inhibitor p-chlorophenylalanine (pCPA) was compared, both for spontaneous circling and for circling induced by apomorphine administration. EOS-injected rats exhibited spontaneous contralateral rotation for up to 3 h after such injections. pCPA-injected rats showed only a slight ipsilateral postural asymmetry, both groups of rats elicited ipsilateral rotation upon apomorphine challenge. Increased ipsilateral dopamine release within the corpus striatrum accompanied this response. It is proposed that GABA and 5-HT may be closely related in modifying the nigrostriatal impulse flow. | [
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PMID:24689 | Dopamine-beta-hydroxylase release following acute selective sympathetic nerve stimulation of the heart, spleen and mesentery. | In increase in the concentration of dopamine-beta-hydroxylase (DBH) in the blood perfusates of the heart, spleen and mesentery of pithed rats was detected following selective sympathetic nerve stimulation. Furthermore, the concentration of enzyme released by each organ correlated with the stimulation frequency. The results provide further corroborative evidence that the concentration of DBH in the plasma is related to the degree of sympathetic tone. If the sampling methods described were applied to non-spinalized animals, the concentrations of DBH in organ perfusates could be used as an index of sympathetic nervous tone and thus provide a useful preparation for the study of drugs on the sympathetic nervous system. | Dopamine-beta-hydroxylase release following acute selective sympathetic nerve stimulation of the heart, spleen and mesentery. In increase in the concentration of dopamine-beta-hydroxylase (DBH) in the blood perfusates of the heart, spleen and mesentery of pithed rats was detected following selective sympathetic nerve stimulation. Furthermore, the concentration of enzyme released by each organ correlated with the stimulation frequency. The results provide further corroborative evidence that the concentration of DBH in the plasma is related to the degree of sympathetic tone. If the sampling methods described were applied to non-spinalized animals, the concentrations of DBH in organ perfusates could be used as an index of sympathetic nervous tone and thus provide a useful preparation for the study of drugs on the sympathetic nervous system. | [
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PMID:24690 | Possible differences in alpha-adrenoceptors in rabbit ileum and spleen. | In isolated tissues from reserpinized rabbits (5 mg kg-1, i.m. 20 h before experiment) and in the presence of cocaine (3 x 10(-5) M), corticosterone (2.8 x 10(-5)M), tropolone (3 x 10(-5) M), propranolol (4 x 10(-5)M) and disodium EDTA (3 X 10(-5)M), the potency ratios (relative to (-)-noradrenaline) of (-)adrenaline, (-)-phenylephrine and (+/-)-methoxamine were (m+/-s.e.) 2.03 +/- 0.13, 0.045 +/- 0.003 and 0.0062 +/- 0.0018 respectively in splenic strips and 1.77 +/- 0.41, 0.093 +/- 0.018 and 0.029 +/- 0.004 respectively in isolated ileum. Although the pA2 values for phentolamine and thymoxamine against (-)-noradrenaline in the two tissues were very similar there was a statistically significant difference when using yohimbine as the alpha-adrenoceptor blocking agent (pA2 = 6.80 +/- 0.30 in spleen; 5.60 +/- 0.12 in ileum). These differences suggest that the alpha-adrenoceptor in the two tissues is not identical. The pA2 value of phentolamine in rabbits ileum was not significantly different whether (-)noradrenaline or (+/-) methoxamine was used as agonist (7.91 +/- 0.07 and 7.97 +/- 0.06 respectively) while that of yohimbine was 5.56 +/- 0.10 using (-)noradrenaline and 6.19 +/- 0.12 using (+/-)methoxamine. In the light of this latter result and, considering the scatter of the experimentally determined values, there may be two alpha-adrenoceptors in rabbit ileum and either or both may not be identical in all respects to the alpha-adrenoceptor found in rabbit spleen. | Possible differences in alpha-adrenoceptors in rabbit ileum and spleen. In isolated tissues from reserpinized rabbits (5 mg kg-1, i.m. 20 h before experiment) and in the presence of cocaine (3 x 10(-5) M), corticosterone (2.8 x 10(-5)M), tropolone (3 x 10(-5) M), propranolol (4 x 10(-5)M) and disodium EDTA (3 X 10(-5)M), the potency ratios (relative to (-)-noradrenaline) of (-)adrenaline, (-)-phenylephrine and (+/-)-methoxamine were (m+/-s.e.) 2.03 +/- 0.13, 0.045 +/- 0.003 and 0.0062 +/- 0.0018 respectively in splenic strips and 1.77 +/- 0.41, 0.093 +/- 0.018 and 0.029 +/- 0.004 respectively in isolated ileum. Although the pA2 values for phentolamine and thymoxamine against (-)-noradrenaline in the two tissues were very similar there was a statistically significant difference when using yohimbine as the alpha-adrenoceptor blocking agent (pA2 = 6.80 +/- 0.30 in spleen; 5.60 +/- 0.12 in ileum). These differences suggest that the alpha-adrenoceptor in the two tissues is not identical. The pA2 value of phentolamine in rabbits ileum was not significantly different whether (-)noradrenaline or (+/-) methoxamine was used as agonist (7.91 +/- 0.07 and 7.97 +/- 0.06 respectively) while that of yohimbine was 5.56 +/- 0.10 using (-)noradrenaline and 6.19 +/- 0.12 using (+/-)methoxamine. In the light of this latter result and, considering the scatter of the experimentally determined values, there may be two alpha-adrenoceptors in rabbit ileum and either or both may not be identical in all respects to the alpha-adrenoceptor found in rabbit spleen. | [
