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PMID:11877 | The seeded growth of calcium phosphates. The kinetics of growth of dicalcium phosphate dihydrate on hydroxyapatite. | The kinetics of growth of calcium phosphate on synthetic HAP seed crystals has been studied at 37 degrees C and at pH values of 4.5 and 5.0 by the pH-stat controlled addition of base. Following an induction, DCPD crystal growth takes place with growth kinetics characteristic of the crystallization of pure DCPD seed. The effect of stirring rate, HAP seed concentration and initial degree of supersaturation with respect to DCPD on the kinetics of the growth process have been determined. The crystallization appears to be controlled by a surface reaction and nucleation of DCPD on the HAP substrate is completed in the initial induction period. Formation of DCPD under these conditions closely simulates plaque and calculus production in the mouth. Thus calculus that develops from the initial plaque is known to contain an appreciable amount of DCPD. | The seeded growth of calcium phosphates. The kinetics of growth of dicalcium phosphate dihydrate on hydroxyapatite. The kinetics of growth of calcium phosphate on synthetic HAP seed crystals has been studied at 37 degrees C and at pH values of 4.5 and 5.0 by the pH-stat controlled addition of base. Following an induction, DCPD crystal growth takes place with growth kinetics characteristic of the crystallization of pure DCPD seed. The effect of stirring rate, HAP seed concentration and initial degree of supersaturation with respect to DCPD on the kinetics of the growth process have been determined. The crystallization appears to be controlled by a surface reaction and nucleation of DCPD on the HAP substrate is completed in the initial induction period. Formation of DCPD under these conditions closely simulates plaque and calculus production in the mouth. Thus calculus that develops from the initial plaque is known to contain an appreciable amount of DCPD. | [
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PMID:11878 | Pyrophosphatase and ATPase of isolated cartilage matrix vesicles. | Some of the characteristics of the pyrophosphatase and ATPase activities studied in isolated cartilage matrix vesicles were found to be similar to those already reported for the solubilized and purified bone alkaline phosphatase. Thus, the pH optimum of the pyrophosphatase activity responded similarly to changes in the concentration of Mg2+, Ca2+, and PPi. Further, the ATPase activity was not activated by Ca2+ in the presence of an optimal Mg2+ concentration. It is proposed that a function of the alkaline phosphatase of matrix vesicles in vivo is to hydrolyze the substrates PPi, ADP, and ATP, which are known inhibitors of calcium phosphate precipitation. | Pyrophosphatase and ATPase of isolated cartilage matrix vesicles. Some of the characteristics of the pyrophosphatase and ATPase activities studied in isolated cartilage matrix vesicles were found to be similar to those already reported for the solubilized and purified bone alkaline phosphatase. Thus, the pH optimum of the pyrophosphatase activity responded similarly to changes in the concentration of Mg2+, Ca2+, and PPi. Further, the ATPase activity was not activated by Ca2+ in the presence of an optimal Mg2+ concentration. It is proposed that a function of the alkaline phosphatase of matrix vesicles in vivo is to hydrolyze the substrates PPi, ADP, and ATP, which are known inhibitors of calcium phosphate precipitation. | [
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PMID:11879 | The hydrolytic activity of L-ascorbic acid. | The ability of L-ascorbic acid to catalyze the liberation of 4-methylumbelliferone from 4-methylumbelliferyl-beta-D-N-acetylglucosaminide, 4-methylumbelliferyl-beta-D-glucuronide, 4-methylumbelliferyl-alpha-D-mannoside, and 4-methylumbelliferyl-beta-D-galactoside is documented. There is an apparent metal and oxygen dependency. The cleavage of two lipids was shown in addition to the hydrolysis of these fluorogenic glycosides. Galactose was liberated from galactosyl-6-[3H]ceramide and oleic acid from cholesterol-[1-14C]oleate by L-ascorbic acid under conditins usually used for in vitro incubations. In common with most in vitro systems, only a small percentage of substrate was degraded. | The hydrolytic activity of L-ascorbic acid. The ability of L-ascorbic acid to catalyze the liberation of 4-methylumbelliferone from 4-methylumbelliferyl-beta-D-N-acetylglucosaminide, 4-methylumbelliferyl-beta-D-glucuronide, 4-methylumbelliferyl-alpha-D-mannoside, and 4-methylumbelliferyl-beta-D-galactoside is documented. There is an apparent metal and oxygen dependency. The cleavage of two lipids was shown in addition to the hydrolysis of these fluorogenic glycosides. Galactose was liberated from galactosyl-6-[3H]ceramide and oleic acid from cholesterol-[1-14C]oleate by L-ascorbic acid under conditins usually used for in vitro incubations. In common with most in vitro systems, only a small percentage of substrate was degraded. | [
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PMID:11880 | Adenosine triphosphate - dependent calcium binding and accumulation by guinea pig cardiac sarcolemma. | Sarcolemma isolated from guinea pig heart ventricles possessed ATP-dependent Ca2+ binding and accumulation (+ oxalate) activities which were not inhibited by sodium azide, oligomycin, or ruthenium red. Ca2+ binding and accumulation by sarcolemma were sensitive to pH, the optimum being about pH 6.8. The concentrations of ATP required for half-maximal binding and accumulation were 94.3 and 172 muM, respectively. Mg2+ up to 5 mM significantly enhanced both activities but was inhibitory at higher concentrations (greater than 10 mM). Sarcolemmal Ca2+ binding and accumulation were stimulated 100% by K+, half-maximal enhancement occurring at 5-10 mM K+. Ca2+ binding and accumulation were both saturable processes and the respective apparent Km values for Ca2+ were 16.4 and 14.3 muM. Ca2+ binding by sarcolemma was a rapid process and the bound Ca2+ was released upon depletion of ATP in the medium. It is suggested that the sarcolemmal Ca2+ transport system may well be of significance in regulation of the contraction-relaxation cycle of cardiac muscle. | Adenosine triphosphate - dependent calcium binding and accumulation by guinea pig cardiac sarcolemma. Sarcolemma isolated from guinea pig heart ventricles possessed ATP-dependent Ca2+ binding and accumulation (+ oxalate) activities which were not inhibited by sodium azide, oligomycin, or ruthenium red. Ca2+ binding and accumulation by sarcolemma were sensitive to pH, the optimum being about pH 6.8. The concentrations of ATP required for half-maximal binding and accumulation were 94.3 and 172 muM, respectively. Mg2+ up to 5 mM significantly enhanced both activities but was inhibitory at higher concentrations (greater than 10 mM). Sarcolemmal Ca2+ binding and accumulation were stimulated 100% by K+, half-maximal enhancement occurring at 5-10 mM K+. Ca2+ binding and accumulation were both saturable processes and the respective apparent Km values for Ca2+ were 16.4 and 14.3 muM. Ca2+ binding by sarcolemma was a rapid process and the bound Ca2+ was released upon depletion of ATP in the medium. It is suggested that the sarcolemmal Ca2+ transport system may well be of significance in regulation of the contraction-relaxation cycle of cardiac muscle. | [
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|
PMID:11881 | Histone-histone interactions. II. Structural stability of the histone H3-H4 complex. | The stability of the histone H3-H4 complex toward urea, changes in pH and ionic strength, and certain chemical modifications have been examined by gel electrophoresis anc circular dichronism. When uncomplexed, the two cysteine residues of histone H3 become rapidly oxidized, forming an intramolecular disulfide bridge which apparently blocks complex formation on return to complexing conditions. The complex was found to be unstable toward low values of pH and ionic strength, concentrations of urea exceeding 1 M, modifications of the cysteine residues, and fragmention in which the C terminal portions of either H3 or H4 are removed. A possible structure for this complex is proposed. | Histone-histone interactions. II. Structural stability of the histone H3-H4 complex. The stability of the histone H3-H4 complex toward urea, changes in pH and ionic strength, and certain chemical modifications have been examined by gel electrophoresis anc circular dichronism. When uncomplexed, the two cysteine residues of histone H3 become rapidly oxidized, forming an intramolecular disulfide bridge which apparently blocks complex formation on return to complexing conditions. The complex was found to be unstable toward low values of pH and ionic strength, concentrations of urea exceeding 1 M, modifications of the cysteine residues, and fragmention in which the C terminal portions of either H3 or H4 are removed. A possible structure for this complex is proposed. | [
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PMID:11882 | Circular dichroism studies of sheep beta-lipotropic hormone. | The far ultraviolet circular dichroism spectra of sheep beta-lipotropic hormone (beta-LPH) were recorded under different conditions of pH, temperature, salt concentration, and solvent composition. Results confirm the stability of the hormone in strong basic or acidic solutions; moreover, temperatures up to 50 degrees C do not seem to affect noticeably the conformation of beta-LPH. However, increasing the NaC1 concentration or addition of dioxane in the solution brings about a conformational transition of the chain, interpreted as an increase in the helical content. The method of Yang (Chen, Y.H., Yang, J. T. & Martinez, H. M. (1972) Biochemistry 11, 4120-4131) was used to compute the proportion of helical, beta, and unordered forms of the hormone chain. The proportions are compared with those obtained from Fasman's predictive method (Chou, P. Y & Fasman, G. D. (1974) Biochemistry 13, 211-221 and Chou, P. Y. & Fasman, G. D. (1974) Biochemistry 13, 222-245) based on the known amino acid sequence of beta-LPH. | Circular dichroism studies of sheep beta-lipotropic hormone. The far ultraviolet circular dichroism spectra of sheep beta-lipotropic hormone (beta-LPH) were recorded under different conditions of pH, temperature, salt concentration, and solvent composition. Results confirm the stability of the hormone in strong basic or acidic solutions; moreover, temperatures up to 50 degrees C do not seem to affect noticeably the conformation of beta-LPH. However, increasing the NaC1 concentration or addition of dioxane in the solution brings about a conformational transition of the chain, interpreted as an increase in the helical content. The method of Yang (Chen, Y.H., Yang, J. T. & Martinez, H. M. (1972) Biochemistry 11, 4120-4131) was used to compute the proportion of helical, beta, and unordered forms of the hormone chain. The proportions are compared with those obtained from Fasman's predictive method (Chou, P. Y & Fasman, G. D. (1974) Biochemistry 13, 211-221 and Chou, P. Y. & Fasman, G. D. (1974) Biochemistry 13, 222-245) based on the known amino acid sequence of beta-LPH. | [
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PMID:11883 | The fusion of erythrocytes by treatment with proteolytic enzymes and polyethylene glycol. | Polyethylene glycol (PEG) has been utilized to induce homokaryocyte formation in avian and mammalian erythrocytes previously treated with proteolytic enzymes. PEG of molecular weight 6,000-7,5000 was found superior to 1,500 and 20,000 MW PEG. Cells exposed to protease alone, prior to PEG treatment, fused to a high degree (60-95% multinucleated cells), whereas trypsin or pepsin treatment alone allowed very little fusion (2.5%). Trypsin lowered the effectiveness of protease when used in combination. Cells which were not treated with proteolytic enzymes agglutinated in the presence of PEG but did not fuse to a significant extent (0.01%). Fusion was also markedly dependent upon the rate at which PEG was eluted during the fusion process. Electron microscopy indicated that fusion began during the elution of PEG from the agglutinated cells. | The fusion of erythrocytes by treatment with proteolytic enzymes and polyethylene glycol. Polyethylene glycol (PEG) has been utilized to induce homokaryocyte formation in avian and mammalian erythrocytes previously treated with proteolytic enzymes. PEG of molecular weight 6,000-7,5000 was found superior to 1,500 and 20,000 MW PEG. Cells exposed to protease alone, prior to PEG treatment, fused to a high degree (60-95% multinucleated cells), whereas trypsin or pepsin treatment alone allowed very little fusion (2.5%). Trypsin lowered the effectiveness of protease when used in combination. Cells which were not treated with proteolytic enzymes agglutinated in the presence of PEG but did not fuse to a significant extent (0.01%). Fusion was also markedly dependent upon the rate at which PEG was eluted during the fusion process. Electron microscopy indicated that fusion began during the elution of PEG from the agglutinated cells. | [
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|
PMID:11884 | Congenital retroflexion of the penis and inguinal cryptorchidism in a presumptive bovine twin with a 60,XY/60,XX/61,XX,+cen chromosome constitution. | Studies were carried out on a yearling Holstein with external genitalia resembling those of a freemartin whose birth and developmental history was unknown. Dissection following slaughter showed testes close to the external inguinal rings, an underdeveloped penis coiled up subcutaneously in the perineum and terminating in a deep fossa at the level of the ischial arch and no evidence of a female genital tract. Chromosome analyses showed 60,XY cells in the blood and 60,XX and 61,XX,+cen cells in other tissues. It is postulated that the animal had a basic 60,XX/61,XX+cen mixoploid chromosome constitution, that the centric fragment functioned as a Y chromosome or as an autosomal modifier of the X chromosome in sex determination which accounted for the animal's Klinefelter syndrome-like abnormalities, and that animal was also twin to a bull which accounted for the presence of 60,XY cells in the blood. | Congenital retroflexion of the penis and inguinal cryptorchidism in a presumptive bovine twin with a 60,XY/60,XX/61,XX,+cen chromosome constitution. Studies were carried out on a yearling Holstein with external genitalia resembling those of a freemartin whose birth and developmental history was unknown. Dissection following slaughter showed testes close to the external inguinal rings, an underdeveloped penis coiled up subcutaneously in the perineum and terminating in a deep fossa at the level of the ischial arch and no evidence of a female genital tract. Chromosome analyses showed 60,XY cells in the blood and 60,XX and 61,XX,+cen cells in other tissues. It is postulated that the animal had a basic 60,XX/61,XX+cen mixoploid chromosome constitution, that the centric fragment functioned as a Y chromosome or as an autosomal modifier of the X chromosome in sex determination which accounted for the animal's Klinefelter syndrome-like abnormalities, and that animal was also twin to a bull which accounted for the presence of 60,XY cells in the blood. | [
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PMID:11886 | Isoferritin composition of tissues and serum in human cancers. | A highly sensitive technique for isoferritin detection using 125I-labeled monospecific anti-human liver ferritin antibody for the identification of isoferritins after the analysis of small quantities of ferritin by isoelectric focusing in polyacryl-amide gels was applied to the study of renal, pancreatic, and colonic carcinomas. In all tumors studied, the isoferritin composition differed from that of the corresponding normal tissue; major isoferritins with pl more basic than those of the normal tissues were consistently detected. Composition of purified ferritin from metastases closely resembled the isoferritin composition of the primary tumors. Examination of the serum isoferritin profiles of four patients with cancers did not reveal the presence of any tumor-specific changes in isoferritins. It is suggested that the abnormality in tissue ferritins in the three human cancers studied is the synthesis of major isoferritins in the more basic range, rather than the appearance of tumor-specific isoferritins in the more acidic range. | Isoferritin composition of tissues and serum in human cancers. A highly sensitive technique for isoferritin detection using 125I-labeled monospecific anti-human liver ferritin antibody for the identification of isoferritins after the analysis of small quantities of ferritin by isoelectric focusing in polyacryl-amide gels was applied to the study of renal, pancreatic, and colonic carcinomas. In all tumors studied, the isoferritin composition differed from that of the corresponding normal tissue; major isoferritins with pl more basic than those of the normal tissues were consistently detected. Composition of purified ferritin from metastases closely resembled the isoferritin composition of the primary tumors. Examination of the serum isoferritin profiles of four patients with cancers did not reveal the presence of any tumor-specific changes in isoferritins. It is suggested that the abnormality in tissue ferritins in the three human cancers studied is the synthesis of major isoferritins in the more basic range, rather than the appearance of tumor-specific isoferritins in the more acidic range. | [
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PMID:11887 | Increased guanylate cyclase activity and guanosine 3',5'-monophosphate content in ethionine-induced hepatomas. | Ethionine-induced hepatomas are characterized by high adenylate cyclase activity and cyclic adenosine 3',5'-monophosphate content relative to those of surrounding liver or liver from pair-fed control rats. The present study examined the properties of the guanylate cyclase-cyclic guanosine 3',5'-monophosphate (cGMP) system of these tissues. cGMP levels of the ethionine-induced hepatomas, determined in both specimens quick-forzen in situ and after in vitro incubation of tissue slices, were approximately 2 times higher than those of surrounding liver or controls. Higher cGMP in the tumors was associated with an increase in whole homogenate, soluble, and particulate guanylate cyclase activities, as well as an increase in soluble cGMP-phosphodiesterase activity. 3-Isobutyl-1-methylxanthine, a potent inhibitor of cGMP-phosphodiesterase activity, potentiated the differences in cGMP between slices of the hepatomas and surrounding liver or control, suggesting that the higher steady-state cGMP content of the tumors reflected enhanced basal cGMP synthesis which was partially offset by increased nucleotide degradation. In the hepatomas, a greater proportion of the total guanylate cyclase activity was located in the particulate cell fraction (31%) as compared to the subcellular distribution of enzyme activity in either surrounding liver or controls (15% of total in the particulate fraction). Carbamylcholine, which increased cGMP 3-fold in surrounding liver and controls, failed to alter cGMP levels inslices of hepatoma. Further, the relative changes in both cGMP accumulation and guanylate cyclase activity of the tumors in response to NaN3, NH2OH, and NaNO2 were blunted compared to surrounding liver or controls, although in each instance a response was clearly evident. Ethionine-induced hepatomas are thus characterized by: (a) significant increases in cGMP content and in guanylate cyclase and cGMP-phosphodiesterase activities, (b) a change in the subcellular distribution of guanylate cyclase, and (c) altered responsiveness of the guanylate cyclase-cGMP system to several agonists. | Increased guanylate cyclase activity and guanosine 3',5'-monophosphate content in ethionine-induced hepatomas. Ethionine-induced hepatomas are characterized by high adenylate cyclase activity and cyclic adenosine 3',5'-monophosphate content relative to those of surrounding liver or liver from pair-fed control rats. The present study examined the properties of the guanylate cyclase-cyclic guanosine 3',5'-monophosphate (cGMP) system of these tissues. cGMP levels of the ethionine-induced hepatomas, determined in both specimens quick-forzen in situ and after in vitro incubation of tissue slices, were approximately 2 times higher than those of surrounding liver or controls. Higher cGMP in the tumors was associated with an increase in whole homogenate, soluble, and particulate guanylate cyclase activities, as well as an increase in soluble cGMP-phosphodiesterase activity. 3-Isobutyl-1-methylxanthine, a potent inhibitor of cGMP-phosphodiesterase activity, potentiated the differences in cGMP between slices of the hepatomas and surrounding liver or control, suggesting that the higher steady-state cGMP content of the tumors reflected enhanced basal cGMP synthesis which was partially offset by increased nucleotide degradation. In the hepatomas, a greater proportion of the total guanylate cyclase activity was located in the particulate cell fraction (31%) as compared to the subcellular distribution of enzyme activity in either surrounding liver or controls (15% of total in the particulate fraction). Carbamylcholine, which increased cGMP 3-fold in surrounding liver and controls, failed to alter cGMP levels inslices of hepatoma. Further, the relative changes in both cGMP accumulation and guanylate cyclase activity of the tumors in response to NaN3, NH2OH, and NaNO2 were blunted compared to surrounding liver or controls, although in each instance a response was clearly evident. Ethionine-induced hepatomas are thus characterized by: (a) significant increases in cGMP content and in guanylate cyclase and cGMP-phosphodiesterase activities, (b) a change in the subcellular distribution of guanylate cyclase, and (c) altered responsiveness of the guanylate cyclase-cGMP system to several agonists. | [
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PMID:11888 | Activation of guanylate cyclase by streptozotocin and 1-methyl-1-nitrosourea. | Streptozotocin has been shown to induce the production of a variety of tumors in rats. The present report demonstrates that streptozotocin and 1-methyl-1-nitrosourea, a component of the streptozotocin molecule and a known carcinogen, stimulate the enzyme guanylate cyclase which catalyzes the production of guanosine 3',5'-monophosphate. At a maximal concentration of 3 mg/ml, these agents activated guanylate cyclase approximately 30-fold in liver, 20-fold in kidney, 15-fold in cerebellum. 15- to 30-fold in cerebrum, 4- to 20-fold inheart, 12-fold in brain stem, 10-fold in lung, and 2-fold in pancreas. Since recent evidence suggests a role for guanosine 3',5'-monophosphate in malignant transformation, the data may help explain the tumor-inducing capacity of these agents. | Activation of guanylate cyclase by streptozotocin and 1-methyl-1-nitrosourea. Streptozotocin has been shown to induce the production of a variety of tumors in rats. The present report demonstrates that streptozotocin and 1-methyl-1-nitrosourea, a component of the streptozotocin molecule and a known carcinogen, stimulate the enzyme guanylate cyclase which catalyzes the production of guanosine 3',5'-monophosphate. At a maximal concentration of 3 mg/ml, these agents activated guanylate cyclase approximately 30-fold in liver, 20-fold in kidney, 15-fold in cerebellum. 15- to 30-fold in cerebrum, 4- to 20-fold inheart, 12-fold in brain stem, 10-fold in lung, and 2-fold in pancreas. Since recent evidence suggests a role for guanosine 3',5'-monophosphate in malignant transformation, the data may help explain the tumor-inducing capacity of these agents. | [
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|
PMID:11889 | Use of an elemental diet in animals during treatment with 5-fluorouracil (NSC-19893). | Rats eating a diet containing casein hydrolysate (10% wt/wt)(diet 3) instead of whole casein (diet 1) exhibited increased tolerance to nine consecutive daily injections of 15 mg/kg of 5-fluorouracil (5-FU). The relative nutritional efficiency of diet 3 was significantly higher during 5-FU treatment. Serum albumin levels measured after 5-FU treatment dropped by only 2.7% in diet 3 groups and by 13.5% in diet 1 groups. Serum albumin values for rats on the control diet (Purina lab chow) were comparable to those on diet 1. No 5-FU-related mortality was observed in any of the groups. Intestinal brush border enzymes were determined in a group of rats on diet 1. At the end of 5-FU treatment statistically significant changes were observed: sucrase dropped to 30% of control and leucylnaphthylamide-hydrolyzing activity dropped to 19% of control. The activity of gamma-glutamyltransferase did not change significantly. It is postulated that under these circumstances a mixture with a prevalence of free amino acids (casein hydrolysate) could be more readily absorbed than a corresponding mixture containing a larger proportion of oligopeptides. | Use of an elemental diet in animals during treatment with 5-fluorouracil (NSC-19893). Rats eating a diet containing casein hydrolysate (10% wt/wt)(diet 3) instead of whole casein (diet 1) exhibited increased tolerance to nine consecutive daily injections of 15 mg/kg of 5-fluorouracil (5-FU). The relative nutritional efficiency of diet 3 was significantly higher during 5-FU treatment. Serum albumin levels measured after 5-FU treatment dropped by only 2.7% in diet 3 groups and by 13.5% in diet 1 groups. Serum albumin values for rats on the control diet (Purina lab chow) were comparable to those on diet 1. No 5-FU-related mortality was observed in any of the groups. Intestinal brush border enzymes were determined in a group of rats on diet 1. At the end of 5-FU treatment statistically significant changes were observed: sucrase dropped to 30% of control and leucylnaphthylamide-hydrolyzing activity dropped to 19% of control. The activity of gamma-glutamyltransferase did not change significantly. It is postulated that under these circumstances a mixture with a prevalence of free amino acids (casein hydrolysate) could be more readily absorbed than a corresponding mixture containing a larger proportion of oligopeptides. | [
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|
PMID:11893 | [Daily variations of the level of neural secretions in Owenia fusiformis. Persistance of a daily rhythm in cultured tissues]. | A quantitative study of elementary granules in neuron axons of the nervous chain of Owenia fusiformis (Annelid polychaete) shows that daily variations of the number of secretory granules occur in unamputated animals and in ventral tissue cultures. These variations are correlated with the thythm of the cell cycle. | [Daily variations of the level of neural secretions in Owenia fusiformis. Persistance of a daily rhythm in cultured tissues]. A quantitative study of elementary granules in neuron axons of the nervous chain of Owenia fusiformis (Annelid polychaete) shows that daily variations of the number of secretory granules occur in unamputated animals and in ventral tissue cultures. These variations are correlated with the thythm of the cell cycle. | [
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|
