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PMID:20445
|
Mast cell binding of neurotensin. I. Iodination of neurotensin and characterization of the interaction of neurotensin with mast cell receptor sites.
|
Neurotensin was iodinated at equimolar concentrations of peptide, iodide, and chloramine-T, producing a labeled peptide with a specific activity of 1000 to 2000 Ci/mmol. Rat mast cells specifically and reversibly bound 1.27 pmol of neurotensin/10(6) cells with a reversible affinity, KD, of 154 nM. Optimum specific binding occurred betwen pH 6.8 and 7.2 under hypotonic conditions and dropped sharply as buffer concentration increased beyond 10 mM. The divalent cations Ca2+ and Mg2+ prevented binding with 50% inhibition at 1.5 and 4 mM, respectively. Binding was strongly and equally inhibited by the sodium and potassium salts of chloride, bromide, and iodide, and to a lesser degree by LiCl. Maximum binding of 125I-neurotensin occurred within 10 min at 0 degrees, and within 1.5 to 2 min binding was reduced to half-maximum in the presence of excess unlabeled neurotensin or upon 20-fold dilution in buffer. Both CaCl2 and NaCl were able to dissociate 60% of the total bound neurotensin: half the label bound was removed in 4 to 6 min. EDTA inhibited the binding only at high concentrations and no requirement was found for sulfhydryl groups, ATP, or a glycoprotein in the binding of neurotensin.
|
Mast cell binding of neurotensin. I. Iodination of neurotensin and characterization of the interaction of neurotensin with mast cell receptor sites. Neurotensin was iodinated at equimolar concentrations of peptide, iodide, and chloramine-T, producing a labeled peptide with a specific activity of 1000 to 2000 Ci/mmol. Rat mast cells specifically and reversibly bound 1.27 pmol of neurotensin/10(6) cells with a reversible affinity, KD, of 154 nM. Optimum specific binding occurred betwen pH 6.8 and 7.2 under hypotonic conditions and dropped sharply as buffer concentration increased beyond 10 mM. The divalent cations Ca2+ and Mg2+ prevented binding with 50% inhibition at 1.5 and 4 mM, respectively. Binding was strongly and equally inhibited by the sodium and potassium salts of chloride, bromide, and iodide, and to a lesser degree by LiCl. Maximum binding of 125I-neurotensin occurred within 10 min at 0 degrees, and within 1.5 to 2 min binding was reduced to half-maximum in the presence of excess unlabeled neurotensin or upon 20-fold dilution in buffer. Both CaCl2 and NaCl were able to dissociate 60% of the total bound neurotensin: half the label bound was removed in 4 to 6 min. EDTA inhibited the binding only at high concentrations and no requirement was found for sulfhydryl groups, ATP, or a glycoprotein in the binding of neurotensin.
|
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] |
PMID:20446
|
RNA polymerase activity in bovine spermatozoa.
|
Washed mature spermatozoa from bulls incorporate ribonucleoside triphosphates into RNA using an endogenous template. Maximum incorporation was observed at 31 degrees C in the presence of MgCl2, all four ribonucleoside triphosphates, beta-mercaptoethanol, and glycine sodium hydroxide buffer at pH 9.0. The amount of synthesis was linearly dependent upon the concentration of spermatozoa and continued for at least 4 h. Digestion studies revealed the RNA to be present in a protected (intracellular?) location in the spermatozoa. The RNA synthesis was inhibited by ethidium bromide, rifampicin, acriflavine, actinomycin D, and caffeine, but not by alpha-amanitine or rifamycin SV. Fractionation of the spermatozoa by sonication and separation of the heads and tails by centrifugation through a discontinuous gradient revealed that more than half of the total RNA polymerase activity was associated with the tail fraction.
|
RNA polymerase activity in bovine spermatozoa. Washed mature spermatozoa from bulls incorporate ribonucleoside triphosphates into RNA using an endogenous template. Maximum incorporation was observed at 31 degrees C in the presence of MgCl2, all four ribonucleoside triphosphates, beta-mercaptoethanol, and glycine sodium hydroxide buffer at pH 9.0. The amount of synthesis was linearly dependent upon the concentration of spermatozoa and continued for at least 4 h. Digestion studies revealed the RNA to be present in a protected (intracellular?) location in the spermatozoa. The RNA synthesis was inhibited by ethidium bromide, rifampicin, acriflavine, actinomycin D, and caffeine, but not by alpha-amanitine or rifamycin SV. Fractionation of the spermatozoa by sonication and separation of the heads and tails by centrifugation through a discontinuous gradient revealed that more than half of the total RNA polymerase activity was associated with the tail fraction.
|
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] |
PMID:20447
|
The contractile basis of amoeboid movement. V. The control of gelation, solation, and contraction in extracts from Dictyostelium discoideum.
|
Motile extracts have been prepared from Dictyostelium discoideum by homogenization and differential centrifugation at 4 degrees C in a stabilization solution (60). These extracts gelled on warming to 25 degrees Celsius and contracted in response to micromolar Ca++ or a pH in excess of 7.0. Optimal gelation occurred in a solution containing 2.5 mM ethylene glycol-bis (beta-aminoethyl ether)N,N,N',N'-tetraacetate (EGTA), 2.5 mM piperazine-N-N'-bis [2-ethane sulfonic acid] (PIPES), 1 mM MgC1(2), 1 mM ATP, and 20 mM KCI at ph 7.0 (relaxation solution), while micromolar levels of Ca++ inhibited gelation. Conditions that solated the gel elicited contraction of extracts containing myosin. This was true regardless of whether chemical (micromolar Ca++, pH >7.0, cytochalasin B, elevated concentrations of KCI, MgC1(2), and sucrose) or physical (pressure, mechanical stress, and cold) means were used to induce solation. Myosin was definitely required for contraction. During Ca++-or pH-elicited contraction: (a) actin, myosin, and a 95,000-dalton polypeptide were concentrated in the contracted extract; (b) the gelation activity was recovered in the material sqeezed out the contracting extract;(c) electron microscopy demonstrated that the number of free, recognizable F-actin filaments increased; (d) the actomyosin MgATPase activity was stimulated by 4- to 10-fold. In the absense of myosin the Dictyostelium extract did not contract, while gelation proceeded normally. During solation of the gel in the absense of myosin: (a) electron microscopy demonstrated that the number of free, recognizable F- actin filaments increased; (b) solation-dependent contraction of the extract and the Ca++-stimulated MgATPase activity were reconstituted by adding puried Dictyostelium myosin. Actin purified from the Dictyostelium extract did not gel (at 2 mg/ml), while low concentrations of actin (0.7-2 mg/ml) that contained several contaminating components underwent rapid Ca++ regulated gelation. These results indicated : (a) gelation in Dictyostelium extracts involves a specific Ca++-sensitive interaction between actin and several other components; (b) myosin is an absolute requirement for contraction of the extract; (c) actin-myosin interactions capable of producing force for movement are prevented in the gel, while solation of the gel by either physical or chemical means results in the release of F-actin capable of interaction with myosin and subsequent contraction. The effectiveness of physical agents in producting contraction suggests that the regulation of contraction by the gel is structural in nature.
|
The contractile basis of amoeboid movement. V. The control of gelation, solation, and contraction in extracts from Dictyostelium discoideum. Motile extracts have been prepared from Dictyostelium discoideum by homogenization and differential centrifugation at 4 degrees C in a stabilization solution (60). These extracts gelled on warming to 25 degrees Celsius and contracted in response to micromolar Ca++ or a pH in excess of 7.0. Optimal gelation occurred in a solution containing 2.5 mM ethylene glycol-bis (beta-aminoethyl ether)N,N,N',N'-tetraacetate (EGTA), 2.5 mM piperazine-N-N'-bis [2-ethane sulfonic acid] (PIPES), 1 mM MgC1(2), 1 mM ATP, and 20 mM KCI at ph 7.0 (relaxation solution), while micromolar levels of Ca++ inhibited gelation. Conditions that solated the gel elicited contraction of extracts containing myosin. This was true regardless of whether chemical (micromolar Ca++, pH >7.0, cytochalasin B, elevated concentrations of KCI, MgC1(2), and sucrose) or physical (pressure, mechanical stress, and cold) means were used to induce solation. Myosin was definitely required for contraction. During Ca++-or pH-elicited contraction: (a) actin, myosin, and a 95,000-dalton polypeptide were concentrated in the contracted extract; (b) the gelation activity was recovered in the material sqeezed out the contracting extract;(c) electron microscopy demonstrated that the number of free, recognizable F-actin filaments increased; (d) the actomyosin MgATPase activity was stimulated by 4- to 10-fold. In the absense of myosin the Dictyostelium extract did not contract, while gelation proceeded normally. During solation of the gel in the absense of myosin: (a) electron microscopy demonstrated that the number of free, recognizable F- actin filaments increased; (b) solation-dependent contraction of the extract and the Ca++-stimulated MgATPase activity were reconstituted by adding puried Dictyostelium myosin. Actin purified from the Dictyostelium extract did not gel (at 2 mg/ml), while low concentrations of actin (0.7-2 mg/ml) that contained several contaminating components underwent rapid Ca++ regulated gelation. These results indicated : (a) gelation in Dictyostelium extracts involves a specific Ca++-sensitive interaction between actin and several other components; (b) myosin is an absolute requirement for contraction of the extract; (c) actin-myosin interactions capable of producing force for movement are prevented in the gel, while solation of the gel by either physical or chemical means results in the release of F-actin capable of interaction with myosin and subsequent contraction. The effectiveness of physical agents in producting contraction suggests that the regulation of contraction by the gel is structural in nature.
|
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] |
PMID:20448
|
The stability of tyrosine aminotransferase and other proteins in enucleated rat hepatoma tissue culture cells.
|
The role of the nucleus in bringing about the induction of tyrosine aminotransferase (TAT) by glucocorticosteroid hormone and its deinduction upon steroid removal has been studied in enucleated rat hepatoma tissue culture cells (FU5-5). Both processes require the presence of the nucleus. However, cytoplasts from preinduced cells show an initial rapid decline in enzyme activity immediately after enucleation followed by maintenance of a constant level of activity. This initial decline in enzyme activity can be partially prevented by trypan blue, an inhibitor of lysosomal activity. This suggests that the early fall in enzyme activity could be due to an increase in the level of lysosomal activity immediately after enucleation. The subsequent constant level of activity seems due to maintenance rather than synthesis and degradation since it is not affected by cycloheximide. The absence of degradation applies to other kinds of proteins in enucleated FU5-5 cells and enucleated mouse fibroblast L cells. These experiments suggest that some kind of labile RNA or protein dependent on the presence of the nucleus is required for the degradation of all classes of proteins in different kinds of cells.
|
The stability of tyrosine aminotransferase and other proteins in enucleated rat hepatoma tissue culture cells. The role of the nucleus in bringing about the induction of tyrosine aminotransferase (TAT) by glucocorticosteroid hormone and its deinduction upon steroid removal has been studied in enucleated rat hepatoma tissue culture cells (FU5-5). Both processes require the presence of the nucleus. However, cytoplasts from preinduced cells show an initial rapid decline in enzyme activity immediately after enucleation followed by maintenance of a constant level of activity. This initial decline in enzyme activity can be partially prevented by trypan blue, an inhibitor of lysosomal activity. This suggests that the early fall in enzyme activity could be due to an increase in the level of lysosomal activity immediately after enucleation. The subsequent constant level of activity seems due to maintenance rather than synthesis and degradation since it is not affected by cycloheximide. The absence of degradation applies to other kinds of proteins in enucleated FU5-5 cells and enucleated mouse fibroblast L cells. These experiments suggest that some kind of labile RNA or protein dependent on the presence of the nucleus is required for the degradation of all classes of proteins in different kinds of cells.
|
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] |
PMID:20450
|
Initiation of human parturition. VI. Identification and quantification of progesterone metabolites produced by the components of human fetal membranes.
|
The fetal membranes play a central role in the initiation of human parturition, a role that may be subserved in part by quantitative changes in progesterone metabolism. In order to identify and to quantify the metabolites of progesterone produced by the components of the human fetal membranes, amnion and chorion laeve tissues and homogenates were incubated with [3H]progesterone in the presence of added NADPH. The radioactive metabolites, 20alpha-hydroxy-4-pregnen-3-one, 5alpha-pregnane-3,20-dione and 3beta-hydroxy-5alpha-pregnan-20-one, were isolated and identified by derivative formation, a combination of chromatographic techniques, and by crystallization to constant specific activity after addition of authentic standards. A simplified technique of metabolite quantification was developed that involves one thin layer chromatographic procedure and liquid scintillation assay of radioactivity. The accuracy of this method was established by comparison with data obtained using classical techniques. This study demonstrates the presence of human fetal membranes 5alpha-reductase, 20alpha-hydroxysteroid oxidoreductase, and 3beta-hydroxy-steroid oxidoreductase, and a qualitative difference between amnion and chorion laeve.
|
Initiation of human parturition. VI. Identification and quantification of progesterone metabolites produced by the components of human fetal membranes. The fetal membranes play a central role in the initiation of human parturition, a role that may be subserved in part by quantitative changes in progesterone metabolism. In order to identify and to quantify the metabolites of progesterone produced by the components of the human fetal membranes, amnion and chorion laeve tissues and homogenates were incubated with [3H]progesterone in the presence of added NADPH. The radioactive metabolites, 20alpha-hydroxy-4-pregnen-3-one, 5alpha-pregnane-3,20-dione and 3beta-hydroxy-5alpha-pregnan-20-one, were isolated and identified by derivative formation, a combination of chromatographic techniques, and by crystallization to constant specific activity after addition of authentic standards. A simplified technique of metabolite quantification was developed that involves one thin layer chromatographic procedure and liquid scintillation assay of radioactivity. The accuracy of this method was established by comparison with data obtained using classical techniques. This study demonstrates the presence of human fetal membranes 5alpha-reductase, 20alpha-hydroxysteroid oxidoreductase, and 3beta-hydroxy-steroid oxidoreductase, and a qualitative difference between amnion and chorion laeve.
|
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] |
PMID:20452
|
Chromatographic separations of alphavirus strains by hydroxylapatite.
|
Hydroxylapatite column chromatography methods were developed to characterize selected alphavirus populations. Different conditions of pH and phosphate molarity were required to obtain satisfactory elution profiles and separations for Western equine encephalomyelitis virus strains, compared with Eastern equine encephalomyelitis virus and Semliki Forest virus strains. Raising the pH of the buffers effected earlier elutions of all viruses. Selection of phosphate gradients with more gentle slopes and adjustment to the proper pH effected better separations of virus subpopulations. Elution profiles were not affected by 0.85% NaCl, 10% fetal calf serum, or 1% bovine serum albumin, which are common constituents of virus-stabilizing diluents. Passage of Western equine encephalomyelitis and Semliki Forest viruses in BHK-21, Vero, or duck embryo cell cultures or in suckling mouse brains did not usually affect elution profiles, unless passage also resulted in a shift in the plaque size marker. Essentially all infectious virus applied to the column was recoverable in appropriate fractions. This permitted accurate determinations of heterogeneity within alphavirus populations. For Western equine encephalomyelitis large-plaque (LP) and small-plaque (SP) virus populations, it was possible to detect ratios of 1 LP in a population of 10(6) SP, and 1 SP in 10(3) LP by using linear phosphate gradients. When stepwise elution procedures were used, it was possible to detect ratios of 1 SP in a population of 10(5) LP. Hydroxylapatite column chromatography therefore appears to be a useful tool for characterizing alphaviruses and for isolating minority subpopulations of viruses of biological or epidemiological importance from apparently homogeneous virus stocks.
|
Chromatographic separations of alphavirus strains by hydroxylapatite. Hydroxylapatite column chromatography methods were developed to characterize selected alphavirus populations. Different conditions of pH and phosphate molarity were required to obtain satisfactory elution profiles and separations for Western equine encephalomyelitis virus strains, compared with Eastern equine encephalomyelitis virus and Semliki Forest virus strains. Raising the pH of the buffers effected earlier elutions of all viruses. Selection of phosphate gradients with more gentle slopes and adjustment to the proper pH effected better separations of virus subpopulations. Elution profiles were not affected by 0.85% NaCl, 10% fetal calf serum, or 1% bovine serum albumin, which are common constituents of virus-stabilizing diluents. Passage of Western equine encephalomyelitis and Semliki Forest viruses in BHK-21, Vero, or duck embryo cell cultures or in suckling mouse brains did not usually affect elution profiles, unless passage also resulted in a shift in the plaque size marker. Essentially all infectious virus applied to the column was recoverable in appropriate fractions. This permitted accurate determinations of heterogeneity within alphavirus populations. For Western equine encephalomyelitis large-plaque (LP) and small-plaque (SP) virus populations, it was possible to detect ratios of 1 LP in a population of 10(6) SP, and 1 SP in 10(3) LP by using linear phosphate gradients. When stepwise elution procedures were used, it was possible to detect ratios of 1 SP in a population of 10(5) LP. Hydroxylapatite column chromatography therefore appears to be a useful tool for characterizing alphaviruses and for isolating minority subpopulations of viruses of biological or epidemiological importance from apparently homogeneous virus stocks.
|
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] |
PMID:20453
|
Sonication as a tool for the study of adenylyl cyclase activity.
|
The technique of sonication was applied in studying adenylyl cyclase activity of cultured fibroblasts. Exposure of BHK 21 c/13 to brief periods of low power sonication gives cell preparations with greater basal, fluoride and hormone sensitive adenylyl cyclase activites than those of broken cell preparations of homogenized cells. The sonicated cells provide a convenient method to study adenylyl cyclase since they are added directly to the adenylyl cyclase reaction vessels without further processing. Maximal epinephrine stimulated activity in sonicated cells is nearly equivalent to that activated by sodium fluoride, but the apparent affinity of the enzyme system is similar to that of broken cell preparations. Furthermore, broken cell preparations of sonicated cells possess greater adenylyl cyclase activity than broken cell preparations of unsonicated cells. This procedure may provide a useful tool for the analysis of the hormonal regulation of adenylyl cyclase activity of isolated cells.
|
Sonication as a tool for the study of adenylyl cyclase activity. The technique of sonication was applied in studying adenylyl cyclase activity of cultured fibroblasts. Exposure of BHK 21 c/13 to brief periods of low power sonication gives cell preparations with greater basal, fluoride and hormone sensitive adenylyl cyclase activites than those of broken cell preparations of homogenized cells. The sonicated cells provide a convenient method to study adenylyl cyclase since they are added directly to the adenylyl cyclase reaction vessels without further processing. Maximal epinephrine stimulated activity in sonicated cells is nearly equivalent to that activated by sodium fluoride, but the apparent affinity of the enzyme system is similar to that of broken cell preparations. Furthermore, broken cell preparations of sonicated cells possess greater adenylyl cyclase activity than broken cell preparations of unsonicated cells. This procedure may provide a useful tool for the analysis of the hormonal regulation of adenylyl cyclase activity of isolated cells.
|
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] |
PMID:20454
|
Blood and tissue distribution of gamma glutamyl transferase in the cow.
|
To test the hepatic specificity of gamma-glutamyl transferase in the cow, the tissue distribution of this enzyme was studied in 10 healthy adult cows. It was located mainly in kidney, pancreas, and liver. The relative hepatic specificity of this enzyme indicated that measurement of its activity in serum could be a valuable test of hepatic damage in the cow. The blood activity measured within 54 plasmas and 54 serums of apparently healthy animals was almost the same, 17.7 +/- 5.7 U/liter for plasma and 18.6 +/- 6.2 U/liter for serum.
|
Blood and tissue distribution of gamma glutamyl transferase in the cow. To test the hepatic specificity of gamma-glutamyl transferase in the cow, the tissue distribution of this enzyme was studied in 10 healthy adult cows. It was located mainly in kidney, pancreas, and liver. The relative hepatic specificity of this enzyme indicated that measurement of its activity in serum could be a valuable test of hepatic damage in the cow. The blood activity measured within 54 plasmas and 54 serums of apparently healthy animals was almost the same, 17.7 +/- 5.7 U/liter for plasma and 18.6 +/- 6.2 U/liter for serum.
|
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] |
PMID:20473
|
Myxomatosis on the Western Plains of Victoria.
|
Myxomatosis on the Western Plains is an enzootic disease in contrast with the epizootic pattern which is general in eastern Australia. The most unusual aspects are the presence of significant numbers of diseased rabbits throughout the winter and the continuously low percentage of rabbits with antibodies to myxoma virus. Climatic and topographic conditions are unsuited to the production of the high densities of mosquitoes necessary for widespread epizootics. Under these conditions the effects of less efficient methods of myxomatosis transmission are apparent. The unusual epidemiology of myxomatosis has resulted in selection for virulence of the virus similar to that which has occurred under summer epizootic conditions. All field strains are now in the mid range of virulence.
|
Myxomatosis on the Western Plains of Victoria. Myxomatosis on the Western Plains is an enzootic disease in contrast with the epizootic pattern which is general in eastern Australia. The most unusual aspects are the presence of significant numbers of diseased rabbits throughout the winter and the continuously low percentage of rabbits with antibodies to myxoma virus. Climatic and topographic conditions are unsuited to the production of the high densities of mosquitoes necessary for widespread epizootics. Under these conditions the effects of less efficient methods of myxomatosis transmission are apparent. The unusual epidemiology of myxomatosis has resulted in selection for virulence of the virus similar to that which has occurred under summer epizootic conditions. All field strains are now in the mid range of virulence.
|
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] |
PMID:20478
|
Phenothiazine phototoxicity: toxic chlorpromazine photoproducts.
|
The effect of preirradiated chlorpromazine (CPZ) solutions has been studied in vivo in the guinea pig and in vitro in a yeast culture. With intracutaneous administration of CPZ solutions preirradiated with long-wave ultraviolet radiation (UVA), an inflammatory response was attained equaling the response achieved when nonirradiated solutions were injected and the skin of the animals was subsequently irradiated with UVA. Preirradiated CPZ was not toxic to the yeast culture. During irradiation the pH of the CPZ solution changed, indicating an accumulation of acid photoproducts. Nonphototoxic phenothiazines and related tricyclic drugs failed to change their pH following irradiation whereas phototoxic compounds behaved like CPZ. The low pH alone could not account for the inflammatory response in vivo, as neutralized irradiated CPZ solutions still induced inflammation, and acidified, nonirradiated solutions failed to cause a reaction. CPZ phototoxicity seems mainly to be due to the effect of photoproducts. The nature of these is not known, but it is suggested that the stable free radical of CPZ might be an active principle.
|
Phenothiazine phototoxicity: toxic chlorpromazine photoproducts. The effect of preirradiated chlorpromazine (CPZ) solutions has been studied in vivo in the guinea pig and in vitro in a yeast culture. With intracutaneous administration of CPZ solutions preirradiated with long-wave ultraviolet radiation (UVA), an inflammatory response was attained equaling the response achieved when nonirradiated solutions were injected and the skin of the animals was subsequently irradiated with UVA. Preirradiated CPZ was not toxic to the yeast culture. During irradiation the pH of the CPZ solution changed, indicating an accumulation of acid photoproducts. Nonphototoxic phenothiazines and related tricyclic drugs failed to change their pH following irradiation whereas phototoxic compounds behaved like CPZ. The low pH alone could not account for the inflammatory response in vivo, as neutralized irradiated CPZ solutions still induced inflammation, and acidified, nonirradiated solutions failed to cause a reaction. CPZ phototoxicity seems mainly to be due to the effect of photoproducts. The nature of these is not known, but it is suggested that the stable free radical of CPZ might be an active principle.
|
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] |
PMID:20479
|
Acute bacterial meningitis at Boston City Hospital during 12 selected years, 1935-1972.
|
Data are presented on the occurrence of and mortality rate from acute bacterial meningitis at Boston City Hospital during 12 years between 1935 and 1972 selected in relation to the introduction of potent antibacterial agents. The most frequent causative organisms were Streptococcus pneumoniae, Neisseria meningitidis, and Haemophilus influenzae, but large proportions were caused by other gram-positive cocci and gram-negative bacilli. The greatest reduction in mortality rate after antibiotics became widely used was in patients with meningococcal and influenzal meningitis who were less than or equal to 19 years old. Less striking reductions occurred in cases of other etiologies in patients less than or equal to 59 years old, but in those greater than or equal to 60 years old, the mortality rate remained high, and the proportion of cases of meningitis in that age group more than doubled. Comparisons with similar data on all bacteremic infections are presented.
|
Acute bacterial meningitis at Boston City Hospital during 12 selected years, 1935-1972. Data are presented on the occurrence of and mortality rate from acute bacterial meningitis at Boston City Hospital during 12 years between 1935 and 1972 selected in relation to the introduction of potent antibacterial agents. The most frequent causative organisms were Streptococcus pneumoniae, Neisseria meningitidis, and Haemophilus influenzae, but large proportions were caused by other gram-positive cocci and gram-negative bacilli. The greatest reduction in mortality rate after antibiotics became widely used was in patients with meningococcal and influenzal meningitis who were less than or equal to 19 years old. Less striking reductions occurred in cases of other etiologies in patients less than or equal to 59 years old, but in those greater than or equal to 60 years old, the mortality rate remained high, and the proportion of cases of meningitis in that age group more than doubled. Comparisons with similar data on all bacteremic infections are presented.