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PMID:24691 | The synthesis and urinary estimation of N-hydroxyaprobarbitone. | N-Hydroxyaprobarbitone (I) has been synthesized by oxidation of aprobarbitone, characterized, and a method developed for its estimation in urine. It has shown to be no more than a minor metabolite (less than 4%) in the human metabolism of aprobarbitone. | The synthesis and urinary estimation of N-hydroxyaprobarbitone. N-Hydroxyaprobarbitone (I) has been synthesized by oxidation of aprobarbitone, characterized, and a method developed for its estimation in urine. It has shown to be no more than a minor metabolite (less than 4%) in the human metabolism of aprobarbitone. | [
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PMID:24692 | Aggregation of antidepressant drugs in aqueous solution. | Light scattering, conductivity and pH methods have been used to examine the aggregation in aqueous solution of a series of antidepressant drugs. The drugs investigated included the hydrochlorides of amitriptyline, butriptyline, protriptyline, nortriptyline, imipramine, desipramine, clomipramine, dothiepin, dibenzepin, opipramol, iprindole, doxepin, mianserin and maprotiline. No significant association of dibenzepin, mianserin or maprotiline hydrochlorides could be detected up to their respective solubility limits. A micellar pattern of association was established for all other compounds. Critical micelle concentrations and micellar properties are reported. | Aggregation of antidepressant drugs in aqueous solution. Light scattering, conductivity and pH methods have been used to examine the aggregation in aqueous solution of a series of antidepressant drugs. The drugs investigated included the hydrochlorides of amitriptyline, butriptyline, protriptyline, nortriptyline, imipramine, desipramine, clomipramine, dothiepin, dibenzepin, opipramol, iprindole, doxepin, mianserin and maprotiline. No significant association of dibenzepin, mianserin or maprotiline hydrochlorides could be detected up to their respective solubility limits. A micellar pattern of association was established for all other compounds. Critical micelle concentrations and micellar properties are reported. | [
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PMID:24706 | Formation of liquid crystal and other non-fluid phases in emulsions containing non-ionic surfactants. | The relation between the phase distribution of components and the nature and properties of dispersions formed in a four-component system containing dodecane, water and two homogeneous non-ionic surfactants has been investigated by phase-rule techniques. Phase equilibrium diagrams for the four ternary systems were investigated and the phase boundaries for the various regions formed in these systems were defined by synthetic and analytical methods. In some of the ternary diagrams a narrow three-phase region occurs where 'gel' or liquid crystal as well as two isotropic liquid phases are present. Dispersions containing the gel phase were very stable. The four-component system has not been fully investigated due to the presence of extensive liquid crystal phases and three-phase regions. However, some distribution data for the quaternary system was obtained. In the presence of sufficient amounts of the short chain non-ionic surfactant C10H21(O.CH2.CH2)3OH, a three-liquid phase region occurs where multiple drops readily form. | Formation of liquid crystal and other non-fluid phases in emulsions containing non-ionic surfactants. The relation between the phase distribution of components and the nature and properties of dispersions formed in a four-component system containing dodecane, water and two homogeneous non-ionic surfactants has been investigated by phase-rule techniques. Phase equilibrium diagrams for the four ternary systems were investigated and the phase boundaries for the various regions formed in these systems were defined by synthetic and analytical methods. In some of the ternary diagrams a narrow three-phase region occurs where 'gel' or liquid crystal as well as two isotropic liquid phases are present. Dispersions containing the gel phase were very stable. The four-component system has not been fully investigated due to the presence of extensive liquid crystal phases and three-phase regions. However, some distribution data for the quaternary system was obtained. In the presence of sufficient amounts of the short chain non-ionic surfactant C10H21(O.CH2.CH2)3OH, a three-liquid phase region occurs where multiple drops readily form. | [