PMID:11894 | [Demonstration of different metabolic pathways starting with glucose or fructose in Rhizobium meliloti]. | Rhizobium meliloti can produce many polysaccharides on D-fructose and D-mannitol. In glucose-grown cultures, few polysaccharides are observed and 2 keto-gluconate is accumulated. | [Demonstration of different metabolic pathways starting with glucose or fructose in Rhizobium meliloti]. Rhizobium meliloti can produce many polysaccharides on D-fructose and D-mannitol. In glucose-grown cultures, few polysaccharides are observed and 2 keto-gluconate is accumulated. | [
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PMID:11895 | [Enzymatic characteristics of the guanylate cyclase of KB cells: their change as a function of the development of the cultures]. | We have localized 71% of the guanylate cyclase activity in the (G X 105,000) supernatent fraction of broken KB cells. The reaction follows Michaelis-Menten kinetics, the apparent Km for GTP is 0,5 mM, as long as GTP is lower than a limited concentration, then activity is inhibited. The ion Mn++ is an absolutely required activator, it does not change enzyme-substrate affinity. The enzyme shows several types of binding sites of Mn++. Guanylate cyclase, studied over a period of development of culture, shows, in KB cells without cell contact, an activity higher than that observed in confluent cells. This is not due to the fact of a change in enzyme-substrate affinity but to a modification of Mn++ influence. | [Enzymatic characteristics of the guanylate cyclase of KB cells: their change as a function of the development of the cultures]. We have localized 71% of the guanylate cyclase activity in the (G X 105,000) supernatent fraction of broken KB cells. The reaction follows Michaelis-Menten kinetics, the apparent Km for GTP is 0,5 mM, as long as GTP is lower than a limited concentration, then activity is inhibited. The ion Mn++ is an absolutely required activator, it does not change enzyme-substrate affinity. The enzyme shows several types of binding sites of Mn++. Guanylate cyclase, studied over a period of development of culture, shows, in KB cells without cell contact, an activity higher than that observed in confluent cells. This is not due to the fact of a change in enzyme-substrate affinity but to a modification of Mn++ influence. | [
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|
PMID:11896 | Facial vein in the rabbit. Neurogenic vasodilation mediated by beta-adrenergic receptors. | A segment of the facial vein of the rabbit, that opposite the buccal cavity, responds to norepinephrine (NE) and opposite the buccal cavity, responds to norepinephrine (NE) and transmural nerve stimulation (TNS) by a brisk biphasic dilation. The dilation in response to both procedures is reveresed by prior exposure to propranolol (10(-6)M). Pretreatment with phenoxybenzamine (10(-5)M) increases the size of the neurogenic response and displaces the NE dose-relaxation curve to the left. Histamine causes a constrictor response exclusively. Sympathetic stimulation of a segment of the facial vein proximal to this buccal segment, and also of the external jugular vein, results in constriction. Light microscopy showed no fequtres which can account for the dilation, and fluorescence histochemistry using a modified Flack technique showed a dense adrenergic nerve plexus extending throughout the thickness of the media. We found that frequency-response characteristics and neuronal uptake of 3H-NE were consistent with findings for a blood vessel with a heavy medial innervation. Also, monoamine oxidase and catechol O-methyltransferase activities were similar to those found in other rabbit veins. Furthermore, these results are consistent with an adrenergic neuroeffector organization in which there is a predominance of beta- over alpha-adrenergic receptors. In conclusion, the presence of a dilator response in this buccal segment of the facial vein may be related to its location in the wall of the cheek, where it may be subjected to considerable stretch. | Facial vein in the rabbit. Neurogenic vasodilation mediated by beta-adrenergic receptors. A segment of the facial vein of the rabbit, that opposite the buccal cavity, responds to norepinephrine (NE) and opposite the buccal cavity, responds to norepinephrine (NE) and transmural nerve stimulation (TNS) by a brisk biphasic dilation. The dilation in response to both procedures is reveresed by prior exposure to propranolol (10(-6)M). Pretreatment with phenoxybenzamine (10(-5)M) increases the size of the neurogenic response and displaces the NE dose-relaxation curve to the left. Histamine causes a constrictor response exclusively. Sympathetic stimulation of a segment of the facial vein proximal to this buccal segment, and also of the external jugular vein, results in constriction. Light microscopy showed no fequtres which can account for the dilation, and fluorescence histochemistry using a modified Flack technique showed a dense adrenergic nerve plexus extending throughout the thickness of the media. We found that frequency-response characteristics and neuronal uptake of 3H-NE were consistent with findings for a blood vessel with a heavy medial innervation. Also, monoamine oxidase and catechol O-methyltransferase activities were similar to those found in other rabbit veins. Furthermore, these results are consistent with an adrenergic neuroeffector organization in which there is a predominance of beta- over alpha-adrenergic receptors. In conclusion, the presence of a dilator response in this buccal segment of the facial vein may be related to its location in the wall of the cheek, where it may be subjected to considerable stretch. | [
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PMID:11897 | Diazotization of catecholamines and their analogs and metabolites for urinary screening tests: chemical aspects. | Coupling of diazotized p-nitroaniline to catecholamines and their metabolites in urine has been proposed for use in screening for secreting neuroblastoma in childhood. We have coupled diazotized p-nitroaniline to catecholamines, derivatives, and metabolites and examined the reaction products by thin-layer chromatography and physico-chemical methods (ultraviolet spectra, mass spectroscopy, nuclear magnetic resonance). We conclude that during diazotization, products containing a p-hydroxybenzyl alcohol or a p-hydroxybenzoic acid structure (e.g., vanillic acid, vanilmandelic acid, 3-methoxy-4-hydroxyphenyl-ethyleneglycol, metanephrine, normetanephrine, synephrine, and isoproterenol) react with a diazonium cation, with release of their alcohol or acid moiety. Therefore the mentioned qualitative screening methods are very nonspecific. In contrast, thin-layer chromatographic screening methods provide complete separation and unambiguous identification of those metabolites and are to be preferred for use in detecting secreting neuroblastoma in childhood. | Diazotization of catecholamines and their analogs and metabolites for urinary screening tests: chemical aspects. Coupling of diazotized p-nitroaniline to catecholamines and their metabolites in urine has been proposed for use in screening for secreting neuroblastoma in childhood. We have coupled diazotized p-nitroaniline to catecholamines, derivatives, and metabolites and examined the reaction products by thin-layer chromatography and physico-chemical methods (ultraviolet spectra, mass spectroscopy, nuclear magnetic resonance). We conclude that during diazotization, products containing a p-hydroxybenzyl alcohol or a p-hydroxybenzoic acid structure (e.g., vanillic acid, vanilmandelic acid, 3-methoxy-4-hydroxyphenyl-ethyleneglycol, metanephrine, normetanephrine, synephrine, and isoproterenol) react with a diazonium cation, with release of their alcohol or acid moiety. Therefore the mentioned qualitative screening methods are very nonspecific. In contrast, thin-layer chromatographic screening methods provide complete separation and unambiguous identification of those metabolites and are to be preferred for use in detecting secreting neuroblastoma in childhood. | [
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|
PMID:11898 | A mechanism for the conversion of oxyhemoglobin to methemoglobin by nitrite. | Each mole of oxyhemoglobin iron converted to methemoglobin causes the oxidation of 1.5 mol of nitrite to nitrate and consumes 1 mol of protons. No oxygen is liberated. The overall reaction has two simultaneously occurring parts. In the beginning the rate-limiting reaction converting O2Hb to metHb is directly proportional to H+ and NO2- concentrations and is independent of metHb. The second portion accounts in major part for the stoichiometry and rate of the overall reaction. In this portion O2Hb tetramers and metHbNO2- are the reactants. Essentially no reaction takes place in the presence of CN-, which displaces nitrite from the metHbNO2-, nor in the presence of 0.5 mol/liter Nal, which converts the O2Hb to alphabeta-dimers. The autocatalytic nature of the overall reaction in the presence of excess nitrite is the result of metHb, which is formed in both parts of the reaction, associating with nitrite to increase the concentration of one reactant of the cyanide-sensitive part. The reaction rates at constant pH in excess nitrite are porportional to the product of the O2Hb concentration and the square of the metHb concentration. The rate increases up to about 66% conversion of O2Hb followed by a decrease as the O2Hb becomes limiting. The dissociation constant of metHbNO2- at 25 degrees C and pH = 6.4 was found to be 1.11+/-0.11 mmol/liter. | A mechanism for the conversion of oxyhemoglobin to methemoglobin by nitrite. Each mole of oxyhemoglobin iron converted to methemoglobin causes the oxidation of 1.5 mol of nitrite to nitrate and consumes 1 mol of protons. No oxygen is liberated. The overall reaction has two simultaneously occurring parts. In the beginning the rate-limiting reaction converting O2Hb to metHb is directly proportional to H+ and NO2- concentrations and is independent of metHb. The second portion accounts in major part for the stoichiometry and rate of the overall reaction. In this portion O2Hb tetramers and metHbNO2- are the reactants. Essentially no reaction takes place in the presence of CN-, which displaces nitrite from the metHbNO2-, nor in the presence of 0.5 mol/liter Nal, which converts the O2Hb to alphabeta-dimers. The autocatalytic nature of the overall reaction in the presence of excess nitrite is the result of metHb, which is formed in both parts of the reaction, associating with nitrite to increase the concentration of one reactant of the cyanide-sensitive part. The reaction rates at constant pH in excess nitrite are porportional to the product of the O2Hb concentration and the square of the metHb concentration. The rate increases up to about 66% conversion of O2Hb followed by a decrease as the O2Hb becomes limiting. The dissociation constant of metHbNO2- at 25 degrees C and pH = 6.4 was found to be 1.11+/-0.11 mmol/liter. | [
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|
PMID:11899 | Relation of pH to fluorescence of serotonin, melatonin, and other indole compounds reacted with o-phthaldialdehyde. | We distinguished serotonin, malatonin, and other indole compounds by their markedly different fluorescence behavior when heated with o-phthaldialdehyde in different concentration of HCl, or after the subsequent addition of alkali. Fluorescence may increase, disappear, or change from blue to orange-red. Only melatonin developed significant fluorescence (at less than 0.5 mug/liter) when heated with the reagent in 50 mmol/liter HCl. Serotonin and 5-hydroxyindole compounds with a C-3 aliphatic chain lost their blue fluorescence when brought from strong to weakly acid and alkaline pH. Intense reddish fluorescence developed in strongly alkaline solution. C-5 methoxy derivatives, such as melatonin, maintained their blue fluorescence in alkaline solution. Differences in fluorescence color (and RF) also characterized these compounds on silica gel plates. The use of toluene decreased blank fluorescence, while sample preparation in dim light, as often recommended, resulted in diminished and less-stable fluorescence. | Relation of pH to fluorescence of serotonin, melatonin, and other indole compounds reacted with o-phthaldialdehyde. We distinguished serotonin, malatonin, and other indole compounds by their markedly different fluorescence behavior when heated with o-phthaldialdehyde in different concentration of HCl, or after the subsequent addition of alkali. Fluorescence may increase, disappear, or change from blue to orange-red. Only melatonin developed significant fluorescence (at less than 0.5 mug/liter) when heated with the reagent in 50 mmol/liter HCl. Serotonin and 5-hydroxyindole compounds with a C-3 aliphatic chain lost their blue fluorescence when brought from strong to weakly acid and alkaline pH. Intense reddish fluorescence developed in strongly alkaline solution. C-5 methoxy derivatives, such as melatonin, maintained their blue fluorescence in alkaline solution. Differences in fluorescence color (and RF) also characterized these compounds on silica gel plates. The use of toluene decreased blank fluorescence, while sample preparation in dim light, as often recommended, resulted in diminished and less-stable fluorescence. | [
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|
PMID:11900 | An optimized continuous-monitoring procedure for semiautomated determination of serum acid phosphatase activity. | A continuous-monitoring method for measuring acid phosphatase activity with alpha-naphthyl phosphate as the substrate was critically evaluated and modified. Using partially purified prostatic acid phosphatase, we show that certain conditions for the assay must be satisfied to ensure linearity. These conditions include maintaining the pH between 5.6 and 5.9 and the addition of detergent to sustain linearity. The results obtained with alpha-naphthyl phosphate have been compared with those obtained by using p-nitrophenyl phosphate as substrate. When used with an automatic rate analyzer, the modified method is as sensitive but more reproducible. | An optimized continuous-monitoring procedure for semiautomated determination of serum acid phosphatase activity. A continuous-monitoring method for measuring acid phosphatase activity with alpha-naphthyl phosphate as the substrate was critically evaluated and modified. Using partially purified prostatic acid phosphatase, we show that certain conditions for the assay must be satisfied to ensure linearity. These conditions include maintaining the pH between 5.6 and 5.9 and the addition of detergent to sustain linearity. The results obtained with alpha-naphthyl phosphate have been compared with those obtained by using p-nitrophenyl phosphate as substrate. When used with an automatic rate analyzer, the modified method is as sensitive but more reproducible. | [
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|
PMID:11901 | Enzymatic rate method for measuring cholesterol in serum. | An enzymatic rate assay is described for measuring cholesterol in serum. Cholesterol is analyzed by mixing 5 mul of sample with a reagent consisting of cholesterol esterase, cholesterol oxidase, catalase, acetylacetone, methanol, and hydroxypolyethoxydodecane in a ammonium phosphate buffer at pH 7.0. The rate of increase in absorbance of the dihydrolutidine product is measured at 37 degrees C and 405 nm. The change in absorbance between 4 and 10 min is used to calculate the cholesterol concentrations by using simultaneously determined free cholesterol standards. The change is linearly related to cholesterol concentration up to 4 g/liter. Samples containing bilirubin up to 200 mg/liter, uric acid up to 200 mg/liter, and hemoglobin up to 1 g/liter, or certain drugs (clofibrate, phenobarbital, nicotinic acid, salicylate, Ketochol, and Ovral) gave no interference. Ascorbic acid added to serum caused a positive interference. Lipemic samples gave values that were slightly lower than did the method of Abell et al., used for comparison. Our kinetic assay, compared with the method of Abell et al., the enzymatic assay used with Abbott's Bichromatic Analyzer, and the Technicon SMA 12/60 enzymatic procedure gave correlation coefficients of 0.992, 0.985, and 0.986, respectively. | Enzymatic rate method for measuring cholesterol in serum. An enzymatic rate assay is described for measuring cholesterol in serum. Cholesterol is analyzed by mixing 5 mul of sample with a reagent consisting of cholesterol esterase, cholesterol oxidase, catalase, acetylacetone, methanol, and hydroxypolyethoxydodecane in a ammonium phosphate buffer at pH 7.0. The rate of increase in absorbance of the dihydrolutidine product is measured at 37 degrees C and 405 nm. The change in absorbance between 4 and 10 min is used to calculate the cholesterol concentrations by using simultaneously determined free cholesterol standards. The change is linearly related to cholesterol concentration up to 4 g/liter. Samples containing bilirubin up to 200 mg/liter, uric acid up to 200 mg/liter, and hemoglobin up to 1 g/liter, or certain drugs (clofibrate, phenobarbital, nicotinic acid, salicylate, Ketochol, and Ovral) gave no interference. Ascorbic acid added to serum caused a positive interference. Lipemic samples gave values that were slightly lower than did the method of Abell et al., used for comparison. Our kinetic assay, compared with the method of Abell et al., the enzymatic assay used with Abbott's Bichromatic Analyzer, and the Technicon SMA 12/60 enzymatic procedure gave correlation coefficients of 0.992, 0.985, and 0.986, respectively. | [
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|
PMID:11905 | Erythrocyte glutathione synthetase in 5-oxoprolinuria: kinetic studies of the mutant enzyme and detection of heterozygotes. | The primary metabolic defect in 5-oxoprolinuria is a generalized deficiency of glutathione synthetase. The activity of this enzyme was determined in cell-free extracts of erythrocytes from patients with 5-oxoprolinuria, their parents and a sibling as well as from normal control individuals. The following activities (pkat/mg of hemoglobin) for glutathione synthetase were obtained: homozygotes mean 0.10 (range 0.07-0.12), heterozygotes mean 3.1 (range 2.8-3.7) and control individuals mean 6.1 (range 5.4-6.7). These results indicate that 5-oxoprolinuria, i.e. the defective gluthione synthetase gene(s), is transmitted by autosomal recessive inheritance. Studies of the kinetics of the low remaining activity of erythrocyte glutathione synthetase in patients with 5-oxoprolinuria failed to reveal defective affinity for glycine, gamma-glutamyl-alpha-aminobutyrate, ATP and Mg2+ ions. Furthermore, the pH optimum, time curves and temperature dependence for the mutant enzyme activity did not significantly differ from the corresponding parameters observed with normal enzyme. | Erythrocyte glutathione synthetase in 5-oxoprolinuria: kinetic studies of the mutant enzyme and detection of heterozygotes. The primary metabolic defect in 5-oxoprolinuria is a generalized deficiency of glutathione synthetase. The activity of this enzyme was determined in cell-free extracts of erythrocytes from patients with 5-oxoprolinuria, their parents and a sibling as well as from normal control individuals. The following activities (pkat/mg of hemoglobin) for glutathione synthetase were obtained: homozygotes mean 0.10 (range 0.07-0.12), heterozygotes mean 3.1 (range 2.8-3.7) and control individuals mean 6.1 (range 5.4-6.7). These results indicate that 5-oxoprolinuria, i.e. the defective gluthione synthetase gene(s), is transmitted by autosomal recessive inheritance. Studies of the kinetics of the low remaining activity of erythrocyte glutathione synthetase in patients with 5-oxoprolinuria failed to reveal defective affinity for glycine, gamma-glutamyl-alpha-aminobutyrate, ATP and Mg2+ ions. Furthermore, the pH optimum, time curves and temperature dependence for the mutant enzyme activity did not significantly differ from the corresponding parameters observed with normal enzyme. | [
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|
PMID:11906 | A microfluorometric assay of leukocyte alpha-1,4-glucosidase. | We describe an improved method for detecting deficiency of the acid hydrolase, alpha-1,4-glucosidase in leukocytes, the enzyme defect in glycogen storage disease Type II (Pompe disease). The procedure requires smaller volumes of blood and less time than previous methods. The assay involves the separation of leukocytes by Peter's method for beta-glucosidase and a modification of Salafsky and Nadler's fluorometric method for alpha-glucosidase. | A microfluorometric assay of leukocyte alpha-1,4-glucosidase. We describe an improved method for detecting deficiency of the acid hydrolase, alpha-1,4-glucosidase in leukocytes, the enzyme defect in glycogen storage disease Type II (Pompe disease). The procedure requires smaller volumes of blood and less time than previous methods. The assay involves the separation of leukocytes by Peter's method for beta-glucosidase and a modification of Salafsky and Nadler's fluorometric method for alpha-glucosidase. | [
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PMID:11907 | Studies on gamma-glutamyl transpeptidase of human urine. | A simple and rapid method suitable for clinical laboratories, for the removal of the inhibitors in urine of gamma-glutamyl transpeptidase activity, using an ion-retardation resin, is described. Gel chromatography of urine resulted in appreciable loss of the enzyme activity. Freezing at -5 degrees C and thawing, did not affect the activity of the enzyme. Dimethyl sulfoxide and polyvinyl pyrrolidone up to a creatinine and NaCl at levels found in normal urine did not inhibit the transpeptidase. | Studies on gamma-glutamyl transpeptidase of human urine. A simple and rapid method suitable for clinical laboratories, for the removal of the inhibitors in urine of gamma-glutamyl transpeptidase activity, using an ion-retardation resin, is described. Gel chromatography of urine resulted in appreciable loss of the enzyme activity. Freezing at -5 degrees C and thawing, did not affect the activity of the enzyme. Dimethyl sulfoxide and polyvinyl pyrrolidone up to a creatinine and NaCl at levels found in normal urine did not inhibit the transpeptidase. | [
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|
PMID:11908 | Studies on the multiple forms of gamma-glutamyltransferase. | In sera of 53 patients with highly raised gamma-glutamyltransferase (gamma-GT) (EC 2.3.2.2) activity, the gamma-GT was separated by column chromatography on Sephadex DEAE A-50 into two fractions. In 35 sera only one gamma-GT fraction was found at a conductivity of 2.0--2.5 mS and in 18 sera two gamma-GT fractions were found at a conductivity of 0.2 mS and again at 2.0--2.5 mS. There was no opvious correlation to the underlying disease. In 20 sera the additional parameters, protein, protein electrophoresis, free and esterified cholesterol, bilirubin, triglycerides, lipoprotein pattern and lipoprotein X were examined. We recognized, that the two forms of the gamma-GT are not true isoenzymes but that the separation of the gamma-GT into two fractions is caused by a different distribution of the cholesterol containing lipoprotei,s. | Studies on the multiple forms of gamma-glutamyltransferase. In sera of 53 patients with highly raised gamma-glutamyltransferase (gamma-GT) (EC 2.3.2.2) activity, the gamma-GT was separated by column chromatography on Sephadex DEAE A-50 into two fractions. In 35 sera only one gamma-GT fraction was found at a conductivity of 2.0--2.5 mS and in 18 sera two gamma-GT fractions were found at a conductivity of 0.2 mS and again at 2.0--2.5 mS. There was no opvious correlation to the underlying disease. In 20 sera the additional parameters, protein, protein electrophoresis, free and esterified cholesterol, bilirubin, triglycerides, lipoprotein pattern and lipoprotein X were examined. We recognized, that the two forms of the gamma-GT are not true isoenzymes but that the separation of the gamma-GT into two fractions is caused by a different distribution of the cholesterol containing lipoprotei,s. | [
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|
PMID:11910 | Human leucocyte alpha-L-fucosidase. | 1. Human alpha-L-fucosidase (EC 3.2.1.51) was studied from leucocytes, urine and serum. 2. The leucocyte and urine enzymes are similar in many properties (KM, pH optimum, electrophoretic pattern, heat stability). 3. The serum alpha-L-fucosidase differs from the leucocyte and 4rine enzyme wit,respect to: electrophoretic pattern, pH optimum and heat stapility. 4. The molecular weight of leucocyte alpha-L-fucosidase was determined to be 80 000 +/- 5000. 5. Cu2+, Hg2+ and PCMB are strong inhibitors of leucocyte alpha-L-fucosidase. This inhibition could be completely reversed by beta-mercaptoethanol, indicating that thiol groups are essential for catalytic activity. | Human leucocyte alpha-L-fucosidase. 1. Human alpha-L-fucosidase (EC 3.2.1.51) was studied from leucocytes, urine and serum. 2. The leucocyte and urine enzymes are similar in many properties (KM, pH optimum, electrophoretic pattern, heat stability). 3. The serum alpha-L-fucosidase differs from the leucocyte and 4rine enzyme wit,respect to: electrophoretic pattern, pH optimum and heat stapility. 4. The molecular weight of leucocyte alpha-L-fucosidase was determined to be 80 000 +/- 5000. 5. Cu2+, Hg2+ and PCMB are strong inhibitors of leucocyte alpha-L-fucosidase. This inhibition could be completely reversed by beta-mercaptoethanol, indicating that thiol groups are essential for catalytic activity. | [
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|
PMID:11911 | Characterization of alpha-L-fucosidase from two different families with fucosidosis. | 1. Two different families with a different type of fucosidase deficiency are described. 2. In the first family the activity of alpha-L-fucosidase in leucocytes of two patients with fucosidosis type I was about 4 to 8% of the normal value. The activity of alpha-L-fucosidase in the leucocytes of the father and the mother are in the heterozygote range, while a sister of the propositus showed normal values. 3. The activity of alpha-L-fucosidase of the propositus from urine, serum and liver were also severely decreased. The activity of alpha-L-fucosidase in the urine of the parents and the healthy sister of thr propositus were about 5% of the mean normal value. However in the serum these values were above 50%. 4. The KM value for alpha-L-fucosidase from leucocytes of the patient was increased about 10 times and in serum this value was even higher. The KM values from the enzyme of the parents were in the normal range. 5. The abnormal enzyme from the propositus is unique in its thermal behaviour since after heating its activity increased. 6. In the second fanily the activity of alpha-L-fucosidase in the leucocytes of the patient is about 30% of the mean normal value, while the arylsulphatase A activity is also decreased (25% of the mean normal value). 7. The activity of alpha-L-fucosidase from the leucocytes of the father and the healthy brother are about 50% of the mean normal level, while the enzyme of the mother showed a normal activity. 8. The alpha-L-fucosidase activity in the urine and the liver of the propositus is also decreased. The serum enzyme activity however was in the normal range. 9. The KM value of alpha-L-fucosidase and heat stability of the enzyme of the patient were normal. In the leectrophoretic pattern of the whole family one bond was missing. | Characterization of alpha-L-fucosidase from two different families with fucosidosis. 1. Two different families with a different type of fucosidase deficiency are described. 2. In the first family the activity of alpha-L-fucosidase in leucocytes of two patients with fucosidosis type I was about 4 to 8% of the normal value. The activity of alpha-L-fucosidase in the leucocytes of the father and the mother are in the heterozygote range, while a sister of the propositus showed normal values. 3. The activity of alpha-L-fucosidase of the propositus from urine, serum and liver were also severely decreased. The activity of alpha-L-fucosidase in the urine of the parents and the healthy sister of thr propositus were about 5% of the mean normal value. However in the serum these values were above 50%. 4. The KM value for alpha-L-fucosidase from leucocytes of the patient was increased about 10 times and in serum this value was even higher. The KM values from the enzyme of the parents were in the normal range. 5. The abnormal enzyme from the propositus is unique in its thermal behaviour since after heating its activity increased. 6. In the second fanily the activity of alpha-L-fucosidase in the leucocytes of the patient is about 30% of the mean normal value, while the arylsulphatase A activity is also decreased (25% of the mean normal value). 7. The activity of alpha-L-fucosidase from the leucocytes of the father and the healthy brother are about 50% of the mean normal level, while the enzyme of the mother showed a normal activity. 8. The alpha-L-fucosidase activity in the urine and the liver of the propositus is also decreased. The serum enzyme activity however was in the normal range. 9. The KM value of alpha-L-fucosidase and heat stability of the enzyme of the patient were normal. In the leectrophoretic pattern of the whole family one bond was missing. | [