|
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] |
PMID:20481
|
Some properties of Hb G San Jose (beta7 glu replaced by gly): comparisons with Hb S.
|
Hb G San Jose (beta7 glu leads to gly) was studied with respect to oxygen affinity, Bohr effect, surface activity in dilute aqueous solutions, mechanical precipitability, heat stability and its ability to copolymerize in the deoxy form with Hb S. Oxygen affinity, Bohr effect, and polymerization with Hb S were found to be identical to those of Hb A when studied under the same conditions. However, surface activity and mechanical precipitation rates of the oxyconformers closely resembled those of oxyhemoglobin S. Hb G San Jose was also found to be slightly more unstable with heat than Hb A, although the instability was not detected by the usual incubation method of 1 hr at 50 degrees and higher temperatures were needed to elicit this difference. It is concluded that the ability to polymerize and the presence of increased surface activity are distinct and separable attributes of hemoglobin mutants. The finding that mixtures of Hb S and Hb G San Jose gel like mixtures of Hb S and Hb A supports the conclusion that only one beta 6 Val combining site per tetramer is required for polymer formation.
|
Some properties of Hb G San Jose (beta7 glu replaced by gly): comparisons with Hb S. Hb G San Jose (beta7 glu leads to gly) was studied with respect to oxygen affinity, Bohr effect, surface activity in dilute aqueous solutions, mechanical precipitability, heat stability and its ability to copolymerize in the deoxy form with Hb S. Oxygen affinity, Bohr effect, and polymerization with Hb S were found to be identical to those of Hb A when studied under the same conditions. However, surface activity and mechanical precipitation rates of the oxyconformers closely resembled those of oxyhemoglobin S. Hb G San Jose was also found to be slightly more unstable with heat than Hb A, although the instability was not detected by the usual incubation method of 1 hr at 50 degrees and higher temperatures were needed to elicit this difference. It is concluded that the ability to polymerize and the presence of increased surface activity are distinct and separable attributes of hemoglobin mutants. The finding that mixtures of Hb S and Hb G San Jose gel like mixtures of Hb S and Hb A supports the conclusion that only one beta 6 Val combining site per tetramer is required for polymer formation.
|
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] |
PMID:20482
|
Effect of dietary fat saturation on acylcoenzyme A:-cholesterol acyltransferase activity of Ehrlich cell microsomes.
|
Ehrlich cells grown in mice fed coconut oil diets (highly saturated) contain about twice as much cholesteryl ester as those grown in mice fed sunflower oil diets (highly polyunsaturated). Acylcoenzyme A: cholesterol acyltransferase (ACAT) activity was 30-100% higher in microsomes prepared from the cells grown on coconut oil (M(c)) than in those prepared from the cells grown on sunflower oil (M(s)). Increased ACAT activity was noted in M(c) with either [1-(14)C]palmitoyl CoA or [1,2-(3)H]cholesterol as the labeled substrate. This occurred at all acyl CoA concentrations tested and, in the [1,2-(3)H]cholesterol assay, with palmitoyl, oleoyl, or linoleoyl CoA as the substrate. The pH optimum for ACAT activity was the same with M(c) and M(s), pH 7.0. ACAT activity obeyed Michaelis-Menten kinetics at palmitoyl CoA concentrations between 1 and 10 micro M. Substrate inhibition occurred at higher concentrations. Kinetic analysis with [1-(14)C]palmitoyl CoA as the substrate indicated that the apparent K(m) for M(c) was 33% smaller than for M(s). There was no difference, however, in apparent V(max) values. The cholesterol and phospholipid contents of M(c) and M(s) were similar, but their fatty acid compositions differed considerably. M(c) contained 2.7 times more monoenoic fatty acid and only half as much polyenoic fatty acid as M(s). Our results indicate that dietary modification of the microsomal fatty acid composition is associated with alterations in the activity of ACAT, an enzyme that is tightly bound to the microsomes. These changes in ACAT activity may be partly responsible for the differences in cholesteryl ester contents of Ehrlich cells grown in mice fed the coconut and sunflower oil diets.
|
Effect of dietary fat saturation on acylcoenzyme A:-cholesterol acyltransferase activity of Ehrlich cell microsomes. Ehrlich cells grown in mice fed coconut oil diets (highly saturated) contain about twice as much cholesteryl ester as those grown in mice fed sunflower oil diets (highly polyunsaturated). Acylcoenzyme A: cholesterol acyltransferase (ACAT) activity was 30-100% higher in microsomes prepared from the cells grown on coconut oil (M(c)) than in those prepared from the cells grown on sunflower oil (M(s)). Increased ACAT activity was noted in M(c) with either [1-(14)C]palmitoyl CoA or [1,2-(3)H]cholesterol as the labeled substrate. This occurred at all acyl CoA concentrations tested and, in the [1,2-(3)H]cholesterol assay, with palmitoyl, oleoyl, or linoleoyl CoA as the substrate. The pH optimum for ACAT activity was the same with M(c) and M(s), pH 7.0. ACAT activity obeyed Michaelis-Menten kinetics at palmitoyl CoA concentrations between 1 and 10 micro M. Substrate inhibition occurred at higher concentrations. Kinetic analysis with [1-(14)C]palmitoyl CoA as the substrate indicated that the apparent K(m) for M(c) was 33% smaller than for M(s). There was no difference, however, in apparent V(max) values. The cholesterol and phospholipid contents of M(c) and M(s) were similar, but their fatty acid compositions differed considerably. M(c) contained 2.7 times more monoenoic fatty acid and only half as much polyenoic fatty acid as M(s). Our results indicate that dietary modification of the microsomal fatty acid composition is associated with alterations in the activity of ACAT, an enzyme that is tightly bound to the microsomes. These changes in ACAT activity may be partly responsible for the differences in cholesteryl ester contents of Ehrlich cells grown in mice fed the coconut and sunflower oil diets.
|
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] |
PMID:20483
|
Effect of sodium valproate on plasma protein binding of diphenylhydantoin.
|
In vivo and in vitro experimental data are presented in support of the hypothesis that sodium valproate displaces diphenylhydantoin (DPH) from plasma protein binding sites. This interaction could have important practical implications in the management of patients on combined therapy with these two drugs. Acute neurological features of DPH intoxication may be precipitated as a result of an increased free (pharmacologically active) fraction, which would not be detected by routine plasma DPH estimations since these reflects largely the bound fraction.
|
Effect of sodium valproate on plasma protein binding of diphenylhydantoin. In vivo and in vitro experimental data are presented in support of the hypothesis that sodium valproate displaces diphenylhydantoin (DPH) from plasma protein binding sites. This interaction could have important practical implications in the management of patients on combined therapy with these two drugs. Acute neurological features of DPH intoxication may be precipitated as a result of an increased free (pharmacologically active) fraction, which would not be detected by routine plasma DPH estimations since these reflects largely the bound fraction.
|
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] |
PMID:20485
|
Mouse spinal cord in cell culture. III. Neuronal chemosensitivity and its relationship to synaptic activity.
|
1. Mouse spinal cord (SC) cells in dissociated cell cultures showed strong electrophysiologic responses to glutamate, gamma-aminobutyric acid (GABA), and glycine when these were iontophoretically applied to the neurons. 2. The extrapolated reversal potential for the glutamate response was 20-30 mV negative in contrast to the positive extrapolated reversal potential for the SC-SC excitatory postsynaptic potential. The data are interpreted as indicating different ionic mechanisms for the glutamate response and the EPSP. 3. The reversal potentials for the glycine and GABA responses were similar to one another and to the IPSP reversal potential. The time course of the glycine and GABA responses were quite different from each other, however. 4. While some SC cells showed a relatively uniform sensitivity over their surfaces to iontophoretically applied glutamate, discrete regions of higher sensitivity occurred on most cells. 5. Release of excitatory and inhibitory transmitter could be elicited by focal application of glutamate and, in favorable instances, this could be shown to be due to the sensitivity of presynaptic terminals to the applied glutamate. Considerable spatial resolution of regions from which transmitter release could be elicited was achieved by this technique. Some correspondence between glutamate "hot spots" and such release sites was found.
|
Mouse spinal cord in cell culture. III. Neuronal chemosensitivity and its relationship to synaptic activity. 1. Mouse spinal cord (SC) cells in dissociated cell cultures showed strong electrophysiologic responses to glutamate, gamma-aminobutyric acid (GABA), and glycine when these were iontophoretically applied to the neurons. 2. The extrapolated reversal potential for the glutamate response was 20-30 mV negative in contrast to the positive extrapolated reversal potential for the SC-SC excitatory postsynaptic potential. The data are interpreted as indicating different ionic mechanisms for the glutamate response and the EPSP. 3. The reversal potentials for the glycine and GABA responses were similar to one another and to the IPSP reversal potential. The time course of the glycine and GABA responses were quite different from each other, however. 4. While some SC cells showed a relatively uniform sensitivity over their surfaces to iontophoretically applied glutamate, discrete regions of higher sensitivity occurred on most cells. 5. Release of excitatory and inhibitory transmitter could be elicited by focal application of glutamate and, in favorable instances, this could be shown to be due to the sensitivity of presynaptic terminals to the applied glutamate. Considerable spatial resolution of regions from which transmitter release could be elicited was achieved by this technique. Some correspondence between glutamate "hot spots" and such release sites was found.
|
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] |
PMID:20486
|
Early effects of hypervitaminosis A on gluconeogenic activity and amino acid metabolizing enzymes of rat liver.
|
In an earlier report from this laboratory, one of the early manifestations of hypervitaminosis A was shown to be a marked stimulation of hepatic gluconeogenesis. In the present study, effects of feeding 30,000 IU of retinyl palmitate to young rats (80-100 g), once daily, for 2 days on the incorporation of 14C-labeled precursors into glucose and glycogen by liver slices, levels of amino acids in blood and tissues, and activities of some important amino acid catabolizing enzymes in the liver were investigated. A stimulation of hepatic gluconeogenesis in hypervitaminosis A was indicated by the increased incorporation of 14C-labeled alanine and bicarbonate into glucose and glycogen by liver slices. Excessive intake of retinol caused a marked increase in the activities of hepatic alanine aminotransferase and ornithine aminotransferase and a decrease in that of tryptophan pyrrolase, without affecting those of tyrosine aminotransferase and serine dehydratase. The ratio of NADH:NAD in the livers of rats fed excess retinol was significantly increased. It is suggested that enhancement of glucoeogenesis in hypervitaminosis A is caused by a stimulation of gluconeogenic activity of the liver.
|
Early effects of hypervitaminosis A on gluconeogenic activity and amino acid metabolizing enzymes of rat liver. In an earlier report from this laboratory, one of the early manifestations of hypervitaminosis A was shown to be a marked stimulation of hepatic gluconeogenesis. In the present study, effects of feeding 30,000 IU of retinyl palmitate to young rats (80-100 g), once daily, for 2 days on the incorporation of 14C-labeled precursors into glucose and glycogen by liver slices, levels of amino acids in blood and tissues, and activities of some important amino acid catabolizing enzymes in the liver were investigated. A stimulation of hepatic gluconeogenesis in hypervitaminosis A was indicated by the increased incorporation of 14C-labeled alanine and bicarbonate into glucose and glycogen by liver slices. Excessive intake of retinol caused a marked increase in the activities of hepatic alanine aminotransferase and ornithine aminotransferase and a decrease in that of tryptophan pyrrolase, without affecting those of tyrosine aminotransferase and serine dehydratase. The ratio of NADH:NAD in the livers of rats fed excess retinol was significantly increased. It is suggested that enhancement of glucoeogenesis in hypervitaminosis A is caused by a stimulation of gluconeogenic activity of the liver.
|
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] |
PMID:20487
|
Sites of organic acid production and patterns of digesta movement in the gastrointestinal tract of the rock hyrax.
|
The rock hyrax, smallest of the present day ungulates and a near relative of the elephant, has a gastrointestinal tract of such complex nature that it may be considered ccomparable to both the simple and complex stomach mammals, and also to birds. They have a stomach to body weight capacity equal to that of sheep. But for all its complexity, the passage of ingesta is reasonably rapid. However, there are three major sites of digesta retention and microbial fermentation of the ingested material. The cranial stomach provides the first major site of fermentation with VFA levels comparable to those observed in the rumen (125-145 mM/liter). Quantities of lactic acid are also produced in the foregut where the pH may vary from 2 to 5.5. The ingested particulate material is exclusively retained in the cranial half of the stomach for 4 hours after consumption, and partially retained in the entire stomach for greater than 24 hours. The mid-gut sacculation provides the second major site of fermentation with the retention of the digesta in this gut segment for up to 16 hours. The two ceca provide the third site. Concentrations of VFA in these latter two gut segments approaches 80 mM/liter, while the near stable and neutral pH corresponds to the absence of lactic acid production.
|
Sites of organic acid production and patterns of digesta movement in the gastrointestinal tract of the rock hyrax. The rock hyrax, smallest of the present day ungulates and a near relative of the elephant, has a gastrointestinal tract of such complex nature that it may be considered ccomparable to both the simple and complex stomach mammals, and also to birds. They have a stomach to body weight capacity equal to that of sheep. But for all its complexity, the passage of ingesta is reasonably rapid. However, there are three major sites of digesta retention and microbial fermentation of the ingested material. The cranial stomach provides the first major site of fermentation with VFA levels comparable to those observed in the rumen (125-145 mM/liter). Quantities of lactic acid are also produced in the foregut where the pH may vary from 2 to 5.5. The ingested particulate material is exclusively retained in the cranial half of the stomach for 4 hours after consumption, and partially retained in the entire stomach for greater than 24 hours. The mid-gut sacculation provides the second major site of fermentation with the retention of the digesta in this gut segment for up to 16 hours. The two ceca provide the third site. Concentrations of VFA in these latter two gut segments approaches 80 mM/liter, while the near stable and neutral pH corresponds to the absence of lactic acid production.
|
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] |
PMID:20491
|
Vehicle effects on ocular drug bioavailability II: Evaluation of pilocarpine.
|
The influence of vehicle composition on ocular penetration of pilocarpine was studied in the albino rabbit. Increasing the pH of a vehicle promoted increased corneal penetration for pilocarpine, in accordance with the pH-partition hypothesis, but a similar series of experiments with a nonionizable drug, glycerin, gave similar results. The extent of pH-induced lacrimation by the vehicle and its effect on precorneal drug concentration also was determined. Increased pilocarpine absorption at neutral to slightly alkaline pH was due primarily to its peculiar solubility characteristics coupled with less irritation and lacrimation rather than a direct pH effect on the molecule. Incorporation of pilocarpine into a petrolatum-based ointment vehicle resulted in increased aqueous humor pilocarpine levels above those provided by an equivalent dose of aqueous solution. The mechanism of this increase was determined to be a higher effective concentration of pilocarpine in the ointment vehicle coupled with an increase in contact time of the dose. The ointment system also exerted an unusual form of vehicle control in that it promoted the corneal penetration of pilocarpine while impeding uniform mixing of the dose with tears and thereby imposed a restriction on the amount of pilocarpine available to the ocular tissues.
|
Vehicle effects on ocular drug bioavailability II: Evaluation of pilocarpine. The influence of vehicle composition on ocular penetration of pilocarpine was studied in the albino rabbit. Increasing the pH of a vehicle promoted increased corneal penetration for pilocarpine, in accordance with the pH-partition hypothesis, but a similar series of experiments with a nonionizable drug, glycerin, gave similar results. The extent of pH-induced lacrimation by the vehicle and its effect on precorneal drug concentration also was determined. Increased pilocarpine absorption at neutral to slightly alkaline pH was due primarily to its peculiar solubility characteristics coupled with less irritation and lacrimation rather than a direct pH effect on the molecule. Incorporation of pilocarpine into a petrolatum-based ointment vehicle resulted in increased aqueous humor pilocarpine levels above those provided by an equivalent dose of aqueous solution. The mechanism of this increase was determined to be a higher effective concentration of pilocarpine in the ointment vehicle coupled with an increase in contact time of the dose. The ointment system also exerted an unusual form of vehicle control in that it promoted the corneal penetration of pilocarpine while impeding uniform mixing of the dose with tears and thereby imposed a restriction on the amount of pilocarpine available to the ocular tissues.
|
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] |
PMID:20492
|
Everted rat intestinal sacs as an in vitro model for assessing absorptivity of new drugs.
|
An in vitro model that utilizes everted rat intestinal sacs was evaluated for assessing the absorptivity of several analogs of potential drug substances prior to formulation work and clinical trials. This model not only is a useful qualitative tool for assessing absorptivity of structurally related compounds but also yields some insight into the process involved in drug absorption. Notwithstanding the complexities involved in the absorption processes, the data support the hypothesis that the absorption of organic electrolytes mainly takes place by the partitioning of the unionized species into the lipoidal membranes and then diffusion.
|
Everted rat intestinal sacs as an in vitro model for assessing absorptivity of new drugs. An in vitro model that utilizes everted rat intestinal sacs was evaluated for assessing the absorptivity of several analogs of potential drug substances prior to formulation work and clinical trials. This model not only is a useful qualitative tool for assessing absorptivity of structurally related compounds but also yields some insight into the process involved in drug absorption. Notwithstanding the complexities involved in the absorption processes, the data support the hypothesis that the absorption of organic electrolytes mainly takes place by the partitioning of the unionized species into the lipoidal membranes and then diffusion.
|
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] |
PMID:20494
|
Characteristics of medicated and unmedicated microglobules recovered from complex coacervates of gelatin-acacia.
|
Medicated and unmedicated microglobules prepared from complex coacervates of Type A gelatin and acacia were recovered as water-insoluble powders consisting of discrete units, which were spontaneously revertible to highly disperse systems when suspended in water or physiological electrolyte solutions. Spherical microglobules containing up to 15% (w/w) sulfamerazine had a nominal diameter of 30 micron in aqueous suspension. Larger but irregularly shaped products containing up to 45% (w/w) sulfamerazine were also recovered. The relationships of the weight of sulfamerazine added per coacervate batch to weight yield and percent (w/w) included sulfamerazine of the microglobules were both linear.
|
Characteristics of medicated and unmedicated microglobules recovered from complex coacervates of gelatin-acacia. Medicated and unmedicated microglobules prepared from complex coacervates of Type A gelatin and acacia were recovered as water-insoluble powders consisting of discrete units, which were spontaneously revertible to highly disperse systems when suspended in water or physiological electrolyte solutions. Spherical microglobules containing up to 15% (w/w) sulfamerazine had a nominal diameter of 30 micron in aqueous suspension. Larger but irregularly shaped products containing up to 45% (w/w) sulfamerazine were also recovered. The relationships of the weight of sulfamerazine added per coacervate batch to weight yield and percent (w/w) included sulfamerazine of the microglobules were both linear.
|
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] |
PMID:20495
|
In vitro adsorption of doxorubicin hydrochloride on insoluble calcium phosphate.
|
The adsorption of doxorubicin hydrochloride, a potent antitumor agent, on solid tribasic calcium phosphate was studied in vitro. A Langmuir adsorption isotherm at pH 7.4 and the maximum adsorption capacity of tribasic calcium phosphate were established. Tribasic calcium phosphate was chosen as a model for solid bone samples, which are stained with doxorubicin in patients who have received long-term doxorubicin therapy.
|
In vitro adsorption of doxorubicin hydrochloride on insoluble calcium phosphate. The adsorption of doxorubicin hydrochloride, a potent antitumor agent, on solid tribasic calcium phosphate was studied in vitro. A Langmuir adsorption isotherm at pH 7.4 and the maximum adsorption capacity of tribasic calcium phosphate were established. Tribasic calcium phosphate was chosen as a model for solid bone samples, which are stained with doxorubicin in patients who have received long-term doxorubicin therapy.
|
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] |
PMID:20497
|
Mechanism of isoproterenol-induced desensitization of tracheal smooth muscle.
|
Isolated rat tracheal smooth muscle became considerably less sensitive to the relaxing action of isoproterenol after being incubated with 5 x 10(-6) M isoproterenol for 30 minutes. Pretreatment of the tissue with propranolol, but not with methylprednisolone, clearly reduced the isoproterenol-induced desensitization. This suggested that propranolol by occupying the beta adrenergic receptor prevented isoproterenol from binding to this receptor, thereby preventing the isoproterenol-induced desensitization. Furthermore, an isoproterenol-desensitized tracheal preparation exhibited a diminished sensitivity to other beta agonists, but not to the spasmolytic actions of D600, hydralazine, sodium nitrite and aminophylline. These results suggested that the beta receptor is specifically involved in the desensitization induced by isoproterenol. A highly desensitized tissue could always be made to undergo complete relaxation by exposing it to sufficiently high concentrations of isoproterenol. Thus, there appeared to be no positive indication of a very large change in the apparent intrinsic activity of the isoproterenol in the desensitized tissue. However, the dissociation constant for the propranolol-beta receptor complex in the desensitized tissue was shown to be 180-fold larger than that in the normal tissue. These findings provide strong evidence that one demonstrable cellular change that occurs in the desensitized tissue is a pronounced reduction in the affinity of the beta receptors for isoproterenol.
|
Mechanism of isoproterenol-induced desensitization of tracheal smooth muscle. Isolated rat tracheal smooth muscle became considerably less sensitive to the relaxing action of isoproterenol after being incubated with 5 x 10(-6) M isoproterenol for 30 minutes. Pretreatment of the tissue with propranolol, but not with methylprednisolone, clearly reduced the isoproterenol-induced desensitization. This suggested that propranolol by occupying the beta adrenergic receptor prevented isoproterenol from binding to this receptor, thereby preventing the isoproterenol-induced desensitization. Furthermore, an isoproterenol-desensitized tracheal preparation exhibited a diminished sensitivity to other beta agonists, but not to the spasmolytic actions of D600, hydralazine, sodium nitrite and aminophylline. These results suggested that the beta receptor is specifically involved in the desensitization induced by isoproterenol. A highly desensitized tissue could always be made to undergo complete relaxation by exposing it to sufficiently high concentrations of isoproterenol. Thus, there appeared to be no positive indication of a very large change in the apparent intrinsic activity of the isoproterenol in the desensitized tissue. However, the dissociation constant for the propranolol-beta receptor complex in the desensitized tissue was shown to be 180-fold larger than that in the normal tissue. These findings provide strong evidence that one demonstrable cellular change that occurs in the desensitized tissue is a pronounced reduction in the affinity of the beta receptors for isoproterenol.
|
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] |
PMID:20498
|
Effect of dipivalyl derivatives of catecholamines on cardiovascular function in the conscious dog.
|
Dipivalyl derivatives of epinephrine, norepinephrine and isoproterenol were injected intravenously into conscious dogs while cardiovascular variables were monitored. The dipivalyl compounds produced cardiovascular effects that were comparable to those produced by their respective parent catecholamines except that the responses had a delayed onset and a prolonged duration. The catecholamines were more potent than their derivatives; norepinephrine was 28 times, epinephrine 15 times and isoproterenol 1.7 times as effective as their respective dipivalyl derivatives. Alpha or beta adrenergic responses to the dipivalyl compounds were attenuated or abolished after alpha adrenergic blockade (phenoxybenzamine or phentolamine), or beta adrenergic blockade (propranolol), respectively, or combined alpha and beta blockade. Since other evidence indicates that the dipivalyl derivatives themselves are inactive, our results suggest that these compounds act as prodrugs that exert their cardiovascular effects only after they are biotransformed to catecholamines.
|
Effect of dipivalyl derivatives of catecholamines on cardiovascular function in the conscious dog. Dipivalyl derivatives of epinephrine, norepinephrine and isoproterenol were injected intravenously into conscious dogs while cardiovascular variables were monitored. The dipivalyl compounds produced cardiovascular effects that were comparable to those produced by their respective parent catecholamines except that the responses had a delayed onset and a prolonged duration. The catecholamines were more potent than their derivatives; norepinephrine was 28 times, epinephrine 15 times and isoproterenol 1.7 times as effective as their respective dipivalyl derivatives. Alpha or beta adrenergic responses to the dipivalyl compounds were attenuated or abolished after alpha adrenergic blockade (phenoxybenzamine or phentolamine), or beta adrenergic blockade (propranolol), respectively, or combined alpha and beta blockade. Since other evidence indicates that the dipivalyl derivatives themselves are inactive, our results suggest that these compounds act as prodrugs that exert their cardiovascular effects only after they are biotransformed to catecholamines.
|
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] |
PMID:20499
|
Effect of changing the composition of the bathing solutions upon the isometric tension-pCa relationship in bundles of crustacean myofibrils.