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PMID:24707 | Factors affecting the disintegration and dissolution of chloroquine phosphate/starch tablets. | A study has been made of the effects produced on the disintegration and dissolution times of chloroquine phosphate tablets by varying their moisture and starch contents and the distribution of the starch in the formulation. 3 to 5% w/w of moisture produces a maximum in the disintegration and dissolution times. Starch added externally as a powder acts only as a disintegrating agent for the tablets, but starch added internally as a paste during granulation acts both as a binding agent and a disintegrant. | Factors affecting the disintegration and dissolution of chloroquine phosphate/starch tablets. A study has been made of the effects produced on the disintegration and dissolution times of chloroquine phosphate tablets by varying their moisture and starch contents and the distribution of the starch in the formulation. 3 to 5% w/w of moisture produces a maximum in the disintegration and dissolution times. Starch added externally as a powder acts only as a disintegrating agent for the tablets, but starch added internally as a paste during granulation acts both as a binding agent and a disintegrant. | [
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PMID:24708 | Control of the composition of gentamicin sulphate by proton magnetic resonance spectrometry. | The proportions of the main components present in gentamicin sulphate complex, gentamicins C1, C1a and C2, can be monitored by 1H nuclear magnetic resonance (nmr) spectrometry. The method depends on measurement of the peak heights of signals for N-methyl and C-methyl groups present in all three components and of those present in C1 and C2 only, followed by calculation of peak height ratios to control composition within acceptable limits. The precision and reproducibility of the method have been established through two collaborative studies each involving seven laboratories. In the second study, with an improved procedure, the mean variance between laboratories with 10 samples was 3.4 X 10(-4) for the N-methyl ratio of the peak at delta 2.75 to that at delta 2.95, and 1.25 X 10(-3) for the C-methyl ratio of the peak at delta 1.25 to that at delta 1.35. Within laboratories the mean variance for triplicate determinations was 7.4 X 10(-5) and 8.9 X 10(-5) respectively. The data presented here form the experimental basis for the test controlling the composition of gentamicin sulphate in the British Pharmacopoeia 1973: Addendum 1975, and for the introduction into the British Pharmacopoeia of nmr spectrometry as an analytical technique. The reference standards and all batches of gentamicin sulphate intended for therapeutic use in the United Kingdom examined by this procedure comply with the limits laid down. | Control of the composition of gentamicin sulphate by proton magnetic resonance spectrometry. The proportions of the main components present in gentamicin sulphate complex, gentamicins C1, C1a and C2, can be monitored by 1H nuclear magnetic resonance (nmr) spectrometry. The method depends on measurement of the peak heights of signals for N-methyl and C-methyl groups present in all three components and of those present in C1 and C2 only, followed by calculation of peak height ratios to control composition within acceptable limits. The precision and reproducibility of the method have been established through two collaborative studies each involving seven laboratories. In the second study, with an improved procedure, the mean variance between laboratories with 10 samples was 3.4 X 10(-4) for the N-methyl ratio of the peak at delta 2.75 to that at delta 2.95, and 1.25 X 10(-3) for the C-methyl ratio of the peak at delta 1.25 to that at delta 1.35. Within laboratories the mean variance for triplicate determinations was 7.4 X 10(-5) and 8.9 X 10(-5) respectively. The data presented here form the experimental basis for the test controlling the composition of gentamicin sulphate in the British Pharmacopoeia 1973: Addendum 1975, and for the introduction into the British Pharmacopoeia of nmr spectrometry as an analytical technique. The reference standards and all batches of gentamicin sulphate intended for therapeutic use in the United Kingdom examined by this procedure comply with the limits laid down. | [
