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PMID:19164
|
Temperature dependence of the absorbance of alkaline solutions of 4-nitrophenyl phosphate--a potential source of error in the measurement of alkaline phosphatase activity.
|
The absorbance of an alkaline solution of 4-nitrophenyl phosphate is a function of temperature. Quantitative evaluation of this phenomenon indicates that it (a) depends on the concentration of the compound and is independent of source, buffer concentration, and pH above 9.0; (b) is reversible; (c) is not a result of alkaline hydrolysis or 4-nitrophenol contamination; and (d) correlates with a temperature-induced shift of its absorbance spectrum. The phenomenon may represent a potential analytical problem in methods for alkaline phosphatase in which this compound is the substrate. If thermal equilibrium is not reached and maintained during an alkaline phosphatase assay, the thermochromic response will be included in the measured rate. The magnitude of this error depends on the thermal response and control characteristics of each particular instrument and the reaction conditions under which such an analysis is performed.
|
Temperature dependence of the absorbance of alkaline solutions of 4-nitrophenyl phosphate--a potential source of error in the measurement of alkaline phosphatase activity. The absorbance of an alkaline solution of 4-nitrophenyl phosphate is a function of temperature. Quantitative evaluation of this phenomenon indicates that it (a) depends on the concentration of the compound and is independent of source, buffer concentration, and pH above 9.0; (b) is reversible; (c) is not a result of alkaline hydrolysis or 4-nitrophenol contamination; and (d) correlates with a temperature-induced shift of its absorbance spectrum. The phenomenon may represent a potential analytical problem in methods for alkaline phosphatase in which this compound is the substrate. If thermal equilibrium is not reached and maintained during an alkaline phosphatase assay, the thermochromic response will be included in the measured rate. The magnitude of this error depends on the thermal response and control characteristics of each particular instrument and the reaction conditions under which such an analysis is performed.
|
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] |
PMID:19165
|
Continuous-flow analysis for glucose, triglycerides, and ATP with immobilized enzymes in tubular form.
|
Small-bore ("Autozyme") tubes with immobilized enzymes at the inner wall have been developed and studied for application in the Technicon "SMAC" high-speed continuous-flow biochemical analyzer. Tubes coated with glucose oxidase (D-glucose:oxygen oxidoreductase, EC 1.1.3.4) have been prepared for the assay of glucose, with colorimetric assay of the hydrogen peroxide produced; tubes coated with glycerol kinase (ATP:glycerol phosphotransferase, EC 2.7.1.30) for the enzymatic assay of triglycerides; tubes coated with hexokinase (ATP:D-hexose-6-phosphotransferase, EC 2.7.1.1) and glucose-6-phosphate dehydrogenase (D-glucose-6-phosphate:NAD+ oxidoreductase EC 1.1.1.49) for the measurement of ATP, an intermediate product in assays for creatine kinase. With use of 10-15 cm lengths of Autozyme tube and SMAC hydraulics (150 samples per hour), assay sensitivity and carryover were similar to values for the corresponding free-enzyme methods. These immobilized enzymes were sufficiently stable for one to eight weeks of continuous use before replacemnt. We conclude that suitable bound-enzyme tubes can replace either single or multiple free-enzyme reagents in many continuous-flow assays at high sampling rates.
|
Continuous-flow analysis for glucose, triglycerides, and ATP with immobilized enzymes in tubular form. Small-bore ("Autozyme") tubes with immobilized enzymes at the inner wall have been developed and studied for application in the Technicon "SMAC" high-speed continuous-flow biochemical analyzer. Tubes coated with glucose oxidase (D-glucose:oxygen oxidoreductase, EC 1.1.3.4) have been prepared for the assay of glucose, with colorimetric assay of the hydrogen peroxide produced; tubes coated with glycerol kinase (ATP:glycerol phosphotransferase, EC 2.7.1.30) for the enzymatic assay of triglycerides; tubes coated with hexokinase (ATP:D-hexose-6-phosphotransferase, EC 2.7.1.1) and glucose-6-phosphate dehydrogenase (D-glucose-6-phosphate:NAD+ oxidoreductase EC 1.1.1.49) for the measurement of ATP, an intermediate product in assays for creatine kinase. With use of 10-15 cm lengths of Autozyme tube and SMAC hydraulics (150 samples per hour), assay sensitivity and carryover were similar to values for the corresponding free-enzyme methods. These immobilized enzymes were sufficiently stable for one to eight weeks of continuous use before replacemnt. We conclude that suitable bound-enzyme tubes can replace either single or multiple free-enzyme reagents in many continuous-flow assays at high sampling rates.
|
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] |
PMID:19166
|
Creatine kinase: re-examination of optimum reaction conditions.
|
I have re-examined optimum reaction conditions for measurement of creatine kinase (EC 2.7.3.2). The optimum pH is 6.45, and 2,2-bis(hydroxymethyl)-2,2',2''-nitrotriethanol acetate, 200 mmol/liter, is the buffer of choice. Thioglycerol, 20 mmol/liter, is superior for both in-assay reactivation and for storage stability of sera. Fluoride, 25 mmol/liter, a broad inactivator of adenylate kinase (EC 2.7.4.3), has little effect on creatine kinase and is superior to AMP for adenylate kinase inhibition in the assay of creatine kinase. Magnesium ion, ADP, and buffer concentrations are interdependent and their optima must be determined together. The hexokinase/glucose-6-phosphate dehydrogenase activity ratio should not exceed 1.6. The range of linearity is limited by the glucose-6-phosphate dehydrogenase and NAD+ concentrations. Glucose-6-phosphate dehydrogenase, ADP, and NAD+ are the constituents most likely to result in unacceptable blanks. Creatine kinase is inhibited noncompetitively by anions: acetate and fluoride inhibit slightly, but sulfates, nitrates, and excessive chlorides should be avoided.
|
Creatine kinase: re-examination of optimum reaction conditions. I have re-examined optimum reaction conditions for measurement of creatine kinase (EC 2.7.3.2). The optimum pH is 6.45, and 2,2-bis(hydroxymethyl)-2,2',2''-nitrotriethanol acetate, 200 mmol/liter, is the buffer of choice. Thioglycerol, 20 mmol/liter, is superior for both in-assay reactivation and for storage stability of sera. Fluoride, 25 mmol/liter, a broad inactivator of adenylate kinase (EC 2.7.4.3), has little effect on creatine kinase and is superior to AMP for adenylate kinase inhibition in the assay of creatine kinase. Magnesium ion, ADP, and buffer concentrations are interdependent and their optima must be determined together. The hexokinase/glucose-6-phosphate dehydrogenase activity ratio should not exceed 1.6. The range of linearity is limited by the glucose-6-phosphate dehydrogenase and NAD+ concentrations. Glucose-6-phosphate dehydrogenase, ADP, and NAD+ are the constituents most likely to result in unacceptable blanks. Creatine kinase is inhibited noncompetitively by anions: acetate and fluoride inhibit slightly, but sulfates, nitrates, and excessive chlorides should be avoided.
|
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0.019542047753930092,
0.08753152936697006,
0.014482393860816956
] |
PMID:19167
|
pH and the activity of porphobilinogen synthase in human blood.
|
For determination of the activity of this enzyme (EC 4.2.1.24) in blood, pH dependency was evaluated by using whole blood and washed erythrocytes. The effects of assay methods, original and final pH, and use of sodium or potassium phosphate buffer were examined. The pattern of pH dependency differed for whole blood and washed erythrocytes, whether blood from normal or lead-exposed subjects was examined. The pH optimum for the activity in erythrocytes from lead-exposed subjects was the same as for the normal persons if the concentration of lead in blood did not exceed 80 microgram/100 g of whole blood. Hemolyzing blood by placing it on solid CO2 decreases the activity.
|
pH and the activity of porphobilinogen synthase in human blood. For determination of the activity of this enzyme (EC 4.2.1.24) in blood, pH dependency was evaluated by using whole blood and washed erythrocytes. The effects of assay methods, original and final pH, and use of sodium or potassium phosphate buffer were examined. The pattern of pH dependency differed for whole blood and washed erythrocytes, whether blood from normal or lead-exposed subjects was examined. The pH optimum for the activity in erythrocytes from lead-exposed subjects was the same as for the normal persons if the concentration of lead in blood did not exceed 80 microgram/100 g of whole blood. Hemolyzing blood by placing it on solid CO2 decreases the activity.
|
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] |
PMID:19168
|
Evaluation of ampouled tonometered buffer solutions as a quality-control system for pH, pCO2, and pO2 measurement.
|
In response to the need for an adequate quality-control system for blood-pH and blood-gas analyzers, we investigated the practical application of ampouled phosphate-bicarbonate-chloride solutions tonometered with mixtures of carbon dioxide, oxygen, and nitrogen. This system offers three discrete sets of pH, pCO2, AND PO2 values, which are consistent with normal and pathophysiologically high and low values. The stated values were based on the U.S. National Bureau of Standards scale for pH and on gas analysis for pCO2 and pO2. Influence of temperature, air contact, calibration gas, and storage was established. Internal and external quality control by means of these ampoules is presented. The system is stable, accurate, precise, and suitable for simultaneous quality control of pH, pCO2, and pO2 measurements.
|
Evaluation of ampouled tonometered buffer solutions as a quality-control system for pH, pCO2, and pO2 measurement. In response to the need for an adequate quality-control system for blood-pH and blood-gas analyzers, we investigated the practical application of ampouled phosphate-bicarbonate-chloride solutions tonometered with mixtures of carbon dioxide, oxygen, and nitrogen. This system offers three discrete sets of pH, pCO2, AND PO2 values, which are consistent with normal and pathophysiologically high and low values. The stated values were based on the U.S. National Bureau of Standards scale for pH and on gas analysis for pCO2 and pO2. Influence of temperature, air contact, calibration gas, and storage was established. Internal and external quality control by means of these ampoules is presented. The system is stable, accurate, precise, and suitable for simultaneous quality control of pH, pCO2, and pO2 measurements.
|
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] |
PMID:19169
|
Sensitive, optimized assay for serum AMP deaminase.
|
A sensitive, optimized assay for serum AMP deaminase (EC 3.5.4.6) is presented, which is based on a colorimetric indophenol determination of ammonia liberated during the enzyme reaction. An average enzyme activity in serum of 2.7 U/liter (n = 36, range 0-6.2) for healthy adults was established. Both the enzyme and colorimetric reactions were optimized to give an assay sensitivity of 0.2 U/liter of serum, a 98-99% analytical recovery in the normal serum enzyme concentration range, and a CV of 7.9% in-run and 10.6% between-run. This optimization included studies on specific buffer, pH, ionic strength, and potential activators. The Km for the enzyme in serum was determinated to be 1.4 X 10(-3) mol/liter, which agrees well with the value reported for human skeletal muscle enzyme. Above-normal serum AMP deaminase activity may be present in various myopathies and could be useful in detecting carriers of the X-linked dystrophies.