|
1. The relative isometric tension-pCa relationship has been determined for isolated bundles of barnacle myofibrils under a variety of ionic conditions using [Ca(2+)]-buffered solutions which also contained an ATP regenerating system (creatine phosphate and creatine kinase).2. The results are in better agreement with the ;consecutive' scheme of reaction rather than with the ;independent' alternative (Ashley & Moisescu, 1972) for the co-operative action of two Ca(2+) ions in the process of tension activation in crustacean skeletal muscle.3. Variations in the pH of the activating solutions did have a marked effect on the relative tension-Ca curve, although no effect was observed on the absolute maximum value for isometric tension. A shift in pH by 0.5 u. in the range 6.6-7.6 shifted the Ca(2+)-activation curve by 0.5 log u. towards lower free Ca(2+) concentrations.4. Changes in the free Mg(2+) concentration of the activating solutions in the millimolar range produced a pronounced shift of the relative tension-pCa curve along the pCa axis. Increasing [Mg(2+)] from 1 to 5 mM shifted the curve by about 0.7 log u. to higher free Ca(2+) concentrations, without significantly modifying its steepness.5. Changes in the MgATP concentration of the activating solutions in the range of 1-13 mM had no significant effect on the relative tension-pCa relationship.6. Varying the K(+) concentration in the activating solutions was also observed to have a marked effect upon the tension-pCa relationship in barnacle. An increase in the K(+) concentration from 90 to 170 mM shifted the curve by some 0.6 log u. towards higher free Ca(2+) concentrations.7. Cooling the standard activating solutions from room temperature to +4 degrees C made no apparent difference to the relative tension-pCa relationship, but decreased significantly the absolute tension responses.8. The results presented show that tonicity by itself has a marked effect upon the absolute steady-state tension levels in isolated bundles of myofibrils.9. Maximum isometric tension in this preparation was not simply related to ionic strength, or to the monovalent cation concentration, but it depended, as well, upon the anionic composition of the activating solution. In addition, a change in ionic strength of 25 mM over the range of 245-270 mM did not appear to modify the relative tension-pCa relationship.10. The effect of the physiologically occurring cations H(+), K(+), Mg(2+) upon the relative isometric tension-pCa relationship can be accounted for on the basis of a model of competitive inhibition between these cations and Ca(2+) for the functional unit for tension. This inhibitory effect appears to involve at least one H(+), one Mg(2+) and two K(+) per each Ca(2+) ion participating in the activation process of the functional unit for tension.
|
Effect of changing the composition of the bathing solutions upon the isometric tension-pCa relationship in bundles of crustacean myofibrils. 1. The relative isometric tension-pCa relationship has been determined for isolated bundles of barnacle myofibrils under a variety of ionic conditions using [Ca(2+)]-buffered solutions which also contained an ATP regenerating system (creatine phosphate and creatine kinase).2. The results are in better agreement with the ;consecutive' scheme of reaction rather than with the ;independent' alternative (Ashley & Moisescu, 1972) for the co-operative action of two Ca(2+) ions in the process of tension activation in crustacean skeletal muscle.3. Variations in the pH of the activating solutions did have a marked effect on the relative tension-Ca curve, although no effect was observed on the absolute maximum value for isometric tension. A shift in pH by 0.5 u. in the range 6.6-7.6 shifted the Ca(2+)-activation curve by 0.5 log u. towards lower free Ca(2+) concentrations.4. Changes in the free Mg(2+) concentration of the activating solutions in the millimolar range produced a pronounced shift of the relative tension-pCa curve along the pCa axis. Increasing [Mg(2+)] from 1 to 5 mM shifted the curve by about 0.7 log u. to higher free Ca(2+) concentrations, without significantly modifying its steepness.5. Changes in the MgATP concentration of the activating solutions in the range of 1-13 mM had no significant effect on the relative tension-pCa relationship.6. Varying the K(+) concentration in the activating solutions was also observed to have a marked effect upon the tension-pCa relationship in barnacle. An increase in the K(+) concentration from 90 to 170 mM shifted the curve by some 0.6 log u. towards higher free Ca(2+) concentrations.7. Cooling the standard activating solutions from room temperature to +4 degrees C made no apparent difference to the relative tension-pCa relationship, but decreased significantly the absolute tension responses.8. The results presented show that tonicity by itself has a marked effect upon the absolute steady-state tension levels in isolated bundles of myofibrils.9. Maximum isometric tension in this preparation was not simply related to ionic strength, or to the monovalent cation concentration, but it depended, as well, upon the anionic composition of the activating solution. In addition, a change in ionic strength of 25 mM over the range of 245-270 mM did not appear to modify the relative tension-pCa relationship.10. The effect of the physiologically occurring cations H(+), K(+), Mg(2+) upon the relative isometric tension-pCa relationship can be accounted for on the basis of a model of competitive inhibition between these cations and Ca(2+) for the functional unit for tension. This inhibitory effect appears to involve at least one H(+), one Mg(2+) and two K(+) per each Ca(2+) ion participating in the activation process of the functional unit for tension.
|
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] |
PMID:20500
|
Daily changes in the blood of conscious pigs with catheters in foetal and uterine vessels during late gestation.
|
1. Catheters were inserted into the foetal carotid artery and maternal middle uterine artery and vein in twenty foetuses from fifteen sows, 91-105 days pregnant. Blood samples were collected from foetal and maternal circulations for periods of 7-21 days after surgery.2. Blood gas tensions, pH, packed cell volume (PCV) and the levels of glucose, fructose and lactic acid in conscious pigs were followed in foetal and maternal bloods during late pregnancy.3. Foetal blood gas tensions, pH, PCV, lactic acid and glucose levels did not change markedly during the last 20-25 days of gestation. Blood fructose concentration fell during this period with the greatest change occurring after 100 days gestation. Foetal and maternal blood pH values were higher than those reported in the sheep, cow, mare and rhesus monkey.4. Small changes in P(CO2) and pH in foetal carotid blood were associated with similar changes in maternal uterine venous blood, so that gradients between sow and foetus remained relatively constant.5. Daily changes in maternal blood glucose levels were associated with smaller changes in foetal blood glucose and fructose levels. Blood glucose concentrations in foetal blood were generally 40-70% of maternal concentrations. Over-all relationships between maternal blood glucose and foetal blood fructose concentrations were masked by the independent fall in foetal fructose levels with age.6. The present findings have been compared with previous observations on acute, anaesthetized preparations in pigs, and chronic preparations in other domestic animals.
|
Daily changes in the blood of conscious pigs with catheters in foetal and uterine vessels during late gestation. 1. Catheters were inserted into the foetal carotid artery and maternal middle uterine artery and vein in twenty foetuses from fifteen sows, 91-105 days pregnant. Blood samples were collected from foetal and maternal circulations for periods of 7-21 days after surgery.2. Blood gas tensions, pH, packed cell volume (PCV) and the levels of glucose, fructose and lactic acid in conscious pigs were followed in foetal and maternal bloods during late pregnancy.3. Foetal blood gas tensions, pH, PCV, lactic acid and glucose levels did not change markedly during the last 20-25 days of gestation. Blood fructose concentration fell during this period with the greatest change occurring after 100 days gestation. Foetal and maternal blood pH values were higher than those reported in the sheep, cow, mare and rhesus monkey.4. Small changes in P(CO2) and pH in foetal carotid blood were associated with similar changes in maternal uterine venous blood, so that gradients between sow and foetus remained relatively constant.5. Daily changes in maternal blood glucose levels were associated with smaller changes in foetal blood glucose and fructose levels. Blood glucose concentrations in foetal blood were generally 40-70% of maternal concentrations. Over-all relationships between maternal blood glucose and foetal blood fructose concentrations were masked by the independent fall in foetal fructose levels with age.6. The present findings have been compared with previous observations on acute, anaesthetized preparations in pigs, and chronic preparations in other domestic animals.
|
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] |
PMID:20501
|
Continuous direct measurement of intracellular chloride and pH in frog skeletal muscle.
|
1. Ion-sensitive electrodes (made with a chloride-sensitive ion-exchange resin) were used to measure the internal chloride activity (a(i) (Cl)) of frog sartorius fibres at 25 degrees C.2. The internal pH (pH(i)) of other sartorius fibres was measured with a recessed tip pH-sensitive electrode (made with pH-sensitive glass).3. In normal bicarbonate-free solution (containing 2.5 mM potassium), the average chloride equilibrium potential, E(Cl) (calculated from a(i) (Cl) and the measured chloride activity of the external solution (a(o) (Cl)) was 87.7 +/- 1.7 mV (mean +/- S.E.; n = 16) in fibres where the average membrane potential, E(m), was 88.3 +/- 1.5 mV (mean +/- S.E.; n = 16). In experiments where a(i) (Cl) was varied between about 1 and 10 mM (which corresponds to values of E(m) between about -105 and -50 mV) E(Cl) was within 1-3 mV of E(m) at equilibrium. These measurements of a(i) (Cl) were obtained from the potential difference between the chloride-sensitive electrode and an intracellular indifferent micro-electrode filled with potassium chloride. If a potassium sulphate-filled indifferent micro-electrode was used, then values of a(i) (Cl) below about 5 mM were erroneously high, probably due to interference from other sarcoplasmic ions at the indifferent electrode.4. In solutions containing 15 mM bicarbonate and gassed with 5% CO(2), pH(i) was 6.9, corresponding to an internal bicarbonate concentration of 7.6 mM. E(Cl) measured in this solution was some 4 mV positive to E(m). Most of the difference between E(Cl) and E(m) could be ascribed to interference by sarcoplasmic bicarbonate on the basis of selectivity measurements of chloride against bicarbonate made on the ion-exchange resin in the relevant range of a(Cl).5. If bicarbonate/CO(2) in the external solution was replaced by HEPES/pure O(2) at constant pH, then pH(i) rose from 6.88 +/- 0.02 (mean +/- S.E.) to 7.05 +/- 0.02. A change in external pH of 1 unit caused pH(i) to change by about 0.02 unit and the intracellular buffering power was calculated to be about 35.6. In solution made hypertonic by the addition of sucrose, E(m) changed little or depolarized and E(Cl) and E(m) remained close. In contrast, in solution made hypertonic by the addition of solid sodium chloride (high-chloride solution) E(Cl) became negative to E(m). Conversely in low chloride solution E(Cl) became positive to E(m).7. When the chloride permeability (P(Cl)) was reduced by the use of acid solution, E(Cl) moved positive to E(m) indicating an accumulation of internal chloride. When P(Cl) was increased again by returning to more alkaline solution, E(m) depolarized to E(Cl).8. The results are consistent with the existence of a small, active movement of chloride, the effects of which are normally obscured by large passive movements of chloride when P(Cl) is large.
|
Continuous direct measurement of intracellular chloride and pH in frog skeletal muscle. 1. Ion-sensitive electrodes (made with a chloride-sensitive ion-exchange resin) were used to measure the internal chloride activity (a(i) (Cl)) of frog sartorius fibres at 25 degrees C.2. The internal pH (pH(i)) of other sartorius fibres was measured with a recessed tip pH-sensitive electrode (made with pH-sensitive glass).3. In normal bicarbonate-free solution (containing 2.5 mM potassium), the average chloride equilibrium potential, E(Cl) (calculated from a(i) (Cl) and the measured chloride activity of the external solution (a(o) (Cl)) was 87.7 +/- 1.7 mV (mean +/- S.E.; n = 16) in fibres where the average membrane potential, E(m), was 88.3 +/- 1.5 mV (mean +/- S.E.; n = 16). In experiments where a(i) (Cl) was varied between about 1 and 10 mM (which corresponds to values of E(m) between about -105 and -50 mV) E(Cl) was within 1-3 mV of E(m) at equilibrium. These measurements of a(i) (Cl) were obtained from the potential difference between the chloride-sensitive electrode and an intracellular indifferent micro-electrode filled with potassium chloride. If a potassium sulphate-filled indifferent micro-electrode was used, then values of a(i) (Cl) below about 5 mM were erroneously high, probably due to interference from other sarcoplasmic ions at the indifferent electrode.4. In solutions containing 15 mM bicarbonate and gassed with 5% CO(2), pH(i) was 6.9, corresponding to an internal bicarbonate concentration of 7.6 mM. E(Cl) measured in this solution was some 4 mV positive to E(m). Most of the difference between E(Cl) and E(m) could be ascribed to interference by sarcoplasmic bicarbonate on the basis of selectivity measurements of chloride against bicarbonate made on the ion-exchange resin in the relevant range of a(Cl).5. If bicarbonate/CO(2) in the external solution was replaced by HEPES/pure O(2) at constant pH, then pH(i) rose from 6.88 +/- 0.02 (mean +/- S.E.) to 7.05 +/- 0.02. A change in external pH of 1 unit caused pH(i) to change by about 0.02 unit and the intracellular buffering power was calculated to be about 35.6. In solution made hypertonic by the addition of sucrose, E(m) changed little or depolarized and E(Cl) and E(m) remained close. In contrast, in solution made hypertonic by the addition of solid sodium chloride (high-chloride solution) E(Cl) became negative to E(m). Conversely in low chloride solution E(Cl) became positive to E(m).7. When the chloride permeability (P(Cl)) was reduced by the use of acid solution, E(Cl) moved positive to E(m) indicating an accumulation of internal chloride. When P(Cl) was increased again by returning to more alkaline solution, E(m) depolarized to E(Cl).8. The results are consistent with the existence of a small, active movement of chloride, the effects of which are normally obscured by large passive movements of chloride when P(Cl) is large.
|
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] |
PMID:20502
|
beta-Adrenergic blocking agents. 16. 1-(Acylaminomethyl-, ureidomethyl-, and ureidoethylphenoxy)-3-amino-2-propanols.
|
The synthesis of a series of 1-(acylaminomethyl-, ureidomethyl-, and ureidoethylphenoxy)-3-amino-2-propanols is described. The compounds were screened as beta-adrenergic receptor antagonists in cats and their partial agonist activity was evaluated in rats depleted of circulating catecholamines. Some of the compounds have a pharmacological profile similar to atenolol. Their structure-activity relationships are discussed.
|
beta-Adrenergic blocking agents. 16. 1-(Acylaminomethyl-, ureidomethyl-, and ureidoethylphenoxy)-3-amino-2-propanols. The synthesis of a series of 1-(acylaminomethyl-, ureidomethyl-, and ureidoethylphenoxy)-3-amino-2-propanols is described. The compounds were screened as beta-adrenergic receptor antagonists in cats and their partial agonist activity was evaluated in rats depleted of circulating catecholamines. Some of the compounds have a pharmacological profile similar to atenolol. Their structure-activity relationships are discussed.
|
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] |
PMID:20503
|
Adrenerigic agents. 7.1 Synthesis and beta-adrenergic agonist activity of several 2-pyridylethanolamines.
|
In a search for new selective bronchodilators, three 2-pyridylethanolamines, i.e., 2-tert-butylamino-1-(5-hydroxy-2-pyridyl)ethanol (2b), a related 6-methylsulfonylmethyl (2c), and, a 6-methyl (2d) derivative, were prepared. These compounds were examined for potential bronchodilator activity in an in vitro test for relaxation of guinea pig tracheal tissue. Potential cardiac stimulant activity was evaluated in vitro by measuring changes in the rate of spontaneously beating guinea pig right atrial muscle. Comparison of potency in the tracheal test relative to that in the atrial procedure provides a measure of selectivity. Results of this study indicate that replacement of the phenyl ring of a para-hydroxylated phenylethanolamine with a 2-pyridyl system generally results in compounds which retain a high order of potency in the tracheal test; however, selectivity for tracheobronchial vs. cardiac tissue is markedly greater for the pyridyl derivatives. The alpha-picoline, 2-tert-butylamino-1-(5-hydroxy-6-methyl-2-pyridyl) ethanol (2d), which bears labile protons at a position meta to the ethanolamine side chain, was about equipotent with the corresponding 6-unsubstituted relative 2b. The reason for the failure of these apparently appropriately located labile protons to enhance beta-adrenoreceptor agonist activity is uncertain.
|
Adrenerigic agents. 7.1 Synthesis and beta-adrenergic agonist activity of several 2-pyridylethanolamines. In a search for new selective bronchodilators, three 2-pyridylethanolamines, i.e., 2-tert-butylamino-1-(5-hydroxy-2-pyridyl)ethanol (2b), a related 6-methylsulfonylmethyl (2c), and, a 6-methyl (2d) derivative, were prepared. These compounds were examined for potential bronchodilator activity in an in vitro test for relaxation of guinea pig tracheal tissue. Potential cardiac stimulant activity was evaluated in vitro by measuring changes in the rate of spontaneously beating guinea pig right atrial muscle. Comparison of potency in the tracheal test relative to that in the atrial procedure provides a measure of selectivity. Results of this study indicate that replacement of the phenyl ring of a para-hydroxylated phenylethanolamine with a 2-pyridyl system generally results in compounds which retain a high order of potency in the tracheal test; however, selectivity for tracheobronchial vs. cardiac tissue is markedly greater for the pyridyl derivatives. The alpha-picoline, 2-tert-butylamino-1-(5-hydroxy-6-methyl-2-pyridyl) ethanol (2d), which bears labile protons at a position meta to the ethanolamine side chain, was about equipotent with the corresponding 6-unsubstituted relative 2b. The reason for the failure of these apparently appropriately located labile protons to enhance beta-adrenoreceptor agonist activity is uncertain.
|
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] |
PMID:20504
|
Adrenergic agents. 8.1 Synthesis and beta-adrenergic agonist activity of some 3-tert-butylamino-2-(substituted phenyl)-1-propanols.
|
Replacement of the benzylic hydroxyl group of N-tert-butylnorepinephrine with a hydroxymethyl substituent affords a propanolamine homologue which retains a high degree of beta-adrenergic agonist activity. As modification of the meta substituent of catecholic ethanolamines, such as N-tert-butylnorepinephrine, often provides compounds that exert a more pronounced effect in relaxing tracheobronchial smooth muscle (beta2-adrenergic agonist) than in stimulating cardiac muscle (beta1-adrenergic response), a series of 3-tert-butylamino-2-(3-substituted 4-hydroxyphenyl)-1-propanols was prepared. The 3-meta substituents included HOCH2 (1b), H2NCONH (1c), MeSO2NH (1d), H (le), and NH2 (1f). These phenylpropanolamine derivatives were compared with their phenylethanolamine counterparts in in vitro tests that measure the ability of these compounds to relax spontaneously contracted guinea pig tracheal smooth muscle (a measure of potential bronchodilating activity) and to increase the rate of contraction of a spontaneously beating guinea pig right atrial preparation (an indicator of potential cardiac stimulating activity). In these tests all of the propanolamine derivatives included in the study were less potent than their ethanolamine relatives. In both series replacement of the catecholic m-hydroxyl group with the indicated substituents usually resulted in compounds with increased selectivity for tracheobronchial vs. cardiac muscle.
|
Adrenergic agents. 8.1 Synthesis and beta-adrenergic agonist activity of some 3-tert-butylamino-2-(substituted phenyl)-1-propanols. Replacement of the benzylic hydroxyl group of N-tert-butylnorepinephrine with a hydroxymethyl substituent affords a propanolamine homologue which retains a high degree of beta-adrenergic agonist activity. As modification of the meta substituent of catecholic ethanolamines, such as N-tert-butylnorepinephrine, often provides compounds that exert a more pronounced effect in relaxing tracheobronchial smooth muscle (beta2-adrenergic agonist) than in stimulating cardiac muscle (beta1-adrenergic response), a series of 3-tert-butylamino-2-(3-substituted 4-hydroxyphenyl)-1-propanols was prepared. The 3-meta substituents included HOCH2 (1b), H2NCONH (1c), MeSO2NH (1d), H (le), and NH2 (1f). These phenylpropanolamine derivatives were compared with their phenylethanolamine counterparts in in vitro tests that measure the ability of these compounds to relax spontaneously contracted guinea pig tracheal smooth muscle (a measure of potential bronchodilating activity) and to increase the rate of contraction of a spontaneously beating guinea pig right atrial preparation (an indicator of potential cardiac stimulating activity). In these tests all of the propanolamine derivatives included in the study were less potent than their ethanolamine relatives. In both series replacement of the catecholic m-hydroxyl group with the indicated substituents usually resulted in compounds with increased selectivity for tracheobronchial vs. cardiac muscle.
|
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] |
PMID:20508
|
Ion permeability and strength of cell contacts: ion permeability and mechanical properties of cell contacts in small intestine epithelium.
|
The effects of several simple parameters (pH, concentration of bivalent cations, osmotic pressure, and temperature) on the ion permeability and mechanical properties of cell contacts have been investigated. It has been shown that the mechanical properties of a cell contact make it possible to describe it as a viscoelastic system. The main contribution to cell adhesion is made by the tight junction. Two populations of acidic centers have been identified on the cell membrane surface. One population interacts with bivalent cations to assure cell adhesion. The other population of weaker acidic centers regulating ion permeation is involved in the cell membrane's interaction of the repulsion type. An intimate correlation has been established between changes in passive transepithelial ion permeability and cell adhesion in response to changes in pH and in bivalent cation concentration. Such a correlation is possible if the tight junction is the principal contributor to the passive ion permeability and mechanical strength of the cell contacts.
|
Ion permeability and strength of cell contacts: ion permeability and mechanical properties of cell contacts in small intestine epithelium. The effects of several simple parameters (pH, concentration of bivalent cations, osmotic pressure, and temperature) on the ion permeability and mechanical properties of cell contacts have been investigated. It has been shown that the mechanical properties of a cell contact make it possible to describe it as a viscoelastic system. The main contribution to cell adhesion is made by the tight junction. Two populations of acidic centers have been identified on the cell membrane surface. One population interacts with bivalent cations to assure cell adhesion. The other population of weaker acidic centers regulating ion permeation is involved in the cell membrane's interaction of the repulsion type. An intimate correlation has been established between changes in passive transepithelial ion permeability and cell adhesion in response to changes in pH and in bivalent cation concentration. Such a correlation is possible if the tight junction is the principal contributor to the passive ion permeability and mechanical strength of the cell contacts.
|
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] |
PMID:20513
|
Determination of specificity in natural cell-mediated cytotoxicity by natural antibodies.
|
Natural cell-mediated cytotoxicity (NCMC) and antibody-dependent cell-mediated cytotoxicity (ADCC) appeared to involve similar cytotoxic mechanisms; effector N-cells killed target cells with IgG antibodies attached to the Fc receptor determining the specificity of the reaction. In NCMC the wide range of specificities detected by natural antibodies provided an effector system capable of recognizing numerous antigens on cultured target cells. When several target cells were tested concurrently, an apparent nonselective cytotoxicity resulted. The specificity of individual reactions against each of the target cells could be demonstrated by selective inhibition with competitor cells. The inhibition of cytotoxicity by competition and the effect of proteases on the effector cell for NCMC, but not for ADCC, initially suggested an antibody on the surface of the natural cytotoxic effector cell. This suggestion was supported by the loss of activity with treatments that removed immunoglobulins on the effector cell and by the recovery of reactivity with incubation of the cells in normal human serum. Absorption of the reconstituting serum with target cells resulted in loss of activity against that target cell, substantiating the role of natural antibodies.
|
Determination of specificity in natural cell-mediated cytotoxicity by natural antibodies. Natural cell-mediated cytotoxicity (NCMC) and antibody-dependent cell-mediated cytotoxicity (ADCC) appeared to involve similar cytotoxic mechanisms; effector N-cells killed target cells with IgG antibodies attached to the Fc receptor determining the specificity of the reaction. In NCMC the wide range of specificities detected by natural antibodies provided an effector system capable of recognizing numerous antigens on cultured target cells. When several target cells were tested concurrently, an apparent nonselective cytotoxicity resulted. The specificity of individual reactions against each of the target cells could be demonstrated by selective inhibition with competitor cells. The inhibition of cytotoxicity by competition and the effect of proteases on the effector cell for NCMC, but not for ADCC, initially suggested an antibody on the surface of the natural cytotoxic effector cell. This suggestion was supported by the loss of activity with treatments that removed immunoglobulins on the effector cell and by the recovery of reactivity with incubation of the cells in normal human serum. Absorption of the reconstituting serum with target cells resulted in loss of activity against that target cell, substantiating the role of natural antibodies.
|
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] |
PMID:20514
|
In vivo immune responses of mice during carcinogenesis by ultraviolet irradiation.
|
In these experiments, we tested in various in vivo assays the immune responses of inbred C3H/HeN(MTV-) (C3H-) mice during carcinogenesis by chronic exposure to UV irradiation. Although the UV-treated mice were unable to reject syngeneic UV-induced tumor transplants, they rejected H-2-incompatible tumor allografts and H-2-compatible skin allografts. The primary hemagglutinin response to sheep red blood cells was normal in these mice, as were the induction of a local graft-versus-host reaction with lymphoid cells from UV-irradiated donors and the induction of an inflammatory response to dimethyl sulfoxide in the footpads of UV-treated mice. An early transient depression of two reactions in UV-irradiated mice occurred: delayed hypersensitivity to dinitrochlorobenzene measured by footpad swelling and the graft-versus-host reaction in UV-irradiated recipients measured by the use of the popliteal lymph node weight gain assay. Both of these reactions returned to a normal level before the development of primary tumors. We conclude that the inability of UV-irradiated mice to reject syngeneic and autochthonous UV-induced tumors was not due to a generalized immunosuppressive effect of chronic UV irradiation.