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|
PMID:11913 | Rapid colorimetric assay of beta-galactosidase and N-acetyl-beta-glucosaminidase in human urine. | Colorimetric methods using 4-nitrophenyl-glycoside substrates for the assay of beta-galactosidase and N-acetyl-beta-glucosaminidase in human urine are described. Interfering substances were removed by gel-filtration of urine. An unidentified low-molecular-weight inhibitor of N-acetyl-beta-glucosaminidase was found. Increased sensitivity of the methods was achieved by high sample volumes, small volumes of buffer to terminate the enzyme reaction and optimal substrate concentration and buffer pH. Incubation periods were shortened to 15 min. Both methods were designed as single-tube tests in which buffer-substrate solutions are prepared in bulk and aliquots are stored in individual containers at -25 degrees C. Under these conditions reagents were stable for at least six months. Precision of both methods was satisfactory. Estimates of normal limits are reported. | Rapid colorimetric assay of beta-galactosidase and N-acetyl-beta-glucosaminidase in human urine. Colorimetric methods using 4-nitrophenyl-glycoside substrates for the assay of beta-galactosidase and N-acetyl-beta-glucosaminidase in human urine are described. Interfering substances were removed by gel-filtration of urine. An unidentified low-molecular-weight inhibitor of N-acetyl-beta-glucosaminidase was found. Increased sensitivity of the methods was achieved by high sample volumes, small volumes of buffer to terminate the enzyme reaction and optimal substrate concentration and buffer pH. Incubation periods were shortened to 15 min. Both methods were designed as single-tube tests in which buffer-substrate solutions are prepared in bulk and aliquots are stored in individual containers at -25 degrees C. Under these conditions reagents were stable for at least six months. Precision of both methods was satisfactory. Estimates of normal limits are reported. | [
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|
PMID:11914 | Comparative studies on cholesterol oxidases from different sources. | 1. Comparison of the characteristics of cholesterol oxidases from different sources was made by a new polarographic method for measurement of the oxygen-consumption rate. 2. A pH optimum of 7.0 was observed for cholesterol oxidases isolated from Nocardia and Brevibacterium, pH 5.0 for the enzyme from Schizophyllum and pH 7.5 for the enzyme from Streptomyces. 3. In the system used in the present study, Ca2+ and Mg2+ had no effect on these enzyme activities. On the other hand, the Schizophyllum enzyme was strongly inhibited by increasing concentrations of Cu2+, whereas the brevibacterium enzyme was slightly activated by them and Nocardia and Streptomyces enzymes were not affected. Hg2+ strongly inhibited the activities of enzymes the Schizophyllum enzyme. 4. Using serum as substrate, the cholesterol oxidases employed, except for the enzyme from Streptomyces, were not active without detergent in the reaction mixture. Effects of various detergents at various concentrations on the enzyme activities were studied. 5. Results of studies on the reaction of cholesterol oxidases on free cholesterol in low-and high-density lipoproteins were also compared. | Comparative studies on cholesterol oxidases from different sources. 1. Comparison of the characteristics of cholesterol oxidases from different sources was made by a new polarographic method for measurement of the oxygen-consumption rate. 2. A pH optimum of 7.0 was observed for cholesterol oxidases isolated from Nocardia and Brevibacterium, pH 5.0 for the enzyme from Schizophyllum and pH 7.5 for the enzyme from Streptomyces. 3. In the system used in the present study, Ca2+ and Mg2+ had no effect on these enzyme activities. On the other hand, the Schizophyllum enzyme was strongly inhibited by increasing concentrations of Cu2+, whereas the brevibacterium enzyme was slightly activated by them and Nocardia and Streptomyces enzymes were not affected. Hg2+ strongly inhibited the activities of enzymes the Schizophyllum enzyme. 4. Using serum as substrate, the cholesterol oxidases employed, except for the enzyme from Streptomyces, were not active without detergent in the reaction mixture. Effects of various detergents at various concentrations on the enzyme activities were studied. 5. Results of studies on the reaction of cholesterol oxidases on free cholesterol in low-and high-density lipoproteins were also compared. | [
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PMID:11915 | Catecholamine-sensitive adenylate cyclase of human fat cell ghosts. Characteristics of the GMP(PNP)-liganded state. | The effects of 5'-guanylyl-imidodiphosphate (GMP(PNP)) on the catecholamine-sensitive adenylate cyclase of human fat cell ghosts were studied. The compound increased basal and epinephrine-stimulated enzyme activity by about 300%; in addition GMP(PNP) increased hormone sensitivity by reducing the epinephrine concentration required, to produce half maximal stimulation. The rate of GMP(PNP)-induced activation was slow in onset and could be enhanced by epinephrine. The GMP(PNP)-activated state was resistant to thermal inactivation and could not be reversed by extensive washing. The application of this compound in clinical studies may be useful because of its stimulating and stabilizing action. | Catecholamine-sensitive adenylate cyclase of human fat cell ghosts. Characteristics of the GMP(PNP)-liganded state. The effects of 5'-guanylyl-imidodiphosphate (GMP(PNP)) on the catecholamine-sensitive adenylate cyclase of human fat cell ghosts were studied. The compound increased basal and epinephrine-stimulated enzyme activity by about 300%; in addition GMP(PNP) increased hormone sensitivity by reducing the epinephrine concentration required, to produce half maximal stimulation. The rate of GMP(PNP)-induced activation was slow in onset and could be enhanced by epinephrine. The GMP(PNP)-activated state was resistant to thermal inactivation and could not be reversed by extensive washing. The application of this compound in clinical studies may be useful because of its stimulating and stabilizing action. | [
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|
PMID:11916 | Galactose-1-phosphate uridyltransferase in cultured cells. | A method to measure the enzyme galactose-1-phosphate uridyltransferase in cultured cells is described. The optimun pH was 8.7 and no enzyme activity was found without preincubation with dithiothreitol. The KM values for Gal-1-P and UDPG for the wild type enzyme were found to be 0.2 mM and 0.08 mM, respectively. Values for the Duarte variant were found to be identical. No significant change in enzyme activity with time after subculture was found in either cultured skin fibroblasts or cultured amniotic fluid cells. Different transferase genotypes were clearly distinguished and reference range established. Transferase levels found in normal cultured amniotic fluid cells were the same as in normal cultured skin fibroblasts. The results of a prenatal diagnosis was obtained within 3 weeks of amniocentesis. | Galactose-1-phosphate uridyltransferase in cultured cells. A method to measure the enzyme galactose-1-phosphate uridyltransferase in cultured cells is described. The optimun pH was 8.7 and no enzyme activity was found without preincubation with dithiothreitol. The KM values for Gal-1-P and UDPG for the wild type enzyme were found to be 0.2 mM and 0.08 mM, respectively. Values for the Duarte variant were found to be identical. No significant change in enzyme activity with time after subculture was found in either cultured skin fibroblasts or cultured amniotic fluid cells. Different transferase genotypes were clearly distinguished and reference range established. Transferase levels found in normal cultured amniotic fluid cells were the same as in normal cultured skin fibroblasts. The results of a prenatal diagnosis was obtained within 3 weeks of amniocentesis. | [
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PMID:11959 | Herbicides and soil microorganisms. | The production and usage of herbicides have increased dramatically in recent years. Consequently there is growing concern about the effects of these chemicals on the environment, particularly the possible long-term effects on soil fertility which may result from disturbance of the soil microflora. This review considers the recent developments in the study of the interactions between herbicides and microorganisms, and discusses the problems of evaluating the results of such studies with a view to determining or predicting possible short-term or long-term effects which may be of practical significance. | Herbicides and soil microorganisms. The production and usage of herbicides have increased dramatically in recent years. Consequently there is growing concern about the effects of these chemicals on the environment, particularly the possible long-term effects on soil fertility which may result from disturbance of the soil microflora. This review considers the recent developments in the study of the interactions between herbicides and microorganisms, and discusses the problems of evaluating the results of such studies with a view to determining or predicting possible short-term or long-term effects which may be of practical significance. | [
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|
PMID:11960 | Evaluation of anxiolytic and amnesic effects of intramuscular lorazepam as a pre-operative medication. | In a double-blind investigation in 21 patients undergoing minor plastic surgery the effects of 4 mg lorazepam or a mixture of 20 mg papaveretum/400 mug hyoscine given intramuscularly as premedicants were studied. Lorazepam produced superior preoperative and post-operative anterograde amnesia, with respect to both incidence and duration. There was no difference in the pre-operative anxiolytic and soporofic effects of the two regimens and post-operatively, analgesic requirements and the incidence of vomitting and restlessness were the same in both groups of patients. | Evaluation of anxiolytic and amnesic effects of intramuscular lorazepam as a pre-operative medication. In a double-blind investigation in 21 patients undergoing minor plastic surgery the effects of 4 mg lorazepam or a mixture of 20 mg papaveretum/400 mug hyoscine given intramuscularly as premedicants were studied. Lorazepam produced superior preoperative and post-operative anterograde amnesia, with respect to both incidence and duration. There was no difference in the pre-operative anxiolytic and soporofic effects of the two regimens and post-operatively, analgesic requirements and the incidence of vomitting and restlessness were the same in both groups of patients. | [
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|
PMID:11961 | Lorazepam in the treatment of neurosis. | An open study was carried out in 40 ambulatory neurotic patients with classical symptoms of anxiety to assess which of their symptoms responded most readily and favourably to treatment with lorazepam. Doses ranged from 2 mg to 15 mg daily and were adjusted to individual patient needs. The study period lasted 4 weeks and patients' individual symptoms were assessed weekly on a 4-point severity rating scale. The results showed a statistically significant and marked improvement during the first week for the majority of the 16 symptoms assessed, particularly emotional tension, irritability and apprehension. Symptoms with a cognitive element of anxiety were controlled to a lesser extent and were slower to respond. The few side-effects which were reported, mainly somnolence, appeared to be dose related and usually occurred during the first few days of treatment. | Lorazepam in the treatment of neurosis. An open study was carried out in 40 ambulatory neurotic patients with classical symptoms of anxiety to assess which of their symptoms responded most readily and favourably to treatment with lorazepam. Doses ranged from 2 mg to 15 mg daily and were adjusted to individual patient needs. The study period lasted 4 weeks and patients' individual symptoms were assessed weekly on a 4-point severity rating scale. The results showed a statistically significant and marked improvement during the first week for the majority of the 16 symptoms assessed, particularly emotional tension, irritability and apprehension. Symptoms with a cognitive element of anxiety were controlled to a lesser extent and were slower to respond. The few side-effects which were reported, mainly somnolence, appeared to be dose related and usually occurred during the first few days of treatment. | [
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|
PMID:11963 | The role of HCO3-stimulated ATPase in buffer transport. | An ATPase stimulated by HCO-3ions and other oxybases and inhibited by SCN- has been found in main excretory duct of rat submaxillary gland, a tissue, capable of actively secreting HCO-3ions. No such ATPase was found in the rabbit duct, which normally does not secrete HCO-3. The HCO-3ATPase was localized in the plasma membrane fraction of the homogenate, as evidenced by the marker 5'nucleotidase. The activities of the HCO-3ATPase increased in metabolic alkalosis and decreased in metabolic acidosis in parallel to secretion of HCO-3 and K+ ions by the rat salivary duct epithelium. In renal cortex tissue, where HCO-3 is actively reabsorbed respectively H+ is secreted, there was also found a parallel change in the activity of the HCO-3ATPase and the rate of active H+ secretion. These findings provide further evidence that the membrane-bound HCO-3ATPase is involved in active H+/HCO-3 transport. The HCO-3ATPase is not only stimulated by HCO-3 but also by other non transportable oxybases, a finding which indicates H+ rather than HCO-3 being the actively transported component of the buffer system. Small concentrations of K+ ions decrease the Km for HCO-3 and thus yield stimulation of the HCO-3-ATPase. Thport changing in parallel with that of H+/HCO-3 may be taken as indicative for a coupled K+-H+-exchange mechanism to which the HCO-3ATPase is linked. | The role of HCO3-stimulated ATPase in buffer transport. An ATPase stimulated by HCO-3ions and other oxybases and inhibited by SCN- has been found in main excretory duct of rat submaxillary gland, a tissue, capable of actively secreting HCO-3ions. No such ATPase was found in the rabbit duct, which normally does not secrete HCO-3. The HCO-3ATPase was localized in the plasma membrane fraction of the homogenate, as evidenced by the marker 5'nucleotidase. The activities of the HCO-3ATPase increased in metabolic alkalosis and decreased in metabolic acidosis in parallel to secretion of HCO-3 and K+ ions by the rat salivary duct epithelium. In renal cortex tissue, where HCO-3 is actively reabsorbed respectively H+ is secreted, there was also found a parallel change in the activity of the HCO-3ATPase and the rate of active H+ secretion. These findings provide further evidence that the membrane-bound HCO-3ATPase is involved in active H+/HCO-3 transport. The HCO-3ATPase is not only stimulated by HCO-3 but also by other non transportable oxybases, a finding which indicates H+ rather than HCO-3 being the actively transported component of the buffer system. Small concentrations of K+ ions decrease the Km for HCO-3 and thus yield stimulation of the HCO-3-ATPase. Thport changing in parallel with that of H+/HCO-3 may be taken as indicative for a coupled K+-H+-exchange mechanism to which the HCO-3ATPase is linked. | [
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PMID:11964 | Polarity of proximal tubular epithelial cells in relation to transepithelial transport. | Transport properties of brush border microvilli and basal-lateral plasma membranes isolated from rat kidney cortex were studied by a millipore filtration technique. Brush border microvilli but not basal-lateral plasma membranes contain sodium dependent stereospecific transport system for D-glucose, L-phenylalanine and inorganic phosphate as indicated by saturability, countertransport and inhibition by structurally related compounds. Reduction of equilbrium uptake by increasing medium osmolarity suggests transport into an osmotically reactive space rather than binding to the membranes. Electrogenecity of the sodium-sugar and sodium-amino-acid cotransport system was established by their dependence on artificially imposed diffusion potentials. Also a NA+/H+ antiport system can be demonstrated in microvilli vesicles by demonstrating counterflow of both ions under short circuit conditions. Basal-lateral plasma membranes contain sodium independent stereospecific transport systems for sugars and amino acids. These results demonstrate a marked functional polarity of the cell membranes in respect to sodium dependent and sodium independent transport systems. This polarity in conjunction with the asymmetrical distribution of sodium between the intra- and extracellular space seems to enable the proximal tubule epithelial cells to perform active transepithelial transport. | Polarity of proximal tubular epithelial cells in relation to transepithelial transport. Transport properties of brush border microvilli and basal-lateral plasma membranes isolated from rat kidney cortex were studied by a millipore filtration technique. Brush border microvilli but not basal-lateral plasma membranes contain sodium dependent stereospecific transport system for D-glucose, L-phenylalanine and inorganic phosphate as indicated by saturability, countertransport and inhibition by structurally related compounds. Reduction of equilbrium uptake by increasing medium osmolarity suggests transport into an osmotically reactive space rather than binding to the membranes. Electrogenecity of the sodium-sugar and sodium-amino-acid cotransport system was established by their dependence on artificially imposed diffusion potentials. Also a NA+/H+ antiport system can be demonstrated in microvilli vesicles by demonstrating counterflow of both ions under short circuit conditions. Basal-lateral plasma membranes contain sodium independent stereospecific transport systems for sugars and amino acids. These results demonstrate a marked functional polarity of the cell membranes in respect to sodium dependent and sodium independent transport systems. This polarity in conjunction with the asymmetrical distribution of sodium between the intra- and extracellular space seems to enable the proximal tubule epithelial cells to perform active transepithelial transport. | [
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|
PMID:11965 | Properties of rat kidney glutaminase enzymes and their role in renal ammoniagenesis. | Rat kidney contains two distinct glutaminase activities; the mitochondrial phosphate-dependent glutaminase and a second glutaminase activity associated with the brush border membrane which is maleate-activated and phosphate-independent. It has recently been shown that the phosphate-independent glutaminase is a partial reaction of gamma-glutamyl transpeptidase and that maleate activates this enzyme by blocking transpeptidation. The gamma-glutamyl transpeptidase in other rat tissues is also affected by maleate. This enzyme has at least a 100-fold greater affinity for glutathione or for glutathione derivatives than for glutamine, suggesting that under physiological conditions glutathione is the preferred substrate. With either type of substrate, maleate affects the Vmax of the reaction but not the Km. These findings suggest that this enzyme probably contributes very little to renal ammoniagenesis. In contrast, the phosphate-dependent glutaminase, whose activity increases 20 to 30-fold in the proximal convoluted tubule cells in response to metabolic acidosis, probably contributes significantly to renal ammoniagenesis. We have purified the rat kidney phosphate-dependent glutaminase and compared the phosphate activation and the phosphate-induced dimerization of the Tris form of this enzyme. There is an excellent correlation between increased activity and extent of dimerization as phosphate concentration is increased. The molecular weights of the Tris form are 1600000 and 316000 in the absence and presence of -1 M NaPO4, respectively. At saturating concentration of phosphate, increasing concentrations of chloride ion similarly reverse both activation and dimerization. These observations suggest that only the dimer form of the Tris enzyme is active. | Properties of rat kidney glutaminase enzymes and their role in renal ammoniagenesis. Rat kidney contains two distinct glutaminase activities; the mitochondrial phosphate-dependent glutaminase and a second glutaminase activity associated with the brush border membrane which is maleate-activated and phosphate-independent. It has recently been shown that the phosphate-independent glutaminase is a partial reaction of gamma-glutamyl transpeptidase and that maleate activates this enzyme by blocking transpeptidation. The gamma-glutamyl transpeptidase in other rat tissues is also affected by maleate. This enzyme has at least a 100-fold greater affinity for glutathione or for glutathione derivatives than for glutamine, suggesting that under physiological conditions glutathione is the preferred substrate. With either type of substrate, maleate affects the Vmax of the reaction but not the Km. These findings suggest that this enzyme probably contributes very little to renal ammoniagenesis. In contrast, the phosphate-dependent glutaminase, whose activity increases 20 to 30-fold in the proximal convoluted tubule cells in response to metabolic acidosis, probably contributes significantly to renal ammoniagenesis. We have purified the rat kidney phosphate-dependent glutaminase and compared the phosphate activation and the phosphate-induced dimerization of the Tris form of this enzyme. There is an excellent correlation between increased activity and extent of dimerization as phosphate concentration is increased. The molecular weights of the Tris form are 1600000 and 316000 in the absence and presence of -1 M NaPO4, respectively. At saturating concentration of phosphate, increasing concentrations of chloride ion similarly reverse both activation and dimerization. These observations suggest that only the dimer form of the Tris enzyme is active. | [
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PMID:11966 | On the role of gamma-glutamyltransferase in renal tubular amino acid reabsorption. | The degradation of glutathione in the kidney of the rat was investigated in vivo and in vitro. When radioactive glutathione or its analogue ophthalmic acid was administered intravenously to mice or rats, the tripeptides were rapidly and completely degraded. Within the organs, no radioactive glutathione, but only labelled glycine was found. The main part of the degrading activity was localized in the kidney. Kidney homogenate degraded glutathione at a rate of 46.5 nmoles/min per mg of protein. This could be inhibited by the gamma-glutamyltransferase inhibitor serine-borate. Isolated renal tubules degraded the tripeptide at a rate of 18 nmoles/min/mg; this reaction was also inhibited by serine-borate. The whole activity was found in the particulate fraction (100000 xg). Glycine and gamma-glutamylglycine were identified as the radio-active products. The results indicate that gamma-glutamyltransferase is able to split glutathione extracellularly in the lumen of the tubule at a very high rate. It is concluded that the enzyme faces the luminal side of the brush border membrane with respect to its substrate gluthathione. This seems to be incompatible with a basic topological prerequisite for the in vivo function of the gamma-glutamyl cycle in renal tubular amino acid reabsorption. | On the role of gamma-glutamyltransferase in renal tubular amino acid reabsorption. The degradation of glutathione in the kidney of the rat was investigated in vivo and in vitro. When radioactive glutathione or its analogue ophthalmic acid was administered intravenously to mice or rats, the tripeptides were rapidly and completely degraded. Within the organs, no radioactive glutathione, but only labelled glycine was found. The main part of the degrading activity was localized in the kidney. Kidney homogenate degraded glutathione at a rate of 46.5 nmoles/min per mg of protein. This could be inhibited by the gamma-glutamyltransferase inhibitor serine-borate. Isolated renal tubules degraded the tripeptide at a rate of 18 nmoles/min/mg; this reaction was also inhibited by serine-borate. The whole activity was found in the particulate fraction (100000 xg). Glycine and gamma-glutamylglycine were identified as the radio-active products. The results indicate that gamma-glutamyltransferase is able to split glutathione extracellularly in the lumen of the tubule at a very high rate. It is concluded that the enzyme faces the luminal side of the brush border membrane with respect to its substrate gluthathione. This seems to be incompatible with a basic topological prerequisite for the in vivo function of the gamma-glutamyl cycle in renal tubular amino acid reabsorption. | [
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|