|
Sensitive, optimized assay for serum AMP deaminase. A sensitive, optimized assay for serum AMP deaminase (EC 3.5.4.6) is presented, which is based on a colorimetric indophenol determination of ammonia liberated during the enzyme reaction. An average enzyme activity in serum of 2.7 U/liter (n = 36, range 0-6.2) for healthy adults was established. Both the enzyme and colorimetric reactions were optimized to give an assay sensitivity of 0.2 U/liter of serum, a 98-99% analytical recovery in the normal serum enzyme concentration range, and a CV of 7.9% in-run and 10.6% between-run. This optimization included studies on specific buffer, pH, ionic strength, and potential activators. The Km for the enzyme in serum was determinated to be 1.4 X 10(-3) mol/liter, which agrees well with the value reported for human skeletal muscle enzyme. Above-normal serum AMP deaminase activity may be present in various myopathies and could be useful in detecting carriers of the X-linked dystrophies.
|
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] |
PMID:19170
|
Semiautomated spectrophotometry of total phospholipids in plasma.
|
A one-stage semiautomated method for estimating total phospholipids in plasma is reported. The phospholipids are extracted into aqueous ethanol, combined with a Lewis acid, and the change in pH is monitored colorimetrically. The method, compared with the acid digestion method of Gomori [J. Lab. Clin. Med. 27, 955 (1942)], gives results that are about 0.25 mmol/liter lower, but seems to be a suitable and less hazardous alternative.
|
Semiautomated spectrophotometry of total phospholipids in plasma. A one-stage semiautomated method for estimating total phospholipids in plasma is reported. The phospholipids are extracted into aqueous ethanol, combined with a Lewis acid, and the change in pH is monitored colorimetrically. The method, compared with the acid digestion method of Gomori [J. Lab. Clin. Med. 27, 955 (1942)], gives results that are about 0.25 mmol/liter lower, but seems to be a suitable and less hazardous alternative.
|
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] |
PMID:19171
|
Gas chromatographic and mass spectrometric studies on urinary organic acids in a patient with congenital lactic acidosis due to pyruvate decarboxylase deficiency.
|
Detailed studies, using gas chromatography and mass spectrometric methods, of the urinary organic acids excreted by a patient with proven pyruvate decarboxylase deficiency are reported. In addition to the greatly-increased levels of lactate and pyruvate, marked elevation in the levels of 2-oxoglutaric, malic, and isocitric acids were observed, with associated increases 2-hydroxyglutaric, fumaric, succinic, and glyceric acids, and reduced citric acid excretion. The levels of excretion during clinically static and acute periods are compared to those in a normal neonate and normal infants. The metabolites observed indicate a probable defect in the oxidation of pyruvate by pyruvate dehydrogenase and suggest the presence of secondary defects in the tricarboxylic acid cycle. Studies of this type may enable the relatively rapid identification of the probable underlying enzyme deficiency in cases of congenital lactic acidosis, prior to confirmatory enzyme studies.
|
Gas chromatographic and mass spectrometric studies on urinary organic acids in a patient with congenital lactic acidosis due to pyruvate decarboxylase deficiency. Detailed studies, using gas chromatography and mass spectrometric methods, of the urinary organic acids excreted by a patient with proven pyruvate decarboxylase deficiency are reported. In addition to the greatly-increased levels of lactate and pyruvate, marked elevation in the levels of 2-oxoglutaric, malic, and isocitric acids were observed, with associated increases 2-hydroxyglutaric, fumaric, succinic, and glyceric acids, and reduced citric acid excretion. The levels of excretion during clinically static and acute periods are compared to those in a normal neonate and normal infants. The metabolites observed indicate a probable defect in the oxidation of pyruvate by pyruvate dehydrogenase and suggest the presence of secondary defects in the tricarboxylic acid cycle. Studies of this type may enable the relatively rapid identification of the probable underlying enzyme deficiency in cases of congenital lactic acidosis, prior to confirmatory enzyme studies.
|
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] |
PMID:19172
|
A gel filament as an analytical tool: the testing of an affinity chromatographic, product-immobilizing linear gel for determination of enzymatic activity.
|
A new technique for quantitative analysis is proposed. An underfined volume of the test solution is exposed to a reagent in the form of a gel filament of capillary dimensions. The linear progress in the gel of the compound in question is followed with the aid of the coloured reaction products deposited along its path. With the use of the enzyme gamma-glutamyl transpeptidase it is shown that the velocity in the system of the component to be analysed, as determined by measurement of its coloured track, is related to its activity in the test solution.
|
A gel filament as an analytical tool: the testing of an affinity chromatographic, product-immobilizing linear gel for determination of enzymatic activity. A new technique for quantitative analysis is proposed. An underfined volume of the test solution is exposed to a reagent in the form of a gel filament of capillary dimensions. The linear progress in the gel of the compound in question is followed with the aid of the coloured reaction products deposited along its path. With the use of the enzyme gamma-glutamyl transpeptidase it is shown that the velocity in the system of the component to be analysed, as determined by measurement of its coloured track, is related to its activity in the test solution.
|
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] |
PMID:19173
|
An automated continuous-monitoring procedure for the determination of acid phosphatase activity in serum.
|
The method of Hillmann, in which hydrolysis of alpha-naphthyl phosphate by acid phosphatase is coupled to the formation of an alpha-naphthol-Fast Red TR azo-compound, has been adapted for use with the LKB Produkter AB 8600 reaction rate analyzer. Factors which affect the reproducibility of the method are described and its performance is shown to be superior to that of a manual phenyl-phosphate procedure.
|
An automated continuous-monitoring procedure for the determination of acid phosphatase activity in serum. The method of Hillmann, in which hydrolysis of alpha-naphthyl phosphate by acid phosphatase is coupled to the formation of an alpha-naphthol-Fast Red TR azo-compound, has been adapted for use with the LKB Produkter AB 8600 reaction rate analyzer. Factors which affect the reproducibility of the method are described and its performance is shown to be superior to that of a manual phenyl-phosphate procedure.
|
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] |
PMID:19174
|
A method for visual assessment of the performance of components in a continuous flow system.
|
When two reagents are mixed in a continuous flow system it is difficult to detect variations in their proportions in the individual liquid segments because such variations produce only small changes in the final colour intensity. A method is described whereby these differences in reagent proportion produce a different pH in each segment and, by the addition of an appropriate pH indicator solution, a different colour for each segment. This method may be used to demonstrate the eveness of reagent injection, reagent mixing and segment/segment interaction associated with continuous flow systems.
|
A method for visual assessment of the performance of components in a continuous flow system. When two reagents are mixed in a continuous flow system it is difficult to detect variations in their proportions in the individual liquid segments because such variations produce only small changes in the final colour intensity. A method is described whereby these differences in reagent proportion produce a different pH in each segment and, by the addition of an appropriate pH indicator solution, a different colour for each segment. This method may be used to demonstrate the eveness of reagent injection, reagent mixing and segment/segment interaction associated with continuous flow systems.
|
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] |
PMID:19175
|
Calcium in bile and calcium salts in gallstones.
|
In gallbladder and common duct bile from patients undergoing cholecystectomy, usually because of gallstones, calcium was found to exist in at least 2 forms. Ultrafiltration showed some calcium was bound to substances with a molecular weight greater than 10 000, and the chief binding agent is likely to be the mixed micelle. Bound calcium was significantly less in common duct bile than in bile from functioning gallbladders, but the amount of ultrafiltrable calcium was the same. Furthermore, ultrafiltrable calcium in gallbladder bile from patients with cholesterol or some calcium carbonate in their gallstones was almost constant for a range of total calcium concentrations of 2.40--9.70 mmol/l. Comparison of ultrafiltrable and total calcium values for the different types of stone-formers showed that the deposition of calcium carbonate in gallstones was not related to any calcium measurement made. However, the presence of calcium phosphate and/or calcium bilirubinate in gallstones could be related to a significant increase in ultrafiltrable calcium in gallbladder bile.
|
Calcium in bile and calcium salts in gallstones. In gallbladder and common duct bile from patients undergoing cholecystectomy, usually because of gallstones, calcium was found to exist in at least 2 forms. Ultrafiltration showed some calcium was bound to substances with a molecular weight greater than 10 000, and the chief binding agent is likely to be the mixed micelle. Bound calcium was significantly less in common duct bile than in bile from functioning gallbladders, but the amount of ultrafiltrable calcium was the same. Furthermore, ultrafiltrable calcium in gallbladder bile from patients with cholesterol or some calcium carbonate in their gallstones was almost constant for a range of total calcium concentrations of 2.40--9.70 mmol/l. Comparison of ultrafiltrable and total calcium values for the different types of stone-formers showed that the deposition of calcium carbonate in gallstones was not related to any calcium measurement made. However, the presence of calcium phosphate and/or calcium bilirubinate in gallstones could be related to a significant increase in ultrafiltrable calcium in gallbladder bile.
|
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] |
PMID:19176
|
A method for enhancing the stability of thiol-containing enzymes immobilised on nylon.
|
The limited stability of immobilised enzymes is a major factor restricting their use in clinical chemistry. The immobilisation procedure described here affords a simple method of enhancing the stability of certain enzymes immobilised on nylon. Studies with alcohol dehydrogenase and glyceraldehyde-3-phosphate dehydrogenase have indicated that a significant increase in stability can be obtained using this immobilisation procedure as compared with conventional procedures. As anticipated the immobilisation procedure was unsatisfactory for oxidases such as glucose oxidase.
|
A method for enhancing the stability of thiol-containing enzymes immobilised on nylon. The limited stability of immobilised enzymes is a major factor restricting their use in clinical chemistry. The immobilisation procedure described here affords a simple method of enhancing the stability of certain enzymes immobilised on nylon. Studies with alcohol dehydrogenase and glyceraldehyde-3-phosphate dehydrogenase have indicated that a significant increase in stability can be obtained using this immobilisation procedure as compared with conventional procedures. As anticipated the immobilisation procedure was unsatisfactory for oxidases such as glucose oxidase.
|
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] |
PMID:19177
|
The activity of gamma-glutamyl transferase in the serum of multiple sclerosis and other neurological diseases.
|
1. In the sera of 142 neurological patients, including 35 cases with multiple sclerosis (MS), and 20 normal controls the gamma-glutamyltransferase (gamma-GT) activity was determined. The result of this investigation was compared with the "combined hippuric acid test" formely often used in MS patients. 2. The MS patients are the only group which significantly differs from the normal controls. There are also small differences between them and some of the other neurological patient groups. About one-third of the MS patients show moderately pathological serum activities. On the other hand, patients with a definite liver affection, especially chronic alcoholics, regularly show an even more important elevation of the values, which makes this test a sensitive tool for the screening of alcohol-induced liver intoxications. 3. Although the gamma-GT test in not quite as sensitive in indicating organic defects in MS patients, it seems to reflected a disturbed liver function in the same way as the more complicated "combined hippuric acid test". It may therefore be used as a simple tool of finding and following up organic defects in MS patients.