|
In vivo immune responses of mice during carcinogenesis by ultraviolet irradiation. In these experiments, we tested in various in vivo assays the immune responses of inbred C3H/HeN(MTV-) (C3H-) mice during carcinogenesis by chronic exposure to UV irradiation. Although the UV-treated mice were unable to reject syngeneic UV-induced tumor transplants, they rejected H-2-incompatible tumor allografts and H-2-compatible skin allografts. The primary hemagglutinin response to sheep red blood cells was normal in these mice, as were the induction of a local graft-versus-host reaction with lymphoid cells from UV-irradiated donors and the induction of an inflammatory response to dimethyl sulfoxide in the footpads of UV-treated mice. An early transient depression of two reactions in UV-irradiated mice occurred: delayed hypersensitivity to dinitrochlorobenzene measured by footpad swelling and the graft-versus-host reaction in UV-irradiated recipients measured by the use of the popliteal lymph node weight gain assay. Both of these reactions returned to a normal level before the development of primary tumors. We conclude that the inability of UV-irradiated mice to reject syngeneic and autochthonous UV-induced tumors was not due to a generalized immunosuppressive effect of chronic UV irradiation.
|
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] |
PMID:20515
|
Stimulation of cellular thymidine kinases by human cytomegalovirus.
|
Thymidine kinase (TK) activity in WI-38 and MRC-5 human fibroblasts was analyzed by discontinuous polyacrylamide gel electrophoresis (disc-PAGE) and discontinuous glycerol gradient electrophoresis (disc-GEP) after subculture or human cytomegalovirus (HCMV) infection. Two peaks of TK activity with different relative fraction-of-migration (R(f)) values were resolved by disc-PAGE or disc-GEP in extracts from log-phase and infected cells. Growing WI-38 cells expressed a slowly migrating (R(f) = 0.14 PAGE, R(f) = 0.4 GEP) peak of TK activity, which was partially inhibited by 1.0 mM dCTP, but which retained little activity at pH 4.5. Growing MRC cells also displayed a slowly migrating peak (R(f) = 0.10 PAGE) with similar properties. Both cell types expressed a faster-migrating TK activity (R(f) = 0.45 PAGE, R(f) = 0.7 GEP) in the growing and resting state that was strongly inhibited by 1 mM dCTP but retained 50% activity at pH 4.5. When either cell type was infected with HCMV, there was a rapid and high-level stimulation of the slowly migrating form of TK and a slight stimulation of the faster-migrating form. Two strains of HCMV (AD169 and Town) failed to produce an electrophoretically distinct virus TK in either cell type after infection. TK enzymes were partially purified by disc-GEP from extracts of log-phase WI-38 or AD169-infected WI-38 cells. Characterization of these enzymes with respect to phosphate donor specificity, pH optima, thermostability, and salt inhibition showed the HCMV-stimulated TKs to be of cellular origin.
|
Stimulation of cellular thymidine kinases by human cytomegalovirus. Thymidine kinase (TK) activity in WI-38 and MRC-5 human fibroblasts was analyzed by discontinuous polyacrylamide gel electrophoresis (disc-PAGE) and discontinuous glycerol gradient electrophoresis (disc-GEP) after subculture or human cytomegalovirus (HCMV) infection. Two peaks of TK activity with different relative fraction-of-migration (R(f)) values were resolved by disc-PAGE or disc-GEP in extracts from log-phase and infected cells. Growing WI-38 cells expressed a slowly migrating (R(f) = 0.14 PAGE, R(f) = 0.4 GEP) peak of TK activity, which was partially inhibited by 1.0 mM dCTP, but which retained little activity at pH 4.5. Growing MRC cells also displayed a slowly migrating peak (R(f) = 0.10 PAGE) with similar properties. Both cell types expressed a faster-migrating TK activity (R(f) = 0.45 PAGE, R(f) = 0.7 GEP) in the growing and resting state that was strongly inhibited by 1 mM dCTP but retained 50% activity at pH 4.5. When either cell type was infected with HCMV, there was a rapid and high-level stimulation of the slowly migrating form of TK and a slight stimulation of the faster-migrating form. Two strains of HCMV (AD169 and Town) failed to produce an electrophoretically distinct virus TK in either cell type after infection. TK enzymes were partially purified by disc-GEP from extracts of log-phase WI-38 or AD169-infected WI-38 cells. Characterization of these enzymes with respect to phosphate donor specificity, pH optima, thermostability, and salt inhibition showed the HCMV-stimulated TKs to be of cellular origin.
|
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] |
PMID:20516
|
Properties of "diplophage": a lipid-containing bacteriophage.
|
We describe the purification and properties of Dp-1, a bacteriophage isolated from Diplococcus pneumoniae. The phage was sensitive to the organic solvents deoxycholate and Sarkosyl, and its infectivity was reduced by treatment with phospholipase C. Electron microscopy indicated the presence of a double-layered coat around the phage particles. Purified phage preparations contained lipid amounting to about 8.5% of the dry weight of the phage, and thin-layer chromatography resolved the lipids into four components. The phage had a buoyant density in CsCl of 1.47 g/cm3, and a sedimentation constant in 0.1 M NaCl of 313S. Analysis in acrylamide gel electrophoresis indicated the presence of three major proteins. Dp-1 DNA shows a density of 1.681 g/cm3. Neutralizing antisera against the phage have a low potency (K less than 120/min).
|
Properties of "diplophage": a lipid-containing bacteriophage. We describe the purification and properties of Dp-1, a bacteriophage isolated from Diplococcus pneumoniae. The phage was sensitive to the organic solvents deoxycholate and Sarkosyl, and its infectivity was reduced by treatment with phospholipase C. Electron microscopy indicated the presence of a double-layered coat around the phage particles. Purified phage preparations contained lipid amounting to about 8.5% of the dry weight of the phage, and thin-layer chromatography resolved the lipids into four components. The phage had a buoyant density in CsCl of 1.47 g/cm3, and a sedimentation constant in 0.1 M NaCl of 313S. Analysis in acrylamide gel electrophoresis indicated the presence of three major proteins. Dp-1 DNA shows a density of 1.681 g/cm3. Neutralizing antisera against the phage have a low potency (K less than 120/min).
|
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] |
PMID:20517
|
Specific binding sites for a parvovirus, minute virus of mice, on cultured mouse cells.
|
The early interactions between parvoviruses and host cells have not been extensively described previously. In this study we have characterized some aspects of viral binding to the cell surface and demonstrated the existence of specific cellular receptor sites for minute virus of mice (MVM) on two murine cell lines that are permissive for viral growth. The interaction had a pH optimum of 7.0 to 7.2, and both the rate and extent of the reactions were slightly affected by temperature. Mouse A-9 cells (L-cell derivative) had approximately 5 X 10(5) specific MVM binding sites per cell, and Friend erythroleukemia cells had 1.5 X 10(5) MVM sites per cell. In contrast, the nonpermissive mouse lymphoid cell line L1210 lacked specific viral receptors. Also, cloned lines of A-9 cells resistant to viral infection have been isolated. One of these lines lacked the "specific" virus attachment sites but exhibited low levels of nonsaturable virus binding. Based on these examples, infectivity is correlated with the presence of specific viral receptors on the cell surface.
|
Specific binding sites for a parvovirus, minute virus of mice, on cultured mouse cells. The early interactions between parvoviruses and host cells have not been extensively described previously. In this study we have characterized some aspects of viral binding to the cell surface and demonstrated the existence of specific cellular receptor sites for minute virus of mice (MVM) on two murine cell lines that are permissive for viral growth. The interaction had a pH optimum of 7.0 to 7.2, and both the rate and extent of the reactions were slightly affected by temperature. Mouse A-9 cells (L-cell derivative) had approximately 5 X 10(5) specific MVM binding sites per cell, and Friend erythroleukemia cells had 1.5 X 10(5) MVM sites per cell. In contrast, the nonpermissive mouse lymphoid cell line L1210 lacked specific viral receptors. Also, cloned lines of A-9 cells resistant to viral infection have been isolated. One of these lines lacked the "specific" virus attachment sites but exhibited low levels of nonsaturable virus binding. Based on these examples, infectivity is correlated with the presence of specific viral receptors on the cell surface.
|
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] |
PMID:20518
|
Division of spermatic vessels in orchiopexy: radionuclide evidence of preservation of testicular circulation.
|
Satisfactory intrascrotal position of the testicles was achieved by dividing the spermatic vessels, leaving the vas deferens intact, in 3 patients with undescended testes. A postoperative scrotal scan provided objective means of evaluating the adequacy of testicular circulation. To determine the efficacy of various procedures for orchiopexy for undescended testes we suggest that postoperative evaluation include radionuclide imaging of the scrotum.
|
Division of spermatic vessels in orchiopexy: radionuclide evidence of preservation of testicular circulation. Satisfactory intrascrotal position of the testicles was achieved by dividing the spermatic vessels, leaving the vas deferens intact, in 3 patients with undescended testes. A postoperative scrotal scan provided objective means of evaluating the adequacy of testicular circulation. To determine the efficacy of various procedures for orchiopexy for undescended testes we suggest that postoperative evaluation include radionuclide imaging of the scrotum.
|
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] |
PMID:20519
|
The medical management of angina pectoris.
|
The availability of excellent short-acting and long-acting drugs for the treatment of angina pectoris needs to be emphasized. Properly used in conjunction with other measures such as the treatment of hypertension and a graded exercise routine, they provide, for most patients with angina, a tested therapeutic program that is remarkably effective, well-tolerated, appropriate for long-term outpatient use, and quite inexpensive.
|
The medical management of angina pectoris. The availability of excellent short-acting and long-acting drugs for the treatment of angina pectoris needs to be emphasized. Properly used in conjunction with other measures such as the treatment of hypertension and a graded exercise routine, they provide, for most patients with angina, a tested therapeutic program that is remarkably effective, well-tolerated, appropriate for long-term outpatient use, and quite inexpensive.
|
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] |
PMID:20520
|
Systemic vasculitis with coexistent large and small vessel involvement. A classification dilemma.
|
A patient with systemic vasculitis had the palpable purpuric lower extremity lesions of leukocytoclastic vasculitis and the renal aneurysms of polyarteritis nodosa, features thought to clearly separate these two syndromes. Questions are raised pertinent to the classification, diagnosis, treatment, and prognosis of systemic vasculitis.
|
Systemic vasculitis with coexistent large and small vessel involvement. A classification dilemma. A patient with systemic vasculitis had the palpable purpuric lower extremity lesions of leukocytoclastic vasculitis and the renal aneurysms of polyarteritis nodosa, features thought to clearly separate these two syndromes. Questions are raised pertinent to the classification, diagnosis, treatment, and prognosis of systemic vasculitis.
|
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] |
PMID:20523
|
Antihypertensive effect of combined treatment with alpha- and beta-adrenergic blockers in the spontaneously hypertensive rat.
|
Using a continuous systolic monitor, effects of oral administration of three alpha-adrenergic blockers, i.e. phenoxybenzamine, phentolamine and a new quinazoline compound (2-[4-(n-butyryl)-homopiperazine-1-yl]-4-amino-6, 7-dimethoxy-quinazoline; E-643), on blood pressure and heart rate in normotensive Wistar rats (NWR) and spontaneously hypertensive rats (SHR). 1) An oral dose of 1 mg/kg phenoxybenzamine or E-643 almost completely reversed pressor response to adrenaline (2 microgram/kg i.v). Phentolamine was 3 to 5 times less effective than phenoxybenzamine and E-643. These alpha-blockers reduced pressor response to noradrenaline (2 microgram/kg i.v.) merely to one half at an oral dose of 100 mg/kg. 2) All these alpha-blockers did not reduce blood pressure but markedly increased heart rate in NWR. On the other hand, they reduced blood pressure of SHR without marked increase in heart rate. Although the alpha-blocking and cardiac stimulating effects of phenoxybenzamine lasted more than several days, its hypotensive effect in SHR disappeared within 24 hours. 3) An oral dose of 30 mg/kg propranolol did not reduce blood pressure in both NWR and SHR but slightly decreased heart rate. The combined treatment of these alpha-blockers with propranolol completely abolished the cardiac stimulating effect of the alpha-blockers and resulted in a definite reduction in blood pressure of NWR and potentiated the hypotensive effect of alpha-blockers in SHR.
|
Antihypertensive effect of combined treatment with alpha- and beta-adrenergic blockers in the spontaneously hypertensive rat. Using a continuous systolic monitor, effects of oral administration of three alpha-adrenergic blockers, i.e. phenoxybenzamine, phentolamine and a new quinazoline compound (2-[4-(n-butyryl)-homopiperazine-1-yl]-4-amino-6, 7-dimethoxy-quinazoline; E-643), on blood pressure and heart rate in normotensive Wistar rats (NWR) and spontaneously hypertensive rats (SHR). 1) An oral dose of 1 mg/kg phenoxybenzamine or E-643 almost completely reversed pressor response to adrenaline (2 microgram/kg i.v). Phentolamine was 3 to 5 times less effective than phenoxybenzamine and E-643. These alpha-blockers reduced pressor response to noradrenaline (2 microgram/kg i.v.) merely to one half at an oral dose of 100 mg/kg. 2) All these alpha-blockers did not reduce blood pressure but markedly increased heart rate in NWR. On the other hand, they reduced blood pressure of SHR without marked increase in heart rate. Although the alpha-blocking and cardiac stimulating effects of phenoxybenzamine lasted more than several days, its hypotensive effect in SHR disappeared within 24 hours. 3) An oral dose of 30 mg/kg propranolol did not reduce blood pressure in both NWR and SHR but slightly decreased heart rate. The combined treatment of these alpha-blockers with propranolol completely abolished the cardiac stimulating effect of the alpha-blockers and resulted in a definite reduction in blood pressure of NWR and potentiated the hypotensive effect of alpha-blockers in SHR.
|
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] |
PMID:20527
|
Response of type B and E Botulinum toxins to purified sulfhydryl-dependent protease produced by Clostridium botulinum type F.
|
A sulfhydryl-dependent protease (SHP) was purified from a culture of Clostridium botulinum type F. The enzyme can activate type E progenitor toxin completely but type B progenitor toxin only partially. This may suggest that SHP by itself could completely activate the toxin of proteolytic C. botulinum types A and F in culture. The toxicity of type E progenitor toxin potentiated by the treatment with SHP persisted, whereas that of derivative toxin decreased rapidly by further incubation with SHP. This may indicate that only the progenitor toxin, the complex of the toxic and nontoxic components, activated by SHP withstands the subsequent exposure to the enzyme in cultures of proteolytic C. botulinum.
|
Response of type B and E Botulinum toxins to purified sulfhydryl-dependent protease produced by Clostridium botulinum type F. A sulfhydryl-dependent protease (SHP) was purified from a culture of Clostridium botulinum type F. The enzyme can activate type E progenitor toxin completely but type B progenitor toxin only partially. This may suggest that SHP by itself could completely activate the toxin of proteolytic C. botulinum types A and F in culture. The toxicity of type E progenitor toxin potentiated by the treatment with SHP persisted, whereas that of derivative toxin decreased rapidly by further incubation with SHP. This may indicate that only the progenitor toxin, the complex of the toxic and nontoxic components, activated by SHP withstands the subsequent exposure to the enzyme in cultures of proteolytic C. botulinum.
|
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] |
PMID:20529
|
[Prevention and therapy of chronic cor pulmonale (author's transl)].
|
First prophylactic and therapeutic possibilities in cor pulmonale are shown on the basis of its pathogenesis. Our own results illustrate the effect of therapy of the underlying lung disease and of the concomitant respiratory insufficiency on the pulmonary arterial hypertension. The relationship between pulmonary artery pressure (PAp), arterial oxygen tension, forced expiratory volume of 1 second (FEV1%VC) and slow inspired viral capacity (VC) is analysed. In obstructive respiratory disorders the PAp rises when FEV1 falls below 40% of VC, in restrictive disorders when VC falls below 70% of predicted rate. 27 patients with chronic obstructive lung disease were treated with bronchodilator aerosols by intermittent positive pressure breathing (IPPB) during 2 years after a control period of 2 years: VC and FEV1 improved, the increase of total lung capacity and the deterioration of arterial blood gases came to a halt. The elevated PAp was always significantly reduced by oxygen therapy or IPPB or the combination of both. Finally, the rationale for avoiding physical stress in established cor pulmonale is illustrated: in healthy men PAp increases by less than 20% when cardiac output is doubled. In patients with cor pulmonale PAp rises to three times the initial value under the same conditions.
|
[Prevention and therapy of chronic cor pulmonale (author's transl)]. First prophylactic and therapeutic possibilities in cor pulmonale are shown on the basis of its pathogenesis. Our own results illustrate the effect of therapy of the underlying lung disease and of the concomitant respiratory insufficiency on the pulmonary arterial hypertension. The relationship between pulmonary artery pressure (PAp), arterial oxygen tension, forced expiratory volume of 1 second (FEV1%VC) and slow inspired viral capacity (VC) is analysed. In obstructive respiratory disorders the PAp rises when FEV1 falls below 40% of VC, in restrictive disorders when VC falls below 70% of predicted rate. 27 patients with chronic obstructive lung disease were treated with bronchodilator aerosols by intermittent positive pressure breathing (IPPB) during 2 years after a control period of 2 years: VC and FEV1 improved, the increase of total lung capacity and the deterioration of arterial blood gases came to a halt. The elevated PAp was always significantly reduced by oxygen therapy or IPPB or the combination of both. Finally, the rationale for avoiding physical stress in established cor pulmonale is illustrated: in healthy men PAp increases by less than 20% when cardiac output is doubled. In patients with cor pulmonale PAp rises to three times the initial value under the same conditions.
|
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] |
PMID:20532
|
[Indices of human blood acid-base equilibrium and enzymatic activity in short-term antiorthostatic hypokinesia].
|
Before and after a 5 day head-down tilt of 4.5 degrees blood samples were withdrawn from different areas of the cardiovascular system of healthy male volunteers. Blood was sampled by means of selective catheterization. Parameters of acid-base equilibrium and activities of aminotransferases and alkaline phosphatase were measured. After immobilization the most significant changes were observed in the blood outflowing from the brain in which significant metabolic acidosis and increased activity of aspartate aminotransferase occurred. A slight increase in the activity of aspartate aminotransferase was also observed in the blood outflowing from the liver and kidneys. It is concluded that brain is the organ which is subjected to short-term simulated weightlessness in the highest degree.
|
[Indices of human blood acid-base equilibrium and enzymatic activity in short-term antiorthostatic hypokinesia]. Before and after a 5 day head-down tilt of 4.5 degrees blood samples were withdrawn from different areas of the cardiovascular system of healthy male volunteers. Blood was sampled by means of selective catheterization. Parameters of acid-base equilibrium and activities of aminotransferases and alkaline phosphatase were measured. After immobilization the most significant changes were observed in the blood outflowing from the brain in which significant metabolic acidosis and increased activity of aspartate aminotransferase occurred. A slight increase in the activity of aspartate aminotransferase was also observed in the blood outflowing from the liver and kidneys. It is concluded that brain is the organ which is subjected to short-term simulated weightlessness in the highest degree.
|
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] |
PMID:20533
|
[Skeletal muscle lactate dehydrogenase isoenzymes in rats after space flight and in hypokinesia].
|
Carbohydrate metabolism of soleus (red) and plantaris (mixed) muscles of rats exposed to space flight and hypokinesia were studied on a comparative basis. The red and mixed muscles responded in a different way to the exposures, as followed from the changes of the isoenzymic spectrum of LDH. The soleus muscle showed the greatest changes in the LDH spectrum on the 15th hypokinetic day. The changes were similar to those found in the soleus muscle of flight rats. The plantaris muscle showed the most marked changes on the 60th hypokinetic day. Alterations in the LDH spectrum of the plantaris muscle were not found in flight or control rats. The findings indicate the major contribution of changes in carbohydrate metabolism to atrophic and dystrophic developments in skeletal muscles under functional load.
|
[Skeletal muscle lactate dehydrogenase isoenzymes in rats after space flight and in hypokinesia]. Carbohydrate metabolism of soleus (red) and plantaris (mixed) muscles of rats exposed to space flight and hypokinesia were studied on a comparative basis. The red and mixed muscles responded in a different way to the exposures, as followed from the changes of the isoenzymic spectrum of LDH. The soleus muscle showed the greatest changes in the LDH spectrum on the 15th hypokinetic day. The changes were similar to those found in the soleus muscle of flight rats. The plantaris muscle showed the most marked changes on the 60th hypokinetic day. Alterations in the LDH spectrum of the plantaris muscle were not found in flight or control rats. The findings indicate the major contribution of changes in carbohydrate metabolism to atrophic and dystrophic developments in skeletal muscles under functional load.
|
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] |
PMID:20538
|
General principles of antimicrobial therapy.
|
In the initial therapy of life-threatening infections in which a bacterial cause is suspected, the emphasis should be on broad antibiotic coverage in contrast to definitive therapy, which is dependent on microbial isolation and, when indicated, in vitro susceptibility tests. In severe infections, antimicrobial agents should be given parenterally, at least initially. The need for optimal dosage is emphasized. This is particularly important when aminoglycosides are administered, for there is a tendency to use inadequate dosage because of concern for potential side effects with these agents. The problems leading to recurrence and persistence of fever during antimicrobial therapy include failure to diagnose and drain abscesses, superinfection, drug fever, and clinical or microbiologic errors. Combinations of antibiotics are indicated in severe infections in severe infections due to Pseudomonas aeruginosa, enterococcal group D streptococci, Klebsiella pneumoniae, and Cryptococcus neoformans. Laboratory aid for the selection of antimicrobial therapy can be of great value but need not always be done, because certain microorganisms have stable, predictable susceptibilities, for example, Streptococcus pneumoniae and Streptococcus pyogenes. Cautious conservatism is advocated with regard to the use of new antimicrobial agents.
|
General principles of antimicrobial therapy. In the initial therapy of life-threatening infections in which a bacterial cause is suspected, the emphasis should be on broad antibiotic coverage in contrast to definitive therapy, which is dependent on microbial isolation and, when indicated, in vitro susceptibility tests. In severe infections, antimicrobial agents should be given parenterally, at least initially. The need for optimal dosage is emphasized. This is particularly important when aminoglycosides are administered, for there is a tendency to use inadequate dosage because of concern for potential side effects with these agents. The problems leading to recurrence and persistence of fever during antimicrobial therapy include failure to diagnose and drain abscesses, superinfection, drug fever, and clinical or microbiologic errors. Combinations of antibiotics are indicated in severe infections in severe infections due to Pseudomonas aeruginosa, enterococcal group D streptococci, Klebsiella pneumoniae, and Cryptococcus neoformans. Laboratory aid for the selection of antimicrobial therapy can be of great value but need not always be done, because certain microorganisms have stable, predictable susceptibilities, for example, Streptococcus pneumoniae and Streptococcus pyogenes. Cautious conservatism is advocated with regard to the use of new antimicrobial agents.
|
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] |
PMID:20550
|
Catecholamine-sensitive adenylate cyclase of human fat cell ghosts: a comparative study using different beta-adrenergic agents.
|
Some of the effects of beta-adrenergic agonists and antagonists on the adenylate cyclase system of human fat cell ghosts were studied. Isoproterenol, by causing about a fourfold increase of enzyme activity, was more potent than epinephrine and norepinephrine (about 2.5--3.0-fold stimulation). The beta2-adrenergic agonists salbutamol, terbutalin, and fenoterol were considerably less effective than the naturally occurring catecholamines. The stimulatory actions of isoproterenol and beta2-adrenergic agonists were competitively inhibited by the beta-blocking agent propranolol. Isoproterenol stimulation was also inhibited by the selective beta1-adrenergic antagonist practolol. This compound, however, was less potent than propranolol. The results are suggestive for an adenylate cyclase system in human fat cell ghosts coupled to beta1-adrenergic receptor sites. These receptors differ from the cardiac beta receptors with respect to practolol affinity.
|
Catecholamine-sensitive adenylate cyclase of human fat cell ghosts: a comparative study using different beta-adrenergic agents. Some of the effects of beta-adrenergic agonists and antagonists on the adenylate cyclase system of human fat cell ghosts were studied. Isoproterenol, by causing about a fourfold increase of enzyme activity, was more potent than epinephrine and norepinephrine (about 2.5--3.0-fold stimulation). The beta2-adrenergic agonists salbutamol, terbutalin, and fenoterol were considerably less effective than the naturally occurring catecholamines. The stimulatory actions of isoproterenol and beta2-adrenergic agonists were competitively inhibited by the beta-blocking agent propranolol. Isoproterenol stimulation was also inhibited by the selective beta1-adrenergic antagonist practolol. This compound, however, was less potent than propranolol. The results are suggestive for an adenylate cyclase system in human fat cell ghosts coupled to beta1-adrenergic receptor sites. These receptors differ from the cardiac beta receptors with respect to practolol affinity.