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PMID:24709 | Isolation and identification of the fluorescent degradation product of some beta-lactam antibiotics. | A fluorescent degradation product of cephalexin, recently employed in the determination of this antibiotic in aqueous solution and human plasma, has been isolated and identified as 2-hydroxy-3-phenyl-6-methylpyrazine. Spectroscopically and chromatographically the product is indistinguishable from that obtained by the same method from a series of other cephalosporins and penicillins bearing an alpha-amino group on the side-chain, and from the pyrazine structure produced by synthesis. Two reported methods for the fluorimetric determination of ampicillin have also been found to yield the same degradation product and not the suggested diketopiperazine structure. | Isolation and identification of the fluorescent degradation product of some beta-lactam antibiotics. A fluorescent degradation product of cephalexin, recently employed in the determination of this antibiotic in aqueous solution and human plasma, has been isolated and identified as 2-hydroxy-3-phenyl-6-methylpyrazine. Spectroscopically and chromatographically the product is indistinguishable from that obtained by the same method from a series of other cephalosporins and penicillins bearing an alpha-amino group on the side-chain, and from the pyrazine structure produced by synthesis. Two reported methods for the fluorimetric determination of ampicillin have also been found to yield the same degradation product and not the suggested diketopiperazine structure. | [
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PMID:24710 | Survival of plasmid-containing strains of Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus in phenylmercuric nitrate and thiomersal. | Strains of Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus harbouring plasmids that confer mercury resistance grew in nutrient broth containing concentrations of phenylmercuric nitrate (PMN) that were inhibitory to isogenic plasmid-less strains. Decimal reduction times of P. aeruginosa and S. aureus in aqueous PMN solution were also increased by the presence of plasmids. The viable count of a plasmid-containing P. aeruginosa strain in Davis and Mingioli's minimal medium (DM) containing 10 microgram ml-1 PMN fell by approximately 99% after 5 h. The count then remained constant for two weeks, when growth recommenced. This pattern of death followed by growth was also observed with the P. aeruginosa strain in DM + 10 microgram ml-1 thiomersal. | Survival of plasmid-containing strains of Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus in phenylmercuric nitrate and thiomersal. Strains of Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus harbouring plasmids that confer mercury resistance grew in nutrient broth containing concentrations of phenylmercuric nitrate (PMN) that were inhibitory to isogenic plasmid-less strains. Decimal reduction times of P. aeruginosa and S. aureus in aqueous PMN solution were also increased by the presence of plasmids. The viable count of a plasmid-containing P. aeruginosa strain in Davis and Mingioli's minimal medium (DM) containing 10 microgram ml-1 PMN fell by approximately 99% after 5 h. The count then remained constant for two weeks, when growth recommenced. This pattern of death followed by growth was also observed with the P. aeruginosa strain in DM + 10 microgram ml-1 thiomersal. | [
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PMID:24711 | Melanocyte-stimulating hormone--mimetic action of the phenothiazines. | We have compared the melanophore-stimulating action of four phenothiazines, trifluoperazine, perphenazine, chlorpromazine, and prochlorperazine, with alpha-MSH on the skin of the lizard Anolis carolinensis, using a new rate method of bioassay. The dose-response curves for the phenothiazines were parallel to that of alpha-MSH, and when given together alpha-MSH and chlorpromazine were additive. The phenothiazines may therefore stimulate melanosome dispersion in the lizard skin by the same mechanism as alpha-MSH; a MSH-mimetic action of phenothiazines may similarly explain their pigmentary action in man. The pigmentary potency of the phenothiazines corresponded with their therapeutic potency in man; this is in keeping with a neuro-regulatory role for MSH peptides and suggests a possible therapeutic use for them. | Melanocyte-stimulating hormone--mimetic action of the phenothiazines. We have compared the melanophore-stimulating action of four phenothiazines, trifluoperazine, perphenazine, chlorpromazine, and prochlorperazine, with alpha-MSH on the skin of the lizard Anolis carolinensis, using a new rate method of bioassay. The dose-response curves for the phenothiazines were parallel to that of alpha-MSH, and when given together alpha-MSH and chlorpromazine were additive. The phenothiazines may therefore stimulate melanosome dispersion in the lizard skin by the same mechanism as alpha-MSH; a MSH-mimetic action of phenothiazines may similarly explain their pigmentary action in man. The pigmentary potency of the phenothiazines corresponded with their therapeutic potency in man; this is in keeping with a neuro-regulatory role for MSH peptides and suggests a possible therapeutic use for them. | [