PMID:11967 | The influence of branched chain aminoacids and their ketoderivatives on renal gluconeogenesis. | Isolated kidney tubules served as a model to investigate the direct effect of branched chain aminoacids and their ketoderivatives on gluconeogenesis. The data presented in this paper demonstrate that the ketoderivatives rather than the branched chain aminoacids themselves inhibit renal glucosesynthesis from various precursors entering the glucogenic pathway at different levels. Though the point of the inhibitory attack of ketoacids could not be localized, an impairement of the kidney cortex to respond to metabolic acidosis with an increase of gluconeogenesis is evident from the present data. The relevance of the presented data concerning the production of hypoglycemia and metabolic acidosis in leucine induced hypoglycemia and maple syrup urine disease is discussed. | The influence of branched chain aminoacids and their ketoderivatives on renal gluconeogenesis. Isolated kidney tubules served as a model to investigate the direct effect of branched chain aminoacids and their ketoderivatives on gluconeogenesis. The data presented in this paper demonstrate that the ketoderivatives rather than the branched chain aminoacids themselves inhibit renal glucosesynthesis from various precursors entering the glucogenic pathway at different levels. Though the point of the inhibitory attack of ketoacids could not be localized, an impairement of the kidney cortex to respond to metabolic acidosis with an increase of gluconeogenesis is evident from the present data. The relevance of the presented data concerning the production of hypoglycemia and metabolic acidosis in leucine induced hypoglycemia and maple syrup urine disease is discussed. | [
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|
PMID:11969 | Human mitochondrial NADP-dependent isocitrate dehydrogenase in man-mouse somatic cell hybrids. | Human mitochondrial NADP-dependent isocitrate dehydrogenase (IDH-2) is expressed in man-mouse somatic cell hybrids as a dimeric molecule. The gene specifing this enzyme was observed to be syntenic with the mannose phosphate isomerase locus in the 56 primary man-mouse clones in this series. The human IDH-2 locus, therefore, may be assigned to chromosome 15. | Human mitochondrial NADP-dependent isocitrate dehydrogenase in man-mouse somatic cell hybrids. Human mitochondrial NADP-dependent isocitrate dehydrogenase (IDH-2) is expressed in man-mouse somatic cell hybrids as a dimeric molecule. The gene specifing this enzyme was observed to be syntenic with the mannose phosphate isomerase locus in the 56 primary man-mouse clones in this series. The human IDH-2 locus, therefore, may be assigned to chromosome 15. | [
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|
PMID:11970 | Immediate hypersensitivity to tuberculin. In vivo and in vitro studies. | Immediate responses of hypersensitivity to skin testing with purified derivative of tuberculin (PPD) were observed in 2.3 percent of 3,248 patients seen in an allergy clinic, and the relationship to delayed responses was questioned. Immediate cutaneous reactions to testing with PPD appeared in all age groups and occurred in nonatopic patients but were more common in atopic patients (p less than 0.005). Delayed cutaneous reactions to testing with PPD occurred in only three out of 76 patients with immediate reactivity. Antihistaminic suppression of immediate reactivity was not followed by evidence of delayed cutaneous reactivity. In vitro tests of lymphocytic stimulation revealed indices of stimulation with PPD to be similar both in patients with immediate and delayed cutaneous reactivity. Failure to manifest delayed cutaneous reactivity following immediate cutaneous reactions alone may be explained by antigen-antibody binding and phagocytosis, by suppressor T-lymphocytes, or by impaired release or lack of response to T-lymphocytic mediators. Adverse reactions to administration of BCG vaccine in patients with immediate cutaneous reactivity might be anticipated. | Immediate hypersensitivity to tuberculin. In vivo and in vitro studies. Immediate responses of hypersensitivity to skin testing with purified derivative of tuberculin (PPD) were observed in 2.3 percent of 3,248 patients seen in an allergy clinic, and the relationship to delayed responses was questioned. Immediate cutaneous reactions to testing with PPD appeared in all age groups and occurred in nonatopic patients but were more common in atopic patients (p less than 0.005). Delayed cutaneous reactions to testing with PPD occurred in only three out of 76 patients with immediate reactivity. Antihistaminic suppression of immediate reactivity was not followed by evidence of delayed cutaneous reactivity. In vitro tests of lymphocytic stimulation revealed indices of stimulation with PPD to be similar both in patients with immediate and delayed cutaneous reactivity. Failure to manifest delayed cutaneous reactivity following immediate cutaneous reactions alone may be explained by antigen-antibody binding and phagocytosis, by suppressor T-lymphocytes, or by impaired release or lack of response to T-lymphocytic mediators. Adverse reactions to administration of BCG vaccine in patients with immediate cutaneous reactivity might be anticipated. | [
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PMID:11971 | Organ and species differences in microsomal activation of methyldopa. | The covalent binding of 3H-methyldopa to microsomal protein in the presence of NADHP and oxygen was studied in various microsomal preparations. Rat and mouse liver microsomes showed high binding, hamster and guinea pig liver microsomes gave intermediate values, whereas no binding was seen with rabbit liver microsomes. No activation of methyldopa was detected with kidney microsomes. Lung microsomes from rats, guinea pigs, and rabbits were quite active with respect to methyldopa binding, and the reactions were totally blocked by superoxide dismutase. There was no sex difference in the binding of methyldopa in liver microsomes from adult rats. No methyldopa activation could be detected in fetal liver microsomes, whereas a rapid increase in activity to above adult levels occurred during the first 2 days after birth. | Organ and species differences in microsomal activation of methyldopa. The covalent binding of 3H-methyldopa to microsomal protein in the presence of NADHP and oxygen was studied in various microsomal preparations. Rat and mouse liver microsomes showed high binding, hamster and guinea pig liver microsomes gave intermediate values, whereas no binding was seen with rabbit liver microsomes. No activation of methyldopa was detected with kidney microsomes. Lung microsomes from rats, guinea pigs, and rabbits were quite active with respect to methyldopa binding, and the reactions were totally blocked by superoxide dismutase. There was no sex difference in the binding of methyldopa in liver microsomes from adult rats. No methyldopa activation could be detected in fetal liver microsomes, whereas a rapid increase in activity to above adult levels occurred during the first 2 days after birth. | [
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PMID:11972 | Xenobiotic metabolism and enzyme induction in isolated rat intestinal microsomes. | Intestinal microsomes were isolated from rats pretreated with 3-methylcholanthrene (MC), phenobarbital (PB), and pregnenolone-16alpha-carbonitrile (PCN). The metabolism of benzo[a]pyrene (BP), 7-ethoxyresorufin, 7-ethoxycoumarin, and biphenyl were examined. MC induces a 30-fold increase in BP metabolism. With control microsomes, BP metabolism is inhibited by metyrapone, SKF 525-A, and antimycin A, but is stimulated 4.5-fold by rotenone. With microsomes from MC-treated rats, BP metabolism is inhibited by metyrapone, SKF 525-A, antimycin A, and rotenone. MC pretreatment increases 7-ethoxyresorufin de-ethylase by almost 20-fold and ethoxycoumarin de-ethylase by almost 18 fold. PB pretreatment produces less than a 2-fold increase in both de-ethylases. PCN pretreatment inhibits 7-ethoxyresorufin and 7-ethoxycoumarin de-ethylases. In rats fasted for 2 days, neither de-ethylase could be detected in intestinal microsomes. Biphenyl 2- and 4-hydroxylase activities were induced less than 4-fold by MC or PB pretreatment. | Xenobiotic metabolism and enzyme induction in isolated rat intestinal microsomes. Intestinal microsomes were isolated from rats pretreated with 3-methylcholanthrene (MC), phenobarbital (PB), and pregnenolone-16alpha-carbonitrile (PCN). The metabolism of benzo[a]pyrene (BP), 7-ethoxyresorufin, 7-ethoxycoumarin, and biphenyl were examined. MC induces a 30-fold increase in BP metabolism. With control microsomes, BP metabolism is inhibited by metyrapone, SKF 525-A, and antimycin A, but is stimulated 4.5-fold by rotenone. With microsomes from MC-treated rats, BP metabolism is inhibited by metyrapone, SKF 525-A, antimycin A, and rotenone. MC pretreatment increases 7-ethoxyresorufin de-ethylase by almost 20-fold and ethoxycoumarin de-ethylase by almost 18 fold. PB pretreatment produces less than a 2-fold increase in both de-ethylases. PCN pretreatment inhibits 7-ethoxyresorufin and 7-ethoxycoumarin de-ethylases. In rats fasted for 2 days, neither de-ethylase could be detected in intestinal microsomes. Biphenyl 2- and 4-hydroxylase activities were induced less than 4-fold by MC or PB pretreatment. | [
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PMID:11973 | The rat brain as a "deep compartment" in the kinetics of a common metabolite of prochlorperazine and perphenazine. | N-[gamma-(2-Chlorophenothiazin-10-yl)propyl]ethylenediamine (Cl-PPED), a common metabolite of prochlorperazine and perphenazine, was orally administered to male rats at a dose of 50 mg/kg, and its concentrations in plasma, liver, lung, kidney, and brain were followed from 6 hr to 40 days after dosage. Small quantities were measured by two-dimensional thin-layer chromatography of a fluorescent derivative and in situ fluorometry of the chromatograms. The kinetic behavior of Cl-PPED in brain differed from that in other tissues in that the concentration maximum was achieved later and the elimination half-life was distinctly longer (8 days vs. 1.2-3 days). Cl-PPED was evenly among the three major brain regions. Subcellular fractionation of liver and brain revealed preferential localization in mitochondria. | The rat brain as a "deep compartment" in the kinetics of a common metabolite of prochlorperazine and perphenazine. N-[gamma-(2-Chlorophenothiazin-10-yl)propyl]ethylenediamine (Cl-PPED), a common metabolite of prochlorperazine and perphenazine, was orally administered to male rats at a dose of 50 mg/kg, and its concentrations in plasma, liver, lung, kidney, and brain were followed from 6 hr to 40 days after dosage. Small quantities were measured by two-dimensional thin-layer chromatography of a fluorescent derivative and in situ fluorometry of the chromatograms. The kinetic behavior of Cl-PPED in brain differed from that in other tissues in that the concentration maximum was achieved later and the elimination half-life was distinctly longer (8 days vs. 1.2-3 days). Cl-PPED was evenly among the three major brain regions. Subcellular fractionation of liver and brain revealed preferential localization in mitochondria. | [
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|
PMID:11974 | Localization of nicotine-14C, cotinine-14C, and nicotine-1'-N-oxide-14C in tissues of the mouse. | The distrubutions of nicotine-14C (2'- or methyl-labeled), cotinine-14C, and nicotine-1'-N-oxide-14C were studied by whole-body autoradiography in mice and in perfused rabbit lung. The compounds were administered either iv, sc, ip, or by inhalation. Blank sections were also incubated with nicotine-14C and cotinine-14C in vitro. The distributions were compared in a black (C57BL/6J), brown (C57L/J), and albino (A/HeJ) strain and in CD-1 germ-free mice. After administration of nicotine-14C in vivo, radioactivity was localized in all strains in bronchi, nasal mucosa, salivary gland, Harder's gland, liver, kidney, stomach, spleen, pancreas, intestine, bone, gallbladder, and adrenal medulla. In the pigmented strains, it was also localized in melanin in the eye, brain, and hair. Radioactivity did not accumulate in the bronchi of late-term fetuses or the 1-day-old newborn but was present in the 2-day-old and older. Cotinine-14C and nicotine-1'-N-oxide-14C, after iv administration, did not localize in bronchi or nasal mucosa at short time intervals after injection. Frozen sections incubated in vitro did not accumulate either nicotine-14C or cotinine-14C in the bronchi; whereas the affinity for melanin was unchanged. Incubation of fresh, nonfrozen mouse lung and perfusion of rabbit lung in vitro with nicotine-14C produced the same localization of radioactivity in bronchi as that seen after in vivo administration. Preheating the frozen sections or lungs in a microwave oven before incubation did not change the localization of radioactivity in the bronchi or other tissues. The localization of radioactivity in bronchi may be due to metabolism, active transport, or binding of nicotine; this mechanism is destroyed by freezing the tissue. | Localization of nicotine-14C, cotinine-14C, and nicotine-1'-N-oxide-14C in tissues of the mouse. The distrubutions of nicotine-14C (2'- or methyl-labeled), cotinine-14C, and nicotine-1'-N-oxide-14C were studied by whole-body autoradiography in mice and in perfused rabbit lung. The compounds were administered either iv, sc, ip, or by inhalation. Blank sections were also incubated with nicotine-14C and cotinine-14C in vitro. The distributions were compared in a black (C57BL/6J), brown (C57L/J), and albino (A/HeJ) strain and in CD-1 germ-free mice. After administration of nicotine-14C in vivo, radioactivity was localized in all strains in bronchi, nasal mucosa, salivary gland, Harder's gland, liver, kidney, stomach, spleen, pancreas, intestine, bone, gallbladder, and adrenal medulla. In the pigmented strains, it was also localized in melanin in the eye, brain, and hair. Radioactivity did not accumulate in the bronchi of late-term fetuses or the 1-day-old newborn but was present in the 2-day-old and older. Cotinine-14C and nicotine-1'-N-oxide-14C, after iv administration, did not localize in bronchi or nasal mucosa at short time intervals after injection. Frozen sections incubated in vitro did not accumulate either nicotine-14C or cotinine-14C in the bronchi; whereas the affinity for melanin was unchanged. Incubation of fresh, nonfrozen mouse lung and perfusion of rabbit lung in vitro with nicotine-14C produced the same localization of radioactivity in bronchi as that seen after in vivo administration. Preheating the frozen sections or lungs in a microwave oven before incubation did not change the localization of radioactivity in the bronchi or other tissues. The localization of radioactivity in bronchi may be due to metabolism, active transport, or binding of nicotine; this mechanism is destroyed by freezing the tissue. | [
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|
PMID:11975 | Effect of sex hormones on the disposition in rats of 1-aminocyclohexane carboxylic acid, a metabolite of semisynthetic penicillin. | The renal clearance of 1-aminocyclohexanecarboxylic acid (ACHC), a metabolite of the semisynthetic penicillin, cyclacillin, is about 10 times faster in female than in male rats. The slower clearance in males is attributed to a higher net rate of reabsorption of the compound from the tubule of the kidney. Because ACHC is not metabolized, it is apparently continuously recirculated through the kidney of the male, resulting in the longer half-life. The sex-related disposition of the metabolite can be modified by gonadectomy and/or treatment with sex hormones. Castrated males show increased urinary excretion and decreased plasma half-life of ACHC relative to intact males. In ovariectomized females, less ACHC is excreted and the half-life is longer than in intact females. Thus, in both sexes, gonadectomy shifts the excretion and the residence time in plasma toward the values of these parameters for the opposite sex. Treatment of castrated males with estradiol markedly enhances the effect of castration, but treatment of ovariectomized females with testosterone propionate has little or no additional effect over ovariectomy. Treatment of intact males with estradiol modifies both excretion and residence time in plasma to a great extent, but treatment of intact females with testosterone has a lesser effect on the disposition of ACHC. These results indicate that excretion and residence time of ACHC in both male and female rats are influenced by sex hormones. The described effect is an example of the action of sex hormones on the transport of foreign compounds in this species. Its mechanism is quite different from the well known influence of sex hormones on the microsomal metabolism of foreign compounds in rats. | Effect of sex hormones on the disposition in rats of 1-aminocyclohexane carboxylic acid, a metabolite of semisynthetic penicillin. The renal clearance of 1-aminocyclohexanecarboxylic acid (ACHC), a metabolite of the semisynthetic penicillin, cyclacillin, is about 10 times faster in female than in male rats. The slower clearance in males is attributed to a higher net rate of reabsorption of the compound from the tubule of the kidney. Because ACHC is not metabolized, it is apparently continuously recirculated through the kidney of the male, resulting in the longer half-life. The sex-related disposition of the metabolite can be modified by gonadectomy and/or treatment with sex hormones. Castrated males show increased urinary excretion and decreased plasma half-life of ACHC relative to intact males. In ovariectomized females, less ACHC is excreted and the half-life is longer than in intact females. Thus, in both sexes, gonadectomy shifts the excretion and the residence time in plasma toward the values of these parameters for the opposite sex. Treatment of castrated males with estradiol markedly enhances the effect of castration, but treatment of ovariectomized females with testosterone propionate has little or no additional effect over ovariectomy. Treatment of intact males with estradiol modifies both excretion and residence time in plasma to a great extent, but treatment of intact females with testosterone has a lesser effect on the disposition of ACHC. These results indicate that excretion and residence time of ACHC in both male and female rats are influenced by sex hormones. The described effect is an example of the action of sex hormones on the transport of foreign compounds in this species. Its mechanism is quite different from the well known influence of sex hormones on the microsomal metabolism of foreign compounds in rats. | [
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|
PMID:11976 | Species differences in the metabolism and disposition of spironolactone. | The absorption, excretion and metabolism of [22-14C]spironolactone was compared in Charles River rats, beagle dogs and rhesus monkeys. The drug was administered at the fixed dose of 5 mg/kg po and iv. From the po/iv ratios of the areas under the plasma radioactivity-time curves, the gastrointestinal absorption of the drug was estimated to be 82% in the rat, 62% in the dog, and 103% in the monkey. The absolute bioavailability of a pharmacologically active metabolite, canrenone, was 57% in the dog and 48% in the monkey. Spironolactone was extensively metabolized in all three species and differences existed in the composition of the metabolites in their plasma, urine, and feces. The amount of radioactivity that was excreted in the urine and feces of all three species was similar after either po or iv administration of the drug. The cumulative average excretion of radioactivity in the urine as percentage of the po dose in 6 days was 4.69% in the rat (N = 5), 18.5% in the dog (N = 3), and 46.0% in the monkey (N = 3). In the feces, the corresponding excretion values were 74.2, 69.3 and 40.1%, respectively. Canrenone excretion in the urine constituted 0.65% of the po dose in the rat, 0.82% in the dog, and 5.86% in the monkey, whereas the excretion of total fluorogenic metabolites constituted 1.1, 1.9, and 12.1% respectively. Comparison of animal data with those published for humans indicated that the disposition and metabolism of spironolactone in the rhesus monkey, rather than those in the rat or the dog, was closest to that in man. | Species differences in the metabolism and disposition of spironolactone. The absorption, excretion and metabolism of [22-14C]spironolactone was compared in Charles River rats, beagle dogs and rhesus monkeys. The drug was administered at the fixed dose of 5 mg/kg po and iv. From the po/iv ratios of the areas under the plasma radioactivity-time curves, the gastrointestinal absorption of the drug was estimated to be 82% in the rat, 62% in the dog, and 103% in the monkey. The absolute bioavailability of a pharmacologically active metabolite, canrenone, was 57% in the dog and 48% in the monkey. Spironolactone was extensively metabolized in all three species and differences existed in the composition of the metabolites in their plasma, urine, and feces. The amount of radioactivity that was excreted in the urine and feces of all three species was similar after either po or iv administration of the drug. The cumulative average excretion of radioactivity in the urine as percentage of the po dose in 6 days was 4.69% in the rat (N = 5), 18.5% in the dog (N = 3), and 46.0% in the monkey (N = 3). In the feces, the corresponding excretion values were 74.2, 69.3 and 40.1%, respectively. Canrenone excretion in the urine constituted 0.65% of the po dose in the rat, 0.82% in the dog, and 5.86% in the monkey, whereas the excretion of total fluorogenic metabolites constituted 1.1, 1.9, and 12.1% respectively. Comparison of animal data with those published for humans indicated that the disposition and metabolism of spironolactone in the rhesus monkey, rather than those in the rat or the dog, was closest to that in man. | [
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|
PMID:11977 | The formation of cytochrome P-450-metabolic intermediate complexes in microsomal fractions from extrahepatic tissues of the rabbit. | Cytochrome P-450-metabolic intermediate complexes were formed from N-hydroxyamphetamine, benzphetamine, norbenzphetamine, and d- and l-amphetamine in lung microsomal fractions and from N-hydroxyamphetamine in small intestinal mucosa microsomal fractions. Complexes were not formed in either tissue from SKF 525-A or propoxyphene. The rates of metabolic intermediate complex formation, per mole of cytochrome P-450, were higher in lung than in liver or small intestine. In contrast to that in liver, metabolic intermediate complex formation in the extrahepatic tissues was not dramatically affected by phenobarbital or 3-methylcholanthrene treatment. | The formation of cytochrome P-450-metabolic intermediate complexes in microsomal fractions from extrahepatic tissues of the rabbit. Cytochrome P-450-metabolic intermediate complexes were formed from N-hydroxyamphetamine, benzphetamine, norbenzphetamine, and d- and l-amphetamine in lung microsomal fractions and from N-hydroxyamphetamine in small intestinal mucosa microsomal fractions. Complexes were not formed in either tissue from SKF 525-A or propoxyphene. The rates of metabolic intermediate complex formation, per mole of cytochrome P-450, were higher in lung than in liver or small intestine. In contrast to that in liver, metabolic intermediate complex formation in the extrahepatic tissues was not dramatically affected by phenobarbital or 3-methylcholanthrene treatment. | [
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|
PMID:11978 | Identification of basic metabolites of 4-[4-(p-chlo- robenzoyl)piperidino]-4'-fluorobutyrophenone, an experimental neuroleptic agent. | Two metabolites of 4-[4-(p-(chlorobenzoyl)piperidino]-4'-fluorobutyrophenone (RMI 9901) as well as unchanged drug, have been identified in the urine of rats following oral administration of the drug. Analysis of basic urine extracts by combined gas chromatography-mass spectrometry was employed for metabolite identification. N-dealkylation appears to be a major metabolic pathway and results in formation of 4-(p-chlorobenzoyl)piperidine (I). Subsequent oxidation of this metabolite results in the formation of 4-(p-chlorobenzoyl)-2-piperidone (II), and represents rather unusual metabolic pathway for compounds of this chemical class. | Identification of basic metabolites of 4-[4-(p-chlo- robenzoyl)piperidino]-4'-fluorobutyrophenone, an experimental neuroleptic agent. Two metabolites of 4-[4-(p-(chlorobenzoyl)piperidino]-4'-fluorobutyrophenone (RMI 9901) as well as unchanged drug, have been identified in the urine of rats following oral administration of the drug. Analysis of basic urine extracts by combined gas chromatography-mass spectrometry was employed for metabolite identification. N-dealkylation appears to be a major metabolic pathway and results in formation of 4-(p-chlorobenzoyl)piperidine (I). Subsequent oxidation of this metabolite results in the formation of 4-(p-chlorobenzoyl)-2-piperidone (II), and represents rather unusual metabolic pathway for compounds of this chemical class. | [
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|
PMID:11982 | [Serum-gastrin levels and gastric-acid secretion in infants (author's transl)]. | Gastric acid secretion was measured in 20 infants aged 6-438 days. The values for the basal acid output and that after stimulation with 6 mug/kg pentagastrin subcutaneously were found to be related to age, body weight and body surface area. But these correlations were not comparable to those in adults. Standard values for different age groups in childhood must therefore be established. Furthermore, the results indicate parietal-cell immaturity during the first six months of life. Measurement of fasting serum-gastrin concentration by radioimmunoassay in 74 infants, aged 1-438 days, and 154 adults as controls revealed a high serum-gastrin level in infants, with an exponential decrease during the first year of life. Despite comparable pH values in gastric juice at one year of life, the gastrin concentrations were higher than those in adults (at a statistically significant level). On the other hand, normal serum-gastrin concentrations were found in ten pregnant women just before delivery. The results suggest a negative feed-back mechanism between gastric-acid secretion and fasting serum-gastrin levels, but such mechanism probably being limited by extragastric gastrin secretion. | [Serum-gastrin levels and gastric-acid secretion in infants (author's transl)]. Gastric acid secretion was measured in 20 infants aged 6-438 days. The values for the basal acid output and that after stimulation with 6 mug/kg pentagastrin subcutaneously were found to be related to age, body weight and body surface area. But these correlations were not comparable to those in adults. Standard values for different age groups in childhood must therefore be established. Furthermore, the results indicate parietal-cell immaturity during the first six months of life. Measurement of fasting serum-gastrin concentration by radioimmunoassay in 74 infants, aged 1-438 days, and 154 adults as controls revealed a high serum-gastrin level in infants, with an exponential decrease during the first year of life. Despite comparable pH values in gastric juice at one year of life, the gastrin concentrations were higher than those in adults (at a statistically significant level). On the other hand, normal serum-gastrin concentrations were found in ten pregnant women just before delivery. The results suggest a negative feed-back mechanism between gastric-acid secretion and fasting serum-gastrin levels, but such mechanism probably being limited by extragastric gastrin secretion. | [
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|