|
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] |
PMID:20551
|
New evidence for growth hormone modulation by the alpha-adrenergic system in man.
|
The effect of BS 100-141, N-amidino 2-(2.6 dichlorophenyl) acetamide hydrochloride, a new drug with central alpha-adrenoceptor activity, on growth hormone (GH) secretion was investigated in 24 young, healthy volunteers. Six normal volunteers were treated with 1 mg and 8 with 2 mg BS 100-141; 8 of these 14 were treated with placebo, using single oral doses. Ten normal volunteers received single oral doses of 2 and 4 mg BS 100-141, 0.15 and 0.30 mg clonidine in a randomized, crossover study. GH secretion was significantly increased after 2 and 4 mg BS 100-141 and 0.30 mg clonidine. Prolactin and insulin plasma levels, glycerol, and blood sugar were not significantly influenced by BS 100-141. The results give new evidence for an alpha-adrenoceptor mechanism modulating GH secretion in normal young men.
|
New evidence for growth hormone modulation by the alpha-adrenergic system in man. The effect of BS 100-141, N-amidino 2-(2.6 dichlorophenyl) acetamide hydrochloride, a new drug with central alpha-adrenoceptor activity, on growth hormone (GH) secretion was investigated in 24 young, healthy volunteers. Six normal volunteers were treated with 1 mg and 8 with 2 mg BS 100-141; 8 of these 14 were treated with placebo, using single oral doses. Ten normal volunteers received single oral doses of 2 and 4 mg BS 100-141, 0.15 and 0.30 mg clonidine in a randomized, crossover study. GH secretion was significantly increased after 2 and 4 mg BS 100-141 and 0.30 mg clonidine. Prolactin and insulin plasma levels, glycerol, and blood sugar were not significantly influenced by BS 100-141. The results give new evidence for an alpha-adrenoceptor mechanism modulating GH secretion in normal young men.
|
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] |
PMID:20557
|
Adverse reactions after smallpox vaccination.
|
Nine hundred and thirty-eight reports of adverse reactions of smallpox vaccination in Australia between 1960 and 1976 have been analysed according to the type of reaction, and the age and sex of vaccinee. In an estimated 5,000,000 vaccinations, the reaction rate was 188/million, and the death rate 1-5/million. Generalized vaccinia was the most common reaction. The more serious reactions--eczema vaccinatum, progressive vaccinia, and neurological and cardiac complications--accounted for 7-4% of the reports. A small number of rarely reported non-specific inflammatory reactions is also included. There was a marked difference in the number of reactions reported in females and males (the female-male ratio was 1-6:1), and this difference increased with age. Paradoxically, of eight reports of cardiac complications, seven concerned males. The administration of vaccinial immune globulin was usually followed by a rapid resolution of the adverse reactions.
|
Adverse reactions after smallpox vaccination. Nine hundred and thirty-eight reports of adverse reactions of smallpox vaccination in Australia between 1960 and 1976 have been analysed according to the type of reaction, and the age and sex of vaccinee. In an estimated 5,000,000 vaccinations, the reaction rate was 188/million, and the death rate 1-5/million. Generalized vaccinia was the most common reaction. The more serious reactions--eczema vaccinatum, progressive vaccinia, and neurological and cardiac complications--accounted for 7-4% of the reports. A small number of rarely reported non-specific inflammatory reactions is also included. There was a marked difference in the number of reactions reported in females and males (the female-male ratio was 1-6:1), and this difference increased with age. Paradoxically, of eight reports of cardiac complications, seven concerned males. The administration of vaccinial immune globulin was usually followed by a rapid resolution of the adverse reactions.
|
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] |
PMID:20554
|
[Hydrogenase activity of the thermophilic hydrogen-oxidizing bacterium Pseudomonas thermophila].
|
The hydrogenase activity of the intact cells of a thermophilic hydrogen-oxidizing bacterium Pseudomonas thermophila K-2 was determined using methylene blue; it was several times higher than the rate of hydrogen uptake in the presence of oxygene and carbon dioxide. The activity of membrane-associated hydrogenase was assayed with the aid of phenazine methosulphate and 2,6-dichlorphenolindophenol as a cascade electron carrier. The enzyme is sufficiently stable in the air. The stability increases in the atmosphere of hydrogen. The membrane-bound enzyme was activated by Mn2+ ions. The pH-optimum of the enzyme activity in 0.1 M Tris-HCI buffer was 8,5-9,0. Natural electron acceptors tested, such as NAD, FMN, riboflavin, and cytochrome c, had no effect on the reaction rate. The enzyme is relatively thermostable: its activity was halved after heating at 78 degrees C for 10 min or at 80 degrees C for 8 min. Energy of activation was calculated. It was 14.5 kcal-mol-1 within the range of 23-40 degrees C and 10.3 kcal-mol-1 within the range of 40-60 degrees C.
|
[Hydrogenase activity of the thermophilic hydrogen-oxidizing bacterium Pseudomonas thermophila]. The hydrogenase activity of the intact cells of a thermophilic hydrogen-oxidizing bacterium Pseudomonas thermophila K-2 was determined using methylene blue; it was several times higher than the rate of hydrogen uptake in the presence of oxygene and carbon dioxide. The activity of membrane-associated hydrogenase was assayed with the aid of phenazine methosulphate and 2,6-dichlorphenolindophenol as a cascade electron carrier. The enzyme is sufficiently stable in the air. The stability increases in the atmosphere of hydrogen. The membrane-bound enzyme was activated by Mn2+ ions. The pH-optimum of the enzyme activity in 0.1 M Tris-HCI buffer was 8,5-9,0. Natural electron acceptors tested, such as NAD, FMN, riboflavin, and cytochrome c, had no effect on the reaction rate. The enzyme is relatively thermostable: its activity was halved after heating at 78 degrees C for 10 min or at 80 degrees C for 8 min. Energy of activation was calculated. It was 14.5 kcal-mol-1 within the range of 23-40 degrees C and 10.3 kcal-mol-1 within the range of 40-60 degrees C.
|
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] |
PMID:20555
|
[Effect of p-chloromercuribenzoate on glutamine synthetase in the food yeast Candida tropicalis].
|
The effect of p-CMB on the activity of glutamine synthetase in the fodder yeast Candida tropicalis was studied in the synthetase reaction in Mg-, Mn-, and Co-activated systems and in the transferase reaction. The activity of glutamine synthetase was inhibited by pCMB, the degree of inhibition depending on the presence of bivalent cations of metals. Preliminay incubation of the enzyme with p-CMB stimulated the action of the latter, whereas metals increased the stability of the enzyme to pCMB. If the enzyme preparation was purified, it became more susceptible to the action of p-CMB. The transferase activity was also inhibited by p-CMB but to a less extent that the synthetase reaction.
|
[Effect of p-chloromercuribenzoate on glutamine synthetase in the food yeast Candida tropicalis]. The effect of p-CMB on the activity of glutamine synthetase in the fodder yeast Candida tropicalis was studied in the synthetase reaction in Mg-, Mn-, and Co-activated systems and in the transferase reaction. The activity of glutamine synthetase was inhibited by pCMB, the degree of inhibition depending on the presence of bivalent cations of metals. Preliminay incubation of the enzyme with p-CMB stimulated the action of the latter, whereas metals increased the stability of the enzyme to pCMB. If the enzyme preparation was purified, it became more susceptible to the action of p-CMB. The transferase activity was also inhibited by p-CMB but to a less extent that the synthetase reaction.
|
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] |
PMID:20556
|
[Oospora lactis strain actively producing lipase].
|
Morphological, cultural and physiological properties of the fungus Oospora lactis producing lipase are described. The activity of lipase depends on the composition of a growth medium, pH, temperature, the duration of cultivation. Optimum conditions for lipase biosynthesis by the fungus have been found. The strain produces extracellular and endocellular lipases during submerged cultivation in the optimum growth medium at a high rate.
|
[Oospora lactis strain actively producing lipase]. Morphological, cultural and physiological properties of the fungus Oospora lactis producing lipase are described. The activity of lipase depends on the composition of a growth medium, pH, temperature, the duration of cultivation. Optimum conditions for lipase biosynthesis by the fungus have been found. The strain produces extracellular and endocellular lipases during submerged cultivation in the optimum growth medium at a high rate.
|
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] |
PMID:20561
|
[Determination of acid-base parameters by means of programmable pocket calculators (author's transl)].
|
The possibility to calculate parameters of acid-base status derived by common laboratory devices without built-in computers is described. The calculation is carried out faster and more exactly than it is possible by nomograms, which is especially suitable when a great quantity of dates occurs. The number of programmable steps in some inexpensive pocket calculators is sufficient for this purpose; this type of "microcomputers" offers advantages of economy and small size so that calculations can be carried out immediately at the site where measurements are taken.
|
[Determination of acid-base parameters by means of programmable pocket calculators (author's transl)]. The possibility to calculate parameters of acid-base status derived by common laboratory devices without built-in computers is described. The calculation is carried out faster and more exactly than it is possible by nomograms, which is especially suitable when a great quantity of dates occurs. The number of programmable steps in some inexpensive pocket calculators is sufficient for this purpose; this type of "microcomputers" offers advantages of economy and small size so that calculations can be carried out immediately at the site where measurements are taken.
|
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] |
PMID:20568
|
Purification and properties of alpha-amylase from chicken (Gallus gallus L.) pancreas.
|
Amylase from chicken pancreas was purified by an affinity method involving filtering a crude extract from pancreas through a Sepharose-wheat albumin column and eluting the retained enzyme with maltose. The purified amylase showed two active bands upon polyacrylamide electrophoresis in an alkaline buffer system and only one band in an acidic buffer system. The enzyme is a Ca2+-glycoprotein which behaves as a typical alpha-amylase. It consists of a single polypeptide chain with molecular weight 53,000 and contains 5.3 moles of reducing sugars per mole of protein. Optimal conditions of pH and temperature for the enzymic activity are 7.5 and 37 degrees C. The enzyme is irreversibly inactivated by removal of Ca2+ by exhaustive dialysis and is activated by the presence in the assay mixture of Cl-; other halides are less effective than Cl- in activating the enzyme.
|
Purification and properties of alpha-amylase from chicken (Gallus gallus L.) pancreas. Amylase from chicken pancreas was purified by an affinity method involving filtering a crude extract from pancreas through a Sepharose-wheat albumin column and eluting the retained enzyme with maltose. The purified amylase showed two active bands upon polyacrylamide electrophoresis in an alkaline buffer system and only one band in an acidic buffer system. The enzyme is a Ca2+-glycoprotein which behaves as a typical alpha-amylase. It consists of a single polypeptide chain with molecular weight 53,000 and contains 5.3 moles of reducing sugars per mole of protein. Optimal conditions of pH and temperature for the enzymic activity are 7.5 and 37 degrees C. The enzyme is irreversibly inactivated by removal of Ca2+ by exhaustive dialysis and is activated by the presence in the assay mixture of Cl-; other halides are less effective than Cl- in activating the enzyme.
|
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] |
PMID:20569
|
Aminoacyl-tRNA synthetases from calf liver: optimized assay conditions and kinetic properties.
|
Kinetic studies have been performed on the "family" of aminoacyl synthetases from calf liver. All assays were based on the esterification of amino acids to tRNA. Optimized reaction conditions for each synthetase are reported. Most of the synthetases show hyperbolic kinetics with respect to both amino acid and tRNA concentration, however a few show sigmoidal kinetics with respect to one substrate. Arginine, methionine and proline synthetases show sigmoidal kinetics with respect to mixed tRNA solutions and have Hill coefficients of 1.30, 1.10 and 1.20 respectively. Alanine and isoleucine synthetases show sigmoidal kinetics with respect to amino acid concentration and have Hill coefficients of 1.21 and 1.40 respectively.
|
Aminoacyl-tRNA synthetases from calf liver: optimized assay conditions and kinetic properties. Kinetic studies have been performed on the "family" of aminoacyl synthetases from calf liver. All assays were based on the esterification of amino acids to tRNA. Optimized reaction conditions for each synthetase are reported. Most of the synthetases show hyperbolic kinetics with respect to both amino acid and tRNA concentration, however a few show sigmoidal kinetics with respect to one substrate. Arginine, methionine and proline synthetases show sigmoidal kinetics with respect to mixed tRNA solutions and have Hill coefficients of 1.30, 1.10 and 1.20 respectively. Alanine and isoleucine synthetases show sigmoidal kinetics with respect to amino acid concentration and have Hill coefficients of 1.21 and 1.40 respectively.
|
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] |
PMID:20570
|
[Recent progress in research on the central regulation of growth hormone secretion (author's transl)].
|
The secretion of pituitary and peripheral hormones is regulated by a chain of reactions in the central nervous system. According to current knowledge this chain consists of the following links: external and internal stimuli leads to mesencephalon leads to limbic system leads to monoaminergic neurones leads to hypothalamic nuclei leads to liberines (releasing hormones) and inhibines (inhibiting horomones) leads to portal system leads to pituitary leads to peripheral endocrine gland. Long-loop and short-loop feedbacks regulate the activity of this relay system. The monoaminergic neurones secrete neurotransmitters, of which three are presently known: norepinephrine, dopamine, and serotonin. Most likely there also exists a cholinergic neurotransmitter. The secretion of neurotransmitters can be increased or reduced by specific drugs. On this basis, certain neuroendocrinopathies, such as deprivation dwarfism, hypothalamic acromegaly, and anorexia nervosa, may be treated speciafically. The pituitary secretion of growth hormone (GH) takes place mainly at night during deep sleep (slow-wave-sleep). This secretory pattern develops during the first year of life. It reaches its peak during puberty. At that time GH-spikes also occur during daytime. During adulthood GH secretion slowly diminishes again and at senescence it corresponds to early childhood.
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[Recent progress in research on the central regulation of growth hormone secretion (author's transl)]. The secretion of pituitary and peripheral hormones is regulated by a chain of reactions in the central nervous system. According to current knowledge this chain consists of the following links: external and internal stimuli leads to mesencephalon leads to limbic system leads to monoaminergic neurones leads to hypothalamic nuclei leads to liberines (releasing hormones) and inhibines (inhibiting horomones) leads to portal system leads to pituitary leads to peripheral endocrine gland. Long-loop and short-loop feedbacks regulate the activity of this relay system. The monoaminergic neurones secrete neurotransmitters, of which three are presently known: norepinephrine, dopamine, and serotonin. Most likely there also exists a cholinergic neurotransmitter. The secretion of neurotransmitters can be increased or reduced by specific drugs. On this basis, certain neuroendocrinopathies, such as deprivation dwarfism, hypothalamic acromegaly, and anorexia nervosa, may be treated speciafically. The pituitary secretion of growth hormone (GH) takes place mainly at night during deep sleep (slow-wave-sleep). This secretory pattern develops during the first year of life. It reaches its peak during puberty. At that time GH-spikes also occur during daytime. During adulthood GH secretion slowly diminishes again and at senescence it corresponds to early childhood.
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] |
PMID:20571
|
[Special problems of asthma bronchiale in childhood (author's transl)].
|
Asthma bronchiale is one of the most common chronic diseases in childhood. The hyperreactivity of the bronchial system, the stimulation of the cholinergic receptors and the blockade of the beta-adrenergic receptors in the bronchial mucosa play a predominant role in the pathogenesis. These proceedings cause bronchial smooth muscle contraction in the larger airways and mucosal edema and mucus hypersecretion in the smaller airways. The diagnosis may be made on the basis of the recurring signs: cough, wheezing, and labored breathing with prolonged expiration. Asthma may be treated by therapy directed at its cause and if necessary by bronchodilators, mucolytic agents and corticosteroids.
|
[Special problems of asthma bronchiale in childhood (author's transl)]. Asthma bronchiale is one of the most common chronic diseases in childhood. The hyperreactivity of the bronchial system, the stimulation of the cholinergic receptors and the blockade of the beta-adrenergic receptors in the bronchial mucosa play a predominant role in the pathogenesis. These proceedings cause bronchial smooth muscle contraction in the larger airways and mucosal edema and mucus hypersecretion in the smaller airways. The diagnosis may be made on the basis of the recurring signs: cough, wheezing, and labored breathing with prolonged expiration. Asthma may be treated by therapy directed at its cause and if necessary by bronchodilators, mucolytic agents and corticosteroids.
|
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] |
PMID:20572
|
Comparative effects of antacids, cimetidine and enteric coating on the therapeutic response to oral enzymes in severe pancreatic insufficiency.
|
To provide a rational basis for pancreatic enzyme replacement therapy, we evaluated, in six patients with advanced pancreatic insufficiency, the effects of various treatment regimens on fecal fat and nitrogen balance and on duodenal recovery of ingested pancreatic enzymes after a solid test meal. The combination of cimetidine (an H2-receptor antagonist) and pancreatin, each given by mouth, produced significantly higher postprandial duodenal recoveries and concentrations of trypsin and lipase (P less than 0.05). Steatorrhea was reduced in all patients and abolished in four of the six. In the dosages used, neither enteric-coated enzymes nor supplemental neutralizing antacids were more effective than pancreatin alone in decreasing steatorrhea or improving duodenal enzyme delivery. Cimetidine may be a useful adjunct to oral pancreatic extract therapy in some patients with severe pancreatic insufficiency who fail to respond to pancreatic enzyme replacement alone.
|
Comparative effects of antacids, cimetidine and enteric coating on the therapeutic response to oral enzymes in severe pancreatic insufficiency. To provide a rational basis for pancreatic enzyme replacement therapy, we evaluated, in six patients with advanced pancreatic insufficiency, the effects of various treatment regimens on fecal fat and nitrogen balance and on duodenal recovery of ingested pancreatic enzymes after a solid test meal. The combination of cimetidine (an H2-receptor antagonist) and pancreatin, each given by mouth, produced significantly higher postprandial duodenal recoveries and concentrations of trypsin and lipase (P less than 0.05). Steatorrhea was reduced in all patients and abolished in four of the six. In the dosages used, neither enteric-coated enzymes nor supplemental neutralizing antacids were more effective than pancreatin alone in decreasing steatorrhea or improving duodenal enzyme delivery. Cimetidine may be a useful adjunct to oral pancreatic extract therapy in some patients with severe pancreatic insufficiency who fail to respond to pancreatic enzyme replacement alone.
|
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] |
PMID:20575
|
Polyvalent pneumococcal-polysaccharide immunization of patients with sickle-cell anemia and patients with splenectomy.
|
To reduce the risk of infection from Streptococcus pneumoniae in hyposplenic patients we administered octavalent pneumococcal vaccine to 77 patients with sickle-cell disease and 19 asplenic persons and compared their response with 82 controls (38 age-matched normal persons and 44 normal black African children). Fifty micrograms each of pneumococcal-polysaccharide Types 1, 3, 6, 7, 14, 18, 19, and 23 were administered subcutaneously. Post-immunization serums (three to four weeks) were available from 52 of 77 patients with sickle-cell disease; the percent responding and the magnitude of the indirect hemagglutination response were comparable to those of the controls. Within two years after immunization we observed eight Str. pneumoniae infections in 106 age-matched unimmunized patients with sickle-cell disease, but none in the 77 immunized (P less than 0.025). We conclude that pneumococcal polysaccharides are immunogenic in hyposplenic patients and may protect against systemic Str. pneumoniae infection.
|
Polyvalent pneumococcal-polysaccharide immunization of patients with sickle-cell anemia and patients with splenectomy. To reduce the risk of infection from Streptococcus pneumoniae in hyposplenic patients we administered octavalent pneumococcal vaccine to 77 patients with sickle-cell disease and 19 asplenic persons and compared their response with 82 controls (38 age-matched normal persons and 44 normal black African children). Fifty micrograms each of pneumococcal-polysaccharide Types 1, 3, 6, 7, 14, 18, 19, and 23 were administered subcutaneously. Post-immunization serums (three to four weeks) were available from 52 of 77 patients with sickle-cell disease; the percent responding and the magnitude of the indirect hemagglutination response were comparable to those of the controls. Within two years after immunization we observed eight Str. pneumoniae infections in 106 age-matched unimmunized patients with sickle-cell disease, but none in the 77 immunized (P less than 0.025). We conclude that pneumococcal polysaccharides are immunogenic in hyposplenic patients and may protect against systemic Str. pneumoniae infection.
|
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] |
PMID:20582
|
The facilitatory actions of aminopyridines and tetraethylammonium on neuromuscular transmission and muscle contractility in avian muscle.
|
The actions of 3,4-diaminopyridine, 4-aminopyridine and tetraethylammonium were studied on the chick biventer cervicis muscle preparation. All three compounds produced a greater augmentation of indirectly elicited twitches than of directly elicited twitches. The compounds did not restore transmission in OmMCa2+ solutions but rather produced contractures that were inhibited by acetylcholine receptor antagonists. The compounds restored twitch height in one-tenth normal Ca2+ solutions and induced spontaneous muscle twitching. The compounds reversed dantrolene-induced block of directly elicited twitches. Interactions between tetraethylammonium and 3,4-diaminopyridine were also studied. In indirectly stimulated preparations, the combined effects of the two compounds were more than additive at one concentration level only. In directly stimulated preparations, the effects of 3,4-diaminopyridine were greatly enhanced by tetraethylammonium pretreatment. 3,4-Diaminopyridine pretreatment produced less synergism than tetraethylammonium pretreatment. It is concluded that the actions of the aminopyridines and tetraethylammonium on transmitter release and muscle contractility are essentially similar. These actions are postulated to arise from an inhibitory action on potassium conductance and on an ability to release calcium from nerve and muscle membranes. On the basis of the interaction studies, it is suggested that the compounds possess different binding capacities for two different sites on the potassium conducting channel.
|
The facilitatory actions of aminopyridines and tetraethylammonium on neuromuscular transmission and muscle contractility in avian muscle. The actions of 3,4-diaminopyridine, 4-aminopyridine and tetraethylammonium were studied on the chick biventer cervicis muscle preparation. All three compounds produced a greater augmentation of indirectly elicited twitches than of directly elicited twitches. The compounds did not restore transmission in OmMCa2+ solutions but rather produced contractures that were inhibited by acetylcholine receptor antagonists. The compounds restored twitch height in one-tenth normal Ca2+ solutions and induced spontaneous muscle twitching. The compounds reversed dantrolene-induced block of directly elicited twitches. Interactions between tetraethylammonium and 3,4-diaminopyridine were also studied. In indirectly stimulated preparations, the combined effects of the two compounds were more than additive at one concentration level only. In directly stimulated preparations, the effects of 3,4-diaminopyridine were greatly enhanced by tetraethylammonium pretreatment. 3,4-Diaminopyridine pretreatment produced less synergism than tetraethylammonium pretreatment. It is concluded that the actions of the aminopyridines and tetraethylammonium on transmitter release and muscle contractility are essentially similar. These actions are postulated to arise from an inhibitory action on potassium conductance and on an ability to release calcium from nerve and muscle membranes. On the basis of the interaction studies, it is suggested that the compounds possess different binding capacities for two different sites on the potassium conducting channel.
|
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-0.05079758167266846,
-0.005320263095200062,
0.036136869341135025,
0.06874420493841171
] |
PMID:20583
|
Effects of db cAMP on tyrosine hydroxylase activity of ganglia and nerve endings.
|
Preincubation of intact superior cervical ganglia or nictitating membrane for 2 h with dibutyryl cyclic AMP (db cAMP) increased the hydroxylation of tyrosine. This effect was not blocked by the protein synthesis inhibitor, cycloheximide. The Km of tyrosine hydroxylase for the substrate, tyrosine, and for the cofactor, reduced pteridine, were decreased by db cAMP. There were no changes in the Vmax of the enzyme. The inhibitory potency of noradrenaline on the hydroxylation of tyrosine was also decreased. Thus an inductive effect may be ruled out. The activation of the enzyme was only observed when the tissues were preincubated with the db cAMP and not when the cyclic nucleotide was added to the isolated enzyme. Preincubation of cervical ganglia for 4 h with db cAMP increased activity of decarboxylase and monoamine oxidase in tissue homogenates without changing the tyrosine hydroxylase activity.