|
The activity of gamma-glutamyl transferase in the serum of multiple sclerosis and other neurological diseases. 1. In the sera of 142 neurological patients, including 35 cases with multiple sclerosis (MS), and 20 normal controls the gamma-glutamyltransferase (gamma-GT) activity was determined. The result of this investigation was compared with the "combined hippuric acid test" formely often used in MS patients. 2. The MS patients are the only group which significantly differs from the normal controls. There are also small differences between them and some of the other neurological patient groups. About one-third of the MS patients show moderately pathological serum activities. On the other hand, patients with a definite liver affection, especially chronic alcoholics, regularly show an even more important elevation of the values, which makes this test a sensitive tool for the screening of alcohol-induced liver intoxications. 3. Although the gamma-GT test in not quite as sensitive in indicating organic defects in MS patients, it seems to reflected a disturbed liver function in the same way as the more complicated "combined hippuric acid test". It may therefore be used as a simple tool of finding and following up organic defects in MS patients.
|
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] |
PMID:19179
|
Determination of enteropeptidase activity in human duodenal aspirates.
|
A sensitive procedure is described for the determination in duodenal aspirates of enteropeptidase activity based on the activation of trypsinogen and the estimation of trypsin formed with benzoyl-arginine-p-nitroanilide. Using the recovery approach where a known amount of purified human enteropeptidase is diluted in duodenal fluid and the recoverable activity determined, this method was shown to give a sensitive and reliable estimate of the enteropeptidase activity in duodenal fluid although it was shown that the enzyme was subject to a 10% activation by components in the duodenal fluid. The reported 5-fold stimulation of enteropeptidase activity by bile salts could not be demonstrated.
|
Determination of enteropeptidase activity in human duodenal aspirates. A sensitive procedure is described for the determination in duodenal aspirates of enteropeptidase activity based on the activation of trypsinogen and the estimation of trypsin formed with benzoyl-arginine-p-nitroanilide. Using the recovery approach where a known amount of purified human enteropeptidase is diluted in duodenal fluid and the recoverable activity determined, this method was shown to give a sensitive and reliable estimate of the enteropeptidase activity in duodenal fluid although it was shown that the enzyme was subject to a 10% activation by components in the duodenal fluid. The reported 5-fold stimulation of enteropeptidase activity by bile salts could not be demonstrated.
|
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] |
PMID:19180
|
Bile lysosomal enzymes: characteristics and pathological significance for various hepatobiliary disorders.
|
The activities of several glycosidases (beta-galactosidase, beta-glucosidase, N-acetyl-beta-glucosaminidase) were demonstrated in human bile. The enzyme activities are increased about 100 times after exclusion of bile salts and other small molecular compounds by Sephadex G-50 gel filtration. The use of 4-methylumbelliferyl derivatives as substrates was useful as measurement of the bile enzyme activities are not altered in the presence of bile pigments. Enzyme characteristics of bile glycosidases were determined: pH optimum and isoelectric point. The bile glycosidase activities were also measured in various hepatobiliary disorders (cholelithiasis, cancer of gallbladder, acute hepatitis, liver cirrhosis and fatty liver). The glycosidase activities in bile from patients with liver diseases, as well as with cholelithiasis, were generally decreased. Isoelectric focusing patterns of biliary glycosidases were similar for specimens from patients with hepatobiliary disorders as compared to normal.
|
Bile lysosomal enzymes: characteristics and pathological significance for various hepatobiliary disorders. The activities of several glycosidases (beta-galactosidase, beta-glucosidase, N-acetyl-beta-glucosaminidase) were demonstrated in human bile. The enzyme activities are increased about 100 times after exclusion of bile salts and other small molecular compounds by Sephadex G-50 gel filtration. The use of 4-methylumbelliferyl derivatives as substrates was useful as measurement of the bile enzyme activities are not altered in the presence of bile pigments. Enzyme characteristics of bile glycosidases were determined: pH optimum and isoelectric point. The bile glycosidase activities were also measured in various hepatobiliary disorders (cholelithiasis, cancer of gallbladder, acute hepatitis, liver cirrhosis and fatty liver). The glycosidase activities in bile from patients with liver diseases, as well as with cholelithiasis, were generally decreased. Isoelectric focusing patterns of biliary glycosidases were similar for specimens from patients with hepatobiliary disorders as compared to normal.
|
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] |
PMID:19181
|
[Glycogen synthase activity in parodontal disease (author's transl)].
|
A biochemical study of an enzyme participating in the synthesis of glycogen is presented, with particular regard to the fluctuations in the amounts of this polysaccharide in human gingival epithelium, during inflammation. The increase in the activity of UDPglucose : glycogen glucosyltransferase can be related to the accumulation of glycogen. Some kinetic parameters of this enzyme are described.
|
[Glycogen synthase activity in parodontal disease (author's transl)]. A biochemical study of an enzyme participating in the synthesis of glycogen is presented, with particular regard to the fluctuations in the amounts of this polysaccharide in human gingival epithelium, during inflammation. The increase in the activity of UDPglucose : glycogen glucosyltransferase can be related to the accumulation of glycogen. Some kinetic parameters of this enzyme are described.
|
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] |
PMID:19183
|
Suitability of commercial enzyme control sera for the quality control of activity determinations of L-aspartate aminotransferase and L-alanine aminotransferase in human serum.
|
Details of a systematic approach to suitability testing of commercial control sera are given for substrate optimized L-aspartate aminotransferase and L-alanine aminotransferase methods at 37 degrees C. Their acceptability for control purposes of standardized methods depends on: (1) the range of control values in relation to borderline values, (2) stability, (3) aspect, clarity, (4) NADH consumption in preincubation time, (5) blank activities, (6) kinetic data as half saturation constants and saturation curves, (7) influence of effectors, (8) isoenzyme pattern. These evaluation criteria are proposed for suitability testing. The term "representativeness" should be introduced as a special criterion for main characteristics of control materials. The authors want to point out the close connection with standardization of methods.
|
Suitability of commercial enzyme control sera for the quality control of activity determinations of L-aspartate aminotransferase and L-alanine aminotransferase in human serum. Details of a systematic approach to suitability testing of commercial control sera are given for substrate optimized L-aspartate aminotransferase and L-alanine aminotransferase methods at 37 degrees C. Their acceptability for control purposes of standardized methods depends on: (1) the range of control values in relation to borderline values, (2) stability, (3) aspect, clarity, (4) NADH consumption in preincubation time, (5) blank activities, (6) kinetic data as half saturation constants and saturation curves, (7) influence of effectors, (8) isoenzyme pattern. These evaluation criteria are proposed for suitability testing. The term "representativeness" should be introduced as a special criterion for main characteristics of control materials. The authors want to point out the close connection with standardization of methods.
|
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] |
PMID:19184
|
A sensitive fluorescence assay for the simultaneous and separate determination of arylsulphatases A and B.
|
A sensitive fluorometric assay utilizing 4-methylumbelliferyl sulphate has been developed for the simultaneous determination of arylsulphatases A and B from leucocytes, based on the differential effect of silver ions on the two enzymes. The procedure allows discrimination between normal cases and those with metachromatic leucodystrophy.
|
A sensitive fluorescence assay for the simultaneous and separate determination of arylsulphatases A and B. A sensitive fluorometric assay utilizing 4-methylumbelliferyl sulphate has been developed for the simultaneous determination of arylsulphatases A and B from leucocytes, based on the differential effect of silver ions on the two enzymes. The procedure allows discrimination between normal cases and those with metachromatic leucodystrophy.
|
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] |
PMID:19187
|
Clinical consequences of polymorphic acetylation of basic drugs.
|
The clinical consequences (therapeutic and toxic) of drug acetylation polymorphism are reviewed for procainamide, hydralazine, phenelzine, isoniazid, and salicylazosulfapyridine. Genetic slow acetylators are more likely than rapid acetylators to experience the following adverse drug reactions: (1) earlier development of procainamide-induced antinuclear antibody; (2) earlier and more frequent development of procainamide-induced systemic lupus erythematosus (SLE); (3) hydralazine-induced SLE; (4) spontaneous SLE; (5) drowsiness and nausea from phenelzine; (6) cyanosis, hemolysis, and transient reticulocytosis from salicylazosulfapyridine; and (7) polyneuropathy after isoniazid therapy. The incidence of isoniazid hepatitis may, however, be more common in rapid than than in slow acetylators. Genetic slow acetylators are also more likely than rapid acetylators to experience greater therapeutic responses from similar doses of the following: phenelzine, hydralazine provided beta blockers are concurrently used, and isoniazid if once weekly therapy is used. Thus, knowledge of the acetylator phenotype of a patient can help determine the relative risk for some drug-related toxic and therapeutic responses.
|
Clinical consequences of polymorphic acetylation of basic drugs. The clinical consequences (therapeutic and toxic) of drug acetylation polymorphism are reviewed for procainamide, hydralazine, phenelzine, isoniazid, and salicylazosulfapyridine. Genetic slow acetylators are more likely than rapid acetylators to experience the following adverse drug reactions: (1) earlier development of procainamide-induced antinuclear antibody; (2) earlier and more frequent development of procainamide-induced systemic lupus erythematosus (SLE); (3) hydralazine-induced SLE; (4) spontaneous SLE; (5) drowsiness and nausea from phenelzine; (6) cyanosis, hemolysis, and transient reticulocytosis from salicylazosulfapyridine; and (7) polyneuropathy after isoniazid therapy. The incidence of isoniazid hepatitis may, however, be more common in rapid than than in slow acetylators. Genetic slow acetylators are also more likely than rapid acetylators to experience greater therapeutic responses from similar doses of the following: phenelzine, hydralazine provided beta blockers are concurrently used, and isoniazid if once weekly therapy is used. Thus, knowledge of the acetylator phenotype of a patient can help determine the relative risk for some drug-related toxic and therapeutic responses.
|
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] |
PMID:19188
|
Effect of antacids on absorption of clorazepate.
|
The effect of a magnesia and alumina antacid suspension on the absorption of clorazepate dipotassium was studied in 15 normal healthy adult subjects who ingested a 15-mg dose of clorazepate alone or with single or multiple doses of antacid. The results of this three-period randomized complete crossover study showed a trend of initially slower absorption and lower peak nordiazepam plasma levels when administered with the antacid suspension. However, there were no significant differences among treatments in the extent of absorption as measured by the area under the plasma level-time curves. Clorazepate plasma levels were of relatively short duration and similar for all treatments. The urinary excretion pattern was likewise comparable with conjugated oxazepam, the major species measured. Plasma elimination half-lives of nordiazepam and clorazepate were not affected by the antacid treatments.