PMID:11979 | Metabolism of methoxyphenamine in man and in monkey. | Metabolites of methoxyphenamine in the urine of human subjects and monkeys were separated by gas-liquid chromatography and identified by comparison of their chromatographic and mass spectrometric behavior with those of synthetic compounds. Aromatic O-demethylation, aromatic ring hydroxylation (both followed by glucuronide conjugation), and N-demethylation were shown to occur in man as well as in monkey. In man these were the principal metabolites, whereas in the monkey three additional unidentified major metabolites were formed. | Metabolism of methoxyphenamine in man and in monkey. Metabolites of methoxyphenamine in the urine of human subjects and monkeys were separated by gas-liquid chromatography and identified by comparison of their chromatographic and mass spectrometric behavior with those of synthetic compounds. Aromatic O-demethylation, aromatic ring hydroxylation (both followed by glucuronide conjugation), and N-demethylation were shown to occur in man as well as in monkey. In man these were the principal metabolites, whereas in the monkey three additional unidentified major metabolites were formed. | [
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|
PMID:11980 | A method for the measurement of L-phenylalanine mustard in the mouse and dog by high-pressure liquid chromatography. | The distribution of L-phenylalanine mustard (L-PAM) was studied in dogs and mice by high-pressure liquid chromatography. Separation of L-PAM from its products of hydrolysis was accomplished with a mu-Bondapak C18 column, a solvent system composed of 2-methoxyethanol/0.1% acetic acid, and solvent programming with a step gradient. Complete separation was effected in less than 15 min. The half-life for disappearance of L-PAM from mouse blood was 41 min, whereas that from dog blood was 29 min. The monohydroxy derivative of L-PAM, L-MOH, disappeared from dog serum with a half-life of 32 min. L-MOH was not detectable in mouse tissue other than blood at times greater than 15 min after injection. In the dog at 4 hr after injection, the tissue/serum concentration ratios were greater than 1 for liver, spleen, intestine, skeletal muscle, urinary bladder and gallbladder. The concentration of L-PAM in the bile was approximately 500 times higher than that in serum. | A method for the measurement of L-phenylalanine mustard in the mouse and dog by high-pressure liquid chromatography. The distribution of L-phenylalanine mustard (L-PAM) was studied in dogs and mice by high-pressure liquid chromatography. Separation of L-PAM from its products of hydrolysis was accomplished with a mu-Bondapak C18 column, a solvent system composed of 2-methoxyethanol/0.1% acetic acid, and solvent programming with a step gradient. Complete separation was effected in less than 15 min. The half-life for disappearance of L-PAM from mouse blood was 41 min, whereas that from dog blood was 29 min. The monohydroxy derivative of L-PAM, L-MOH, disappeared from dog serum with a half-life of 32 min. L-MOH was not detectable in mouse tissue other than blood at times greater than 15 min after injection. In the dog at 4 hr after injection, the tissue/serum concentration ratios were greater than 1 for liver, spleen, intestine, skeletal muscle, urinary bladder and gallbladder. The concentration of L-PAM in the bile was approximately 500 times higher than that in serum. | [
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|
PMID:11984 | [Lactic acidosis in diabetics on biguanides (author's transl)]. | A systematic search for cases of lactic acidosis among diabetics on biguanides revealed ten during an eight-month period, while in the preceding ten years not a single case had been definitely diagnosed. This represents a prevelance of 1 in 2000 patients admitted to hospital. All ten were over 60 years old and had previously been treated for heart failure. Most of them had suffered from renal insufficiency for some time. There was no case of biguanide overdosage. Criteria for the diagnosis of lactic acidosis are lactic concentration in blood averaging 18.4 mmol/l and a low pH, averaging 6.9. Serum-biguanide concentration (measured by radioimmuno-assay) was markedly increased. Most of the patients were in circulatory shock on admission or soon after and all of them had a history of gastro-intestinal complaints. Despite intensive treatment with insulin and glucose and careful correction of the acidosis with bicarbonate four of the ten patients died. | [Lactic acidosis in diabetics on biguanides (author's transl)]. A systematic search for cases of lactic acidosis among diabetics on biguanides revealed ten during an eight-month period, while in the preceding ten years not a single case had been definitely diagnosed. This represents a prevelance of 1 in 2000 patients admitted to hospital. All ten were over 60 years old and had previously been treated for heart failure. Most of them had suffered from renal insufficiency for some time. There was no case of biguanide overdosage. Criteria for the diagnosis of lactic acidosis are lactic concentration in blood averaging 18.4 mmol/l and a low pH, averaging 6.9. Serum-biguanide concentration (measured by radioimmuno-assay) was markedly increased. Most of the patients were in circulatory shock on admission or soon after and all of them had a history of gastro-intestinal complaints. Despite intensive treatment with insulin and glucose and careful correction of the acidosis with bicarbonate four of the ten patients died. | [
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|
PMID:11986 | [Alcohol-psychotropic drug interactions]. | The problem of ethanol interactions with psychotropic drugs involves various factors whose complex action is examined in this article. Great care should be taken with regard to the alcohol consumption among patients treated with sedatives; the depressive action of ethanol on the central nervous system could be potentiated especially at the beginning of the treatment, as it seems that a certain habituation to the interactions may exist. However, one should be aware that the personality of the patient is perhaps more important to take into account that the consumption of drugs, at least as regards to the factors of risk in traffic accidents. Parallel to the interactions involving the receptors of the central nervous system, metabolic interactions are possible, however more difficult to describe: they affect drugs absorption, their hepatic metabolism and seem to act especially in chronic alcoholism or during barbiturates consumption. | [Alcohol-psychotropic drug interactions]. The problem of ethanol interactions with psychotropic drugs involves various factors whose complex action is examined in this article. Great care should be taken with regard to the alcohol consumption among patients treated with sedatives; the depressive action of ethanol on the central nervous system could be potentiated especially at the beginning of the treatment, as it seems that a certain habituation to the interactions may exist. However, one should be aware that the personality of the patient is perhaps more important to take into account that the consumption of drugs, at least as regards to the factors of risk in traffic accidents. Parallel to the interactions involving the receptors of the central nervous system, metabolic interactions are possible, however more difficult to describe: they affect drugs absorption, their hepatic metabolism and seem to act especially in chronic alcoholism or during barbiturates consumption. | [
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|
PMID:11987 | Developmental changes in serum luteinizing hormone, follicle stimulating hormone and androgen levels in males of two inbred mouse strains. | A comprehensive developmental study of serum LH, FSH and androgen concentrations was carried out in male mice of two inbred strains. Gonadotropic hormone (GTH) levels rose 10-15 days prior to the pubertal increase in serum androgen with FSH preceding LH in this regard. In both strains GTH titer rose to a peak at 30 or 35 days and then steadily declined to adult levels which were strain-specific. Serum androgen was detected at low, relatively steady levels until the pubertal increase between 30 and 50 days postpartum. The results are interpreted as supporting the hypothesis that a shift in feedback sensitivity, occurring at about 20 days of age, may be involved in the onset of puberty in the male mouse. | Developmental changes in serum luteinizing hormone, follicle stimulating hormone and androgen levels in males of two inbred mouse strains. A comprehensive developmental study of serum LH, FSH and androgen concentrations was carried out in male mice of two inbred strains. Gonadotropic hormone (GTH) levels rose 10-15 days prior to the pubertal increase in serum androgen with FSH preceding LH in this regard. In both strains GTH titer rose to a peak at 30 or 35 days and then steadily declined to adult levels which were strain-specific. Serum androgen was detected at low, relatively steady levels until the pubertal increase between 30 and 50 days postpartum. The results are interpreted as supporting the hypothesis that a shift in feedback sensitivity, occurring at about 20 days of age, may be involved in the onset of puberty in the male mouse. | [
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|
PMID:11988 | Corticotropin releasing factor distribution in normal and Brattleboro rat brain, and effect of deafferentation, hypophysectomy and steroid treatment in normal animals. | Corticotropin-Releasing Factor (CRF) activity was determined (dispersed pituitary cell assay) in rat median eminence (ME), various hypothalamic nuclei, as well as in entire median basal hypothalamus (MBH) and extra-hypothalamic areas. Highest concentrations were seen in ME, with decreased concentrations noted proceeding dorsally and cephalad from ME. Potency (NIAMDD HE-RP-1, ME reference extract, equivalent to 1.0) estimates were: ME-2.2; arcuate n.-0.88; dorsomedial n.-041; ventromedial n.-0.35; periventricular n.-0.24; hypothalamus-0.05; thalamus-0.01; cortex-0.005. Measurable, but lesser amounts, than in the above cited nuclei, were present in supraoptic and paraventricular nuclei. CRF activity was not measurable in preoptic area, septum, olfactory bulb, striatum, mesencephalon, pons, medulla or cerebellum. Complete hypothalamic deafferentation was accompanied by an increase in CRF activity/mug protein in ME and MBH, associated with decreased AM plasma ACTH and corticosterone concentrations. CRF-like activity in ME and MBH increased following hypophysectomy and after dexamethasone pretreatment. These findings indicate that CRF is mainly synthesized in the ME and surrounding area, and this source of CRF is sensitive to feedback effects and that extrahypothalamic inputs affect CRF release. Female animals had higher ME CRF content than did male animals. Homozygous and heterozygous Brattleboro rats had significantly less CRF in ME and MBH than did control animals, with significant differences also noted between homozygous and heterozygous animals. | Corticotropin releasing factor distribution in normal and Brattleboro rat brain, and effect of deafferentation, hypophysectomy and steroid treatment in normal animals. Corticotropin-Releasing Factor (CRF) activity was determined (dispersed pituitary cell assay) in rat median eminence (ME), various hypothalamic nuclei, as well as in entire median basal hypothalamus (MBH) and extra-hypothalamic areas. Highest concentrations were seen in ME, with decreased concentrations noted proceeding dorsally and cephalad from ME. Potency (NIAMDD HE-RP-1, ME reference extract, equivalent to 1.0) estimates were: ME-2.2; arcuate n.-0.88; dorsomedial n.-041; ventromedial n.-0.35; periventricular n.-0.24; hypothalamus-0.05; thalamus-0.01; cortex-0.005. Measurable, but lesser amounts, than in the above cited nuclei, were present in supraoptic and paraventricular nuclei. CRF activity was not measurable in preoptic area, septum, olfactory bulb, striatum, mesencephalon, pons, medulla or cerebellum. Complete hypothalamic deafferentation was accompanied by an increase in CRF activity/mug protein in ME and MBH, associated with decreased AM plasma ACTH and corticosterone concentrations. CRF-like activity in ME and MBH increased following hypophysectomy and after dexamethasone pretreatment. These findings indicate that CRF is mainly synthesized in the ME and surrounding area, and this source of CRF is sensitive to feedback effects and that extrahypothalamic inputs affect CRF release. Female animals had higher ME CRF content than did male animals. Homozygous and heterozygous Brattleboro rats had significantly less CRF in ME and MBH than did control animals, with significant differences also noted between homozygous and heterozygous animals. | [
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|
PMID:11989 | The insulin receptor of the turkey erythrocyte: similarity to mammalian insulin receptors. | Avian erythrocytes possess insulin receptors which have binding properties that are virtually identical to those of the well studied mammalian insulin receptors. The affinity for porcine insulin was identical for the turkey and mammalian receptors over the entire range of insulin concentrations, as was the affinity of each of four insulin analogues which differed 300-fold in biological potency. Insulin induced acceleration of dissociation (i.e., the negatively cooperativite site-site interaction) was indistinguishable over a 10(6) range of insulin concentrations. Sharp pH dependence of binding was identical for turkey and mammalian receptors. The effects of temperature on association, dissociation and steady state binding were also identical. Thus, although birds and mammals have evolved separately for 300 million years there has been little change in the properties of the insulin receptor over this time period. | The insulin receptor of the turkey erythrocyte: similarity to mammalian insulin receptors. Avian erythrocytes possess insulin receptors which have binding properties that are virtually identical to those of the well studied mammalian insulin receptors. The affinity for porcine insulin was identical for the turkey and mammalian receptors over the entire range of insulin concentrations, as was the affinity of each of four insulin analogues which differed 300-fold in biological potency. Insulin induced acceleration of dissociation (i.e., the negatively cooperativite site-site interaction) was indistinguishable over a 10(6) range of insulin concentrations. Sharp pH dependence of binding was identical for turkey and mammalian receptors. The effects of temperature on association, dissociation and steady state binding were also identical. Thus, although birds and mammals have evolved separately for 300 million years there has been little change in the properties of the insulin receptor over this time period. | [
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|
PMID:11990 | Pyridine nucleotides in pancreatic islets during inhibition of insulin release by exogenous insulin. | In vitro addition of rat insulin (200, 400 or 800 muU/ml) to collagenase-isolated pancreatic islets of adult rats diminished glucose (3 mg/ml)-induced insulin release which was correlated with a decrease of the ratio of total NADPH/NADP and inhibition of glucose oxidation via the pentose phosphate shunt (PPS). NADH and NAD levels were not affected. It is suggested that exogenous insulin diminishes the islet total NADPH/NADP ratio by a direct or indirect decrease in PPS activity. However, it is also conceivable that insulin decreases this ratio through another mechanism than PPS. It is possible that inhibition of insulin secretion by exogenous insulin is at least in part due to the decrease of the NADPH/NADP ratio. | Pyridine nucleotides in pancreatic islets during inhibition of insulin release by exogenous insulin. In vitro addition of rat insulin (200, 400 or 800 muU/ml) to collagenase-isolated pancreatic islets of adult rats diminished glucose (3 mg/ml)-induced insulin release which was correlated with a decrease of the ratio of total NADPH/NADP and inhibition of glucose oxidation via the pentose phosphate shunt (PPS). NADH and NAD levels were not affected. It is suggested that exogenous insulin diminishes the islet total NADPH/NADP ratio by a direct or indirect decrease in PPS activity. However, it is also conceivable that insulin decreases this ratio through another mechanism than PPS. It is possible that inhibition of insulin secretion by exogenous insulin is at least in part due to the decrease of the NADPH/NADP ratio. | [
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|
PMID:11991 | Binding of nuclear triiodothyronine (T3) binding protein-T3 complex to chromatin. | Nuclear tirrodothyronine (T3) binding protein (NTBP)-T3 complex, prepared from liver nuclei of rats given [125I]T3 in vovo, rebinds to rat nuclear chromatin at pH 7.4 and at low, but not high, KCl concentrations. Liver NTBP-T3 complex binds to chromatin from liver, kidney, heart, brain, testis, and spleen. Binding was depressed at pH8, by addition of 10 mm CaCl2 or 100 mM MgCl2, and by 1 mM GTP or UTP. Although heart chromatin bound the most NTBP-T3 complex and brain the least, there is no clear separation of binding activity on comparison of three T3-responsive tissues (heart, liver, kidney) to the T3 insensitive tissues. Under the conditions of these experiments, there was no evident competition for binding sites on any of the six chromatins tested. | Binding of nuclear triiodothyronine (T3) binding protein-T3 complex to chromatin. Nuclear tirrodothyronine (T3) binding protein (NTBP)-T3 complex, prepared from liver nuclei of rats given [125I]T3 in vovo, rebinds to rat nuclear chromatin at pH 7.4 and at low, but not high, KCl concentrations. Liver NTBP-T3 complex binds to chromatin from liver, kidney, heart, brain, testis, and spleen. Binding was depressed at pH8, by addition of 10 mm CaCl2 or 100 mM MgCl2, and by 1 mM GTP or UTP. Although heart chromatin bound the most NTBP-T3 complex and brain the least, there is no clear separation of binding activity on comparison of three T3-responsive tissues (heart, liver, kidney) to the T3 insensitive tissues. Under the conditions of these experiments, there was no evident competition for binding sites on any of the six chromatins tested. | [
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PMID:11992 | Teratopsychogenetic effects apparently produced by nonphysiological neurotransmitter concentrations during brain differentiation. | In male rats treated with pargyline, reserpine or pyridostigmine during neonatal life significant permanent changes of sexual behaviour and conditioned learning behaviour were observed in juvenile and/or adult life. Male sexual activity and learning capacity were permanently decreased in neonatally pargyline- or reserpine-treated animals, but permanently increased in neonatally pyridostigmine-treated rats. These findings suggest that nonphysiological concentrations and/or turnover rates of neurotransmitters, if produced during a critical period of brain differentiation, are able to induce lifelond effective behavioural changes, i.e. teratopsychogenetic effects. | Teratopsychogenetic effects apparently produced by nonphysiological neurotransmitter concentrations during brain differentiation. In male rats treated with pargyline, reserpine or pyridostigmine during neonatal life significant permanent changes of sexual behaviour and conditioned learning behaviour were observed in juvenile and/or adult life. Male sexual activity and learning capacity were permanently decreased in neonatally pargyline- or reserpine-treated animals, but permanently increased in neonatally pyridostigmine-treated rats. These findings suggest that nonphysiological concentrations and/or turnover rates of neurotransmitters, if produced during a critical period of brain differentiation, are able to induce lifelond effective behavioural changes, i.e. teratopsychogenetic effects. | [
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|
PMID:11993 | [Plasma testosterone values in patients with somatosexual development disturbances from 11 years old to adulthood]. | The testosterone plasma level was determined in 5 groups: 1. in 69 normal juveniles and 85 fertile males at the age of 11 to 45 years, 2. in 42 patients with hypospadia or epispadia aged 11 to 25 years, 3. in 72 males with unilateral cryptorchidism at the age of 11 to 45 years, 4. in 83 males with bilateral cryptorchidism aged 11 to 45 years and 5. in 106 patients with Klinefelter's syndrome at the age of 16 to 45 years. A pubertal increase of the testosterone plasma level was found to begin in subjects with cryptorchidism or Klinefelter's syndrome at a similar age as in the control group. However, as early as at the age of 13 to 14 years decreased testosterone values were found in the patients as compared to normal juveniles. Between 19 and 20 years, the plasma testosterone level was significantly decreased in all patient-groups as compared to the controls of similar age. In adulthood, plasma testosterone concentrations in the patient groups were observed to be 4 to 6 ng/ml without significant age-dependent changes, which are characteristic of normospermic males. Different degrees of clinical symptoms indicating androgen deficiency found in various patient groups despite similar androgen levels in adulthood suggest a different responsiveness of their target organs to androgens. | [Plasma testosterone values in patients with somatosexual development disturbances from 11 years old to adulthood]. The testosterone plasma level was determined in 5 groups: 1. in 69 normal juveniles and 85 fertile males at the age of 11 to 45 years, 2. in 42 patients with hypospadia or epispadia aged 11 to 25 years, 3. in 72 males with unilateral cryptorchidism at the age of 11 to 45 years, 4. in 83 males with bilateral cryptorchidism aged 11 to 45 years and 5. in 106 patients with Klinefelter's syndrome at the age of 16 to 45 years. A pubertal increase of the testosterone plasma level was found to begin in subjects with cryptorchidism or Klinefelter's syndrome at a similar age as in the control group. However, as early as at the age of 13 to 14 years decreased testosterone values were found in the patients as compared to normal juveniles. Between 19 and 20 years, the plasma testosterone level was significantly decreased in all patient-groups as compared to the controls of similar age. In adulthood, plasma testosterone concentrations in the patient groups were observed to be 4 to 6 ng/ml without significant age-dependent changes, which are characteristic of normospermic males. Different degrees of clinical symptoms indicating androgen deficiency found in various patient groups despite similar androgen levels in adulthood suggest a different responsiveness of their target organs to androgens. | [
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|
PMID:11995 | In vivo measurements of oxyhaemoglobin saturation by a fiberoptic catheter. The effects of variations in pH and haematocrit. | The development of plastic fiberoptic catheters, cheaper and less fragile than glass ones, has enabled the more widespread use of oxyhaemoglobin saturation (SO2) monitoring. They allow direct determinations of SO2 to be made, using reflection spectrophotometry. The purpose of the present work was to evaluate the accuracy of SO2 measurements as provided by these catheters. The studies were performed in dogs, with large variations of SO2, acid-base balance and haematocrit levels. | In vivo measurements of oxyhaemoglobin saturation by a fiberoptic catheter. The effects of variations in pH and haematocrit. The development of plastic fiberoptic catheters, cheaper and less fragile than glass ones, has enabled the more widespread use of oxyhaemoglobin saturation (SO2) monitoring. They allow direct determinations of SO2 to be made, using reflection spectrophotometry. The purpose of the present work was to evaluate the accuracy of SO2 measurements as provided by these catheters. The studies were performed in dogs, with large variations of SO2, acid-base balance and haematocrit levels. | [
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|
PMID:11996 | The action of bombesin on gastric secretion of the chicken. | The natural tetradecapeptide bombesin at a threshold dose of 3--5 ng/kg/min i.v. stimulates gastric secretion in the chicken with either an acute or a chronic proventriculus fistula; this effect is blocked by atropine. The occurrence of tachyphylaxis in the acute, but not in the chronic preparation, and the inhibition of the response by proventriculus acidification, in the presence of an intact sensitivity to caerulein -- a directly stimulating agent -- support the hypothesis of an indirect mechanism of action of bombesin, namely the release of endogenous gastrin, as well as of the existence of gastrin also in the chicken. | The action of bombesin on gastric secretion of the chicken. The natural tetradecapeptide bombesin at a threshold dose of 3--5 ng/kg/min i.v. stimulates gastric secretion in the chicken with either an acute or a chronic proventriculus fistula; this effect is blocked by atropine. The occurrence of tachyphylaxis in the acute, but not in the chronic preparation, and the inhibition of the response by proventriculus acidification, in the presence of an intact sensitivity to caerulein -- a directly stimulating agent -- support the hypothesis of an indirect mechanism of action of bombesin, namely the release of endogenous gastrin, as well as of the existence of gastrin also in the chicken. | [
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|
PMID:11997 | Further evidence implicating E-type prostaglandins in the patency of the lamb ductus arteriosus. | A blocker of prostaglandin (PG) snythesis, indomethacin, given to pregnant ewes near term produces constriction of the lamb ductus arteriosus in utero. The result supports the hypothesis formulated previously that E-type prostaglandins, which are potent relaxant on the ductus, are responsible for maintaining the patency of the vessel during foetal life. | Further evidence implicating E-type prostaglandins in the patency of the lamb ductus arteriosus. A blocker of prostaglandin (PG) snythesis, indomethacin, given to pregnant ewes near term produces constriction of the lamb ductus arteriosus in utero. The result supports the hypothesis formulated previously that E-type prostaglandins, which are potent relaxant on the ductus, are responsible for maintaining the patency of the vessel during foetal life. | [
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|