|
Effects of db cAMP on tyrosine hydroxylase activity of ganglia and nerve endings. Preincubation of intact superior cervical ganglia or nictitating membrane for 2 h with dibutyryl cyclic AMP (db cAMP) increased the hydroxylation of tyrosine. This effect was not blocked by the protein synthesis inhibitor, cycloheximide. The Km of tyrosine hydroxylase for the substrate, tyrosine, and for the cofactor, reduced pteridine, were decreased by db cAMP. There were no changes in the Vmax of the enzyme. The inhibitory potency of noradrenaline on the hydroxylation of tyrosine was also decreased. Thus an inductive effect may be ruled out. The activation of the enzyme was only observed when the tissues were preincubated with the db cAMP and not when the cyclic nucleotide was added to the isolated enzyme. Preincubation of cervical ganglia for 4 h with db cAMP increased activity of decarboxylase and monoamine oxidase in tissue homogenates without changing the tyrosine hydroxylase activity.
|
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] |
PMID:20584
|
Naloxone-precipitated withdrawal reveals sensitization to neurotransmitters in morphine tolerant/dependent rats.
|
Morphine tolerant/dependent rats were tested for their sensitivity to putative neurotransmitters or other receptor agonists injected intracerebroventricularly (i.c.v.) during naloxone-precipitated withdrawal. Dopamine, apomorphine, clonidine and serotonin were found to reinitiate withdrawal jumping behaviour when injected 30 min after naloxone. Dopamine and apomorphine also reinitiated jumping, but of a lesser intensity, when injected 3 h after naloxone-precipitated withdrawal. I.c.v. injection of acetylcholine or prostaglandin E1 failed to reinitiate withdrawal jumping. In addition, all the above substances failed to induce jumping behaviour in naive rats or in morphine tolerant/dependent rats before naloxone-precipitated withdrawal. Morphine tolerance and dependence therefore appears to be associated with changes in the sensitivity of the CNS to putative neurotransmitter substances. These changes are best demonstrated during the sudden termination of opiate action that is caused by administration of naloxone.
|
Naloxone-precipitated withdrawal reveals sensitization to neurotransmitters in morphine tolerant/dependent rats. Morphine tolerant/dependent rats were tested for their sensitivity to putative neurotransmitters or other receptor agonists injected intracerebroventricularly (i.c.v.) during naloxone-precipitated withdrawal. Dopamine, apomorphine, clonidine and serotonin were found to reinitiate withdrawal jumping behaviour when injected 30 min after naloxone. Dopamine and apomorphine also reinitiated jumping, but of a lesser intensity, when injected 3 h after naloxone-precipitated withdrawal. I.c.v. injection of acetylcholine or prostaglandin E1 failed to reinitiate withdrawal jumping. In addition, all the above substances failed to induce jumping behaviour in naive rats or in morphine tolerant/dependent rats before naloxone-precipitated withdrawal. Morphine tolerance and dependence therefore appears to be associated with changes in the sensitivity of the CNS to putative neurotransmitter substances. These changes are best demonstrated during the sudden termination of opiate action that is caused by administration of naloxone.
|
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] |
PMID:20586
|
Enhanced sympathetic activity in young spontaneously hypertensive rats is not the trigger mechanism for genetic hypertension.
|
In young, spontaneously hypertensive rats (SHR), a preganglionic, nerve-dependent, elevation of choline acetyltransferase (ChAc) and tyrosine hydroxylase (TH) activities was found in celiac ganglia as compared with those in young, normotensive Kyoto Wistar rats, that was not present in superior cervical ganglia, stellate ganglia and adrenal glands. The rise in both enzyme activities in the celiac ganglion disappeared in adult SHR. An elevation of plasma norepinephrine and dopamine beta-hydroxylase levels found in prehypertensive SHR, a probable indication of peripheral sympathetic activation, disappeared after the bilateral removal of the celiac ganglion. However, ganglionectomy did not change the subsequent development of hypertension. These results indicate that the faster maturation of the celiac ganglion and the end organs it innervates in yount SHR are causally related to the activation of the peripheral sympathetic nervous system. The peripheral sympathetic activation in young SHR is regarded as a warning sign but this does not trigger the development of hypertension.
|
Enhanced sympathetic activity in young spontaneously hypertensive rats is not the trigger mechanism for genetic hypertension. In young, spontaneously hypertensive rats (SHR), a preganglionic, nerve-dependent, elevation of choline acetyltransferase (ChAc) and tyrosine hydroxylase (TH) activities was found in celiac ganglia as compared with those in young, normotensive Kyoto Wistar rats, that was not present in superior cervical ganglia, stellate ganglia and adrenal glands. The rise in both enzyme activities in the celiac ganglion disappeared in adult SHR. An elevation of plasma norepinephrine and dopamine beta-hydroxylase levels found in prehypertensive SHR, a probable indication of peripheral sympathetic activation, disappeared after the bilateral removal of the celiac ganglion. However, ganglionectomy did not change the subsequent development of hypertension. These results indicate that the faster maturation of the celiac ganglion and the end organs it innervates in yount SHR are causally related to the activation of the peripheral sympathetic nervous system. The peripheral sympathetic activation in young SHR is regarded as a warning sign but this does not trigger the development of hypertension.
|
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0.0034781061112880707,
0.06448832899332047,
-0.015706507489085197
] |
PMID:20592
|
[A case of polyarteritis nodosa associated with pulmonary hypertension].
|
A case of polyarteritis nodosa marked by concomitant systemic and pulmonary hypertension is presented. The part played by pulmonary hypertension in this disease is briefly discussed in the light of the necropsy findings and the literature.
|
[A case of polyarteritis nodosa associated with pulmonary hypertension]. A case of polyarteritis nodosa marked by concomitant systemic and pulmonary hypertension is presented. The part played by pulmonary hypertension in this disease is briefly discussed in the light of the necropsy findings and the literature.
|
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] |
PMID:20594
|
[Effect of age of mouse recipients on the functional activity of transplanted hematopoietic and lymphoid cells].
|
When transplanting the bone marrow cells from adult C57BL mice to the lethally irradiated (CBA X C57BL) F1 hybrids of different age, the decrease of the colony forming activity of the stem haemopoietic cells was observed in the spleen of the older recipients, as compared with the 3 months old ones. The joint transplantation of the bone marrow and thymus cells resulted in both the cases in the stimulation of the growth of colonies. The number of endogenous colonies of haemopoietic cells arising in the spleen of animals following the sublethal irradiation was greater in younger hybrids. After the induction of the "transplant versus host" reaction by the lymph node or spleen cells from the CBA mice, the relative weight of spleen and regional lymph node, respectively, in the older recipients exceeded those in the younger ones.
|
[Effect of age of mouse recipients on the functional activity of transplanted hematopoietic and lymphoid cells]. When transplanting the bone marrow cells from adult C57BL mice to the lethally irradiated (CBA X C57BL) F1 hybrids of different age, the decrease of the colony forming activity of the stem haemopoietic cells was observed in the spleen of the older recipients, as compared with the 3 months old ones. The joint transplantation of the bone marrow and thymus cells resulted in both the cases in the stimulation of the growth of colonies. The number of endogenous colonies of haemopoietic cells arising in the spleen of animals following the sublethal irradiation was greater in younger hybrids. After the induction of the "transplant versus host" reaction by the lymph node or spleen cells from the CBA mice, the relative weight of spleen and regional lymph node, respectively, in the older recipients exceeded those in the younger ones.
|
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] |
PMID:20595
|
Familial tyrosinaemia with eye and skin lesions. Presentation of two cases.
|
Two cases of tyrosinaemia with eye and skin lesions typical of the Richner-Hanhart syndrome are described. The patients are a 29- and 26-year-old brother and sister. They do not show neurological abnormalities or mental retardation. Parents are not consanguineous and family history is negative for similar conditions. The diagnosis of type II tyrosinaemia was based upon an increase of blood tyrosine (14-16mg/100 ml), tyrosinuria and absence of liver and kidney abnormalities. The treatment with a low tyrosine phenylalanine diet has resulted in a disappearence of the ocular manifestations while the cutaneous lesions are much improved.
|
Familial tyrosinaemia with eye and skin lesions. Presentation of two cases. Two cases of tyrosinaemia with eye and skin lesions typical of the Richner-Hanhart syndrome are described. The patients are a 29- and 26-year-old brother and sister. They do not show neurological abnormalities or mental retardation. Parents are not consanguineous and family history is negative for similar conditions. The diagnosis of type II tyrosinaemia was based upon an increase of blood tyrosine (14-16mg/100 ml), tyrosinuria and absence of liver and kidney abnormalities. The treatment with a low tyrosine phenylalanine diet has resulted in a disappearence of the ocular manifestations while the cutaneous lesions are much improved.
|
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] |
PMID:20596
|
Prenatal diagnosis of metachromatic leukodystrophy by electrophoretic and immunologic techniques.
|
Electrophoretic examination of extracts of cultured amniotic fluid cells from a pregnancy at risk for metachromatic leukodystrophy (MLD) showed absence of arylsulfatase A (AS-A) activity. Immunodiffusion with anti-human AS-A immune serum failed to show enzymatically active arcs of immune precipitate. Electrophoretic studies and quantitative assay of extracts of organs from the aborted fetus confirmed the diagnosis of MLD. Electrophoresis of amniotic fluid from this and one additional fetus with MLD showed an arylsulfatase pattern qualitatively and quantitatively indistinguishable from normal. In both normal and MLD fluids, the AS-A band was replaced by a band with lower anodal mobility. Only the anodal band of normal amniotic fluid, however, reacted with the anti-AS-A immune serum in immunoelectrophoresis. Assay of amniotic fluic with p-nitrocatechol sulfate (PNCS) as a substrate showed marked deficiency of "AS-A" activity in the fluids from the two MLD pregnancies. An optimal procedure for prenatal detection of MLD should include electrophoresis of extracts of cultured amniotic fluid cells with visual demonstration of absence of AS-A activity. Immunologic techniques applied to cell-free amniotic fluid may be of help in the rapid identification of the fetal genotypes.
|
Prenatal diagnosis of metachromatic leukodystrophy by electrophoretic and immunologic techniques. Electrophoretic examination of extracts of cultured amniotic fluid cells from a pregnancy at risk for metachromatic leukodystrophy (MLD) showed absence of arylsulfatase A (AS-A) activity. Immunodiffusion with anti-human AS-A immune serum failed to show enzymatically active arcs of immune precipitate. Electrophoretic studies and quantitative assay of extracts of organs from the aborted fetus confirmed the diagnosis of MLD. Electrophoresis of amniotic fluid from this and one additional fetus with MLD showed an arylsulfatase pattern qualitatively and quantitatively indistinguishable from normal. In both normal and MLD fluids, the AS-A band was replaced by a band with lower anodal mobility. Only the anodal band of normal amniotic fluid, however, reacted with the anti-AS-A immune serum in immunoelectrophoresis. Assay of amniotic fluic with p-nitrocatechol sulfate (PNCS) as a substrate showed marked deficiency of "AS-A" activity in the fluids from the two MLD pregnancies. An optimal procedure for prenatal detection of MLD should include electrophoresis of extracts of cultured amniotic fluid cells with visual demonstration of absence of AS-A activity. Immunologic techniques applied to cell-free amniotic fluid may be of help in the rapid identification of the fetal genotypes.
|
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] |
PMID:20597
|
A patient with severe neurologic symptoms and acetoacetyl-CoA thiolase deficiency.
|
A detailed kinetic characterization of the cytosolic acetoacetyl-CoA thiolase in liver, subcutaneous adipose tissue, and cultured fibroblasts of a patient with severe neurologic symptoms showed this enzyme to be much more sensitive to inhibition by CoA than that enzyme of normal human liver. The activity of the cytosolic acetoacetyl-CoA of this patient (14.0 mu mol-min-1-g (w/w) was in the lower range of normal (48.7 +/- 20.4 mu mol-min-1 g (w/w), n=6), but the ratio of the mitochondrial thiolase over the cytosoli thiolase was significantly increased (14.9 versus 4.54 +/- 2.92, n =6). The increased inhibition of the cytosolic acetoacetyl-CoA thiolase by CoA shows up as a 2-fold increase of the apparent Km for acetoacetyl-CoA at physiologic concentrations of CoA (64 micrometer for the enzyme of the patient verus 35 micrometer for the normal enzymes), resulting in a decreased utilization of the substrate. Aberrant kinetic behaviour of the mitochondrial thiolases of the patient has also been observed, but this has not been analysed further. Morphologic studies of the liver showed greatly increased deposition of glycogen and the occurrence of lipofuschin-like granules.
|
A patient with severe neurologic symptoms and acetoacetyl-CoA thiolase deficiency. A detailed kinetic characterization of the cytosolic acetoacetyl-CoA thiolase in liver, subcutaneous adipose tissue, and cultured fibroblasts of a patient with severe neurologic symptoms showed this enzyme to be much more sensitive to inhibition by CoA than that enzyme of normal human liver. The activity of the cytosolic acetoacetyl-CoA of this patient (14.0 mu mol-min-1-g (w/w) was in the lower range of normal (48.7 +/- 20.4 mu mol-min-1 g (w/w), n=6), but the ratio of the mitochondrial thiolase over the cytosoli thiolase was significantly increased (14.9 versus 4.54 +/- 2.92, n =6). The increased inhibition of the cytosolic acetoacetyl-CoA thiolase by CoA shows up as a 2-fold increase of the apparent Km for acetoacetyl-CoA at physiologic concentrations of CoA (64 micrometer for the enzyme of the patient verus 35 micrometer for the normal enzymes), resulting in a decreased utilization of the substrate. Aberrant kinetic behaviour of the mitochondrial thiolases of the patient has also been observed, but this has not been analysed further. Morphologic studies of the liver showed greatly increased deposition of glycogen and the occurrence of lipofuschin-like granules.
|
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] |
PMID:20600
|
An emerging zoonosis in Scandinavia -- nephropathia epidemica.
|
The paper reviews as zoonosis the disease called Nephropathia epidemica in north European countries. The etiology is as yet unsolved but a viral one is highly suspected with small rodents (voles and mice) as the source of infection. Based on the epidemiological peculiarities of the disease, the ecology of the agent and hypothetical ways of transmission of infection from small rodents to man are discussed.
|
An emerging zoonosis in Scandinavia -- nephropathia epidemica. The paper reviews as zoonosis the disease called Nephropathia epidemica in north European countries. The etiology is as yet unsolved but a viral one is highly suspected with small rodents (voles and mice) as the source of infection. Based on the epidemiological peculiarities of the disease, the ecology of the agent and hypothetical ways of transmission of infection from small rodents to man are discussed.
|
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] |
PMID:20602
|
[The renin-angiotensin-aldosterone system in hypertensive subjects. III- The use of beta-blockers in reno-vascular hypertension (author's transl)].
|
Six patients with permanent hypertension with renal artery stenosis were treated by conservative reparative surgery appart from one of them (unilateral nephrectomy) and were all seen again at the 8th month at the earliest in the absence of any anti-hypertensive therapy. Study of the renin-angiotensin system carried out on a normal sodium diet, after stopping all anti-hypertensive treatment for at least 15 days, was combined with the anti-hypertensive response under the influence of a beta-blocker. There were two types of response pre-operatively: firstly, with beta-blockers alone a normal blood pressure which remained normal postoperatively; the second group of patients remained hypertensive, requiring the addition of diuretics, and remained hypertensive after surgery. This response, although non-specific, would appear to represent an important element in assessing the curability of reno-vascular hypertension.
|
[The renin-angiotensin-aldosterone system in hypertensive subjects. III- The use of beta-blockers in reno-vascular hypertension (author's transl)]. Six patients with permanent hypertension with renal artery stenosis were treated by conservative reparative surgery appart from one of them (unilateral nephrectomy) and were all seen again at the 8th month at the earliest in the absence of any anti-hypertensive therapy. Study of the renin-angiotensin system carried out on a normal sodium diet, after stopping all anti-hypertensive treatment for at least 15 days, was combined with the anti-hypertensive response under the influence of a beta-blocker. There were two types of response pre-operatively: firstly, with beta-blockers alone a normal blood pressure which remained normal postoperatively; the second group of patients remained hypertensive, requiring the addition of diuretics, and remained hypertensive after surgery. This response, although non-specific, would appear to represent an important element in assessing the curability of reno-vascular hypertension.
|
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] |
PMID:20603
|
Enzymatic synthesis of Q nucleoside containing mannose in the anticodon of tRNA: isolation of a novel mannosyltransferase from a cell-free extract of rat liver.
|
The Q nucleosides isolated from rabbit liver tRNA are known to have sugars (mannose or galactose) linked to their cyclopentene diol moiety. A Q nucleoside containing mannose (manQ) was synthesized by a cell-free system from rat liver, using purified E. coli tRNAAsp as an acceptor and GDP-mannose as a donor molecule. The novel mannosyltransferase catalyzing this reaction was purified from a particulate-free soluble enzyme fraction and found to be strictly specific for tRNAAsp. These results, together with the anomeric configuration of mannose in Q nucleoside, indicate that no lipid intermediate is involved in the biosynthesis of Q nucleoside.
|
Enzymatic synthesis of Q nucleoside containing mannose in the anticodon of tRNA: isolation of a novel mannosyltransferase from a cell-free extract of rat liver. The Q nucleosides isolated from rabbit liver tRNA are known to have sugars (mannose or galactose) linked to their cyclopentene diol moiety. A Q nucleoside containing mannose (manQ) was synthesized by a cell-free system from rat liver, using purified E. coli tRNAAsp as an acceptor and GDP-mannose as a donor molecule. The novel mannosyltransferase catalyzing this reaction was purified from a particulate-free soluble enzyme fraction and found to be strictly specific for tRNAAsp. These results, together with the anomeric configuration of mannose in Q nucleoside, indicate that no lipid intermediate is involved in the biosynthesis of Q nucleoside.
|
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] |
PMID:20604
|
The role of the AT pairs in the acid denaturation of DNA.
|
It has been determined previously that the protonation of the GC pairs induces a DNA conformation change which leads to a "metastable" structure. The role of the AT pairs, however, is no well known because the protonation does not modify their spectral properties. By means of an indirect method based on the binding of proflavine, it has been determined that the AT pairs are protonated before the acid-induced denaturation and that they seem to be unable to assume a conformation change when protonated. These results would indicate that the protonated AT pairs may be responsible for the induction of the acid denaturation and not the GC pairs as it was thought previously.
|
The role of the AT pairs in the acid denaturation of DNA. It has been determined previously that the protonation of the GC pairs induces a DNA conformation change which leads to a "metastable" structure. The role of the AT pairs, however, is no well known because the protonation does not modify their spectral properties. By means of an indirect method based on the binding of proflavine, it has been determined that the AT pairs are protonated before the acid-induced denaturation and that they seem to be unable to assume a conformation change when protonated. These results would indicate that the protonated AT pairs may be responsible for the induction of the acid denaturation and not the GC pairs as it was thought previously.
|
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] |
PMID:20608
|
Participation of 5-hydroxytryptamine in anticonvulsive action of benzodiazepines.
|
The influence of several compounds activating or producing hypofunction of the serotonergic system was studied on the convulsive threshold and anticonvulsive action of benzodiazepines (chlordiazepoxide, diazepam, oxazepam, nitrazepam) in the pentylenetetrazol test. No changes in the convulsive threshold either for clonic, nor for tonic phase were found. However, the anticonvulsive action of benzodiazepines was enhanced by 5-hydroxytryptophan, 5-methoxytryptamine (given together with pargyline), or fenfluramine. p-Chlorophenylalanine and methergoline did not affect, but cyproheptadine enhanced the anticonvulsive action of most of benzodiazepines tested.
|
Participation of 5-hydroxytryptamine in anticonvulsive action of benzodiazepines. The influence of several compounds activating or producing hypofunction of the serotonergic system was studied on the convulsive threshold and anticonvulsive action of benzodiazepines (chlordiazepoxide, diazepam, oxazepam, nitrazepam) in the pentylenetetrazol test. No changes in the convulsive threshold either for clonic, nor for tonic phase were found. However, the anticonvulsive action of benzodiazepines was enhanced by 5-hydroxytryptophan, 5-methoxytryptamine (given together with pargyline), or fenfluramine. p-Chlorophenylalanine and methergoline did not affect, but cyproheptadine enhanced the anticonvulsive action of most of benzodiazepines tested.
|
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] |
PMID:20609
|
The effect of alpha-adrenolytics on central action of agonists and antagonists of dopaminergic system.
|
alpha-Adrenoceptor blocking agents: phenoxybenzamine, phentolamine and aceperone, potentiated and prolonged amphetamine and apomorphine stereotypy in the rat, but inhibited the amphetamine hypermotility. The hypermotility produced by apomorphine was antagonized only by aceperone. Cataleptogenic action of haloperidol and fluphenazine was inhibited by the alpha-adrenoceptor blocking agents. Phenoxybenzamine and phentolamine, and to a smaller degree aceperone, inhibited the depletion of dopamine following pretreatment of rats with alpha-methyltyrosine. The results indicate that the blockade of central alpha-adrenergic receptors changes the reactivity of dopaminergic system on the action of its agonists and antagonists.
|
The effect of alpha-adrenolytics on central action of agonists and antagonists of dopaminergic system. alpha-Adrenoceptor blocking agents: phenoxybenzamine, phentolamine and aceperone, potentiated and prolonged amphetamine and apomorphine stereotypy in the rat, but inhibited the amphetamine hypermotility. The hypermotility produced by apomorphine was antagonized only by aceperone. Cataleptogenic action of haloperidol and fluphenazine was inhibited by the alpha-adrenoceptor blocking agents. Phenoxybenzamine and phentolamine, and to a smaller degree aceperone, inhibited the depletion of dopamine following pretreatment of rats with alpha-methyltyrosine. The results indicate that the blockade of central alpha-adrenergic receptors changes the reactivity of dopaminergic system on the action of its agonists and antagonists.
|
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] |
PMID:20610
|
The effect of beta-adrenergic receptor blocking agents on hypertensive action of noradrenaline injected into the lateral ventricle of rat brain.
|
Noradrenaline (NA), 100 microgram intraventricularly (ivtr), produces an evident hypertension, lasting for up to 15 min. A previous ivtr administration of propranolol (100 and 500 microgram), alprenolol (100 microgram), sotalol (100 microgram), (-)INPEA (100 microgram), (+)INPEA (100 microgram), Ko 1366 (100 and 200 microgram), or practolol (20 microgram) abolishes the hypertensive action of NA. The results indicate that the central nervous system of the rat contains structures similar to the peripheral beta-adrenoceptor, and that they are involved in the central regulation of circulation.
|
The effect of beta-adrenergic receptor blocking agents on hypertensive action of noradrenaline injected into the lateral ventricle of rat brain. Noradrenaline (NA), 100 microgram intraventricularly (ivtr), produces an evident hypertension, lasting for up to 15 min. A previous ivtr administration of propranolol (100 and 500 microgram), alprenolol (100 microgram), sotalol (100 microgram), (-)INPEA (100 microgram), (+)INPEA (100 microgram), Ko 1366 (100 and 200 microgram), or practolol (20 microgram) abolishes the hypertensive action of NA. The results indicate that the central nervous system of the rat contains structures similar to the peripheral beta-adrenoceptor, and that they are involved in the central regulation of circulation.
|
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] |
PMID:20611
|
Kinetics of drug decomposition. Part 45. Logk--pH profile for rolitetracycline degradation.
|
UV spectrophotometry and fluorometry were applied for an examination of rolitetracycline (RT) degradation at pH's ranged from ca 0-8 to 13-0. It was demonstrated that these methods enable to follow the rate of degradation of tetracycline formed in the course of RT hydrolysis. However, they do not allow to monitor the hydrolysis of RT to tetracycline. Application of the so-called "subtraction technique" permits to calculate the rate constants for the total, reversible epimerization reaction and for the subsequent degradation of the resultant products. A rate equation derived for RT degradation in an alkaline medium beginning at pH 10-5 contains a term second-order in the hydroxide ion. Such a relationship was not yet observed in the course of an antibiotic degradation.
|
Kinetics of drug decomposition. Part 45. Logk--pH profile for rolitetracycline degradation. UV spectrophotometry and fluorometry were applied for an examination of rolitetracycline (RT) degradation at pH's ranged from ca 0-8 to 13-0. It was demonstrated that these methods enable to follow the rate of degradation of tetracycline formed in the course of RT hydrolysis. However, they do not allow to monitor the hydrolysis of RT to tetracycline. Application of the so-called "subtraction technique" permits to calculate the rate constants for the total, reversible epimerization reaction and for the subsequent degradation of the resultant products. A rate equation derived for RT degradation in an alkaline medium beginning at pH 10-5 contains a term second-order in the hydroxide ion. Such a relationship was not yet observed in the course of an antibiotic degradation.
|
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] |
PMID:20612
|
Kinetics and mechanism of degradation of some 2-sulfanilamidopyrimidine derivatives. Part III. The use of Hammett equation for kinetic investigation of 2-sulfanilamidopyrimidine derivatives autoxidation.