|
Effect of antacids on absorption of clorazepate. The effect of a magnesia and alumina antacid suspension on the absorption of clorazepate dipotassium was studied in 15 normal healthy adult subjects who ingested a 15-mg dose of clorazepate alone or with single or multiple doses of antacid. The results of this three-period randomized complete crossover study showed a trend of initially slower absorption and lower peak nordiazepam plasma levels when administered with the antacid suspension. However, there were no significant differences among treatments in the extent of absorption as measured by the area under the plasma level-time curves. Clorazepate plasma levels were of relatively short duration and similar for all treatments. The urinary excretion pattern was likewise comparable with conjugated oxazepam, the major species measured. Plasma elimination half-lives of nordiazepam and clorazepate were not affected by the antacid treatments.
|
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] |
PMID:19189
|
Leukocyte cyclic adenosine monophosphate in asthmatic children. Effects of adrenergic therapy.
|
Blood specimens for measurement of leukocyte cyclic adenosine monophosphate (AMP) were obtained at weekly intervals from asthmatic children who were participating in a double-blind, crossover study to compare the effects of two adrenergic agents and a placebo. When patients were treated with the placebo, the basal measurements and the cyclic AMP responses of leukocytes to in vitro stimulation with epinephrine (10(-4) M) were similar to those of normal subjects but within one week after initiation of treatment with an adrenergic bronchodilator, leukocyte cyclic AMP responses to adrenergic stimulation in vitro decreased and remained low during the remainder of the treatment period. Within one week after discontinuation of adrenergic therapy, leukocyte cyclic AMP responses returned to the control level. Our results indicate that the alterations in leukocyte cyclic AMP metabolism which have been observed previously in asthmatic patients may result from medications used for treatment of asthma.
|
Leukocyte cyclic adenosine monophosphate in asthmatic children. Effects of adrenergic therapy. Blood specimens for measurement of leukocyte cyclic adenosine monophosphate (AMP) were obtained at weekly intervals from asthmatic children who were participating in a double-blind, crossover study to compare the effects of two adrenergic agents and a placebo. When patients were treated with the placebo, the basal measurements and the cyclic AMP responses of leukocytes to in vitro stimulation with epinephrine (10(-4) M) were similar to those of normal subjects but within one week after initiation of treatment with an adrenergic bronchodilator, leukocyte cyclic AMP responses to adrenergic stimulation in vitro decreased and remained low during the remainder of the treatment period. Within one week after discontinuation of adrenergic therapy, leukocyte cyclic AMP responses returned to the control level. Our results indicate that the alterations in leukocyte cyclic AMP metabolism which have been observed previously in asthmatic patients may result from medications used for treatment of asthma.
|
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] |
PMID:19186
|
Equine angiotensin converting enzyme: a zinc metalloenzyme.
|
1. Angiotensin I converting enzyme from horse plasma has been extensively purified and shown to be homogeneous by disc-gel electrophoresis. 2. The metal ion involved in the catalytic reaction of the enzyme has been identified for the first time as zinc by atomic absorption spectrometry. 3. A number of other physicochemical properties of the enzyme are described and compared with results obtained by other investigators. The molecular weight was determined by gel filtration to be 113 000 daltons. The pH maximum was found to be 7-4. The chloride activation of the enzyme appears to act by facilitation of substrate binding to the enzyme. 4. By use of enzyme inhibitors, tyrosine has been implicated as a functional residue at the active site of the enzyme. 5. The enzyme shows a fairly high degree of specificity towards its substrates.
|
Equine angiotensin converting enzyme: a zinc metalloenzyme. 1. Angiotensin I converting enzyme from horse plasma has been extensively purified and shown to be homogeneous by disc-gel electrophoresis. 2. The metal ion involved in the catalytic reaction of the enzyme has been identified for the first time as zinc by atomic absorption spectrometry. 3. A number of other physicochemical properties of the enzyme are described and compared with results obtained by other investigators. The molecular weight was determined by gel filtration to be 113 000 daltons. The pH maximum was found to be 7-4. The chloride activation of the enzyme appears to act by facilitation of substrate binding to the enzyme. 4. By use of enzyme inhibitors, tyrosine has been implicated as a functional residue at the active site of the enzyme. 5. The enzyme shows a fairly high degree of specificity towards its substrates.
|
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] |
PMID:19191
|
Evidence that 'inactive' renin is produced outside the kidney of the rat.
|
1. Inactive renin, which can be converted into an active form by acidification to pH 3-3, represents 4-63% of the total renin in rat peripheral plasma. 2. No inactive component could be found in the blood-free venous effluent of the perfused rat kidney before and after stimulation with isoprenaline. 3. This suggests a possible extrarenal source for inactive renin and may explain the presence of this component in anephric patients.
|
Evidence that 'inactive' renin is produced outside the kidney of the rat. 1. Inactive renin, which can be converted into an active form by acidification to pH 3-3, represents 4-63% of the total renin in rat peripheral plasma. 2. No inactive component could be found in the blood-free venous effluent of the perfused rat kidney before and after stimulation with isoprenaline. 3. This suggests a possible extrarenal source for inactive renin and may explain the presence of this component in anephric patients.
|
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] |
PMID:19200
|
Studies of neurotransmitter release in the pathogenesis of hypertension.
|
Studies using a sensitive radioenzymatic assay for plasma noradrenaline suggest there is a selective overactivity of the sympathetic nervous system in essential hypertension. Methodology which allows the study of local sympathetic turnover in CNS nuclei and peripheral blood vessels is described. This approach has been used to study the non-innervated sympathetic turnover phaeochromocytoma. It is suggested that studies of local regulatory mechanism in neurotransmitter release are required to give a greater understanding of the central and peripheral role of the sympathetic nervous system in the pathogenesis of hypertension.
|
Studies of neurotransmitter release in the pathogenesis of hypertension. Studies using a sensitive radioenzymatic assay for plasma noradrenaline suggest there is a selective overactivity of the sympathetic nervous system in essential hypertension. Methodology which allows the study of local sympathetic turnover in CNS nuclei and peripheral blood vessels is described. This approach has been used to study the non-innervated sympathetic turnover phaeochromocytoma. It is suggested that studies of local regulatory mechanism in neurotransmitter release are required to give a greater understanding of the central and peripheral role of the sympathetic nervous system in the pathogenesis of hypertension.
|
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] |
PMID:19202
|
Hypothalamic action of adrenoreceptor blocking agents.
|
In cats anaesthetized with pentobarbital sodium, the posterior hypothalamus was superfused and electrically stimulated with a push-pull cannula. The pressor response to stimulation of this hypothalamic area was inhibited when the hypothalamus was superfused with drugs blocking either alpha-adrenoreceptors (piperoxan, tolazoline), or beta-adrenoreceptors--(+/-)-propranolol, (-)-propranolol, practolol, sotalol, metoprolol. (+)-Propranolol and a concentration of procaine equianaesthetic to propranolol were ineffective. During superfusion with tolazoline in the presence of practolol the inhibition was twice as that when the hypothalamus was superfused with either tolazoline or practolol. In another series of experiments the push-pull cannula was inserted into the anterior hypothalamus. The depressor response to stimulation of this area was inhibited by the hypothalamic superfusion with the alpha-adrenoreceptor blocking drugs phentolamine, tolazoline, piperoxan or yohimbine. Hypothalamic superfusion with phenylephrine abolished the inhibitory effect of phentolamine on the depressor response. The results indicate that adrenoreceptors are present in the hypothalamus and that they are involved in blood pressure changes elicited by hypothalamic stimulation.
|
Hypothalamic action of adrenoreceptor blocking agents. In cats anaesthetized with pentobarbital sodium, the posterior hypothalamus was superfused and electrically stimulated with a push-pull cannula. The pressor response to stimulation of this hypothalamic area was inhibited when the hypothalamus was superfused with drugs blocking either alpha-adrenoreceptors (piperoxan, tolazoline), or beta-adrenoreceptors--(+/-)-propranolol, (-)-propranolol, practolol, sotalol, metoprolol. (+)-Propranolol and a concentration of procaine equianaesthetic to propranolol were ineffective. During superfusion with tolazoline in the presence of practolol the inhibition was twice as that when the hypothalamus was superfused with either tolazoline or practolol. In another series of experiments the push-pull cannula was inserted into the anterior hypothalamus. The depressor response to stimulation of this area was inhibited by the hypothalamic superfusion with the alpha-adrenoreceptor blocking drugs phentolamine, tolazoline, piperoxan or yohimbine. Hypothalamic superfusion with phenylephrine abolished the inhibitory effect of phentolamine on the depressor response. The results indicate that adrenoreceptors are present in the hypothalamus and that they are involved in blood pressure changes elicited by hypothalamic stimulation.
|
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0.024876320734620094,
0.06902710348367691
] |
PMID:19203
|
Catecholamines, cyclic AMP and renin in two contrasting forms of essential hypertension.
|
Essential hypertension (EH) can be subdivided according to the sympathetic and renin activity into two contrasting forms: (1) borderline beta-hyperadrenergic renin hyperresponsive and (2) stable beta-hypoadrenergic renin hyporesponsive EH. These two forms probably represent two expreme poles in the spectrum of EH in which sympathetic and renin hyper- or hyporeactivity cannot be accounted for by catecholamine determinations solely. beta-Adrenergic responsiveness monitored by plasma cyclic AMP determinations revealed plasma cyclic AMP, renin and circulatory hyperresponsiveness to isoproterenol in borderline hyperadrenergic EH while the opposite, cyclic AMP and renin hyporesponsiveness to insulin-induced hypoglycemia have been described in low renin stable EH. The kidney is in the center of the adrenergic abnormality in the two forms of EH with the borderline one excreting into the urine catecholamines not accounted for by their glomerular filtration. Catecholamines solely, however, do not account for the differences in both forms of EH which can probably be attributed to their different beta-adrenergic responsiveness.
|
Catecholamines, cyclic AMP and renin in two contrasting forms of essential hypertension. Essential hypertension (EH) can be subdivided according to the sympathetic and renin activity into two contrasting forms: (1) borderline beta-hyperadrenergic renin hyperresponsive and (2) stable beta-hypoadrenergic renin hyporesponsive EH. These two forms probably represent two expreme poles in the spectrum of EH in which sympathetic and renin hyper- or hyporeactivity cannot be accounted for by catecholamine determinations solely. beta-Adrenergic responsiveness monitored by plasma cyclic AMP determinations revealed plasma cyclic AMP, renin and circulatory hyperresponsiveness to isoproterenol in borderline hyperadrenergic EH while the opposite, cyclic AMP and renin hyporesponsiveness to insulin-induced hypoglycemia have been described in low renin stable EH. The kidney is in the center of the adrenergic abnormality in the two forms of EH with the borderline one excreting into the urine catecholamines not accounted for by their glomerular filtration. Catecholamines solely, however, do not account for the differences in both forms of EH which can probably be attributed to their different beta-adrenergic responsiveness.