PMID:11998 | A comparison of the actions of ICI66082 and propranolol on cardiac and peripheral beta-adrenoceptors. | The relative blocking potencies of ICI66082 and propranolol with respect to heart rate contractility, diastolic blood pressure and peripheral vascular conductance were compared in anaesthetized dogs. Peripheral blood flow was measured with an electromagnetic flow-probe around the descending aorta with retrograde cannulation of the inferior mesenteric artery for intra-arterial injections of isoprenaline. Cardiac and peripheral vascular effects of ICI66082 and propranolol were compared in terms of the shifts in the dose--response curves after i.v. and intra-arterial injections of isoprenaline. Propranolol was twice as potent as an equimolar dose of ICI66082 on cardiac beta-adrenoceptors. It was 70--130 times more potent in its action on the peripheral vascular receptors. Propranolol itself was 3 times more potent in blocking peripheral vascular receptors than cardiac beta-receptors. ICI66082 was 17--21 times more active in blocking the myocardial beta-adrenoceptors than those in the peripheral vessels. Electrophysiological studies showed that ICI66082 is devoid of membrane-depressant properties in concentrations up to 100 mg/l. | A comparison of the actions of ICI66082 and propranolol on cardiac and peripheral beta-adrenoceptors. The relative blocking potencies of ICI66082 and propranolol with respect to heart rate contractility, diastolic blood pressure and peripheral vascular conductance were compared in anaesthetized dogs. Peripheral blood flow was measured with an electromagnetic flow-probe around the descending aorta with retrograde cannulation of the inferior mesenteric artery for intra-arterial injections of isoprenaline. Cardiac and peripheral vascular effects of ICI66082 and propranolol were compared in terms of the shifts in the dose--response curves after i.v. and intra-arterial injections of isoprenaline. Propranolol was twice as potent as an equimolar dose of ICI66082 on cardiac beta-adrenoceptors. It was 70--130 times more potent in its action on the peripheral vascular receptors. Propranolol itself was 3 times more potent in blocking peripheral vascular receptors than cardiac beta-receptors. ICI66082 was 17--21 times more active in blocking the myocardial beta-adrenoceptors than those in the peripheral vessels. Electrophysiological studies showed that ICI66082 is devoid of membrane-depressant properties in concentrations up to 100 mg/l. | [
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|
PMID:11999 | Effect of propranolol on antinociceptive, tolerance- and dependence-producing properties of morphine in rodents and monkeys. | Since an abstinence syndrome may accompany the injection of opioids in addicts pretreated with propranolol the morphine antagonistic properties of this compound were investigated. Racemic propranolol did not significantly affect the antinociceptive ED50 of morphine in rodents and neither precipitated abstinence in morphine-dependent monkeys nor exacerbated the syndrome in 24 hr withdrawn monkeys. Multiple doses of propranolol did not alter the development of physical dependence on morphine in monkeys. Clinical narcotic antagonism would not be predicted from this profile. Evidence for a possible propranolol-morphine interaction came from studies using the mouse tail flick test. Thus, after 8 injections of propranolol (over 4 days) mice were tolerant to normally effective doses of morphine. Concurrent injections of naloxone antagonised this effect. When propranolol and morphine were administered concurrently the morphine ED50 (on day 5) was twice that of the group receiving morphine alone. Similar results were obtained with d-propranolol; practolol had a neutral effect. | Effect of propranolol on antinociceptive, tolerance- and dependence-producing properties of morphine in rodents and monkeys. Since an abstinence syndrome may accompany the injection of opioids in addicts pretreated with propranolol the morphine antagonistic properties of this compound were investigated. Racemic propranolol did not significantly affect the antinociceptive ED50 of morphine in rodents and neither precipitated abstinence in morphine-dependent monkeys nor exacerbated the syndrome in 24 hr withdrawn monkeys. Multiple doses of propranolol did not alter the development of physical dependence on morphine in monkeys. Clinical narcotic antagonism would not be predicted from this profile. Evidence for a possible propranolol-morphine interaction came from studies using the mouse tail flick test. Thus, after 8 injections of propranolol (over 4 days) mice were tolerant to normally effective doses of morphine. Concurrent injections of naloxone antagonised this effect. When propranolol and morphine were administered concurrently the morphine ED50 (on day 5) was twice that of the group receiving morphine alone. Similar results were obtained with d-propranolol; practolol had a neutral effect. | [
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|
PMID:12000 | Separate receptors mediating the positive inotropic and chronotropic effect of histamine in guinea-pig atria. | The direct positive inotropic effect of histamine was studied on paced left atrial preparation from guinea pigs. Histamine (10(-8) to 10(-4) M) increased the maximum tension developed in left atria incubated at 35degreesC and driven at 2 Hz. The maximum increase in tension was 60% of that observed with norepinephrine. Metiamide (3 X 10(-5) M, a specific H2-receptor antagonist) did not alter the inotropic response of left atria to histamine. However, tripelennamine (a typical H1-receptor antagonist) competitively shifted the histamine inotropic dose--response curve to the right at concentrations from 10(-8) to 10(-7) M. Higher concentrations (3 X 10(-7) and 10(-6) M) caused little further additional shift to the right. The positive chronotropic effect of histamine on spontaneously beating atria was competitively antagonized by metiamide (10(-6) and 3 X 10(-6) M). These results demonstrate that in guinea-pig atria histamine increases myocardial contractility by an interaction with receptors closely related to classical H1-receptors while its chronotropic effect is mediated by interaction with H2-receptors. | Separate receptors mediating the positive inotropic and chronotropic effect of histamine in guinea-pig atria. The direct positive inotropic effect of histamine was studied on paced left atrial preparation from guinea pigs. Histamine (10(-8) to 10(-4) M) increased the maximum tension developed in left atria incubated at 35degreesC and driven at 2 Hz. The maximum increase in tension was 60% of that observed with norepinephrine. Metiamide (3 X 10(-5) M, a specific H2-receptor antagonist) did not alter the inotropic response of left atria to histamine. However, tripelennamine (a typical H1-receptor antagonist) competitively shifted the histamine inotropic dose--response curve to the right at concentrations from 10(-8) to 10(-7) M. Higher concentrations (3 X 10(-7) and 10(-6) M) caused little further additional shift to the right. The positive chronotropic effect of histamine on spontaneously beating atria was competitively antagonized by metiamide (10(-6) and 3 X 10(-6) M). These results demonstrate that in guinea-pig atria histamine increases myocardial contractility by an interaction with receptors closely related to classical H1-receptors while its chronotropic effect is mediated by interaction with H2-receptors. | [
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|
PMID:12027 | The effects of altering the pH of seminal fluid on the sex ratio of rabbit offspring. | It has been suggested that the pH of the vagina at the time of fertilization may have a differential effect on X- or Y-bearing sperm and thereby affect the sex of the offspring. To test this postulate, rabbit semen was collected, diluted 1:10 with a buffer of pH 5.4, 6.9, or 9.6, and after 20 minutes 0.5 ml of semen-buffer mixture was used for insemination in an ovulation-induced female. Newborn pups were examined both externally and internally for gender. The females inseminated with acidic semen had 6 litters, 50 offspring, with 48% males; those with neutral semen had 8 litters, 48 offspring, with 63% males; and those with alkaline semen had 7 litters, 49 offspring, with 49% males. There was no significant difference in these sex ratios from the expected 50% males. Motility of rabbit sperm at 23 degrees C in buffers of pH 4.6, 5.4, 6.9, 9.6, and 9.8 was reduced in vitro as the pH deviated from neutrality. Acid conditions were more detrimental than alkaline conditions. Sperm lost their motility more quickly in buffers of 37 degrees C than in buffers of 23 degrees C. It was not possible with scanning electron microscopy to distinguish morphologically between X- and Y-bearing sperm. It seems unlikely that a direct effect of pH on sperm can be a single influence on the sex of offspring. | The effects of altering the pH of seminal fluid on the sex ratio of rabbit offspring. It has been suggested that the pH of the vagina at the time of fertilization may have a differential effect on X- or Y-bearing sperm and thereby affect the sex of the offspring. To test this postulate, rabbit semen was collected, diluted 1:10 with a buffer of pH 5.4, 6.9, or 9.6, and after 20 minutes 0.5 ml of semen-buffer mixture was used for insemination in an ovulation-induced female. Newborn pups were examined both externally and internally for gender. The females inseminated with acidic semen had 6 litters, 50 offspring, with 48% males; those with neutral semen had 8 litters, 48 offspring, with 63% males; and those with alkaline semen had 7 litters, 49 offspring, with 49% males. There was no significant difference in these sex ratios from the expected 50% males. Motility of rabbit sperm at 23 degrees C in buffers of pH 4.6, 5.4, 6.9, 9.6, and 9.8 was reduced in vitro as the pH deviated from neutrality. Acid conditions were more detrimental than alkaline conditions. Sperm lost their motility more quickly in buffers of 37 degrees C than in buffers of 23 degrees C. It was not possible with scanning electron microscopy to distinguish morphologically between X- and Y-bearing sperm. It seems unlikely that a direct effect of pH on sperm can be a single influence on the sex of offspring. | [
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|
PMID:12062 | Properties of prostaglandin F2alpha receptors in bovine corpus luteum cell membranes. | The specific binding of 3H-labeled prostaglandin (PG) F2alpha to bovine corpus luteum cell membranes was a rapid (K1=1.1 X 10(4) M(-1)S(-1) and reversible (K(-1)=3.3 X 10(-4) S(-1)) process at 22 degrees C. The specific binding was also a saturable process exhibiting two classes of receptors with apparent dissociation constants (Kds) of 1.6 X 10(-9) M and 2.4 X 10(-8) M. The heterogenous nature of [3H]PGF2alpha binding does not appear to be due to negative cooperatively but merely to represent the existence of two independent groups of receptor sites with discrete affinities. Free energy changes of +11.9 and +10.3 Kcal/mol were calculated from the Kds of high and low affinity receptors, respectively. The binding of [3H]PGF2alpha to the membranes was not accompanied by any detectable changes in receptor-bound or free [3H]PGF2alpha. Addition of increasing amounts of unlabeled PGF2alpha resulted in a dose-dependent inhibition of [3H]PGF2alpha binding to the membranes, with complete inhibition occurring at 10(-6) M. Other unlabeled PGs such as PGF1alpha, PGE2 (5-fold), PGE1 (120-fold), PGA1 and PGB1 (about 10,000-fold) were less effective when compared to unlabeled PGF2alpha in inhibiting [3H] PGF2alpha binding to the membranes. The metabolites of PGF2alpha, 15-keto-PGF2alpha and 13,14-dihydro-15-keto-PGF2alpha had 100-fold less affinity for PGF2alpha receptors. 15(S)15-Methyl-PGF2alpha, an analogue of PGF2alpha, had a fairly high affinity but lower than its parent molecule. Various unsaturated fatty acids, indomethacin and 7-oxa-13-prostynoic acid had 3,000- to 10,000-fold less affinities for PGF2alpha receptors. Incubation of membranes with various enzymes revealed that PGF2alpha receptor molecules are protein in nature which require membrane lipids and specific phospholipids for binding function. Among the various phospholipids used, sphingomyelin was found to be very effective in restoring the loss of [3H]PGF2alpha binding in phospholipase C-treated membranes. N-Ethylmaleimide, but not other SH group alkylating agents inhibited binding. The binding was also inhibited by tetranitromethane, dinitrofluorobenzene and acetic anhydride. This suggested that tyrosyl, histidyl, tryptophan and amino (any one or all of them) but not SH groups were involved in binding interaction. | Properties of prostaglandin F2alpha receptors in bovine corpus luteum cell membranes. The specific binding of 3H-labeled prostaglandin (PG) F2alpha to bovine corpus luteum cell membranes was a rapid (K1=1.1 X 10(4) M(-1)S(-1) and reversible (K(-1)=3.3 X 10(-4) S(-1)) process at 22 degrees C. The specific binding was also a saturable process exhibiting two classes of receptors with apparent dissociation constants (Kds) of 1.6 X 10(-9) M and 2.4 X 10(-8) M. The heterogenous nature of [3H]PGF2alpha binding does not appear to be due to negative cooperatively but merely to represent the existence of two independent groups of receptor sites with discrete affinities. Free energy changes of +11.9 and +10.3 Kcal/mol were calculated from the Kds of high and low affinity receptors, respectively. The binding of [3H]PGF2alpha to the membranes was not accompanied by any detectable changes in receptor-bound or free [3H]PGF2alpha. Addition of increasing amounts of unlabeled PGF2alpha resulted in a dose-dependent inhibition of [3H]PGF2alpha binding to the membranes, with complete inhibition occurring at 10(-6) M. Other unlabeled PGs such as PGF1alpha, PGE2 (5-fold), PGE1 (120-fold), PGA1 and PGB1 (about 10,000-fold) were less effective when compared to unlabeled PGF2alpha in inhibiting [3H] PGF2alpha binding to the membranes. The metabolites of PGF2alpha, 15-keto-PGF2alpha and 13,14-dihydro-15-keto-PGF2alpha had 100-fold less affinity for PGF2alpha receptors. 15(S)15-Methyl-PGF2alpha, an analogue of PGF2alpha, had a fairly high affinity but lower than its parent molecule. Various unsaturated fatty acids, indomethacin and 7-oxa-13-prostynoic acid had 3,000- to 10,000-fold less affinities for PGF2alpha receptors. Incubation of membranes with various enzymes revealed that PGF2alpha receptor molecules are protein in nature which require membrane lipids and specific phospholipids for binding function. Among the various phospholipids used, sphingomyelin was found to be very effective in restoring the loss of [3H]PGF2alpha binding in phospholipase C-treated membranes. N-Ethylmaleimide, but not other SH group alkylating agents inhibited binding. The binding was also inhibited by tetranitromethane, dinitrofluorobenzene and acetic anhydride. This suggested that tyrosyl, histidyl, tryptophan and amino (any one or all of them) but not SH groups were involved in binding interaction. | [
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|
PMID:12063 | Isozyme patterns of branched-chain amino acid transminase in cultured Morris hepatoma 7316A. | Cultured cells from Morris hepatoma 7316A contained isozymes I and II, but not isozyme III, of branched-chain amino acid transaminase. They also contained tyrosine transaminase. Isozyme II and tyrosine transaminase were induced by addition of cortisol. These findings agree well with in vivo findings. However, prolonged culutre of the cells for over 500 days caused deviation of the chromosomal bumber and activity of both enzymes, and they were no longer affected by cortisol. A tumor formed by back-transplantation of the cells showed the typical isozyme pattern of rapidly growing hepatomas, such as Yoshida ascites hepatomas, i.e., isozymes I and III. These results were discussed in relation to change of gene expression during culture. | Isozyme patterns of branched-chain amino acid transminase in cultured Morris hepatoma 7316A. Cultured cells from Morris hepatoma 7316A contained isozymes I and II, but not isozyme III, of branched-chain amino acid transaminase. They also contained tyrosine transaminase. Isozyme II and tyrosine transaminase were induced by addition of cortisol. These findings agree well with in vivo findings. However, prolonged culutre of the cells for over 500 days caused deviation of the chromosomal bumber and activity of both enzymes, and they were no longer affected by cortisol. A tumor formed by back-transplantation of the cells showed the typical isozyme pattern of rapidly growing hepatomas, such as Yoshida ascites hepatomas, i.e., isozymes I and III. These results were discussed in relation to change of gene expression during culture. | [
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|
PMID:12064 | Serum gamma-glutamyl transpeptidase activity in viral hepatitis: suppression in pregnancy and by birth control pills. | gamma-Glutamyl transpeptidase (GGT) activity in serum was increased in the majority of women with viral hepatitis occurring in the first half of pregnancy. By contrast, GGT activity was abnormal less frequently and the mean value was relatively depressed, even though hepatitis was as severe, in the second half of gestation. Mean GGT activity was also lower, and abnormal values were less frequent, in nonpregnant women with viral hepatitis who were taking birth control pills (BCP). Depressed GGT is not attributable to an inhibitor in serum in women in late pregnancy or taking BCP. The data suggest that estrogen and/or progestational compounds affect liver such that less GGT is released into blood with acute hepatocellular injury. In addition, hyperbilirubinemia was found to be associated with depressed serum GGT activity, and bilirubin added to serum in vitro interfered with measured activity of the enzyme. | Serum gamma-glutamyl transpeptidase activity in viral hepatitis: suppression in pregnancy and by birth control pills. gamma-Glutamyl transpeptidase (GGT) activity in serum was increased in the majority of women with viral hepatitis occurring in the first half of pregnancy. By contrast, GGT activity was abnormal less frequently and the mean value was relatively depressed, even though hepatitis was as severe, in the second half of gestation. Mean GGT activity was also lower, and abnormal values were less frequent, in nonpregnant women with viral hepatitis who were taking birth control pills (BCP). Depressed GGT is not attributable to an inhibitor in serum in women in late pregnancy or taking BCP. The data suggest that estrogen and/or progestational compounds affect liver such that less GGT is released into blood with acute hepatocellular injury. In addition, hyperbilirubinemia was found to be associated with depressed serum GGT activity, and bilirubin added to serum in vitro interfered with measured activity of the enzyme. | [
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|
PMID:12065 | [Effect of O-methylhydroxylamine on extracellular phage cd]. | Study was made of lethal and mutagenic effect of 1 M and 0,5 M O-methylhydroxylamine (OMHA) on extracellular phage Sd. The correlation between chemical changes of the genome and the degree of phage inactivation under the action of OMHA has been established within the range of studied pH (4,5-7,0) of the reaction medium. OMHA in activates the phage at the highest rate at pH 5,0, which agrees with chemical data indicating that the total rate of OMHA modification of cytidine units is maximal at this pH. Inactivation curves of OMHA-treated phage are single-hit at pH investigated, but have a small initial shoulder; at pH 5,0 and 4,5 inactivation curves consist of two exponents, the second exponent having the smallest slope, that is the phage is characterized by an increased resistance to OMHA at this section. The increased phage resistance can be explained by transforming the original product IV (cross-linked with protein) into the product II (N4-methoxy-6-methoxyamine-5,6-dihydrocytidine) which can be repaired in contrast to IV. OMHA has a high mutagenic effect on phage Sd. Under optimal conditions (at pH 4,5) the mutagen induces plaque mutants (up to 6%) among survived phages. The data obtained correlate with the fact that with decreasing pH (from 5,0 to 4,5) the ratio of the "mutagen" unit - N4-methoxycytidine (product III) to the "inactivating" one (product II) increases. The curves of mutation induction under the action of OMHA have a characteristic form with the initial linear section and the maximum or the plateau similar to mutation curves to be observed under the action of radiation and chemical agents. | [Effect of O-methylhydroxylamine on extracellular phage cd]. Study was made of lethal and mutagenic effect of 1 M and 0,5 M O-methylhydroxylamine (OMHA) on extracellular phage Sd. The correlation between chemical changes of the genome and the degree of phage inactivation under the action of OMHA has been established within the range of studied pH (4,5-7,0) of the reaction medium. OMHA in activates the phage at the highest rate at pH 5,0, which agrees with chemical data indicating that the total rate of OMHA modification of cytidine units is maximal at this pH. Inactivation curves of OMHA-treated phage are single-hit at pH investigated, but have a small initial shoulder; at pH 5,0 and 4,5 inactivation curves consist of two exponents, the second exponent having the smallest slope, that is the phage is characterized by an increased resistance to OMHA at this section. The increased phage resistance can be explained by transforming the original product IV (cross-linked with protein) into the product II (N4-methoxy-6-methoxyamine-5,6-dihydrocytidine) which can be repaired in contrast to IV. OMHA has a high mutagenic effect on phage Sd. Under optimal conditions (at pH 4,5) the mutagen induces plaque mutants (up to 6%) among survived phages. The data obtained correlate with the fact that with decreasing pH (from 5,0 to 4,5) the ratio of the "mutagen" unit - N4-methoxycytidine (product III) to the "inactivating" one (product II) increases. The curves of mutation induction under the action of OMHA have a characteristic form with the initial linear section and the maximum or the plateau similar to mutation curves to be observed under the action of radiation and chemical agents. | [
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|
PMID:12067 | Blood pH: a test for assessment of severity in proctocolitis. | Acid base balance was studied in 58 patients with active idiopathic proctocolitis; the condition of 10 of them was complicated by toxic megacolon. Arterial blood pH increased progressively with increased severity of the colitis and as the lesions became more widespread. Statistically significant differences were observed in pH values between the mild/moderate and severe forms and between the severe and complicated forms ('toxic megacolon'). A linear correlation was found between pH and the amount of intestinal gas, pulse rate, and plasma albumin. | Blood pH: a test for assessment of severity in proctocolitis. Acid base balance was studied in 58 patients with active idiopathic proctocolitis; the condition of 10 of them was complicated by toxic megacolon. Arterial blood pH increased progressively with increased severity of the colitis and as the lesions became more widespread. Statistically significant differences were observed in pH values between the mild/moderate and severe forms and between the severe and complicated forms ('toxic megacolon'). A linear correlation was found between pH and the amount of intestinal gas, pulse rate, and plasma albumin. | [
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|
PMID:12068 | Relationships between mucosal hydrolysis and transport of two phenylalanine dipeptides. | In order to investigate the source of free amino acids found in the gut lumen during absorption of dipeptides, as well as evaluating the role of brush border peptidases in the mucosal hydrolysis of dipeptides during absorption, rates of dipeptide disappearance and appearance of hydrolytic products were measured during perfusion of rat jejunum and ileum in vivo with buffered and unbuffered 10 mM solutions of glycl-L-phenylalanine (Gly-Phe) and L-phenylalanyl-glycine (Phe-Gly). Mucosal brush border peptidase activity was then measured in the perfused segments in vitro at luminal pH and at two substrate concentrations. In addition cytosol peptidase activity in the perfused segments was measured at pH 7-4 and at 10 mM substrate concentrations. In the jejunum, there was a relationship between rates of free phenylalanine appearance in vivo (Phe-Gly greater than Gly-Phe) and rates of brush border (Phe-Gly greater than Gly-Phe) rather than cytosol (Gly-Phe greater than Phe-Gly) peptidase activities. No constant relationship between free phenylalanine appearance and hydrolysis of the dipeptides by either brush border or cytosol peptidases was observed in the ileal studies. These findings suggest that, in the jejunum, hydrolytic products originate from the surface of the cell whereas, in the ileum, hydrolytic products originate from both the intracellular compartment as well as from the surface of the mucosal cell. In the jejunum, in vitro rates of brush border hydrolysis of Gly-Phe were always less than in vivo disappearance rates, whereas rates of Phe-Gly brush border hydrolysis always exceeded luminal disappearance rates. These data imply that Gly-Phe is predominantly transported intact and hydrolysed by cytosol peptidases, In contrast, brush border peptidases play an importnat role in the mucosal hydrolysis of Phe-Gly. | Relationships between mucosal hydrolysis and transport of two phenylalanine dipeptides. In order to investigate the source of free amino acids found in the gut lumen during absorption of dipeptides, as well as evaluating the role of brush border peptidases in the mucosal hydrolysis of dipeptides during absorption, rates of dipeptide disappearance and appearance of hydrolytic products were measured during perfusion of rat jejunum and ileum in vivo with buffered and unbuffered 10 mM solutions of glycl-L-phenylalanine (Gly-Phe) and L-phenylalanyl-glycine (Phe-Gly). Mucosal brush border peptidase activity was then measured in the perfused segments in vitro at luminal pH and at two substrate concentrations. In addition cytosol peptidase activity in the perfused segments was measured at pH 7-4 and at 10 mM substrate concentrations. In the jejunum, there was a relationship between rates of free phenylalanine appearance in vivo (Phe-Gly greater than Gly-Phe) and rates of brush border (Phe-Gly greater than Gly-Phe) rather than cytosol (Gly-Phe greater than Phe-Gly) peptidase activities. No constant relationship between free phenylalanine appearance and hydrolysis of the dipeptides by either brush border or cytosol peptidases was observed in the ileal studies. These findings suggest that, in the jejunum, hydrolytic products originate from the surface of the cell whereas, in the ileum, hydrolytic products originate from both the intracellular compartment as well as from the surface of the mucosal cell. In the jejunum, in vitro rates of brush border hydrolysis of Gly-Phe were always less than in vivo disappearance rates, whereas rates of Phe-Gly brush border hydrolysis always exceeded luminal disappearance rates. These data imply that Gly-Phe is predominantly transported intact and hydrolysed by cytosol peptidases, In contrast, brush border peptidases play an importnat role in the mucosal hydrolysis of Phe-Gly. | [
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|
PMID:12069 | Effect of metiamide on basal and stimulated serum cholecystokinin levels in duodenal ulcer patients. | Serum cholecystokinin (CCK) levels were measured in 10 patients with chronic duodenal ulcers, fasting and at intervals after two standard tests meals (300 ml of 40 mmol/1 phenylalanine solution), one given before and one during H2-receptor blockade with metiamide (200 mg four times a day). Fasting serum CCK levels were lower in all patients during treatment with metiamide (the mean level falling from 306-0 +/- 102-0 (SEM) to 82-1 +/- 23-6 pg/ml after treatment (p less than 0-01)). In contrast, peak serum CCK levels after the meal were not significantly different (7400 +/- 1141 pg/ml before treatment and 7569 +/- 1293 pg/ml on metiamide). We conclude that in duodenal ulcer patients CCK secretion under basal condtions may be in part dependent on stimulation of the small intestinal mucosa by gastric acid, but that, after an amino acid meal, gastric acid secretion is less important in determining the amount of CCK released. | Effect of metiamide on basal and stimulated serum cholecystokinin levels in duodenal ulcer patients. Serum cholecystokinin (CCK) levels were measured in 10 patients with chronic duodenal ulcers, fasting and at intervals after two standard tests meals (300 ml of 40 mmol/1 phenylalanine solution), one given before and one during H2-receptor blockade with metiamide (200 mg four times a day). Fasting serum CCK levels were lower in all patients during treatment with metiamide (the mean level falling from 306-0 +/- 102-0 (SEM) to 82-1 +/- 23-6 pg/ml after treatment (p less than 0-01)). In contrast, peak serum CCK levels after the meal were not significantly different (7400 +/- 1141 pg/ml before treatment and 7569 +/- 1293 pg/ml on metiamide). We conclude that in duodenal ulcer patients CCK secretion under basal condtions may be in part dependent on stimulation of the small intestinal mucosa by gastric acid, but that, after an amino acid meal, gastric acid secretion is less important in determining the amount of CCK released. | [