|
General first-order rate constants for autoxidation of sulfadiazine, sulfamerazine, sulfadimidine, sulfaperine and sulfamethoxydiazine in the air oxygen atmosphere, in solutions of pH 4-7, at 403, 411 and 418 K were determined from the absorbance measurements in 0-1 mole/dm3 HCl at 243 or 333 nm, using the so-called "subtraction technique". The thermodynamic parameters of this reaction were determined (deltaHa, deltaH not equal to, deltaS not equal to, deltaG not equal to and logA). The effect of the substituents in positions 4, 5 and 6 of the pyrimidine ring on the rate of autoxidation was interpreted in terms of the Hammett equation.
|
Kinetics and mechanism of degradation of some 2-sulfanilamidopyrimidine derivatives. Part III. The use of Hammett equation for kinetic investigation of 2-sulfanilamidopyrimidine derivatives autoxidation. General first-order rate constants for autoxidation of sulfadiazine, sulfamerazine, sulfadimidine, sulfaperine and sulfamethoxydiazine in the air oxygen atmosphere, in solutions of pH 4-7, at 403, 411 and 418 K were determined from the absorbance measurements in 0-1 mole/dm3 HCl at 243 or 333 nm, using the so-called "subtraction technique". The thermodynamic parameters of this reaction were determined (deltaHa, deltaH not equal to, deltaS not equal to, deltaG not equal to and logA). The effect of the substituents in positions 4, 5 and 6 of the pyrimidine ring on the rate of autoxidation was interpreted in terms of the Hammett equation.
|
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] |
PMID:20615
|
Purification and properties of a ribonuclease in human urine that hydrolyses polycytidylic acid.
|
Human urine RNase was purified about 2000-fold. The preparation is free from phosphatase, phosphodiesterase and DNase activities. On electrophoresis through polyacrylamide gel at pH 8.3, it migrates toward the anode and stains with periodic acid-Schiff reagent, suggesting that it is acidic and glycoprotein in nature. Its isoelectric point is at pH 4.1. It has a molecular weight of about 21,500. It is thermostable at pH 4.2 and thermolabile at pH 8.5. It has a pH optimum at 6.5. It exhibits highest preference for cytidine 3'-phosphate linkages. Its activity on poly (C) is endonucleolytic. It cleaves poly (C) via intramolecular transphosphorylation. It has no action on cytidine 2': 3'-cyclic phosphate or uridine 2':3'-cyclic phosphate. Its rate of hydrolysis of poly (U) is less than 2% of that of poly C). Poly (A) and poly (G) are totally inert to its action. Its action on poly (C) is inhibited by poly (G), poly (A) and poly (U). It differs from bovine pancreatic Rnase A in its physical, chemical and catalytic properties. It is, however, similar to human serum and pancreatic RNase in all its properties, suggesting that pancreas is its likely source.
|
Purification and properties of a ribonuclease in human urine that hydrolyses polycytidylic acid. Human urine RNase was purified about 2000-fold. The preparation is free from phosphatase, phosphodiesterase and DNase activities. On electrophoresis through polyacrylamide gel at pH 8.3, it migrates toward the anode and stains with periodic acid-Schiff reagent, suggesting that it is acidic and glycoprotein in nature. Its isoelectric point is at pH 4.1. It has a molecular weight of about 21,500. It is thermostable at pH 4.2 and thermolabile at pH 8.5. It has a pH optimum at 6.5. It exhibits highest preference for cytidine 3'-phosphate linkages. Its activity on poly (C) is endonucleolytic. It cleaves poly (C) via intramolecular transphosphorylation. It has no action on cytidine 2': 3'-cyclic phosphate or uridine 2':3'-cyclic phosphate. Its rate of hydrolysis of poly (U) is less than 2% of that of poly C). Poly (A) and poly (G) are totally inert to its action. Its action on poly (C) is inhibited by poly (G), poly (A) and poly (U). It differs from bovine pancreatic Rnase A in its physical, chemical and catalytic properties. It is, however, similar to human serum and pancreatic RNase in all its properties, suggesting that pancreas is its likely source.
|
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] |
PMID:20613
|
Kinetics of drug decomposition. Part 48. Specific acid-base hydrolysis of rolitetracycline.
|
Using chromatographic-spectrophotometric method, the effect of H+ and OH- ions on the rate of hydrolysis of rolitetracycline (RT) to tetracycline (T) at 20degreeC was investigated. The advantage of this analytical procedure over the direct fluorescence or UV absorption measurements of RT solutions during the kinetic investigations in HCl medium was demonstrated. The catalytic rate constants (kH+ and kOH-) for the hydrolytic degradation of RT under oxygen-free conditions are given.
|
Kinetics of drug decomposition. Part 48. Specific acid-base hydrolysis of rolitetracycline. Using chromatographic-spectrophotometric method, the effect of H+ and OH- ions on the rate of hydrolysis of rolitetracycline (RT) to tetracycline (T) at 20degreeC was investigated. The advantage of this analytical procedure over the direct fluorescence or UV absorption measurements of RT solutions during the kinetic investigations in HCl medium was demonstrated. The catalytic rate constants (kH+ and kOH-) for the hydrolytic degradation of RT under oxygen-free conditions are given.
|
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] |
PMID:20617
|
[Properties of urease immobilized by chemical binding to a derivatives of cellulose].
|
A comparative study of properties of soluble urease and urease immobilized by its binding to cellulose derivatives was carried out. Upon chemical binding of urease to cellulose derivatives the Mikhaelis constant and pH optimum for the enzyme action changed insignificantly. Maximum enzymic activity of soluble and immobilized urease occurred at 50 degrees C. The energy of activation of urea hydrolysis by urease during enzyme immobilization also changed insignificantly.
|
[Properties of urease immobilized by chemical binding to a derivatives of cellulose]. A comparative study of properties of soluble urease and urease immobilized by its binding to cellulose derivatives was carried out. Upon chemical binding of urease to cellulose derivatives the Mikhaelis constant and pH optimum for the enzyme action changed insignificantly. Maximum enzymic activity of soluble and immobilized urease occurred at 50 degrees C. The energy of activation of urea hydrolysis by urease during enzyme immobilization also changed insignificantly.
|
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] |
PMID:20620
|
Primary photochemical event in vision: proton translocation.
|
Picosecond studies of rhodopsin in low-temperature glasses have been carried out in order to observe directly the risetime of prelumirhodopsin, the first intermediate in the visual pathway. Only at 20 K or below can the risetime of this intermediate be resolved and even at 4 K it is astoundingly rapid, about 36 psec. An examination of the Arrhenius dependence on temperature of the rate of formation of prelumirhodopsin shows a strong deviation from linearity at low temperatures, i.e., non-Arrhenius behavior. This marked non-linear behavior is characteristic of a quantum mechanical tunneling event such as the translocation of hydrogen. An excellent candidate for the tunnelling process is the hydrogen of the protonated Schiff base formed between opsin and its retinal chromophore. Deuterium-exchanged rhodopsin, in which the Schiff base hydrogen is replaced by a deuterium, also shows a marked non-Arrhenius temperature dependence at low temperatures, consistent with tunneling. The rate of formation of prelumirhodopsin in deuterium-exchanged samples is much slower and a deuterium isotope effect kH/kD approximately or equal to 7 is observed. The data support a model in which the formation of prelumirhodopsin involves translocation of a proton toward the Schiff base nitrogen of the retinal chromophore.
|
Primary photochemical event in vision: proton translocation. Picosecond studies of rhodopsin in low-temperature glasses have been carried out in order to observe directly the risetime of prelumirhodopsin, the first intermediate in the visual pathway. Only at 20 K or below can the risetime of this intermediate be resolved and even at 4 K it is astoundingly rapid, about 36 psec. An examination of the Arrhenius dependence on temperature of the rate of formation of prelumirhodopsin shows a strong deviation from linearity at low temperatures, i.e., non-Arrhenius behavior. This marked non-linear behavior is characteristic of a quantum mechanical tunneling event such as the translocation of hydrogen. An excellent candidate for the tunnelling process is the hydrogen of the protonated Schiff base formed between opsin and its retinal chromophore. Deuterium-exchanged rhodopsin, in which the Schiff base hydrogen is replaced by a deuterium, also shows a marked non-Arrhenius temperature dependence at low temperatures, consistent with tunneling. The rate of formation of prelumirhodopsin in deuterium-exchanged samples is much slower and a deuterium isotope effect kH/kD approximately or equal to 7 is observed. The data support a model in which the formation of prelumirhodopsin involves translocation of a proton toward the Schiff base nitrogen of the retinal chromophore.
|
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] |
PMID:20618
|
[Denaturation of the alpha-amylase of Bacillus subtilis in an acid medium].
|
By fractionation of purine compounds (sorbtion on the cation exchange resin KU-2 (H+), extraction, precipitation of purine compounds as Ag-complexes) a "purine fraction" of the culture liquid epiphytic bacteria No. 703 isolated from barley overground organs was obtained. The presence of isopentenyl cytokinins was demonstrated by quality reactions of the purine fraction with aromatic amines and phenols as well as by the values of Rf and UV spectrum of individual compounds examined by paper chromatography of the purine fraction.
|
[Denaturation of the alpha-amylase of Bacillus subtilis in an acid medium]. By fractionation of purine compounds (sorbtion on the cation exchange resin KU-2 (H+), extraction, precipitation of purine compounds as Ag-complexes) a "purine fraction" of the culture liquid epiphytic bacteria No. 703 isolated from barley overground organs was obtained. The presence of isopentenyl cytokinins was demonstrated by quality reactions of the purine fraction with aromatic amines and phenols as well as by the values of Rf and UV spectrum of individual compounds examined by paper chromatography of the purine fraction.
|
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] |
PMID:20621
|
Sodium-stimulated glutamate uptake in membrane vesicles of Escherichia coli: the role of ion gradients.
|
Membrane vesicles prepared from Escherichia coli B/r grown on glutamate as a sole source of carbon and energy require sodium for glutamate accumulation when energized by D-lactate oxidation. Glutamate uptake can also be driven by a prearranged sodium gradient (out to in) in the absence of an energy source or a protonmotive force. Sodium ions are exchanged rapidly in respiring vesicles and the sodium gradient may be large enough under certain conditions to drive glutamate uptake after the protonmotive force is abolished with m-chlorocarbonylcyanide phenylhydrazone. Glutamate uptake due to a prearranged sodium gradient or lactate oxidation is inhibited by monensin but not by nigericin. Transport does not occur in response to valinomycin-induced membrane potential. We interpret these results to indicate that glutamate transport is obligately coupled to sodium transport and can only occur when there is a net flux of sodium ions. This flux is driven by a chemical gradient of sodium that is created by the protonmotive force generated by respiration.
|
Sodium-stimulated glutamate uptake in membrane vesicles of Escherichia coli: the role of ion gradients. Membrane vesicles prepared from Escherichia coli B/r grown on glutamate as a sole source of carbon and energy require sodium for glutamate accumulation when energized by D-lactate oxidation. Glutamate uptake can also be driven by a prearranged sodium gradient (out to in) in the absence of an energy source or a protonmotive force. Sodium ions are exchanged rapidly in respiring vesicles and the sodium gradient may be large enough under certain conditions to drive glutamate uptake after the protonmotive force is abolished with m-chlorocarbonylcyanide phenylhydrazone. Glutamate uptake due to a prearranged sodium gradient or lactate oxidation is inhibited by monensin but not by nigericin. Transport does not occur in response to valinomycin-induced membrane potential. We interpret these results to indicate that glutamate transport is obligately coupled to sodium transport and can only occur when there is a net flux of sodium ions. This flux is driven by a chemical gradient of sodium that is created by the protonmotive force generated by respiration.
|
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] |
PMID:20619
|
[Metabolism of sex steroid hormones in neuroendocrine organs. I. In vitro androgen metabolism in the hypophyses and epiphyses of rats during the puberal period].
|
Metabolism of tritium-labeled testosterone, delta4-androstane-3, 17-dion, dihydrotestosterone. 5alpha-androstane-3alpha, 17beta-diod and 5alpha-androstane-3beta, 17beta-dion by the isolated hypophyses and epiphyses of male and female 30-day rats was studied. The glands were incubated in Eagle's medium containing the mentioned androgens with the concentration of 1-2.10(-8) M for 90 min at 37 degrees C. As revealed, restoration of testosterone and delta4 androstane-3, 17-dion by 5alpha-reductase occurred in the epiphyses and hypophyses; under conditions of incubation androgen hydroxylation was realized only in the 3alpha- and 17beta-positions. Organ and sexual specificity of the androgen metabolism in the hypophyses and the epiphyses was quantitative in character: the metabolism level of all the androgens was greater in female rats than in the male both in the hypophysis and in the epiphyses; the activity of 5-reductase was lower in the epiphyses of animals of the both sexes than in the hypophyses. It appeared that in the process of biotransformation androgens in the organs under study served as precursors of the polar unidentified metabolites poorly retained by the tissues and eliminated from the organs into the incubation medium during the incubation.
|
[Metabolism of sex steroid hormones in neuroendocrine organs. I. In vitro androgen metabolism in the hypophyses and epiphyses of rats during the puberal period]. Metabolism of tritium-labeled testosterone, delta4-androstane-3, 17-dion, dihydrotestosterone. 5alpha-androstane-3alpha, 17beta-diod and 5alpha-androstane-3beta, 17beta-dion by the isolated hypophyses and epiphyses of male and female 30-day rats was studied. The glands were incubated in Eagle's medium containing the mentioned androgens with the concentration of 1-2.10(-8) M for 90 min at 37 degrees C. As revealed, restoration of testosterone and delta4 androstane-3, 17-dion by 5alpha-reductase occurred in the epiphyses and hypophyses; under conditions of incubation androgen hydroxylation was realized only in the 3alpha- and 17beta-positions. Organ and sexual specificity of the androgen metabolism in the hypophyses and the epiphyses was quantitative in character: the metabolism level of all the androgens was greater in female rats than in the male both in the hypophysis and in the epiphyses; the activity of 5-reductase was lower in the epiphyses of animals of the both sexes than in the hypophyses. It appeared that in the process of biotransformation androgens in the organs under study served as precursors of the polar unidentified metabolites poorly retained by the tissues and eliminated from the organs into the incubation medium during the incubation.
|
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] |
PMID:20622
|
Fatty acid synthetase from Brevibacterium ammoniagenes: formation of monounsaturated fatty acids by a multienzyme complex.
|
A multienzyme fatty acid synthetase complex isolated from Brevibacterium ammoniagenes has been purified to a specific activity of 1440 nmol of malonyl-CoA incorporated per min/mg. The enzyme is homogeneous, as judged by gel electrophoresis on agarose gels, and has a molecular weight of 1.2 X 10(6). Both NADPH and NADH are required for activity. In contrast to other fatty acid synthetase complexes, the enzyme catalyzes the synthesis of both long-chain saturated and monounsaturated fatty acids from malonyl-CoA and acetyl-CoA. The formation of unsaturated fatty acids is oxygen-independent and sharply reduced by 3-decynoyl-N-acetylcysteamine, a known inhibitor of Escherchia coli beta-hydroxydecanoyl thioester dehydrase (EC 4.2.1.60).
|
Fatty acid synthetase from Brevibacterium ammoniagenes: formation of monounsaturated fatty acids by a multienzyme complex. A multienzyme fatty acid synthetase complex isolated from Brevibacterium ammoniagenes has been purified to a specific activity of 1440 nmol of malonyl-CoA incorporated per min/mg. The enzyme is homogeneous, as judged by gel electrophoresis on agarose gels, and has a molecular weight of 1.2 X 10(6). Both NADPH and NADH are required for activity. In contrast to other fatty acid synthetase complexes, the enzyme catalyzes the synthesis of both long-chain saturated and monounsaturated fatty acids from malonyl-CoA and acetyl-CoA. The formation of unsaturated fatty acids is oxygen-independent and sharply reduced by 3-decynoyl-N-acetylcysteamine, a known inhibitor of Escherchia coli beta-hydroxydecanoyl thioester dehydrase (EC 4.2.1.60).
|
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] |
PMID:20623
|
Nitric oxide activates guanylate cyclase and increases guanosine 3':5'-cyclic monophosphate levels in various tissue preparations.
|
Nitric oxide gas (NO) increased guanylate cyclase [GTP pyrophosphate-lyase (cyclizing), EC 4.6.1.2] activity in soluble and particulate preparations from various tissues. The effect was dose-dependent and was observed with all tissue preparations examined. The extent of activation was variable among different tissue preparations and was greatest (19- to 33-fold) with supernatant fractions of homogenates from liver, lung, tracheal smooth muscle, heart, kidney, cerebral cortex, and cerebellum. Smaller effects (5- to 14-fold) were observed with supernatant fractions from skeletal muscle, spleen, intestinal muscle, adrenal, and epididymal fat. Activation was also observed with partially purified preparations of guanylate cyclase. Activation of rat liver supernatant preparations was augmented slightly with reducing agents, decreased with some oxidizing agents, and greater in a nitrogen than in an oxygen atmosphere. After activation with NO, guanylate cyclase activity decreased with a half-life of 3-4 at 4 degrees but re-exposure to NO resulted in reactivation of preparations. Sodium azide, sodium nitrite, hydroxylamine, and sodium nitroprusside also increased guanylate cyclase activity as reported previously. NO alone and in combination with these agents produced approximately the same degree of maximal activation, suggesting that all of these agents act through a similar mechanism. NO also increased the accumulation of cyclic GMP but not cyclic AMP in incubations of minces from various rat tissues. We propose that various nitro compounds and those capable of forming NO in incubations activate guanylate cyclase through a similar but undefined mechanism. These effects may explain the high activities of guanylate cyclase in certain tissues (e.g., lung and intestinal mucosa) that are exposed to environmental nitro compounds.
|
Nitric oxide activates guanylate cyclase and increases guanosine 3':5'-cyclic monophosphate levels in various tissue preparations. Nitric oxide gas (NO) increased guanylate cyclase [GTP pyrophosphate-lyase (cyclizing), EC 4.6.1.2] activity in soluble and particulate preparations from various tissues. The effect was dose-dependent and was observed with all tissue preparations examined. The extent of activation was variable among different tissue preparations and was greatest (19- to 33-fold) with supernatant fractions of homogenates from liver, lung, tracheal smooth muscle, heart, kidney, cerebral cortex, and cerebellum. Smaller effects (5- to 14-fold) were observed with supernatant fractions from skeletal muscle, spleen, intestinal muscle, adrenal, and epididymal fat. Activation was also observed with partially purified preparations of guanylate cyclase. Activation of rat liver supernatant preparations was augmented slightly with reducing agents, decreased with some oxidizing agents, and greater in a nitrogen than in an oxygen atmosphere. After activation with NO, guanylate cyclase activity decreased with a half-life of 3-4 at 4 degrees but re-exposure to NO resulted in reactivation of preparations. Sodium azide, sodium nitrite, hydroxylamine, and sodium nitroprusside also increased guanylate cyclase activity as reported previously. NO alone and in combination with these agents produced approximately the same degree of maximal activation, suggesting that all of these agents act through a similar mechanism. NO also increased the accumulation of cyclic GMP but not cyclic AMP in incubations of minces from various rat tissues. We propose that various nitro compounds and those capable of forming NO in incubations activate guanylate cyclase through a similar but undefined mechanism. These effects may explain the high activities of guanylate cyclase in certain tissues (e.g., lung and intestinal mucosa) that are exposed to environmental nitro compounds.
|
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] |
PMID:20624
|
Unequivocal demonstration of fructose-1,6-bisphosphatase in mammalian brain.
|
Fructose-1,6-bisphosphatase (D-fructose-1,6-bisphosphate 1-phosphohydrolase; EC 3.1.3.11) has been found in rat brain and identified unequivocally. The enzyme has been purified to 95% homogeneity by standard procedures, including adsorption to a phosphocellulose column followed by elution with substrate. The purified enzyme exhibits a broad optimum above pH 7.6. Both fructose 1,6-bisphosphate and sedoheptulose 1,7-bisphosphate are substrates of this enzyme; the hydrolysis of the latter occurs at about 20% of the rate of the former, and the Km for fructose 1,6-bisphosphate is approximately 1.32 X 10(-4) M. 5'-AMP, an inhibitor of other mammalian-fructose-1,6-bisphosphatases, is without effect, and in further contrast with the other enzymes there is no metal requirement for activity. Purified brain enzyme fails to crossreact with the antibody prepared against the purified liver fructose-1,6-bisphosphatase. On the other hand, antiserum produced against the brain fructose-1,6-bisphosphatase quantitatively precipitates the enzyme activity and forms precipitin bands with preparations of brain fructose-1,6-bisphosphatase.
|
Unequivocal demonstration of fructose-1,6-bisphosphatase in mammalian brain. Fructose-1,6-bisphosphatase (D-fructose-1,6-bisphosphate 1-phosphohydrolase; EC 3.1.3.11) has been found in rat brain and identified unequivocally. The enzyme has been purified to 95% homogeneity by standard procedures, including adsorption to a phosphocellulose column followed by elution with substrate. The purified enzyme exhibits a broad optimum above pH 7.6. Both fructose 1,6-bisphosphate and sedoheptulose 1,7-bisphosphate are substrates of this enzyme; the hydrolysis of the latter occurs at about 20% of the rate of the former, and the Km for fructose 1,6-bisphosphate is approximately 1.32 X 10(-4) M. 5'-AMP, an inhibitor of other mammalian-fructose-1,6-bisphosphatases, is without effect, and in further contrast with the other enzymes there is no metal requirement for activity. Purified brain enzyme fails to crossreact with the antibody prepared against the purified liver fructose-1,6-bisphosphatase. On the other hand, antiserum produced against the brain fructose-1,6-bisphosphatase quantitatively precipitates the enzyme activity and forms precipitin bands with preparations of brain fructose-1,6-bisphosphatase.
|
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0.027295254170894623,
0.033532846719026566,
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] |
PMID:20625
|
Resonance Raman spectroscopy of arsanilazocarboxypeptidase A: determination of the nature of the azotyrosyl-248-zinc complex.
|
Resonance Raman spectra of arsanilazotryosyl-248 carboxypeptidase A (peptidyl-L-amino-acid hydrolase, EC 3.4.12.2) exhibit only the vibrational bands of its chromophoric azotyrosyl-248 residue uncomplicated by background interference from either water or other components of the protein. The resonance Raman spectra contain multiple, discrete bands which change as a function of pH, thereby demonstrating the existence of interconvertible species of the azotyrosine probe in solution. Spectra of model azophenols and of the apoazoenzyme establish the identity of these species. All conclusions about the azoenzyme based on the resonance Raman spectra, including the apparent pK values for the interconversion of these species, are in complete agreement with those drawn earlier from studies by absorption spectroscopy. In addition, the properties of resonance Raman bands that have been identified with the motions of specific atoms of azotyrosyl-248 provide details of the interactions of specific atoms of this chromophore with the catalytic zinc atom at the active site. In particular, this has allowed elucidation of the structure of the azotyrosyl-248-zinc coordination complex. Such experiments are also providing information on the effects of crystallization on the enzyme and on its interaction with inhibitors. The important potential of resonance Raman spectroscopy for the study of the structure of chromophoric components of active enzymatic sites and of metal complex ions is discussed.
|
Resonance Raman spectroscopy of arsanilazocarboxypeptidase A: determination of the nature of the azotyrosyl-248-zinc complex. Resonance Raman spectra of arsanilazotryosyl-248 carboxypeptidase A (peptidyl-L-amino-acid hydrolase, EC 3.4.12.2) exhibit only the vibrational bands of its chromophoric azotyrosyl-248 residue uncomplicated by background interference from either water or other components of the protein. The resonance Raman spectra contain multiple, discrete bands which change as a function of pH, thereby demonstrating the existence of interconvertible species of the azotyrosine probe in solution. Spectra of model azophenols and of the apoazoenzyme establish the identity of these species. All conclusions about the azoenzyme based on the resonance Raman spectra, including the apparent pK values for the interconversion of these species, are in complete agreement with those drawn earlier from studies by absorption spectroscopy. In addition, the properties of resonance Raman bands that have been identified with the motions of specific atoms of azotyrosyl-248 provide details of the interactions of specific atoms of this chromophore with the catalytic zinc atom at the active site. In particular, this has allowed elucidation of the structure of the azotyrosyl-248-zinc coordination complex. Such experiments are also providing information on the effects of crystallization on the enzyme and on its interaction with inhibitors. The important potential of resonance Raman spectroscopy for the study of the structure of chromophoric components of active enzymatic sites and of metal complex ions is discussed.
|
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] |
PMID:20626
|
Selective inhibition of gamma-glutamyl-cycle enzymes by substrate analogs.
|
Substrate analogs have been obtained that selectively inhibit the reactions of the gamma-glutamyl cycle or that are susceptible to only limited metabolism by the cycle. Thus, glutathione synthesis may be inhibited and analogs of glutathione may be synthesized that do not participate in transpeptidation. Specific inhibitors of gamma-glutamylcyclotransferase and 5-oxoprolinase have been obtained. The findings offer new approaches to the in vivo study of the cycle and also to the design of more specifically directed analogs of inhibitors such as methionine sulfoximine and 6-diazo-5-oxonorleucine.