|
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] |
PMID:19206
|
Suppression of reactions to certain cosmetics.
|
Reactions to hair dyes and bleaches may be "suppressed" with corticosteroids and antihistamines. Reactions to nail polish may be prevented by a "drying" or "polymerizing" technique. Sensitization to certain perfume ingredients may be inhibited by a "quenching" phenomenon.
|
Suppression of reactions to certain cosmetics. Reactions to hair dyes and bleaches may be "suppressed" with corticosteroids and antihistamines. Reactions to nail polish may be prevented by a "drying" or "polymerizing" technique. Sensitization to certain perfume ingredients may be inhibited by a "quenching" phenomenon.
|
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] |
PMID:19208
|
Arteriosclerotic aneurysm of the coronary artery.
|
The findings in a patient with an angiographically proven aneurysm of the coronary artery are described. The case is reviewed in the light of 115 similar cases reported in the literature. The patient had had numerous episodes of variant angina, a feature not previously described in coronary arterial aneurysms, which may be related to embolic showers originating from the aneurysm.
|
Arteriosclerotic aneurysm of the coronary artery. The findings in a patient with an angiographically proven aneurysm of the coronary artery are described. The case is reviewed in the light of 115 similar cases reported in the literature. The patient had had numerous episodes of variant angina, a feature not previously described in coronary arterial aneurysms, which may be related to embolic showers originating from the aneurysm.
|
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] |
PMID:19205
|
Treatment of rheumatoid arthritis with tolmetin: a comparison with alclofenac.
|
A double-blind between-patient trial over 6 weeks to compare the effects of 1.6 g tolmetin daily with 4 g alclofenac daily was completed by 37 out of 44 out-patients with rheumatoid arthritis. Tolmetin proved as effective as alclofenac in relieving pain and in reducing both the articular index score and the number of inflamed joints. However, alclofenac produced a significantly greater reduction in the duration of morning stiffness, which could have been related to the timing of the initial daily dose of tolmetin. Onset of fatigue was significantly improved by both drugs. Neither treatment group exhibited any significant changes in the serum levels of IgG, IgA, and IgM, nor in the latex and sheep-cell agglutination titres. Although neither drug gave rise to serious side-effects, 3 patients (2 on tolmetin, 1 on alclofenac) were withdrawn because of skin rash, 2 (on tolmetin) because of gastro-intestinal upsets, and 1 from each group because of lack of analgesic effects.
|
Treatment of rheumatoid arthritis with tolmetin: a comparison with alclofenac. A double-blind between-patient trial over 6 weeks to compare the effects of 1.6 g tolmetin daily with 4 g alclofenac daily was completed by 37 out of 44 out-patients with rheumatoid arthritis. Tolmetin proved as effective as alclofenac in relieving pain and in reducing both the articular index score and the number of inflamed joints. However, alclofenac produced a significantly greater reduction in the duration of morning stiffness, which could have been related to the timing of the initial daily dose of tolmetin. Onset of fatigue was significantly improved by both drugs. Neither treatment group exhibited any significant changes in the serum levels of IgG, IgA, and IgM, nor in the latex and sheep-cell agglutination titres. Although neither drug gave rise to serious side-effects, 3 patients (2 on tolmetin, 1 on alclofenac) were withdrawn because of skin rash, 2 (on tolmetin) because of gastro-intestinal upsets, and 1 from each group because of lack of analgesic effects.
|
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] |
PMID:19211
|
Metabolism and disposition of l-alpha-acetylmethadol in the rat.
|
The metabolism and disposition of the long-acting narcotic analygesic l-alpha-acetylmethadol (LAAM) were studied in the rat. 3H-LAAM was administered to male and female rats at doses of 5 mg/kg po and iv, and 10 mg/kg po. LAAM was rapidly absorbed and extensively metabolized. Five metabolites-noracetylmethadol, dinoracetylmethadol, methadol, normethadol, and N-acetylnormethadol-were identified in plasma and urine. Feces were the major route of elimination for the parent drug and metabolites. Less than 20% of the administered dose was excreted in the urine and, of this, greater than 90% was in the form of conjugates or polar metabolites. There is an apparent sex-related difference in LAAM disposition in the rat. LAAM and metabolites tended to persist in higher levels in female rats as compared with male rats. Similarly, male rats tended to excrete the drug at a faster rate than did females.
|
Metabolism and disposition of l-alpha-acetylmethadol in the rat. The metabolism and disposition of the long-acting narcotic analygesic l-alpha-acetylmethadol (LAAM) were studied in the rat. 3H-LAAM was administered to male and female rats at doses of 5 mg/kg po and iv, and 10 mg/kg po. LAAM was rapidly absorbed and extensively metabolized. Five metabolites-noracetylmethadol, dinoracetylmethadol, methadol, normethadol, and N-acetylnormethadol-were identified in plasma and urine. Feces were the major route of elimination for the parent drug and metabolites. Less than 20% of the administered dose was excreted in the urine and, of this, greater than 90% was in the form of conjugates or polar metabolites. There is an apparent sex-related difference in LAAM disposition in the rat. LAAM and metabolites tended to persist in higher levels in female rats as compared with male rats. Similarly, male rats tended to excrete the drug at a faster rate than did females.
|
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] |
PMID:19212
|
The metabolism of p-methoxyamphetamine in dog and monkey. O-demethylation as a major route.
|
p-Hydroxyamphetamine (PHA) and p-hydroxybenzoic acid (PHBA) have been isolated by gas-liquid chromatography (GC) as urinary metabolites of p-methoxyamphetamine (PMA) in dogs and rhesus monkeys. The amounts of PMA excreted unchanged in the total 24-hr urine samples in dogs and monkeys were 26% and 3%, respectively, of the total administered doses. The amounts of the pharmacologically active metabolite PHA excreted in 24-hr urine samples were 13% in the dog and 44% in the monkey. In the dog, 72% of the PHA was present as free (unconjugated) PHA, whereas in the monkey the amount of PHA present in the free form was only 14%.
|
The metabolism of p-methoxyamphetamine in dog and monkey. O-demethylation as a major route. p-Hydroxyamphetamine (PHA) and p-hydroxybenzoic acid (PHBA) have been isolated by gas-liquid chromatography (GC) as urinary metabolites of p-methoxyamphetamine (PMA) in dogs and rhesus monkeys. The amounts of PMA excreted unchanged in the total 24-hr urine samples in dogs and monkeys were 26% and 3%, respectively, of the total administered doses. The amounts of the pharmacologically active metabolite PHA excreted in 24-hr urine samples were 13% in the dog and 44% in the monkey. In the dog, 72% of the PHA was present as free (unconjugated) PHA, whereas in the monkey the amount of PHA present in the free form was only 14%.
|
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] |
PMID:19213
|
Determination of optically active thiopental, thiamylal, and their metabolites in human urine.
|
Methods involving solvent extraction and gas-liquid chromatography have been developed for quantitative determination of certain thiobarbiturates and their major metabolites in urine. The structures of the metabolites identified were confirmed by the use of gas chromatography/mass spectrometry and 13C-nuclear magnetic resonance spectroscopy. The present analytic methods were applied to the quantitative determination of (R)-(+)- and (S)-(-)-thiopental as well as (R)-(+)- and (S)-(-)-thiamylal and their major metabolites excreted in human urine. Minor metabolites were isolated and identified.
|
Determination of optically active thiopental, thiamylal, and their metabolites in human urine. Methods involving solvent extraction and gas-liquid chromatography have been developed for quantitative determination of certain thiobarbiturates and their major metabolites in urine. The structures of the metabolites identified were confirmed by the use of gas chromatography/mass spectrometry and 13C-nuclear magnetic resonance spectroscopy. The present analytic methods were applied to the quantitative determination of (R)-(+)- and (S)-(-)-thiopental as well as (R)-(+)- and (S)-(-)-thiamylal and their major metabolites excreted in human urine. Minor metabolites were isolated and identified.
|
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] |
PMID:19214
|
The interrelationship between the metabolism of (S)-continine-N-oxide and (S)-cotinine.
|
The metabolism of (S)-cotinine-N-oxide was studied in the rabbit and the dog. The pattern of Koenig-positive substances in the urine of the animals suggested the presence of cotinine, demethylcotinine, hydroxycotinine, and allohydroxycotinine, compounds already previously identified as metabolites of (S)-cotinine and (S)-nicotine in many mammalian species. In the dog, 34% of the administered oral dose of (S)-cotinine-N-oxide was recovered from the urine, and 21% was recovered from the urine of the rabbit. Confirmation of the presence of (S)-cotinine, (S)-demethulcotinine, hydroxycotinine, and allohydroxycotinine in the urine of the rabbits was obtained by isolation of the metabolites as themselves of as derivatives. The data, although establishing the possibilities of an intermediary role for (S)-cotinine-N-oxide in the metabolism of nicotine, do not clearly indicate whether the metabolites such as demethylcotinine arise via the route (S)-cotinine-N-oxide leads to (S)-cotinine leads to (S)-demethylcotinine or via the alternate route (S)-cotinine-N-oxide leads to (S)-demethylcotinine-N-oxide leads to (S)-demethylcotinine.
|
The interrelationship between the metabolism of (S)-continine-N-oxide and (S)-cotinine. The metabolism of (S)-cotinine-N-oxide was studied in the rabbit and the dog. The pattern of Koenig-positive substances in the urine of the animals suggested the presence of cotinine, demethylcotinine, hydroxycotinine, and allohydroxycotinine, compounds already previously identified as metabolites of (S)-cotinine and (S)-nicotine in many mammalian species. In the dog, 34% of the administered oral dose of (S)-cotinine-N-oxide was recovered from the urine, and 21% was recovered from the urine of the rabbit. Confirmation of the presence of (S)-cotinine, (S)-demethulcotinine, hydroxycotinine, and allohydroxycotinine in the urine of the rabbits was obtained by isolation of the metabolites as themselves of as derivatives. The data, although establishing the possibilities of an intermediary role for (S)-cotinine-N-oxide in the metabolism of nicotine, do not clearly indicate whether the metabolites such as demethylcotinine arise via the route (S)-cotinine-N-oxide leads to (S)-cotinine leads to (S)-demethylcotinine or via the alternate route (S)-cotinine-N-oxide leads to (S)-demethylcotinine-N-oxide leads to (S)-demethylcotinine.
|
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] |
PMID:19215
|
The metabolism of styrene oxide in the isolated perfused rat liver. Identification and quantitation of major metabolites.
|
Isolated perfused rat livers rapidly metabolized 14C-styrene oxide. A large proportion of the administered radioactivity was excreted in the bile as a single compound that was identified as S-(1-phenyl-2-hydroxyethyl)glutathione by comparison with an authentic synthetic standard. The circulating perfusate was found to contain approximately equal amounts of styrene glycol, mandelic acid, and the glutathione derivative.