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|
PMID:12070 | Reversible norepinephrine binding to rabbit myometrium: relationship to sites of known biological significance. | Catecholamines bind specifically to several cell types and subcellular fractions. This binding is not consistent with binding to the adrenergic receptor. Catecholamines would be expected to bind to sites other than the adrenergic receptor (uptake 1, uptake 2, nerve vesicles, catecholamine-O-methyl transferase and serum albumin). We characterized the binding of 3H-norepinephrine to microsomes prepared from rabbit myometrium. In the presence of antioxidant binding was reversible and 3H-norepinephrine could be recovered bound to microsomes or free in incubation media. The binding was not characteristic of binding to any one known biological site. It may represent binding to a mixture of these sites or may be fortuitous binding to sites of unknown biological significance. | Reversible norepinephrine binding to rabbit myometrium: relationship to sites of known biological significance. Catecholamines bind specifically to several cell types and subcellular fractions. This binding is not consistent with binding to the adrenergic receptor. Catecholamines would be expected to bind to sites other than the adrenergic receptor (uptake 1, uptake 2, nerve vesicles, catecholamine-O-methyl transferase and serum albumin). We characterized the binding of 3H-norepinephrine to microsomes prepared from rabbit myometrium. In the presence of antioxidant binding was reversible and 3H-norepinephrine could be recovered bound to microsomes or free in incubation media. The binding was not characteristic of binding to any one known biological site. It may represent binding to a mixture of these sites or may be fortuitous binding to sites of unknown biological significance. | [
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|
PMID:12071 | Occurrence of C1 inactivator and other proteinase inhibitors in euglobulin fractions and their influence on fibrinolytic activity. | Considerable amounts of C1 inactivator and inter-alpha-trypsin inhibitor pecipitate during euglobulin fractionation of human plasma. The amount precipitated depends on the ionic strength and the pH during the fractionation procedure. In contrast, alpha1-anti-trypsin, alpha2-macroglobulinand antithrombin II are present in euglobulin fractions in trace amounts only. The fibrinolytic activity of the euglobulin fractions is inhibited by the endogenous C1 inactivator, particularly as shown by comparison of normal and hereditary angioneurotic edema (HANE) plasma. | Occurrence of C1 inactivator and other proteinase inhibitors in euglobulin fractions and their influence on fibrinolytic activity. Considerable amounts of C1 inactivator and inter-alpha-trypsin inhibitor pecipitate during euglobulin fractionation of human plasma. The amount precipitated depends on the ionic strength and the pH during the fractionation procedure. In contrast, alpha1-anti-trypsin, alpha2-macroglobulinand antithrombin II are present in euglobulin fractions in trace amounts only. The fibrinolytic activity of the euglobulin fractions is inhibited by the endogenous C1 inactivator, particularly as shown by comparison of normal and hereditary angioneurotic edema (HANE) plasma. | [
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|
PMID:12072 | Studies on the dynamics and mechanism of glibenclamide-induced insulin secretion. | Sustained, 60-minute perfusion of glibenclamide (0.5, 1.5 and 10 mug/ml) elicits a one-phase insulin release profile, formed by a rapid secretion peak followed by a second peak with lower insulin levels than the former. Basal insulin secretion values are observed during the period comprised between 13 and 60 minutes of perfusion. Concurrent stimulation with glucose (100, 150, 200 and 300 mg%) plus glibenclamide (1 mug/ml) causes a marked rise in both phases of insulin secretion. The addition of glibenclamide does not modify the biphasic secretion pattern caused by maximal glucose concentration (400 mg%). The maximal values of both phases of secretion in the dose-response curve elicited by different glucose concentrations shift to the left when glibenclamide is added to the perfusate. The increase in insulin secretion caused by glibenclamide is not inhibited by puromycin. Both theophylline and phentolamine modify and increase the glibenclamide-induced insulin release pattern. Propranolol and imidazole inhibit glibenclamide-induced insulin release. Our results suggest that: 1. Glibenclamide increases beta cell sensitivity to glucose stimulation. 2. Glibenclamide and glucose induce secretion of insulin originating in the same compartment. 3. Modification of alpha and beta adrenergic receptors may modify glibodulate the beta cell response to glibenclamide. | Studies on the dynamics and mechanism of glibenclamide-induced insulin secretion. Sustained, 60-minute perfusion of glibenclamide (0.5, 1.5 and 10 mug/ml) elicits a one-phase insulin release profile, formed by a rapid secretion peak followed by a second peak with lower insulin levels than the former. Basal insulin secretion values are observed during the period comprised between 13 and 60 minutes of perfusion. Concurrent stimulation with glucose (100, 150, 200 and 300 mg%) plus glibenclamide (1 mug/ml) causes a marked rise in both phases of insulin secretion. The addition of glibenclamide does not modify the biphasic secretion pattern caused by maximal glucose concentration (400 mg%). The maximal values of both phases of secretion in the dose-response curve elicited by different glucose concentrations shift to the left when glibenclamide is added to the perfusate. The increase in insulin secretion caused by glibenclamide is not inhibited by puromycin. Both theophylline and phentolamine modify and increase the glibenclamide-induced insulin release pattern. Propranolol and imidazole inhibit glibenclamide-induced insulin release. Our results suggest that: 1. Glibenclamide increases beta cell sensitivity to glucose stimulation. 2. Glibenclamide and glucose induce secretion of insulin originating in the same compartment. 3. Modification of alpha and beta adrenergic receptors may modify glibodulate the beta cell response to glibenclamide. | [
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|
PMID:12073 | Plasma testosterone in prepubertal cryptorchid boys under long-term HCG therapy. | Plasma testosterone concentrations were determined before and after 6 weeks of human chorionic gonadotropin treatment in 36 prepubertal boys with bilateral or unilateral cryptorchidism. Mean +/- SD basal and post-treatment values (ng/100 ml) in the bilateral group were: treatment successful (n = 14): 32 +/- 19 and 302 +/- 49, treatment unsuccessful (n = 12): 20 +/- 15 and 176 +/- 73. The figures for the unilateral group were: treatment successful (n = 6): 23 +/- 9 and 244 +/- 41, treatment unsuccessful (n = 5): 22 +/- 11 and 264 +/- 102. In the bilateral group significant differences in the T response emerged when successfully treated boys were compared to unsuccessfully treated ones. It is concluded that Leydig cell function may be impaired in some cases of cryptorchidism. | Plasma testosterone in prepubertal cryptorchid boys under long-term HCG therapy. Plasma testosterone concentrations were determined before and after 6 weeks of human chorionic gonadotropin treatment in 36 prepubertal boys with bilateral or unilateral cryptorchidism. Mean +/- SD basal and post-treatment values (ng/100 ml) in the bilateral group were: treatment successful (n = 14): 32 +/- 19 and 302 +/- 49, treatment unsuccessful (n = 12): 20 +/- 15 and 176 +/- 73. The figures for the unilateral group were: treatment successful (n = 6): 23 +/- 9 and 244 +/- 41, treatment unsuccessful (n = 5): 22 +/- 11 and 264 +/- 102. In the bilateral group significant differences in the T response emerged when successfully treated boys were compared to unsuccessfully treated ones. It is concluded that Leydig cell function may be impaired in some cases of cryptorchidism. | [
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|
PMID:12074 | Effects of D-glyceraldehyde and 3-o-methylglucose upon fluorescence of reduced pyridine nucleotides from perifused isolated pancreatic islets. | In perfused pancreatic islets, the fluorescence of reduced pyridine nucleotides was recorded continuously, D-Glyceraldehyde (5 mM) or 3-o-methylglucose (27.5 mM) never caused a net fluorescence increase. Since stepwise changes of the D-glucose concentration between 0 and 20 mM always induced a fluorescence increase, it is concluded that glucose on the one hand and glyceraldehyde or 3-o-methylglucose on the other hand cause different metabolic states in pancreatic islets. | Effects of D-glyceraldehyde and 3-o-methylglucose upon fluorescence of reduced pyridine nucleotides from perifused isolated pancreatic islets. In perfused pancreatic islets, the fluorescence of reduced pyridine nucleotides was recorded continuously, D-Glyceraldehyde (5 mM) or 3-o-methylglucose (27.5 mM) never caused a net fluorescence increase. Since stepwise changes of the D-glucose concentration between 0 and 20 mM always induced a fluorescence increase, it is concluded that glucose on the one hand and glyceraldehyde or 3-o-methylglucose on the other hand cause different metabolic states in pancreatic islets. | [
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PMID:12075 | Peptidase inactivation of hypothalamic releasing hormones. | With the structural characterization of the hypothalamic hormones, luteinizing hormone-releasing hormone (LH-RH), thyrotrophin-releasing (TRH), melanocyte-stimulating hormone release-inhibiting hormine (MIH), and growth hormone release-inhibiting hormone, (GH-RIH or somatostatin), it has been possible to investigate their enzymic inactivation by peptidases which are present at various sites in the body. Enzymes may play an important part in the control of polypeptide hormone levels and the peptidases acting on these four hypothalamic hormones may regulate the amount of TRH, LH-RH, MIH and somatostatin released from the hypothalamus, or their action at the level of the pituitary and their removal from the circulation. By studying the peptidase enzymes, further information may be obtained on the physiological mechanisms controlling the secretion and actions of hypothalamic hormones, as well as on the design of analogues with increased or competitive activity. | Peptidase inactivation of hypothalamic releasing hormones. With the structural characterization of the hypothalamic hormones, luteinizing hormone-releasing hormone (LH-RH), thyrotrophin-releasing (TRH), melanocyte-stimulating hormone release-inhibiting hormine (MIH), and growth hormone release-inhibiting hormone, (GH-RIH or somatostatin), it has been possible to investigate their enzymic inactivation by peptidases which are present at various sites in the body. Enzymes may play an important part in the control of polypeptide hormone levels and the peptidases acting on these four hypothalamic hormones may regulate the amount of TRH, LH-RH, MIH and somatostatin released from the hypothalamus, or their action at the level of the pituitary and their removal from the circulation. By studying the peptidase enzymes, further information may be obtained on the physiological mechanisms controlling the secretion and actions of hypothalamic hormones, as well as on the design of analogues with increased or competitive activity. | [
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|
PMID:12076 | [The biosynthesis of glutathione in human erythrocytes (author's transl)]. | The concentrations of glutathione precursors in human erythrocytes were investigated. 300muM glutamate, 375 muM glycine, and 10muM cysteine were found by automated amino acid analysis. The concentration of 2-aminobutyrate, the precursor of ophthalmic acid, was 15muM. The influence of the activities of endogenous or added glutamyl-cysteine synthetase and glutathione synthetase on the rate of glutathione biosynthesis was measured in membrane-free hemolysates under physiological conditions. The results show that the rate of the overall biosynthesis mainly depends on the formation of the dipeptide glutamyl-cysteine. The effect of glutathione precursor concentrations on the synthesis of the tripeptide was investigated at constant (endogenous) activities of the synthesizing enzymes. The rate was not enhanced by addition of glutamate and/or glycine unless cysteine or glutamyl-cysteine was also added. It is concluded that the concentration of cysteine limits the actual rate of the glutamyl-cysteine-synthetase reaction in vivo. No cysteine or bis(glutamyl)cystine was detected in human hemolysate; however, these disulfides were converted to glutathione. This indicates that erythrocytes have an appropriate system for their reduction, since the disulfides themselves are not substrates for the glutathione-synthesizing enzymes. Studies with intact human red cells indicate that the uptake of cysteine is the rate-determining step in the biosynthesis of glutathione. | [The biosynthesis of glutathione in human erythrocytes (author's transl)]. The concentrations of glutathione precursors in human erythrocytes were investigated. 300muM glutamate, 375 muM glycine, and 10muM cysteine were found by automated amino acid analysis. The concentration of 2-aminobutyrate, the precursor of ophthalmic acid, was 15muM. The influence of the activities of endogenous or added glutamyl-cysteine synthetase and glutathione synthetase on the rate of glutathione biosynthesis was measured in membrane-free hemolysates under physiological conditions. The results show that the rate of the overall biosynthesis mainly depends on the formation of the dipeptide glutamyl-cysteine. The effect of glutathione precursor concentrations on the synthesis of the tripeptide was investigated at constant (endogenous) activities of the synthesizing enzymes. The rate was not enhanced by addition of glutamate and/or glycine unless cysteine or glutamyl-cysteine was also added. It is concluded that the concentration of cysteine limits the actual rate of the glutamyl-cysteine-synthetase reaction in vivo. No cysteine or bis(glutamyl)cystine was detected in human hemolysate; however, these disulfides were converted to glutathione. This indicates that erythrocytes have an appropriate system for their reduction, since the disulfides themselves are not substrates for the glutathione-synthesizing enzymes. Studies with intact human red cells indicate that the uptake of cysteine is the rate-determining step in the biosynthesis of glutathione. | [
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|
PMID:12077 | [Does a modified gamma-glutamyl cycle exist in human erythrocytes (author's transl)]. | The first step in the biosynthesis of glutathione is the formation of gamma-glutamyl-cysteine by the enzyme glutamyl-cysteine synthetase. Since this enzyme is not specific for cysteine, different gamma-glutamylamino acids may be formed in vivo which represent potential substrates for the enzymes gamma-glutamylcyclotransferase; in this way 5-oxo-L-proline and free amino acid are formed. We investigated in membrane-free hemolysate the competition between the biosynthesis of glutathione or ophthalmic acid and the degradation of gamma-glutamyl peptides by measuring the formation of 5-oxoproline. The endogenous rate of 5-oxoproline production was 0.13 muM/min. This increased to 2muM/min after addition of 2-aminobutyrate, and to 10muM/min after addition of glutamate and 2-aminobutyrate to hemolysate. Addition of cysteine resulted in an increased oxoproline production only under conditions where glutamyl-cysteine accumulated. In addition, it was shown that for glutamyl-2-aminobutyrate the degradation to 5-oxoproline is faster than the utilization for the tripeptide synthesis. This was not the case for glutamyl-cysteine. Since membrane-free hemolysate (which lacks gamma-glutamyltransferase) is able to produce 5-oxoproline starting from glutamate, it is concluded that this 5-oxoprolinent amino acid transport via a modified gamma-glutamyl cycle. | [Does a modified gamma-glutamyl cycle exist in human erythrocytes (author's transl)]. The first step in the biosynthesis of glutathione is the formation of gamma-glutamyl-cysteine by the enzyme glutamyl-cysteine synthetase. Since this enzyme is not specific for cysteine, different gamma-glutamylamino acids may be formed in vivo which represent potential substrates for the enzymes gamma-glutamylcyclotransferase; in this way 5-oxo-L-proline and free amino acid are formed. We investigated in membrane-free hemolysate the competition between the biosynthesis of glutathione or ophthalmic acid and the degradation of gamma-glutamyl peptides by measuring the formation of 5-oxoproline. The endogenous rate of 5-oxoproline production was 0.13 muM/min. This increased to 2muM/min after addition of 2-aminobutyrate, and to 10muM/min after addition of glutamate and 2-aminobutyrate to hemolysate. Addition of cysteine resulted in an increased oxoproline production only under conditions where glutamyl-cysteine accumulated. In addition, it was shown that for glutamyl-2-aminobutyrate the degradation to 5-oxoproline is faster than the utilization for the tripeptide synthesis. This was not the case for glutamyl-cysteine. Since membrane-free hemolysate (which lacks gamma-glutamyltransferase) is able to produce 5-oxoproline starting from glutamate, it is concluded that this 5-oxoprolinent amino acid transport via a modified gamma-glutamyl cycle. | [
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|
PMID:12079 | [Carcinogenic polycyclic aromatic hydrocarbons in petroleum products: possible prevention of mineral oil cancer]. | A real epidemy of cutaneous cancer has appeared from 1954 in a french area specialized in the metal machining industry (valley of the Arve, Haute-Saoie). Two investigations have been carried out: - one, of clinical and epidemiological nature over a period of 15 years, has recorded 133 casesof spinocellular epitheliams often located in the scrotum and has allowed to estimate the hazard for the population of this region at 25 per 100 000 (that is to say 36 times more than for the general population). - the other, of chemical nature, has revealed on one hand that the new cutting oils, such as delivered to the utilizers, rather frequently contain benzo [a] pyrene (concentration : 0,5 to 150 ug/l), and on the other hand that the used oils are inclined to grow richer in carcinogenic hydrocarbons. The prevention must be directed towards a supplementary effort in the refining of industrial oils and proposals for toxicological threshold value, which would allow to supply machine shops with oils no longer presenting a carcinogenic hazards. | [Carcinogenic polycyclic aromatic hydrocarbons in petroleum products: possible prevention of mineral oil cancer]. A real epidemy of cutaneous cancer has appeared from 1954 in a french area specialized in the metal machining industry (valley of the Arve, Haute-Saoie). Two investigations have been carried out: - one, of clinical and epidemiological nature over a period of 15 years, has recorded 133 casesof spinocellular epitheliams often located in the scrotum and has allowed to estimate the hazard for the population of this region at 25 per 100 000 (that is to say 36 times more than for the general population). - the other, of chemical nature, has revealed on one hand that the new cutting oils, such as delivered to the utilizers, rather frequently contain benzo [a] pyrene (concentration : 0,5 to 150 ug/l), and on the other hand that the used oils are inclined to grow richer in carcinogenic hydrocarbons. The prevention must be directed towards a supplementary effort in the refining of industrial oils and proposals for toxicological threshold value, which would allow to supply machine shops with oils no longer presenting a carcinogenic hazards. | [
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|
PMID:12080 | Radiation cancer, safety standards and current levels of exposure. | Cancer can be induced by radiation in any tissue where cancer occurs naturally. The observation that antenatal diagnostic radiography causes a small but definit increas in childhood cancer is as good evidence as could be expected in support of the scientific expectation that there would be no threshold of dose for carcinogenesis. A linear relation between radiation dose and frequency of of induced cancer is a necessary assumption for a system of radiological protection but is not necessarily a reasonable basis for realistic assessments of cancer risk. Indeed there are radiobiological and epidemiological reasons to the contrary. If the linear hypothesis is accepted then at the present time in the UK the routine practice of medicine is of about 2 orders of magnitude more important in causing cancer than environmental pollution by discharge of radio-activity. The acceptability of radiation safety standards for occupational exposure may be justified by comparison of radiation cancer risks with risks from fatal accidents in the safer industries. The acceptability of the corresponding standards for members of the public seems to require more public discussion of the concept of negligible risk. Emotional reactions to uncontrolled releases of radio-activity are based at least in part on a failure to appreciate the hypothesis of linearity. | Radiation cancer, safety standards and current levels of exposure. Cancer can be induced by radiation in any tissue where cancer occurs naturally. The observation that antenatal diagnostic radiography causes a small but definit increas in childhood cancer is as good evidence as could be expected in support of the scientific expectation that there would be no threshold of dose for carcinogenesis. A linear relation between radiation dose and frequency of of induced cancer is a necessary assumption for a system of radiological protection but is not necessarily a reasonable basis for realistic assessments of cancer risk. Indeed there are radiobiological and epidemiological reasons to the contrary. If the linear hypothesis is accepted then at the present time in the UK the routine practice of medicine is of about 2 orders of magnitude more important in causing cancer than environmental pollution by discharge of radio-activity. The acceptability of radiation safety standards for occupational exposure may be justified by comparison of radiation cancer risks with risks from fatal accidents in the safer industries. The acceptability of the corresponding standards for members of the public seems to require more public discussion of the concept of negligible risk. Emotional reactions to uncontrolled releases of radio-activity are based at least in part on a failure to appreciate the hypothesis of linearity. | [
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|
PMID:12081 | [Epidemiological aspects in carcinogenic risk evaluation arising from pollution]. | Descriptive epidemiology in which population groups are studied can be of considerable value in drawing attention to a potential carcinogen. This must be followed by analytical studies which might either lead with a fair degree of certainty to the identification of the responsible factor or at least help to identify the high-risk group who could then be screened for the suspected cancer. Such epidemiological research must always take into account the results of experimental studies. | [Epidemiological aspects in carcinogenic risk evaluation arising from pollution]. Descriptive epidemiology in which population groups are studied can be of considerable value in drawing attention to a potential carcinogen. This must be followed by analytical studies which might either lead with a fair degree of certainty to the identification of the responsible factor or at least help to identify the high-risk group who could then be screened for the suspected cancer. Such epidemiological research must always take into account the results of experimental studies. | [
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|
PMID:12084 | Effect of thiocyanate on nitrite estimation and the cleavage of nitrosamines. | Thiocyanate or bromide increased the colour formed by nitrite reacting with sulfanilic acid and naphthylethylenediamine. If the colour reagents were added together with thiocyanate (final concentration, M/10), the colour intensity was doubled. If sulfanilic acid was added three minutes before addition of naphthylethylenediamine, the relationship between nitrite concentration and colour production was more linear in the presence of thiocyanate. This effect was due to thiocyanate catalysing the diazotization of sulfanilic acid and inhibiting the reaction of nitrite with naphthylethylenediamine. Bromide and thiocyanate are similar in their catalytic effects on nitrosation, and hydrobromic acid in glacial acetic acid is an effective reagent for denitrosation of nitrosamine. Although thiocyanate catalysed denitrosation of nitrosamines, the effect was small except with nitrosomethylaniline, which had also been found to be denitrosated by sulfanilic acid. Thiocyanate could not be used generally for the destruction of nitrosamines; it was also found to be ineffective as an alternative to hydrobromic acid in the estimation of nitrosamines. | Effect of thiocyanate on nitrite estimation and the cleavage of nitrosamines. Thiocyanate or bromide increased the colour formed by nitrite reacting with sulfanilic acid and naphthylethylenediamine. If the colour reagents were added together with thiocyanate (final concentration, M/10), the colour intensity was doubled. If sulfanilic acid was added three minutes before addition of naphthylethylenediamine, the relationship between nitrite concentration and colour production was more linear in the presence of thiocyanate. This effect was due to thiocyanate catalysing the diazotization of sulfanilic acid and inhibiting the reaction of nitrite with naphthylethylenediamine. Bromide and thiocyanate are similar in their catalytic effects on nitrosation, and hydrobromic acid in glacial acetic acid is an effective reagent for denitrosation of nitrosamine. Although thiocyanate catalysed denitrosation of nitrosamines, the effect was small except with nitrosomethylaniline, which had also been found to be denitrosated by sulfanilic acid. Thiocyanate could not be used generally for the destruction of nitrosamines; it was also found to be ineffective as an alternative to hydrobromic acid in the estimation of nitrosamines. | [