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PMID:24712 | The effect of various topical antibiotic and antibacterial agents on the middle and inner ear of the guinea-pig. | The topical application of antibiotics and anti-inflammatory agents is used to reduce inflammation of the middle ear mucosa and irradicate infection. Many of the antibiotic and anti-inflammatory preparations so used contain compounds known to be or are potentially ototoxic. Eighteen antibiotic, antibacterial, antifungal and anti-inflammatory compounds and four commonly used solvents were screened for the absence of ototoxicity and inflammation to the middle ear mucosa. All compounds were injected intra-tympanically and sensory hair cells loss recorded graphically. Inflammation of the middle ear mucosa was assessed macroscopically. Of the 18 compounds only three, penicillin, carbenicillin and nystatin were free of hair cell toxicity and inflammatory effects on the middle ear mucosa. Only one of the commonly used solvents was free of side effects. | The effect of various topical antibiotic and antibacterial agents on the middle and inner ear of the guinea-pig. The topical application of antibiotics and anti-inflammatory agents is used to reduce inflammation of the middle ear mucosa and irradicate infection. Many of the antibiotic and anti-inflammatory preparations so used contain compounds known to be or are potentially ototoxic. Eighteen antibiotic, antibacterial, antifungal and anti-inflammatory compounds and four commonly used solvents were screened for the absence of ototoxicity and inflammation to the middle ear mucosa. All compounds were injected intra-tympanically and sensory hair cells loss recorded graphically. Inflammation of the middle ear mucosa was assessed macroscopically. Of the 18 compounds only three, penicillin, carbenicillin and nystatin were free of hair cell toxicity and inflammatory effects on the middle ear mucosa. Only one of the commonly used solvents was free of side effects. | [
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PMID:24713 | Fate of [14C]warfarin in guinea-pigs: effect of a concomitant single dose of salicylate. | When a single dose of sodium salicylate (177.8 mg kg-1, by mouth) was given with [14C] warfarin (1 mg kg-1, i.p.) to guinea-pigs, the salicylate depressed the blood concentrations of 14C for 6 h. At 1 h, salicylate increased the distribution of 14C in the liver and brain, but at 1 and 6 h it was decreased in the blood and kidney. A significant portion of the 14C was excreted into the bile, but was subject to enterohepatic circulation and then excreted by the kidney. There was an enhancement of the biliary elimination of 14C in the first 5 h after salicylate and a decrease in 14C concentration in blood; the proportion of warfarin to its metabolites excreted in the urine and bile was unchanged. Salicylate displaced serum protein bound [14C]warfarin in vitro. Salicylate increases the initial biliary elimination of warfarin by displacing some of that bound to plasma protein. This facilitated uptake of warfarin by the liver where it was metabolized. This effect of salicylate did not modify the hypoprothrombinaemia produced by warfarin. | Fate of [14C]warfarin in guinea-pigs: effect of a concomitant single dose of salicylate. When a single dose of sodium salicylate (177.8 mg kg-1, by mouth) was given with [14C] warfarin (1 mg kg-1, i.p.) to guinea-pigs, the salicylate depressed the blood concentrations of 14C for 6 h. At 1 h, salicylate increased the distribution of 14C in the liver and brain, but at 1 and 6 h it was decreased in the blood and kidney. A significant portion of the 14C was excreted into the bile, but was subject to enterohepatic circulation and then excreted by the kidney. There was an enhancement of the biliary elimination of 14C in the first 5 h after salicylate and a decrease in 14C concentration in blood; the proportion of warfarin to its metabolites excreted in the urine and bile was unchanged. Salicylate displaced serum protein bound [14C]warfarin in vitro. Salicylate increases the initial biliary elimination of warfarin by displacing some of that bound to plasma protein. This facilitated uptake of warfarin by the liver where it was metabolized. This effect of salicylate did not modify the hypoprothrombinaemia produced by warfarin. | [
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PMID:24714 | Inhibitors of gastric lesions in the rat. | The production by stress of gastric lesions in rats was inhibited by metiamide and by mepyramine. Lesions induced by indomethacin treatment were inhibited by mepyramine but not by metiamide. Those induced by aspirin treatment in pylorus-ligated rats were not affected by either antihistamine drug. Oral glutamine inhibited lesion production in all three systems whereas aspirin orally markedly potentiated it. Sodium salicylate inhibited both indomethacin-induced lesions and those produced by aspirin in pylorus-ligated rats. On the other hand, copper salicylate inhibited stress-induced lesions and it, like copper aspirinate, also markedly reduced the extent of lesions produced by aspirin. On the basis of these results, stress-induced lesion production offers a suitable animal model for testing anti-ulcer drugs as it is, like most human gastric ulcers, inhibited by H2-receptor inhibitors like metiamide. | Inhibitors of gastric lesions in the rat. The production by stress of gastric lesions in rats was inhibited