|
The metabolism of styrene oxide in the isolated perfused rat liver. Identification and quantitation of major metabolites. Isolated perfused rat livers rapidly metabolized 14C-styrene oxide. A large proportion of the administered radioactivity was excreted in the bile as a single compound that was identified as S-(1-phenyl-2-hydroxyethyl)glutathione by comparison with an authentic synthetic standard. The circulating perfusate was found to contain approximately equal amounts of styrene glycol, mandelic acid, and the glutathione derivative.
|
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0.07833948731422424,
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] |
PMID:19216
|
Biliary excretion of iopanoate glucuronide by the rat.
|
Biliary excretion, plasma decay, and tissue distribution of 125I after a single intravenous dose (13 mumol/kg) of 125I-Iopanoate and 125I-iopanoate glucuronide were compared in control and phenobarbital-pretreated rats under either anesthesia. In control animals the plasma decay of iopanoate glucuronide following iopanoate glucuronide administration was significantly greater than that after iopanoate administration. Biliary clearance following iopanoate glucuronide was approximately 20 times greater than that after iopanoate; within 60 min, 82% of the dose of iopanoate glucuronide, compared to 23% of the dose of iopanoate, was recovered in bile. Phenobarbital pretreatment significantly increased the biliary excretion following iopanoate administration but had no effect on that following iopanoate glucuronide. The urinary excretion after both agents was less than 3% of the administered dose within the 60-min experimental period. The data suggest that intracellular metabolism and/or binding may account for the marked difference in the biliary excretion of iopanoate glucuronide following iopanoate as compared to that after administration of iopanoate glucuronide.
|
Biliary excretion of iopanoate glucuronide by the rat. Biliary excretion, plasma decay, and tissue distribution of 125I after a single intravenous dose (13 mumol/kg) of 125I-Iopanoate and 125I-iopanoate glucuronide were compared in control and phenobarbital-pretreated rats under either anesthesia. In control animals the plasma decay of iopanoate glucuronide following iopanoate glucuronide administration was significantly greater than that after iopanoate administration. Biliary clearance following iopanoate glucuronide was approximately 20 times greater than that after iopanoate; within 60 min, 82% of the dose of iopanoate glucuronide, compared to 23% of the dose of iopanoate, was recovered in bile. Phenobarbital pretreatment significantly increased the biliary excretion following iopanoate administration but had no effect on that following iopanoate glucuronide. The urinary excretion after both agents was less than 3% of the administered dose within the 60-min experimental period. The data suggest that intracellular metabolism and/or binding may account for the marked difference in the biliary excretion of iopanoate glucuronide following iopanoate as compared to that after administration of iopanoate glucuronide.
|
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] |
PMID:19218
|
A preliminary pharmacokinetic model for several chlorinated biphenyls in the rat.
|
A mathematical model is presented to describe the kinetics of distribution, metabolism, and excretion of 4-chloro-, 4,4'-dichloro-, 2,2',4,5,5'-pentachloro-, and 2,2',4,4',5,5'-hexachlorobiphenyl in rats given an intravenols dose of 0.6 mg/kg. A modified flow-limited model simulates the penta- and hexachlorobiphenyl data for periods up to 96 hr but underestimates the mono- and dichlorobiphenyl data beyond 48 hr. The rate constant for metabolism by the liver decreases as degree of chlorination increases such that the rate constant is 200 times smaller for the hexachlorobiphenyl than for the monochlorobiphenyl. The value of the biliary clearance of metabolites is nearly the same for each chlorinated biphenyl, whereas the value of the urinary clearance decreases with increasing degree of chlorination, being 10 times smaller for hexachlorobiphenyl than for monochlorophenyl. The distribution coefficients between most tissues and blood are larger for each parent compound than for its metabolites. Hexachlorobiphenyl has the largest distribution coefficient of all the chlorinated biphenyls in each tissue, whereas the mono-, di-, and pentachlorobiphenyls show no consistent variation. For each compound the distribution coefficient is greater in the fat than in any other tissue. Changes in the fat volume of the growing rats were incorporated into the model in order to simulate the hexachlorobiphenyl concentrations in blood and fat for 42 days.
|
A preliminary pharmacokinetic model for several chlorinated biphenyls in the rat. A mathematical model is presented to describe the kinetics of distribution, metabolism, and excretion of 4-chloro-, 4,4'-dichloro-, 2,2',4,5,5'-pentachloro-, and 2,2',4,4',5,5'-hexachlorobiphenyl in rats given an intravenols dose of 0.6 mg/kg. A modified flow-limited model simulates the penta- and hexachlorobiphenyl data for periods up to 96 hr but underestimates the mono- and dichlorobiphenyl data beyond 48 hr. The rate constant for metabolism by the liver decreases as degree of chlorination increases such that the rate constant is 200 times smaller for the hexachlorobiphenyl than for the monochlorobiphenyl. The value of the biliary clearance of metabolites is nearly the same for each chlorinated biphenyl, whereas the value of the urinary clearance decreases with increasing degree of chlorination, being 10 times smaller for hexachlorobiphenyl than for monochlorophenyl. The distribution coefficients between most tissues and blood are larger for each parent compound than for its metabolites. Hexachlorobiphenyl has the largest distribution coefficient of all the chlorinated biphenyls in each tissue, whereas the mono-, di-, and pentachlorobiphenyls show no consistent variation. For each compound the distribution coefficient is greater in the fat than in any other tissue. Changes in the fat volume of the growing rats were incorporated into the model in order to simulate the hexachlorobiphenyl concentrations in blood and fat for 42 days.
|
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] |
PMID:19219
|
Studies on the glucuronidation of 7-hydroxychlorpromazine in vitro.
|
The glucuronide of 7-hydroxychlorpromazine (CPOH), formed by incubation with hepatic microsomes and UDP-[U-14C]glucuronic acid, was isolated by extraction into 1-butanol at pH 1 and assayed by liquid scintillation counting. Although concentrations of CPOH greater than 3 X 10(-4) M stimulated its conjugation, the apparent KM of the transferase in microsomes from guinea pig liver for CPOH was 9.5 X 10(-5) M. The apparent activation of glucuronyltransferase by high concentrations of CPOH was prevented when preparations were maximmally stimulated by Triton X-100. The rate of conjugation of CPOH by preparations of guinea pig liver was almost twice that measured with those of rat liver. However, the maximal stimulation of the conjugation of CPOH by Triton with preparations of rat liver was almost twice that observed with microsomes from guinea pig liver. The conjugation of CPOH by microsomes from rat and guinea pig liver was inhibited about 25% by SKF 525-A (1 X 10(-4) M). The inhibitory action of SKF 525-A decreased at concentrations greater than 5 X 10(-4) M and no inhibition was observed at 1 X 10(-3) M.
|
Studies on the glucuronidation of 7-hydroxychlorpromazine in vitro. The glucuronide of 7-hydroxychlorpromazine (CPOH), formed by incubation with hepatic microsomes and UDP-[U-14C]glucuronic acid, was isolated by extraction into 1-butanol at pH 1 and assayed by liquid scintillation counting. Although concentrations of CPOH greater than 3 X 10(-4) M stimulated its conjugation, the apparent KM of the transferase in microsomes from guinea pig liver for CPOH was 9.5 X 10(-5) M. The apparent activation of glucuronyltransferase by high concentrations of CPOH was prevented when preparations were maximmally stimulated by Triton X-100. The rate of conjugation of CPOH by preparations of guinea pig liver was almost twice that measured with those of rat liver. However, the maximal stimulation of the conjugation of CPOH by Triton with preparations of rat liver was almost twice that observed with microsomes from guinea pig liver. The conjugation of CPOH by microsomes from rat and guinea pig liver was inhibited about 25% by SKF 525-A (1 X 10(-4) M). The inhibitory action of SKF 525-A decreased at concentrations greater than 5 X 10(-4) M and no inhibition was observed at 1 X 10(-3) M.
|
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] |
PMID:19220
|
Effect of experimental diabetes on drug metabolism in the rat.
|
The effect of experimental diabetes on hepatic drug metabolism was studied in male Holtzman rats. Treatment of animals with streptozotocin and 6-aminonicotinamide, both agents which produce an insulin-deficient animal, caused prolongation of hexobarbital sleeping times and inhibition of the rate of metabolism of both hexobarbital and, to a lesser extent, aniline in vitro. Treatment of animals with N-methylacetamide, a diabetogen which does not cause insulin deficiency in the animal but rather produces an insulin-resistant state, did not affect the metabolism in vitro of either hexobarbital or aniline. Neither insulin nor any of the diabetogenic agents had any direct effect on drug metabolism in vitro. Furthermore, hepatic microsomal protein and cytochrome P-450 contents were not significantly different in any of the diabetic animals from those of the control animals. Hyperglycemia produced by glucose infusion did not affect the metabolism of hexobarbital in vitro. The effects of streptozotocin and 6-aminonicotinamide appeared to be at least partially due to the presence of an inhibitor in the liver cytosol which correlated with elevated hepatic cyclic AMP concentrations.
|
Effect of experimental diabetes on drug metabolism in the rat. The effect of experimental diabetes on hepatic drug metabolism was studied in male Holtzman rats. Treatment of animals with streptozotocin and 6-aminonicotinamide, both agents which produce an insulin-deficient animal, caused prolongation of hexobarbital sleeping times and inhibition of the rate of metabolism of both hexobarbital and, to a lesser extent, aniline in vitro. Treatment of animals with N-methylacetamide, a diabetogen which does not cause insulin deficiency in the animal but rather produces an insulin-resistant state, did not affect the metabolism in vitro of either hexobarbital or aniline. Neither insulin nor any of the diabetogenic agents had any direct effect on drug metabolism in vitro. Furthermore, hepatic microsomal protein and cytochrome P-450 contents were not significantly different in any of the diabetic animals from those of the control animals. Hyperglycemia produced by glucose infusion did not affect the metabolism of hexobarbital in vitro. The effects of streptozotocin and 6-aminonicotinamide appeared to be at least partially due to the presence of an inhibitor in the liver cytosol which correlated with elevated hepatic cyclic AMP concentrations.
|
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] |
PMID:19227
|
[The effect of various front tooth filling materials on the in vitro growth of Streptococcus mutans].
|
While silicate cement inhibits bacterial growth, resin materials promote it immediately after mixing. Toxic tissue reactions may be induced by bacterial plaque accumulation. Our testing method complements existing in-vitro tests for recording chemo-toxic activities.
|
[The effect of various front tooth filling materials on the in vitro growth of Streptococcus mutans]. While silicate cement inhibits bacterial growth, resin materials promote it immediately after mixing. Toxic tissue reactions may be induced by bacterial plaque accumulation. Our testing method complements existing in-vitro tests for recording chemo-toxic activities.
|
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] |
PMID:19228
|
[Effect of beta-adrenergic blockaders on the uterine musculature of rats].