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PMID:12085 | Effects of gallic acid and of ethanol on formation of nitrosodiethylamine. | In vitro experiments demonstrate that the polyphenol, gallic acid, can both catalyse and inhibit NDEA formation. The products of reaction depend considerably on pH conditions and relative concentrations of the reactants. Therefore, interpretation of in vitro experiments in terms of possible in vivo effects in a given population must take into consideration detailed information on eating and drinking habits which might affect pH conditions and relative concentrations of the reactants in the digestive tract. In this respect, it is interesting to note that epidemiological studies conducted by the agency (Day 1) indicate that tea, which contains gallotannins, is consumed more frequently but in a more diluted brew in the area of high oesophageal cancer incidence of the Caspian littoral than in the low incidence area. Our preliminary study on ethanol suggests that its effects will be found to lack the complications present with the tannins and would thus be more easily extrapolated to in vivo situations. | Effects of gallic acid and of ethanol on formation of nitrosodiethylamine. In vitro experiments demonstrate that the polyphenol, gallic acid, can both catalyse and inhibit NDEA formation. The products of reaction depend considerably on pH conditions and relative concentrations of the reactants. Therefore, interpretation of in vitro experiments in terms of possible in vivo effects in a given population must take into consideration detailed information on eating and drinking habits which might affect pH conditions and relative concentrations of the reactants in the digestive tract. In this respect, it is interesting to note that epidemiological studies conducted by the agency (Day 1) indicate that tea, which contains gallotannins, is consumed more frequently but in a more diluted brew in the area of high oesophageal cancer incidence of the Caspian littoral than in the low incidence area. Our preliminary study on ethanol suggests that its effects will be found to lack the complications present with the tannins and would thus be more easily extrapolated to in vivo situations. | [
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PMID:12090 | Effects of some inhibitors on the nitrosation of drugs in human gastric juice. | The nitrosation of several drugs under simulated stomach conditions was only weakly inhibited by the beverages studied. With easily and rapidly nitrosatable drugs, such as aminophenazone or piperazine, administered orally, the use of an inhibitor of nitrosation is to be recommended. Of all the substances investigated, ascorbic acid should be regarded as the best inhibitor because of its pronounced activity at the pH values occurring in the stomach and its lack of toxic effects. We would like to propose that the drugs under consideration should be made up to contain a sufficient amount of ascorbic acid. | Effects of some inhibitors on the nitrosation of drugs in human gastric juice. The nitrosation of several drugs under simulated stomach conditions was only weakly inhibited by the beverages studied. With easily and rapidly nitrosatable drugs, such as aminophenazone or piperazine, administered orally, the use of an inhibitor of nitrosation is to be recommended. Of all the substances investigated, ascorbic acid should be regarded as the best inhibitor because of its pronounced activity at the pH values occurring in the stomach and its lack of toxic effects. We would like to propose that the drugs under consideration should be made up to contain a sufficient amount of ascorbic acid. | [
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PMID:12092 | Chemical studies on tobacco smoke. XLII. Nitrosonornicotine: presence in tobacco, formation and carcinogenicity. | NNN is the first organic carcinogen isolated from unburned tobacco. It has been found in smoking tobaccos, chewing tobaccos and in snuff in concentrations between 0.3 and 90.0 mug. This appears to be an unusually high concentration for a nitrosamine in an environmental agent. We have presented data which suggest that NNN, and possibly other unknown nitrosamines, are formed during the curing of tobacco and that the nitrate content of tobacco is an important factor in nitrosamine formation. Studies with N'-methylanabasine applied to tobacco plants are currently under way to test the idea that nicotine rather than nornicotine is the major precursor of NNN in processed tobacco. In mice, NNN induces adenomas of the lung. Bioassays with rats have shown that NNN is carcinogenic to the oesophagus and the nasal cavity. These chemical and biological data are consistent with the observation that tobacco chewers face an increased risk of cancer of the oesophagus. This observation does not, of course, rule out the possibility that other tobacco carcinogens are responsible for the increased cancer risk of tobacco chewers. | Chemical studies on tobacco smoke. XLII. Nitrosonornicotine: presence in tobacco, formation and carcinogenicity. NNN is the first organic carcinogen isolated from unburned tobacco. It has been found in smoking tobaccos, chewing tobaccos and in snuff in concentrations between 0.3 and 90.0 mug. This appears to be an unusually high concentration for a nitrosamine in an environmental agent. We have presented data which suggest that NNN, and possibly other unknown nitrosamines, are formed during the curing of tobacco and that the nitrate content of tobacco is an important factor in nitrosamine formation. Studies with N'-methylanabasine applied to tobacco plants are currently under way to test the idea that nicotine rather than nornicotine is the major precursor of NNN in processed tobacco. In mice, NNN induces adenomas of the lung. Bioassays with rats have shown that NNN is carcinogenic to the oesophagus and the nasal cavity. These chemical and biological data are consistent with the observation that tobacco chewers face an increased risk of cancer of the oesophagus. This observation does not, of course, rule out the possibility that other tobacco carcinogens are responsible for the increased cancer risk of tobacco chewers. | [
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PMID:12111 | Microbicidal mechanisms of human granulocytes: synergistic effects of granulocyte elastase and myeloperoxidase or chymotrypsin-like cationic protein. | The antibacterial activity of a myeloperoxidase (MPO)-glucose oxidase system was found to be greatly increased by granulocyte elastase, present in azurophil granules of human neutrophils. The MPO-H2O3-mediated killing of both Escherichia coli and Staphylococcus aureus was potentiated by granuocyte elastase at an acid pH, whereas at pH 7.4 only killing of E. coli was potentiated. The potentiating effect of elastase was not dependent on the enzymatic properties of the protein since it was not abolished by heating, which destroys the enzymatic activity. A peptide chloromethyl ketone elastase inhibitor abolished both elastolytic activity and the pctentiating effects on MPO-H2-O2-mediated bacterial killing. The antibacterial activity of chymotrypsin-like cationic protein of human neutrophils was also potentiated by elastase. Other degradative enzymes isolated from human granulocytes, e.g., collagenase and lysozyme, did not potentiate MPO-H2O2-mediated or cationic protein-dependent bacterial killing. The present study indicates that a neutrophil constitutent, elastase, which is not microbicidal by itself, can initiate sublethal changes that render some microorganisms more susceptible to the action of microbicidal agents like MPO and chymotrypsin-like cationic protein. | Microbicidal mechanisms of human granulocytes: synergistic effects of granulocyte elastase and myeloperoxidase or chymotrypsin-like cationic protein. The antibacterial activity of a myeloperoxidase (MPO)-glucose oxidase system was found to be greatly increased by granulocyte elastase, present in azurophil granules of human neutrophils. The MPO-H2O3-mediated killing of both Escherichia coli and Staphylococcus aureus was potentiated by granuocyte elastase at an acid pH, whereas at pH 7.4 only killing of E. coli was potentiated. The potentiating effect of elastase was not dependent on the enzymatic properties of the protein since it was not abolished by heating, which destroys the enzymatic activity. A peptide chloromethyl ketone elastase inhibitor abolished both elastolytic activity and the pctentiating effects on MPO-H2-O2-mediated bacterial killing. The antibacterial activity of chymotrypsin-like cationic protein of human neutrophils was also potentiated by elastase. Other degradative enzymes isolated from human granulocytes, e.g., collagenase and lysozyme, did not potentiate MPO-H2O2-mediated or cationic protein-dependent bacterial killing. The present study indicates that a neutrophil constitutent, elastase, which is not microbicidal by itself, can initiate sublethal changes that render some microorganisms more susceptible to the action of microbicidal agents like MPO and chymotrypsin-like cationic protein. | [
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PMID:12113 | Effect of bromazepam on gastric acid secretion related to hypnotically induced anxiety. | The effects of bromazepam (0.1 mg/kg b.w. i.v.) and of placebo on gastric acid secretion related to hypnotically induced anxiety were evaluated in a double blind study, 22 experiments were carried out on 4 healthy volunteers. Drugs were injected after one basal hour. Hypnosis was induced immediately thereafter, and a sequence of anxiety-charged situations out of the subjects past was recalled. After one hour, posthypnotic amnesia was suggested, the subjects awakened and observation continued for another hour. Acid output was measured by means of intragastric titration and a telemetering capsule. During hypnosis and recalling of anxiety in both series, acid output decreased. In the posthypnotic hour there was a significant increase of secretion in the placebo-series, while there was virtually no change in the the bromazepam-series. The pattern of acid output in the placebo-series seems to result from an activation of the sympathetic system under anxiety and a rebound vagal activation in the posthypnotic hour. By contrast, under the sedating effect of bromazepam, no anxiety could be evoked and no rebound vagal activation and thereby no increase of acid secretion resulted. | Effect of bromazepam on gastric acid secretion related to hypnotically induced anxiety. The effects of bromazepam (0.1 mg/kg b.w. i.v.) and of placebo on gastric acid secretion related to hypnotically induced anxiety were evaluated in a double blind study, 22 experiments were carried out on 4 healthy volunteers. Drugs were injected after one basal hour. Hypnosis was induced immediately thereafter, and a sequence of anxiety-charged situations out of the subjects past was recalled. After one hour, posthypnotic amnesia was suggested, the subjects awakened and observation continued for another hour. Acid output was measured by means of intragastric titration and a telemetering capsule. During hypnosis and recalling of anxiety in both series, acid output decreased. In the posthypnotic hour there was a significant increase of secretion in the placebo-series, while there was virtually no change in the the bromazepam-series. The pattern of acid output in the placebo-series seems to result from an activation of the sympathetic system under anxiety and a rebound vagal activation in the posthypnotic hour. By contrast, under the sedating effect of bromazepam, no anxiety could be evoked and no rebound vagal activation and thereby no increase of acid secretion resulted. | [
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|
PMID:12114 | [Effect of fominoben on ventilation, oxygen uptake and blood gases in patients with obstructive ventilation disorders]. | The substituted benzylamin-derivative fominoben (PB 89 Noleptan) was intravenously administered to 12 patients with chronic obstructive lung disease in order to determine, whether an analeptic action on respiration, which had been found by others in animal studies and in healthy subjects, can also be demonstrated in patients with COLD. Time ventilation showed no statistically significant change. Respiratory rate was increased for a short time, alveolar ventilation showed a slight but significant increase 35 minutes after i.v. injection of fominoben, however no significant change in the first 10 minutes after injection.--Arterial pO2 was slightly but not significantly increased in the first 10 minutes after fominoben, while the same patients showed a significant decrease of pO2 after injection of placebo. As alveolar ventilation at this time had not significantly changed, the increase in pO2 can only be explained by an improvement of regional ventilation-perfusion ratio by fominoben. -In conclusion it can be stated, that a marked stimulative action on respiration by fominoben could not be demonstrated. There was, however, no depression of respiration as it is associated with most other caugh medications. As the drug has been shown to be an excellent caugh sedative, lack of respiratory depression can be considered as a considerable advantage. | [Effect of fominoben on ventilation, oxygen uptake and blood gases in patients with obstructive ventilation disorders]. The substituted benzylamin-derivative fominoben (PB 89 Noleptan) was intravenously administered to 12 patients with chronic obstructive lung disease in order to determine, whether an analeptic action on respiration, which had been found by others in animal studies and in healthy subjects, can also be demonstrated in patients with COLD. Time ventilation showed no statistically significant change. Respiratory rate was increased for a short time, alveolar ventilation showed a slight but significant increase 35 minutes after i.v. injection of fominoben, however no significant change in the first 10 minutes after injection.--Arterial pO2 was slightly but not significantly increased in the first 10 minutes after fominoben, while the same patients showed a significant decrease of pO2 after injection of placebo. As alveolar ventilation at this time had not significantly changed, the increase in pO2 can only be explained by an improvement of regional ventilation-perfusion ratio by fominoben. -In conclusion it can be stated, that a marked stimulative action on respiration by fominoben could not be demonstrated. There was, however, no depression of respiration as it is associated with most other caugh medications. As the drug has been shown to be an excellent caugh sedative, lack of respiratory depression can be considered as a considerable advantage. | [
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|
PMID:12115 | Antihypertensive effect of labetalol, a new alpha- and beta-adrenergic blocking agent. | Labetalol (AH5158) a new alpha- and beta-adrenergic blocking agent was given to 12 hypertensive patients for an average of 7 months. Statistically significant and clinically relevant reductions of blood pressure both in the recumbent and standing positions were observed. Side effects were few and the absence of postural hypotension was noted. | Antihypertensive effect of labetalol, a new alpha- and beta-adrenergic blocking agent. Labetalol (AH5158) a new alpha- and beta-adrenergic blocking agent was given to 12 hypertensive patients for an average of 7 months. Statistically significant and clinically relevant reductions of blood pressure both in the recumbent and standing positions were observed. Side effects were few and the absence of postural hypotension was noted. | [
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|
PMID:12116 | Preliminary experience with vascularised Fallopian tube transplants in the human female. | The preoperative screening and postoperative case records of two patients that underwent vascularised Fallopian tube transplants are described. In both cases the operation was followed by immunological rejection. This was chronic and clinically quiescent in the first patient, but an accelerated rejection associated with a pyrexial illness and a rise in serum fibrinogen, blood platelet and serum lactic dehydrogenase levels was observed in the second case. Technical feasibility was demonstrated but further progress in the field of Fallopian tube transplants will be dependent on the development of safer techniques of immunosuppression considered necessary for graft survival. | Preliminary experience with vascularised Fallopian tube transplants in the human female. The preoperative screening and postoperative case records of two patients that underwent vascularised Fallopian tube transplants are described. In both cases the operation was followed by immunological rejection. This was chronic and clinically quiescent in the first patient, but an accelerated rejection associated with a pyrexial illness and a rise in serum fibrinogen, blood platelet and serum lactic dehydrogenase levels was observed in the second case. Technical feasibility was demonstrated but further progress in the field of Fallopian tube transplants will be dependent on the development of safer techniques of immunosuppression considered necessary for graft survival. | [
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PMID:12117 | Completed pregnancy following vascularized heterotopic autotransplantation of the Fallopian tube in the ewe. | Transplantation of the Fallopian tube is a technically feasible operation. Previous authors have reported pregnancies following turbo-ovarian transplants, but there appears to be no record of gestation following transplantation of the isolated oviduct. This communication illustrates the method that we have used to perform Fallopian tube transplants in the ewe and documents our first full-term pregnancy following this operation. It provides further evidence of the technical feasibility of Fallopian tube transplants. | Completed pregnancy following vascularized heterotopic autotransplantation of the Fallopian tube in the ewe. Transplantation of the Fallopian tube is a technically feasible operation. Previous authors have reported pregnancies following turbo-ovarian transplants, but there appears to be no record of gestation following transplantation of the isolated oviduct. This communication illustrates the method that we have used to perform Fallopian tube transplants in the ewe and documents our first full-term pregnancy following this operation. It provides further evidence of the technical feasibility of Fallopian tube transplants. | [
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|
PMID:12118 | Hormone-receptor interactions--basic mechanisms. | An understanding of the basic mechanisms of hormone action is becoming an important part of a clinician's training. With the advent of radioreceptor assays and their comparison with radioimmunoassays, we are becoming increasingly aware that the normal physiological function of a hormone is not necessarily dependent on "normal" levels of hormone being present in the plasma. Even if plasma levels are normal, if a particular target cell lacks the receptor for the particular hormone the target tissue will not respond to the hormone. It has been shown, for example, by many workers that steroid hormones act on their cells via a receptor located in the cytoplasm of the target cells (1-4). Since the presence or absence of such receptors are becoming increasingly important in terms of unexplained infertility, endometrial carcinoma and breast cancer, it is necessary that the practicing clinician be familiar with the concept of receptors and have some understanding of their mode of action. In this brief presentation, I will explain certain terminology and summarize the state of the art so that you can critically read the literature concerning new developments in the area of hormone action which is to become increasingly important in the next few years. I will discuss some of the aspects of the mechanism of peptide hormones such as LH and FSH but will devote most of my attention discussing the details of steroid hormone action since our knowledge in this area is much more complete. I will also explain the terms frequently discussed in the literature concerned with hormone-receptor interactions. | Hormone-receptor interactions--basic mechanisms. An understanding of the basic mechanisms of hormone action is becoming an important part of a clinician's training. With the advent of radioreceptor assays and their comparison with radioimmunoassays, we are becoming increasingly aware that the normal physiological function of a hormone is not necessarily dependent on "normal" levels of hormone being present in the plasma. Even if plasma levels are normal, if a particular target cell lacks the receptor for the particular hormone the target tissue will not respond to the hormone. It has been shown, for example, by many workers that steroid hormones act on their cells via a receptor located in the cytoplasm of the target cells (1-4). Since the presence or absence of such receptors are becoming increasingly important in terms of unexplained infertility, endometrial carcinoma and breast cancer, it is necessary that the practicing clinician be familiar with the concept of receptors and have some understanding of their mode of action. In this brief presentation, I will explain certain terminology and summarize the state of the art so that you can critically read the literature concerning new developments in the area of hormone action which is to become increasingly important in the next few years. I will discuss some of the aspects of the mechanism of peptide hormones such as LH and FSH but will devote most of my attention discussing the details of steroid hormone action since our knowledge in this area is much more complete. I will also explain the terms frequently discussed in the literature concerned with hormone-receptor interactions. | [
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|
PMID:12119 | Effect of caffeine on the motility, viability, oxygen consumption and glycolytic rate of ejaculated human normokinetic and hypokinetic spermatozoa. | In this report the effect of caffeine, IBMX and Cholera toxin on ejaculated human spermatozoa was tested. It was found that caffeine was the most effective drug, and that Cholera toxin was ineffective. Caffeine stimulated motility, preserved viability and increased the glucose, fructose utilization and lactate production of sperm in whole semen and washed sperm. Oxygen consumption however was decreased. These results of the normokinetic ejaculates were defined as a basis for comparison with the response of hypokinetic sperm. A group of 11 hypokinetic ejaculated spermatozoa were treated by caffeine and only five responded to caffeine stimulation by a response, similar to that of normokinetic sperm. They were defined as "responsive." The other group of six patients who did not respond were defined as a "non-responsive" class. The possibility that the in vitro treated sperm of the responsive class ejaculate is suitable for artificial insemination to their wives, is discussed. | Effect of caffeine on the motility, viability, oxygen consumption and glycolytic rate of ejaculated human normokinetic and hypokinetic spermatozoa. In this report the effect of caffeine, IBMX and Cholera toxin on ejaculated human spermatozoa was tested. It was found that caffeine was the most effective drug, and that Cholera toxin was ineffective. Caffeine stimulated motility, preserved viability and increased the glucose, fructose utilization and lactate production of sperm in whole semen and washed sperm. Oxygen consumption however was decreased. These results of the normokinetic ejaculates were defined as a basis for comparison with the response of hypokinetic sperm. A group of 11 hypokinetic ejaculated spermatozoa were treated by caffeine and only five responded to caffeine stimulation by a response, similar to that of normokinetic sperm. They were defined as "responsive." The other group of six patients who did not respond were defined as a "non-responsive" class. The possibility that the in vitro treated sperm of the responsive class ejaculate is suitable for artificial insemination to their wives, is discussed. | [
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PMID:12120 | Blastoid transformation of lymphocytes in response to spermatozoa. | Microlymphocyte transformation tests were done on 33 female rabbits immunized with different components of rabbit ejaculate. In rabbits injected with total ejaculate or washed spermatozoa a specific stimulation of lymphocytes could be demonstrated by exposure to washed spermatozoa. It is shown that it is necessary to notice the length of time for cultivation, the antigen concentration and especially the time-interval after the last immunization. | Blastoid transformation of lymphocytes in response to spermatozoa. Microlymphocyte transformation tests were done on 33 female rabbits immunized with different components of rabbit ejaculate. In rabbits injected with total ejaculate or washed spermatozoa a specific stimulation of lymphocytes could be demonstrated by exposure to washed spermatozoa. It is shown that it is necessary to notice the length of time for cultivation, the antigen concentration and especially the time-interval after the last immunization. | [
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PMID:12121 | GPC diesterase activity in human endometrial secretion. (Its variations under the action of estrogens, clomiphene citrate, D-norgestrel (post-coital and low dose) and intrauterine device (IUD).) | Diesterase activity was studied in human uterine secretions of normal women and in those under treatment for sterility or contraception. Endometrial secretions were obtained from 78 patients and the material divided into four groups: normal women, under different estrogens; with D-Norgestrel treatment (daily and post-coital) and patients with IUD Lippes D. The mean concentration of free choline delivered by the GPC diesterase in the normal group was 777 +/- 128 mug/ml (SD 286). Under hormonal treatment an increase of diesterase activity was observed. D-Norgestrel post-coital produced a fall of the enzymatic activity between 180 to 420 minutes. The uninterrupted use of D-Norgestrel (30 gammas daily) produced a loss of diesterase activity in 80% of cases studied. The use of IUD (Lippes D) did not modify the enzymatic activity in this group. | GPC diesterase activity in human endometrial secretion. (Its variations under the action of estrogens, clomiphene citrate, D-norgestrel (post-coital and low dose) and intrauterine device (IUD).). Diesterase activity was studied in human uterine secretions of normal women and in those under treatment for sterility or contraception. Endometrial secretions were obtained from 78 patients and the material divided into four groups: normal women, under different estrogens; with D-Norgestrel treatment (daily and post-coital) and patients with IUD Lippes D. The mean concentration of free choline delivered by the GPC diesterase in the normal group was 777 +/- 128 mug/ml (SD 286). Under hormonal treatment an increase of diesterase activity was observed. D-Norgestrel post-coital produced a fall of the enzymatic activity between 180 to 420 minutes. The uninterrupted use of D-Norgestrel (30 gammas daily) produced a loss of diesterase activity in 80% of cases studied. The use of IUD (Lippes D) did not modify the enzymatic activity in this group. | [
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