by metiamide and by mepyramine. Lesions induced by indomethacin treatment were inhibited by mepyramine but not by metiamide. Those induced by aspirin treatment in pylorus-ligated rats were not affected by either antihistamine drug. Oral glutamine inhibited lesion production in all three systems whereas aspirin orally markedly potentiated it. Sodium salicylate inhibited both indomethacin-induced lesions and those produced by aspirin in pylorus-ligated rats. On the other hand, copper salicylate inhibited stress-induced lesions and it, like copper aspirinate, also markedly reduced the extent of lesions produced by aspirin. On the basis of these results, stress-induced lesion production offers a suitable animal model for testing anti-ulcer drugs as it is, like most human gastric ulcers, inhibited by H2-receptor inhibitors like metiamide. | [
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PMID:24729 | Effects of catecholamine depleting drugs and d-amphetamine on self-stimulation of the substantia nigra and locus coeruleus. | 6-Hydroxydopamine treatments which preferentially depleted either norepinephrine or dopamine were used to define the importance of these transmitter systems in the behavioral alterations produced by catecholamine synthesis inhibitors and d-amphetamine on self-stimulation of the locus coeruleus and substantia nigra. After chronic reduction of brain dopamine, an acute depression of self-stimulation of both the locus coeruleus and substantia nigra occurred. Preferential depletion of norepinephrine with 6-hydroxydopamine did not result in a significant decrease in self-stimulation of locus coeruleus or substantia nigra. However, a dose of alpha-methyltyrosine wihch had no effect in control rats or in rats with brain norepinephrine depleted caused a significant reduction in responding at both electrode placements in animals depleted of brain dopamine. Administration of U-14,624 affected neither substantia nigra nor locus coeruleus self-stimulation, even though it produced an additional 70% depletion of norepinephrine. When d-amphetamine sulfate was given to 6-hydroxydopamine-treated rats, the facilitation of self-stimulation produced by this compound was significantly attenuated in rats with prior depletion of brain dopamine. Depletion of brain norepinephrine did not affect the actions of d-amphetamine on self-stimulation. In other experiments, the actions of d-amphetamine to increase self-stimulation of animals pretreated with reserpine was found to be antagonized by alpha-methyltyrosine but not by U-14,624. Results suggest that drugs can alter self-stimulation of a site in brain anatomically associated with noradrenergic neural pathways and self-stimulation of a site primarily associated with dopaminergic pathways in a similar manner. These data also provided evidence for the involvement of dopamine fibers in the pharmacological actions of d-amphetamine, reserpine and alpha-methyltyrosine. | Effects of catecholamine depleting drugs and d-amphetamine on self-stimulation of the substantia nigra and locus coeruleus. 6-Hydroxydopamine treatments which preferentially depleted either norepinephrine or dopamine were used to define the importance of these transmitter systems in the behavioral alterations produced by catecholamine synthesis inhibitors and d-amphetamine on self-stimulation of the locus coeruleus and substantia nigra. After chronic reduction of brain dopamine, an acute depression of self-stimulation of both the locus coeruleus and substantia nigra occurred. Preferential depletion of norepinephrine with 6-hydroxydopamine did not result in a significant decrease in self-stimulation of locus coeruleus or substantia nigra. However, a dose of alpha-methyltyrosine wihch had no effect in control rats or in rats with brain norepinephrine depleted caused a significant reduction in responding at both electrode placements in animals depleted of brain dopamine. Administration of U-14,624 affected neither substantia nigra nor locus coeruleus self-stimulation, even though it produced an additional 70% depletion of norepinephrine. When d-amphetamine sulfate was given to 6-hydroxydopamine-treated rats, the facilitation of self-stimulation produced by this compound was significantly attenuated in rats with prior depletion of brain dopamine. Depletion of brain norepinephrine did not affect the actions of d-amphetamine on self-stimulation. In other experiments, the actions of d-amphetamine to increase self-stimulation of animals pretreated with reserpine was found to be antagonized by alpha-methyltyrosine but not by U-14,624. Results suggest that drugs can alter self-stimulation of a site in brain anatomically associated with noradrenergic neural pathways and self-stimulation of a site primarily associated with dopaminergic pathways in a similar manner. These data also provided evidence for the involvement of dopamine fibers in the pharmacological actions of d-amphetamine, reserpine and alpha-methyltyrosine. | [
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