|
The authors carried out a study on beta-adrenergic blockers Proprafnolol, Pindolol and Practolol, on an isolated uterus of a rat, obtained at various phases oestrus. The Beta-adrenergic blocker Pindolol manifested inhibiting effect on the spontaneous contractility of the uterus more marked after elevated level of gestagens. The pharmacological analysis gave foundation to the authors to assume that the relaxation of Pindolol was due to direct myotropic action. Propranolol in large doses and at high gestagenic level manifested antioxytocic activity. Practolol did not affect both spontaneous uterine activity as oxytoxin contraction.
|
[Effect of beta-adrenergic blockaders on the uterine musculature of rats]. The authors carried out a study on beta-adrenergic blockers Proprafnolol, Pindolol and Practolol, on an isolated uterus of a rat, obtained at various phases oestrus. The Beta-adrenergic blocker Pindolol manifested inhibiting effect on the spontaneous contractility of the uterus more marked after elevated level of gestagens. The pharmacological analysis gave foundation to the authors to assume that the relaxation of Pindolol was due to direct myotropic action. Propranolol in large doses and at high gestagenic level manifested antioxytocic activity. Practolol did not affect both spontaneous uterine activity as oxytoxin contraction.
|
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] |
PMID:19229
|
[EEG-restitution and brain metabolism during electroconvulsive treatment in relaxation (author's transl)].
|
Blood-gas (pO2, pCO2) and pH-changes of venous (v.jugularis interna) and arterial (A.femoralis) blood samples, furthermore glucose utilization and lactate-, pyruvate-production of brain were investigated during electroconvulsive treatment in relaxation of 45 psychotic patients. The blood-gas values and substrate concentrations were statistically evaluated and represented in a function of the characteristic phases of the postconvulsive EEG-activity. A correlation was found between the glucose metabolism of the brain and the postconvulsive recovery of EEG. The restitution of postconvulsive brain metabolism runs discontinuously in the first 12 minutes of postconvulsive state. In the phase of electric silence and periodic delta-waves the brain metabolism was shifted to anaerobic direction. During the treatment no anoxic anoxia or acidosis takes place during the seizure activity and restitution, the measurable metabolic changes are moderate and supposedly play no important role in the "effect" of treatment.
|
[EEG-restitution and brain metabolism during electroconvulsive treatment in relaxation (author's transl)]. Blood-gas (pO2, pCO2) and pH-changes of venous (v.jugularis interna) and arterial (A.femoralis) blood samples, furthermore glucose utilization and lactate-, pyruvate-production of brain were investigated during electroconvulsive treatment in relaxation of 45 psychotic patients. The blood-gas values and substrate concentrations were statistically evaluated and represented in a function of the characteristic phases of the postconvulsive EEG-activity. A correlation was found between the glucose metabolism of the brain and the postconvulsive recovery of EEG. The restitution of postconvulsive brain metabolism runs discontinuously in the first 12 minutes of postconvulsive state. In the phase of electric silence and periodic delta-waves the brain metabolism was shifted to anaerobic direction. During the treatment no anoxic anoxia or acidosis takes place during the seizure activity and restitution, the measurable metabolic changes are moderate and supposedly play no important role in the "effect" of treatment.
|
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] |
PMID:19230
|
[Comparative study using double-blind method of sultopride and thioproperazine].
|
This report includes a statistical analysis of the results of a double blind comparative study between sultopride and thioproperazine. Two rating scales were used: the standard B.P.R.S. and a simplified scale including 7 items: agitation--delusion--thought-disorganization--anxiety--aggressivity--hallucinations--autism. The comparison of the mean of the global scores obtained with the two rating scales shows a significant difference in activity in favour of sultopride. The analysis of the individual items shows regular modifications in favour of sultopride but these are not statistically significant. There are, also, no differences in the side-effects observed, particularly extra-pyramidal effects.
|
[Comparative study using double-blind method of sultopride and thioproperazine]. This report includes a statistical analysis of the results of a double blind comparative study between sultopride and thioproperazine. Two rating scales were used: the standard B.P.R.S. and a simplified scale including 7 items: agitation--delusion--thought-disorganization--anxiety--aggressivity--hallucinations--autism. The comparison of the mean of the global scores obtained with the two rating scales shows a significant difference in activity in favour of sultopride. The analysis of the individual items shows regular modifications in favour of sultopride but these are not statistically significant. There are, also, no differences in the side-effects observed, particularly extra-pyramidal effects.
|
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] |
PMID:19231
|
[A new psychotropic drug: carpipramine, intermediate compound between 2 therapeutic classes].
|
An open clinical study was conducted with carpipramine in 100 hospitalized subjects presenting various mental disorders. The therapeutic results on symptoms were assessed both as a whole and with the help of a rating scale. Doses varied from 50 to 400 mg per day. Carpipramine seems to be particularly efficient on schizophrenias, 66 cases of which were tested. The best results were observed in hebephrenic forms and depressive syndroms during the illness; in these indications, carpipramine exerts a clear psychomotor stimulating activity which is useful in decreasing indifference, apathy and ideomotor slowness. Schizophrenias with paranoid delusions or depersonalization anxiety tend to be somehow aggravated. Carpipramine does not seem to be a true antidepressant despite its desinhibitory properties. The compound proves useful in deficits of the psychomotor tone such as those occuring in psychasthenia or the deficit syndrom which follows withdrawal from opiates. Clinical and biological tolerances seem to be excellent and extrapyramidal side effects are exceptional. Carpipramine may be considered as a strongly desinhibitory neuroleptic agent which bears some resemblance to antidepressants because of its psychoanaleptic effect. The authors raise the question of possible antipsychotic properties in higher doses in relation to pharmacological data and a bipolar, antipsychotic and predominantly desinhibitory, therapeutic action.
|
[A new psychotropic drug: carpipramine, intermediate compound between 2 therapeutic classes]. An open clinical study was conducted with carpipramine in 100 hospitalized subjects presenting various mental disorders. The therapeutic results on symptoms were assessed both as a whole and with the help of a rating scale. Doses varied from 50 to 400 mg per day. Carpipramine seems to be particularly efficient on schizophrenias, 66 cases of which were tested. The best results were observed in hebephrenic forms and depressive syndroms during the illness; in these indications, carpipramine exerts a clear psychomotor stimulating activity which is useful in decreasing indifference, apathy and ideomotor slowness. Schizophrenias with paranoid delusions or depersonalization anxiety tend to be somehow aggravated. Carpipramine does not seem to be a true antidepressant despite its desinhibitory properties. The compound proves useful in deficits of the psychomotor tone such as those occuring in psychasthenia or the deficit syndrom which follows withdrawal from opiates. Clinical and biological tolerances seem to be excellent and extrapyramidal side effects are exceptional. Carpipramine may be considered as a strongly desinhibitory neuroleptic agent which bears some resemblance to antidepressants because of its psychoanaleptic effect. The authors raise the question of possible antipsychotic properties in higher doses in relation to pharmacological data and a bipolar, antipsychotic and predominantly desinhibitory, therapeutic action.
|
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] |
PMID:19232
|
[Psychosensorial manifestations observed in psychotic patients treated with betamimetic drugs].
|
We report four observations of women who had acute or subacute psychotic crisis and who were stabilized without any manifestation. In order to delay delivery these women received during pre-partum high doses of beta-mimetic drugs (isoxsuprine and salbutamol). Such a prescription seems to have been responsible of acute and transient psychotic symptoms similar to those observed in drug-induced psychosis (derealisation preceeding depersonalization, visual and auditory hallucinations, anxiety). Recently Gluckman has described a similar case to those reported here. But in his case the conditions of coming out were different and there was a lack of psychotic antecedents. Previous works concerning "psychotic-like" effects of beta-blokers rise the question of possible psychodysleptic effects of beta-mimetic drugs.
|
[Psychosensorial manifestations observed in psychotic patients treated with betamimetic drugs]. We report four observations of women who had acute or subacute psychotic crisis and who were stabilized without any manifestation. In order to delay delivery these women received during pre-partum high doses of beta-mimetic drugs (isoxsuprine and salbutamol). Such a prescription seems to have been responsible of acute and transient psychotic symptoms similar to those observed in drug-induced psychosis (derealisation preceeding depersonalization, visual and auditory hallucinations, anxiety). Recently Gluckman has described a similar case to those reported here. But in his case the conditions of coming out were different and there was a lack of psychotic antecedents. Previous works concerning "psychotic-like" effects of beta-blokers rise the question of possible psychodysleptic effects of beta-mimetic drugs.
|
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] |
PMID:19233
|
Estradiol binding capacity in the cryptorchid rat testis.
|
Mature male Sprague-Dawley rats were made surgically cryptorchid at 47--50 days of age. Animals were sacrificed as controls and at 7, 14, and 21 days post-surgery. The testes were removed and utilized to determine the effects of cryptorchidism on tissue weight loss, histological changes, and cytoplasmic estradiol binding capacity. Testis weight decreased from a mean +/- SE of 1.1 +/- 0.09 to .41 +/- 0.02 g during the 21 day period, a 66% decrease in weight. Light microscopy revealed that the process of spermatogenesis was inhibited, the germinal epithelium had degenerated, and the tubules had become smaller in diameter. The interstitial tissue volume progressively increased through day 21 post-surgery. The cytoplasmic estradiol binding capacity, expressed as fmoles [3H]-estradiol/mg cytosol protein, increased markedly from a control of 16 +/- 1.1 to 36 +/- 8.0 at day 7 after surgery; this represented a 126% increase in binding capacity in one week. Binding capacity was equivalent to 54 +/- 7.4 and 58 +/- 10.5 fmol/mg on days 14 and 21 post-surgery, respectively. Testicular content of cytoplasmic estrogen receptors was shown to increase from 349 to a maximum of 575 fmoles/testis at day 7. Receptor content then declined slowly to a value of 481 at day 21. Testicular content of receptor is presumed to reflect an alteration in the Leydig cells induced by cryptorchidism.
|
Estradiol binding capacity in the cryptorchid rat testis. Mature male Sprague-Dawley rats were made surgically cryptorchid at 47--50 days of age. Animals were sacrificed as controls and at 7, 14, and 21 days post-surgery. The testes were removed and utilized to determine the effects of cryptorchidism on tissue weight loss, histological changes, and cytoplasmic estradiol binding capacity. Testis weight decreased from a mean +/- SE of 1.1 +/- 0.09 to .41 +/- 0.02 g during the 21 day period, a 66% decrease in weight. Light microscopy revealed that the process of spermatogenesis was inhibited, the germinal epithelium had degenerated, and the tubules had become smaller in diameter. The interstitial tissue volume progressively increased through day 21 post-surgery. The cytoplasmic estradiol binding capacity, expressed as fmoles [3H]-estradiol/mg cytosol protein, increased markedly from a control of 16 +/- 1.1 to 36 +/- 8.0 at day 7 after surgery; this represented a 126% increase in binding capacity in one week. Binding capacity was equivalent to 54 +/- 7.4 and 58 +/- 10.5 fmol/mg on days 14 and 21 post-surgery, respectively. Testicular content of cytoplasmic estrogen receptors was shown to increase from 349 to a maximum of 575 fmoles/testis at day 7. Receptor content then declined slowly to a value of 481 at day 21. Testicular content of receptor is presumed to reflect an alteration in the Leydig cells induced by cryptorchidism.