|
Selective inhibition of gamma-glutamyl-cycle enzymes by substrate analogs. Substrate analogs have been obtained that selectively inhibit the reactions of the gamma-glutamyl cycle or that are susceptible to only limited metabolism by the cycle. Thus, glutathione synthesis may be inhibited and analogs of glutathione may be synthesized that do not participate in transpeptidation. Specific inhibitors of gamma-glutamylcyclotransferase and 5-oxoprolinase have been obtained. The findings offer new approaches to the in vivo study of the cycle and also to the design of more specifically directed analogs of inhibitors such as methionine sulfoximine and 6-diazo-5-oxonorleucine.
|
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] |
PMID:20627
|
Quantum efficiency of photosynthetic energy conversion.
|
The quantum efficiency of photosynthetic energy conversion was investigated in isolated spinach chloroplasts by measurements of the quantum requirements of ATP formation by cyclic and noncyclic photophosphorylation catalyzed by ferredoxin. ATP formation had a requirement of about 2 quanta per 1 ATP at 715 nm (corresponding to a requirement of 1 quantum per electron) and a requirement of 4 quanta per ATP (corresponding to a requirement of 2 quanta per electron) at 554 nm. When cyclic and noncyclic photophosphorylation were operating concurrently at 554 nm, a total of about 12 quanta was required to generate the two NADPH and three ATP needed for the assimilation of one CO2 to the level of glucose.
|
Quantum efficiency of photosynthetic energy conversion. The quantum efficiency of photosynthetic energy conversion was investigated in isolated spinach chloroplasts by measurements of the quantum requirements of ATP formation by cyclic and noncyclic photophosphorylation catalyzed by ferredoxin. ATP formation had a requirement of about 2 quanta per 1 ATP at 715 nm (corresponding to a requirement of 1 quantum per electron) and a requirement of 4 quanta per ATP (corresponding to a requirement of 2 quanta per electron) at 554 nm. When cyclic and noncyclic photophosphorylation were operating concurrently at 554 nm, a total of about 12 quanta was required to generate the two NADPH and three ATP needed for the assimilation of one CO2 to the level of glucose.
|
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] |
PMID:20628
|
Embryologic development of a mouse sympathetic ganglion in vivo and in vitro.
|
The morphologic and biochemical development of the embryonic mouse superior cervical ganglion was characterized in vivo and in tissue culture. From 13 days of gestation, when the superior cervical ganglion was first visible, to birth at 19 days, tyrosine hydroxylase [tyrosine 3-monooxygenase; L-tyrosine, tetrahydropteridine:oxygen oxidoreductase (3-hydroxylating); EC 1.14.16.2] activity increased 100-fold in vivo. Explants of ganglia from 14-day embryos exhibited abundant neurite outgrowth in basal medium without added nerve growth factor (NGF), and increases in tyrosine hydroxylase activity paralleled that observed in vivo. Ganglia from 14-day embryos elaborated neurites and exhibited 3-fold increases in enzyme activity in vitro in the presence of antiserum to NGF (anti-NGF) or NGF + anti-NGF. In direct contrast, ganglia from 18-day fetuses failed to grow without added NGF or in medium containing anti-NGF or NGF + anti-NGF: virtually no axon outgrowth occurred and tyrosine hydroxylase activity decreased by half. These observations suggest that developmental regulatory mechanisms change radically during embryologic and fetal life of mammalian superior cervical ganglion.
|
Embryologic development of a mouse sympathetic ganglion in vivo and in vitro. The morphologic and biochemical development of the embryonic mouse superior cervical ganglion was characterized in vivo and in tissue culture. From 13 days of gestation, when the superior cervical ganglion was first visible, to birth at 19 days, tyrosine hydroxylase [tyrosine 3-monooxygenase; L-tyrosine, tetrahydropteridine:oxygen oxidoreductase (3-hydroxylating); EC 1.14.16.2] activity increased 100-fold in vivo. Explants of ganglia from 14-day embryos exhibited abundant neurite outgrowth in basal medium without added nerve growth factor (NGF), and increases in tyrosine hydroxylase activity paralleled that observed in vivo. Ganglia from 14-day embryos elaborated neurites and exhibited 3-fold increases in enzyme activity in vitro in the presence of antiserum to NGF (anti-NGF) or NGF + anti-NGF. In direct contrast, ganglia from 18-day fetuses failed to grow without added NGF or in medium containing anti-NGF or NGF + anti-NGF: virtually no axon outgrowth occurred and tyrosine hydroxylase activity decreased by half. These observations suggest that developmental regulatory mechanisms change radically during embryologic and fetal life of mammalian superior cervical ganglion.
|
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] |
PMID:20629
|
Thyrotropin-releasing hormone-like material in the rat retina: changes due to environmental lighting.
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Material reacting with an antibody to thyrotropin-releasing hormone (TRH) has been found to be present in the rat retina. The compound present in the retina cochromatographed with authentic TRH and most of its activity was lost when incubated with pyroglutamate aminopeptidase (L-pyroglutamyl-peptide hydrolase, EC 3.4.11.8), an enzyme that degrades TRH. The TRH-like activity in the rat retina was low during the night and high during the day. There was a 4-hr lag period after the lights were turned on before peak TRH levels were attained. A decrease in TRH was seen after 2 hr of darkness and the level of TRH was lowest after 4 hr of darkness. Retinal TRH is elevated by environmental lighting regardless of the time of the day. These findings suggest that TRH may be involved in retinal photorecptive mechanisms.
|
Thyrotropin-releasing hormone-like material in the rat retina: changes due to environmental lighting. Material reacting with an antibody to thyrotropin-releasing hormone (TRH) has been found to be present in the rat retina. The compound present in the retina cochromatographed with authentic TRH and most of its activity was lost when incubated with pyroglutamate aminopeptidase (L-pyroglutamyl-peptide hydrolase, EC 3.4.11.8), an enzyme that degrades TRH. The TRH-like activity in the rat retina was low during the night and high during the day. There was a 4-hr lag period after the lights were turned on before peak TRH levels were attained. A decrease in TRH was seen after 2 hr of darkness and the level of TRH was lowest after 4 hr of darkness. Retinal TRH is elevated by environmental lighting regardless of the time of the day. These findings suggest that TRH may be involved in retinal photorecptive mechanisms.
|
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] |
PMID:20630
|
Stimulation of human platelet guanylate cyclase by unsaturated fatty acid peroxides.
|
Guanylate cyclase [GTP pyrophosphate-lyase (cyclizing), EC 4.6.1.2] activity of human platelet homogenates was stimulated by the addition of phospholipase A2 or unsaturated fatty acids such as oleic, vaccenic, linoleic, linolenic, eicosenoic, eicosadienoic, and arachidonic acids. The addition of lipoxidase potentiated the fatty acid-induced stimulation of guanylate cyclase purified by DEAE-cellulose column chromatography. The extent of the stimulation was dependent on the concentration of the oxidized form of these fatty acids (peroxides). Saturated fatty acids such as stearic and arachidic acids had no effect on the guanylate cyclase activity in the presence or absence of lipoxidase, indicating that human plateletguanylate cyclase is stimulated by unsaturated fatty acid peroxides rather than by fatty acids. Hemoglobin prevented the enzyme stimulation produced by low concentrations of fatty acid peroxides, but enhanced stimulation of the enzyme activity with high concentrations of fatty acid peroxides. 2-Mercaptoethanol, dithiothreitol, and N-ethylmaleimide inhibited the guanylate cyclase activities both in the presence and absence of unsaturated fatty acidperoxide. The stimulation of guanylate cyclase activity by unsaturated fatty acid peroxidesis attributed to oxidation of sulfhydryl residues of the enzyme protein.
|
Stimulation of human platelet guanylate cyclase by unsaturated fatty acid peroxides. Guanylate cyclase [GTP pyrophosphate-lyase (cyclizing), EC 4.6.1.2] activity of human platelet homogenates was stimulated by the addition of phospholipase A2 or unsaturated fatty acids such as oleic, vaccenic, linoleic, linolenic, eicosenoic, eicosadienoic, and arachidonic acids. The addition of lipoxidase potentiated the fatty acid-induced stimulation of guanylate cyclase purified by DEAE-cellulose column chromatography. The extent of the stimulation was dependent on the concentration of the oxidized form of these fatty acids (peroxides). Saturated fatty acids such as stearic and arachidic acids had no effect on the guanylate cyclase activity in the presence or absence of lipoxidase, indicating that human plateletguanylate cyclase is stimulated by unsaturated fatty acid peroxides rather than by fatty acids. Hemoglobin prevented the enzyme stimulation produced by low concentrations of fatty acid peroxides, but enhanced stimulation of the enzyme activity with high concentrations of fatty acid peroxides. 2-Mercaptoethanol, dithiothreitol, and N-ethylmaleimide inhibited the guanylate cyclase activities both in the presence and absence of unsaturated fatty acidperoxide. The stimulation of guanylate cyclase activity by unsaturated fatty acid peroxidesis attributed to oxidation of sulfhydryl residues of the enzyme protein.
|
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0.007236348930746317,
-0.0007033174624666572,
0.06287981569766998,
0.024702690541744232,
0.011831489391624928,
-0.033226702362298965,
-0.06865101307630539,
-0.023596322163939476,
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0.03148718178272247,
0.038957323879003525
] |
PMID:20631
|
Purification and properties of human erythrocyte pyrimidine 5'-nucleotidase.
|
A 250,000-fold purification of pyrimidine 5'-nucleotidase from human erythrocytes has been achieved using a combination of DEAE-cellulose chromatography, ammonium sulfate fractionation, gel filtration, and isoelectric focusing. Polyacrylamide disc and starch gel electrophoresis of the purified material show two strong protein bands. On starch gel these bands exhibited pyrimidine 5'-nucleotidase activity. Two faint protein bands devoid of enzyme activity were also found in the case of polyacrylamide electrophoresis. The enzyme has a pH optimum at 7.5 and is most stable between pH 6 and 7.5. The enzyme has pI of 5.0 and a molecular weight of 28,000 by gel filtration. The Km of the purified enzyme was 10 muM, compared to 40 muM when measured in hemolysate. The higher Km in the hemolysate is due to the presence of an inhibitor. Inorganic phosphate was shown to be a competitive inhibitor of pyrimidine 5'-nucleotidase and inorganic phosphate in the hemolysate may be responsible for increasing the Km of the enzyme for the substrate cytidine monophosphate.
|
Purification and properties of human erythrocyte pyrimidine 5'-nucleotidase. A 250,000-fold purification of pyrimidine 5'-nucleotidase from human erythrocytes has been achieved using a combination of DEAE-cellulose chromatography, ammonium sulfate fractionation, gel filtration, and isoelectric focusing. Polyacrylamide disc and starch gel electrophoresis of the purified material show two strong protein bands. On starch gel these bands exhibited pyrimidine 5'-nucleotidase activity. Two faint protein bands devoid of enzyme activity were also found in the case of polyacrylamide electrophoresis. The enzyme has a pH optimum at 7.5 and is most stable between pH 6 and 7.5. The enzyme has pI of 5.0 and a molecular weight of 28,000 by gel filtration. The Km of the purified enzyme was 10 muM, compared to 40 muM when measured in hemolysate. The higher Km in the hemolysate is due to the presence of an inhibitor. Inorganic phosphate was shown to be a competitive inhibitor of pyrimidine 5'-nucleotidase and inorganic phosphate in the hemolysate may be responsible for increasing the Km of the enzyme for the substrate cytidine monophosphate.
|
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] |
PMID:20632
|
Specific benzodiazepine receptors in rat brain characterized by high-affinity (3H)diazepam binding.
|
[3H]Diazepam appears to bind specifically to a single, saturable, binding site located on rat brain membranes, with an affinity constant near 3 nM at pH 7.4. Specific binding constitutes more than 90% of total binding at 0 degrees and less than 10% of total binding at 37 degrees. Arrhenius plots suggest a sharp conformational change in the diazepam receptor near 18 degrees. Mitochondrial fractions from rat kidney, liver, and lung exhibit some [3H]diazepam binding that can be displaced by nonradioactive diazepam and several other benzodiazepines. However, Ro-4864, which is almost inactive in displacing [3H]diazepam from brain membranes, is extremely potent in displacing it from kidney mitochondria. Conversely, clonazepam, the most potent inhibitor of brain binding, is an extremely weak inhibitor of kidney binding. Furthermore, diazepam binding to kidney mitochondria has an affinity constantof 40 nM, about 15 times higher than that in brain. No specific diazepam binding was detected in intestine or skeletal muscle. Thus, specific [3H]diazepam binding to membranes appears to be restricted to brain, where it is unevenly distributed: the density of diazepam receptors is about five times higher in cortex (the highest density) than in pons-meddula (lowest density). Trypsin and chymotrypsin completely abolished specific [3H]diazepambinding in brain and kidney.
|
Specific benzodiazepine receptors in rat brain characterized by high-affinity (3H)diazepam binding. [3H]Diazepam appears to bind specifically to a single, saturable, binding site located on rat brain membranes, with an affinity constant near 3 nM at pH 7.4. Specific binding constitutes more than 90% of total binding at 0 degrees and less than 10% of total binding at 37 degrees. Arrhenius plots suggest a sharp conformational change in the diazepam receptor near 18 degrees. Mitochondrial fractions from rat kidney, liver, and lung exhibit some [3H]diazepam binding that can be displaced by nonradioactive diazepam and several other benzodiazepines. However, Ro-4864, which is almost inactive in displacing [3H]diazepam from brain membranes, is extremely potent in displacing it from kidney mitochondria. Conversely, clonazepam, the most potent inhibitor of brain binding, is an extremely weak inhibitor of kidney binding. Furthermore, diazepam binding to kidney mitochondria has an affinity constantof 40 nM, about 15 times higher than that in brain. No specific diazepam binding was detected in intestine or skeletal muscle. Thus, specific [3H]diazepam binding to membranes appears to be restricted to brain, where it is unevenly distributed: the density of diazepam receptors is about five times higher in cortex (the highest density) than in pons-meddula (lowest density). Trypsin and chymotrypsin completely abolished specific [3H]diazepambinding in brain and kidney.
|
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] |
PMID:20633
|
Kinetics of the alkaline tetramer leads to dimer dissociation in liganded human hemoglobin: a laser light-scattering stopped-flow study.
|
The first-order dissociation of tetrameric HbCO to the dimer has been studied overthe pH range 10.30-11.57 in a light-scattering stopped-flow apparatus using argon-ion laser excitation. The first-order dissociation rate constant varies from 0.25 sec-1 to 24.0 sec-1over this pH interval. A semilogarithmic plot of k versus pH has a slope of 2.56 at pH 11.07, the midpoint. The pH dependence of the dissociation of the tetramer is consistent with progressive titration of alpha1-alpha2 and beta1-beta2 salt bridges. At pH 10.66, the dissociation rates of HbO2, HbCO, methemoglobin, and HbCN vary less than 20% from their mean value. A study of the dissociation kinetics as a function of protein concentration allows one to obtain both association and dissociation rate constants, and hence equilibrium constants, for the tetramer in equilibrium dimer reaction. In this manner, equilibrium constants were obtained on protein solutions with less than 15 sec of exposure to dissociating conditions.
|
Kinetics of the alkaline tetramer leads to dimer dissociation in liganded human hemoglobin: a laser light-scattering stopped-flow study. The first-order dissociation of tetrameric HbCO to the dimer has been studied overthe pH range 10.30-11.57 in a light-scattering stopped-flow apparatus using argon-ion laser excitation. The first-order dissociation rate constant varies from 0.25 sec-1 to 24.0 sec-1over this pH interval. A semilogarithmic plot of k versus pH has a slope of 2.56 at pH 11.07, the midpoint. The pH dependence of the dissociation of the tetramer is consistent with progressive titration of alpha1-alpha2 and beta1-beta2 salt bridges. At pH 10.66, the dissociation rates of HbO2, HbCO, methemoglobin, and HbCN vary less than 20% from their mean value. A study of the dissociation kinetics as a function of protein concentration allows one to obtain both association and dissociation rate constants, and hence equilibrium constants, for the tetramer in equilibrium dimer reaction. In this manner, equilibrium constants were obtained on protein solutions with less than 15 sec of exposure to dissociating conditions.
|
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] |
PMID:20647
|
Antimuscarinic properties of antidepressants: dibenzepin (Noveril).
|
The antimuscarinic potency of dibenzepin (Noveril) was estimated by measuring (a) central in vivo effects in mice (antihypothermia and antitremor, both induced by oxotremorine), (b) peripheral in vivo activity (mydriasis caused by systemic administration of the drug), (c) the effects of dibenzepin on isolated smooth muscle from guinea pig ileum, and (d) in vitro determination of the affinity constant of dibezepine toward the muscarinic binding sites in whole mouse-brain homogenate. The data allowed the construction of a normalized antimuscarinic potency scale for some of the common tricyclic antidepressants. With a value of 1 for scopolamine, the following relative anticholinergic potencies were calculated: dibenzepin--1/600, nortriptylne--1/300, imipramine - 1/200, and amitriptyline - 1/75. These values suggest an explanation for the absence of clinically detectable anticholinergic side effects during treatment of depression with high doses of dibenzepin. Structural and spatial interrelations among various tricyclic antidepressants and scopolamine are discussed.
|
Antimuscarinic properties of antidepressants: dibenzepin (Noveril). The antimuscarinic potency of dibenzepin (Noveril) was estimated by measuring (a) central in vivo effects in mice (antihypothermia and antitremor, both induced by oxotremorine), (b) peripheral in vivo activity (mydriasis caused by systemic administration of the drug), (c) the effects of dibenzepin on isolated smooth muscle from guinea pig ileum, and (d) in vitro determination of the affinity constant of dibezepine toward the muscarinic binding sites in whole mouse-brain homogenate. The data allowed the construction of a normalized antimuscarinic potency scale for some of the common tricyclic antidepressants. With a value of 1 for scopolamine, the following relative anticholinergic potencies were calculated: dibenzepin--1/600, nortriptylne--1/300, imipramine - 1/200, and amitriptyline - 1/75. These values suggest an explanation for the absence of clinically detectable anticholinergic side effects during treatment of depression with high doses of dibenzepin. Structural and spatial interrelations among various tricyclic antidepressants and scopolamine are discussed.
|
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] |
PMID:20648
|
[Stability of tetracycline during fermentation].
|
The stability of tetracycline was studied at pH 6.0 and 1.5. The thermal inactivation constant at the lower pH was found to vary between 1.62 min-1 (40 degrees C) and 65.83 min-1 (100 degrees C). A degradation mechanism was proposed in the light of the variation of entropy and steric factors.
|
[Stability of tetracycline during fermentation]. The stability of tetracycline was studied at pH 6.0 and 1.5. The thermal inactivation constant at the lower pH was found to vary between 1.62 min-1 (40 degrees C) and 65.83 min-1 (100 degrees C). A degradation mechanism was proposed in the light of the variation of entropy and steric factors.
|
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] |
PMID:20652
|
beta-blocking agents and human fat cell adenylate cyclase.
|
The effects of various beta-blocking agents upon the catecholamine-activated adenylate cyclase of human fat cell ghosts were studied. Both non-selective and cardio-selective beta-blocking drugs are capable in inhibiting the human enzyme system. The concentrations of the non-selective beta-blockers (bupranolol, alprenolol, propranolol, 4-hydroxypropranolol, prindolol and oxyprenolol) required to produce half maximal inhibition of isoproterenol-stimulated rates of cAMP-formation were found to be in a narrow range (5X10(-7) M-3X10(-6) M).The cardioselective beta-blocking agents, however, were only 1/100 (methypranol) to 1/1000 (atenolol, practolol) as potent as the non-selective drugs.
|
beta-blocking agents and human fat cell adenylate cyclase. The effects of various beta-blocking agents upon the catecholamine-activated adenylate cyclase of human fat cell ghosts were studied. Both non-selective and cardio-selective beta-blocking drugs are capable in inhibiting the human enzyme system. The concentrations of the non-selective beta-blockers (bupranolol, alprenolol, propranolol, 4-hydroxypropranolol, prindolol and oxyprenolol) required to produce half maximal inhibition of isoproterenol-stimulated rates of cAMP-formation were found to be in a narrow range (5X10(-7) M-3X10(-6) M).The cardioselective beta-blocking agents, however, were only 1/100 (methypranol) to 1/1000 (atenolol, practolol) as potent as the non-selective drugs.
|
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-0.031617600470781326,
-0.02999778836965561,
0.1108376607298851,
0.04424498975276947
] |
PMID:20653
|
[The effect of sympathomimetric drugs on exocrine pancreatic function. Studies in the isolated cat pancreas (author's transl)].
|
This study reports on effects of beta-sympathomimetic drugs on the exocrine function of the isolated cat pancreas. The injection of these drugs resulted in an increase of pancreatic protein and enzyme secretion, the excretion of calcium into the juice and the rate of volume output were not changed. The enzyme secretion was inhibited by different adrenolytic drugs (phenoxybenzamine, practolol, butoxamine), by tetracain, hemicholinium, and atropine. The pancreatic function of cats pretreated by reserpine or 6-hydroxydopamine was not altered. It is concluded, that beta-sympathomimetic drugs influence the pancreatic protein and enzyme secretion in a cholinergic way. These results are consistent with those of other investigators showing a suppressibility of the isoproterenol-induced pancreatic amylase secretion by atropine.
|
[The effect of sympathomimetric drugs on exocrine pancreatic function. Studies in the isolated cat pancreas (author's transl)]. This study reports on effects of beta-sympathomimetic drugs on the exocrine function of the isolated cat pancreas. The injection of these drugs resulted in an increase of pancreatic protein and enzyme secretion, the excretion of calcium into the juice and the rate of volume output were not changed. The enzyme secretion was inhibited by different adrenolytic drugs (phenoxybenzamine, practolol, butoxamine), by tetracain, hemicholinium, and atropine. The pancreatic function of cats pretreated by reserpine or 6-hydroxydopamine was not altered. It is concluded, that beta-sympathomimetic drugs influence the pancreatic protein and enzyme secretion in a cholinergic way. These results are consistent with those of other investigators showing a suppressibility of the isoproterenol-induced pancreatic amylase secretion by atropine.
|
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] |
PMID:20654
|
Quantifying drugs influence on cardio-respiratory synchronism.
|
The paper concerns the situation in which, for a considerable time, the onset of each respiratory cycle coincides with one and the same section of a cardiac cycle. A standardized procedure is described with which, in anesthetized rabbits, the potentiality of the biological mechanisms responsible for the phenomenon may be quantified. The procedure consists of progressively prolonging the cardiac cycles by selectively cooling the sinus node. Care is taken that during such procedure a situation is passed in which the respiratory cycles are a multiple of the cardiac cycles. The synchronizing kybernetic system, if any, must click in, then. As the prolonging of the cardiac cycles continues, the synchrony will get lost the sooner the weaker the potentialities of the kybernetic system and vice versa. Four drugs are tested as to their potential action on the kybernetic system. Nikethamide and methylphenidate tend to decrease its efficacy, allobarbital and benzoctamine tend to increase it, none of them, however, being conclusively effective.
|
Quantifying drugs influence on cardio-respiratory synchronism. The paper concerns the situation in which, for a considerable time, the onset of each respiratory cycle coincides with one and the same section of a cardiac cycle. A standardized procedure is described with which, in anesthetized rabbits, the potentiality of the biological mechanisms responsible for the phenomenon may be quantified. The procedure consists of progressively prolonging the cardiac cycles by selectively cooling the sinus node. Care is taken that during such procedure a situation is passed in which the respiratory cycles are a multiple of the cardiac cycles. The synchronizing kybernetic system, if any, must click in, then. As the prolonging of the cardiac cycles continues, the synchrony will get lost the sooner the weaker the potentialities of the kybernetic system and vice versa. Four drugs are tested as to their potential action on the kybernetic system. Nikethamide and methylphenidate tend to decrease its efficacy, allobarbital and benzoctamine tend to increase it, none of them, however, being conclusively effective.
|
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] |
PMID:20656
|
A placebo-controlled, double-blind trial of Benorylate tablets in the treatment of bursitis and synovitis due to sports injury.
|
In this two-week study, Benoral tablets, at a 4.5 g daily dosage were compared with a matching placebo in 20 patients suffering from sports injury. Four assessments were made: pain at rest, pain on movement, tenderness and soft tissue swelling. In each case the active treatment group (benorylate) was statistically greater than that in the placebo group at one week and after two weeks' treatment.
|
A placebo-controlled, double-blind trial of Benorylate tablets in the treatment of bursitis and synovitis due to sports injury. In this two-week study, Benoral tablets, at a 4.5 g daily dosage were compared with a matching placebo in 20 patients suffering from sports injury. Four assessments were made: pain at rest, pain on movement, tenderness and soft tissue swelling. In each case the active treatment group (benorylate) was statistically greater than that in the placebo group at one week and after two weeks' treatment.
|
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] |
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