|
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] |
PMID:19235
|
Possible role of calcium-dependent protein phosphorylation in mediating neurotransmitter release and anticonvulsant action.
|
The results demonstrate that calcium and phenytoin have antagonistic actions on the net level of endogenous phosphorylation of specific rat brain proteins. These specific phosphoproteins were shown to be present in synaptosome preparations, and evidence is presented for the localization of these phosphoproteins within the presynaptic nerve terminal. The data are compatible with the hypothesis that the antagonistic actions of calcium and phenytoin on the phosphorylation of synaptosome proteins may be the underlying molecular mechanism mediating the opposing actions of these agents on the release of neurotransmitter from the presynaptic nerve terminal.
|
Possible role of calcium-dependent protein phosphorylation in mediating neurotransmitter release and anticonvulsant action. The results demonstrate that calcium and phenytoin have antagonistic actions on the net level of endogenous phosphorylation of specific rat brain proteins. These specific phosphoproteins were shown to be present in synaptosome preparations, and evidence is presented for the localization of these phosphoproteins within the presynaptic nerve terminal. The data are compatible with the hypothesis that the antagonistic actions of calcium and phenytoin on the phosphorylation of synaptosome proteins may be the underlying molecular mechanism mediating the opposing actions of these agents on the release of neurotransmitter from the presynaptic nerve terminal.
|
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] |
PMID:19236
|
gamma-Glutamyltranspeptidase activity in newborn rat kidney brush border.
|
gamma-Glutamyltranspeptidase, known to be localized in the proximal tubule cell brush border in the rat, is a membrane-bound enzyme which transfers the gamma-glutamyl moiety of glutathione or its analogue gamma-glutamyl-p-nitroanilide to an amino acid or dipeptide acceptor. Brush borders were isolated from the kidneys of newborn and adult Sprague-Dawley rats and assayed for gamma-glutamyltranspeptidase activity. There is an increase in specific activity in the brush border with maturation. Newborn and adult brush border preparations exhibit similar pH optima, substrate affinities, apparent Km values, patterns of heat inactivation, inhibition by glutathione, and migration on polyacrylamide gels. Polyacrylamide gel electrophoresis of a deoxycholate extract of brush border proteins and subsequent reaction with substrate within the gel reveal the presence of two bands, suggesting the presence of two forms of gamma-glutamyltranspeptidase in the rat kidney brush border.
|
gamma-Glutamyltranspeptidase activity in newborn rat kidney brush border. gamma-Glutamyltranspeptidase, known to be localized in the proximal tubule cell brush border in the rat, is a membrane-bound enzyme which transfers the gamma-glutamyl moiety of glutathione or its analogue gamma-glutamyl-p-nitroanilide to an amino acid or dipeptide acceptor. Brush borders were isolated from the kidneys of newborn and adult Sprague-Dawley rats and assayed for gamma-glutamyltranspeptidase activity. There is an increase in specific activity in the brush border with maturation. Newborn and adult brush border preparations exhibit similar pH optima, substrate affinities, apparent Km values, patterns of heat inactivation, inhibition by glutathione, and migration on polyacrylamide gels. Polyacrylamide gel electrophoresis of a deoxycholate extract of brush border proteins and subsequent reaction with substrate within the gel reveal the presence of two bands, suggesting the presence of two forms of gamma-glutamyltranspeptidase in the rat kidney brush border.
|
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] |
PMID:19237
|
The optimum pH of renal adenosine triphosphatase and its variation with the type of ATP.
|
The in vitro optimum pH of renal (Na+, K+)-ATPase, assumed to be 7.8, depends on the type of ATP used. The accepted value is obtained with 'grade II' ATP but with purer ATP ('Sigma grade') it lies close to the intracellular pH in rat medulla and cortex (pH 7.0, 7.2). Values were identical in rats subjected to dietary potassium depletion for 2-4 weeks. The optimal Mg2+ concentration was also influenced by the type of ATP but Mg ATPase activity was unaffected. Both types of ATP were hydrolysed by trichloracetic acid. Where the purer ATP is used the assay conditions need to be modified accordingly.
|
The optimum pH of renal adenosine triphosphatase and its variation with the type of ATP. The in vitro optimum pH of renal (Na+, K+)-ATPase, assumed to be 7.8, depends on the type of ATP used. The accepted value is obtained with 'grade II' ATP but with purer ATP ('Sigma grade') it lies close to the intracellular pH in rat medulla and cortex (pH 7.0, 7.2). Values were identical in rats subjected to dietary potassium depletion for 2-4 weeks. The optimal Mg2+ concentration was also influenced by the type of ATP but Mg ATPase activity was unaffected. Both types of ATP were hydrolysed by trichloracetic acid. Where the purer ATP is used the assay conditions need to be modified accordingly.
|
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] |
PMID:19238
|
Lack of hepatic enzymatic adaptation to low and high levels of dietary protein in the adult cat.
|
The activities of three urea cycle enzymes, several nitrogen catabolic, gluconeogenic, and lipogenic enzymes were measured in the liver of adult cats fed: a commercial kibble; a 17.5 or 70% protein purified diet, or starved for 5 days. Except for an increase in tyrosine transaminase (EC 2.6.1.5) after feeding the high protein diet, there were no changes in the activities of the hepatic enzymes as influenced by dietary protein level. Likewise, starvation had a minimal effect on the activities of these enzymes as compared to that found in similar experiments in rats. These results indicate that the cat may have only minimal capabilities for enzyme adaptation as compared to that found in many herbivores and omnivores and may provide an explanation as to why cats have an unusually high protein requirement as compared to many other mammals.
|
Lack of hepatic enzymatic adaptation to low and high levels of dietary protein in the adult cat. The activities of three urea cycle enzymes, several nitrogen catabolic, gluconeogenic, and lipogenic enzymes were measured in the liver of adult cats fed: a commercial kibble; a 17.5 or 70% protein purified diet, or starved for 5 days. Except for an increase in tyrosine transaminase (EC 2.6.1.5) after feeding the high protein diet, there were no changes in the activities of the hepatic enzymes as influenced by dietary protein level. Likewise, starvation had a minimal effect on the activities of these enzymes as compared to that found in similar experiments in rats. These results indicate that the cat may have only minimal capabilities for enzyme adaptation as compared to that found in many herbivores and omnivores and may provide an explanation as to why cats have an unusually high protein requirement as compared to many other mammals.
|
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] |
PMID:19240
|
Dissociation of small-intestinal sucrase - isomaltase complex into enzymatically active subunits.
|
1. The sucrase - isomaltase complex from rabbit small intestine dissociated into its subunits upon reaction with citraconic anhydride. They can recombine after deacylation under mild acidic conditions. 2. When citraconylated, the subunits could be separated and isolated in a catalytically active form. 3. The previously reported procedure for separation of the subunits by alkaline treatment at pH 9.6 is apparently not due to contaminating degradative enzymes (possibly still present at undetectable levels in the isolated sucrase - isomaltase complex) but to the action of alkali.
|
Dissociation of small-intestinal sucrase - isomaltase complex into enzymatically active subunits. 1. The sucrase - isomaltase complex from rabbit small intestine dissociated into its subunits upon reaction with citraconic anhydride. They can recombine after deacylation under mild acidic conditions. 2. When citraconylated, the subunits could be separated and isolated in a catalytically active form. 3. The previously reported procedure for separation of the subunits by alkaline treatment at pH 9.6 is apparently not due to contaminating degradative enzymes (possibly still present at undetectable levels in the isolated sucrase - isomaltase complex) but to the action of alkali.
|
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] |
PMID:19242
|
On the pH-dependence of the reaction of hemoglobin with carbon monoxide.
|
Flash-photolysis experiments were performed on solutions of carbonmonoxy hemoglobin (human Hb A) as function as pH. The fraction of fast reaction and the amount of photodissociation as produced by a given amount of light quanta has been analyzed in terms of the allosteric model of ligand binding by Monod, Wyman and Changeux. It is shown how the switch-over point of the allosteric transition from the T or R state is controlled by the protons, which act as allosteric effectors.
|
On the pH-dependence of the reaction of hemoglobin with carbon monoxide. Flash-photolysis experiments were performed on solutions of carbonmonoxy hemoglobin (human Hb A) as function as pH. The fraction of fast reaction and the amount of photodissociation as produced by a given amount of light quanta has been analyzed in terms of the allosteric model of ligand binding by Monod, Wyman and Changeux. It is shown how the switch-over point of the allosteric transition from the T or R state is controlled by the protons, which act as allosteric effectors.
|
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] |
PMID:19243
|
The role of an essential histidine residue of yeast alcohol dehydrogenase.
|
1. Inactivation of yeast alcohol dehydrogenase for diethyl pyrocarbonate indicates that one histidine residue per enzyme subunit is necessary for enzymic activity. The inactivated enzyme regains its activity over a period of days. 2. Enzyme modified by diethyl pyrocarbonate can form the binary enzyme - NADH complex with the same maximum NADH-binding capacity as that of native enzyme. Modified enzyme cannot form normal ternary complexes of the type enzyme - NADH - acetamide and enzyme - NAD+ - pyrazole, which are characteristic of native enzyme. 3. The rate constant for the reaction of enzyme with diethyl pyrocarbonate has been determined over the pH range 5.5--9. The histidine residue involved has approximately the same pKa as free histidine, but is 10-fold more reactive than free histidine.
|
The role of an essential histidine residue of yeast alcohol dehydrogenase. 1. Inactivation of yeast alcohol dehydrogenase for diethyl pyrocarbonate indicates that one histidine residue per enzyme subunit is necessary for enzymic activity. The inactivated enzyme regains its activity over a period of days. 2. Enzyme modified by diethyl pyrocarbonate can form the binary enzyme - NADH complex with the same maximum NADH-binding capacity as that of native enzyme. Modified enzyme cannot form normal ternary complexes of the type enzyme - NADH - acetamide and enzyme - NAD+ - pyrazole, which are characteristic of native enzyme. 3. The rate constant for the reaction of enzyme with diethyl pyrocarbonate has been determined over the pH range 5.5--9. The histidine residue involved has approximately the same pKa as free histidine, but is 10-fold more reactive than free histidine.
|
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] |
PMID:19244
|
The effect of macromolecular polyanions on the functional properties of human hemoglobin.
|
The binding of dextran sulphate and heparin to human hemoglobin and their effect on the properties of gas transport have been investigated. Both dextran sulphate and heparin are strongly bound by oxy-hemoglobin as well as deoxyhemoglobin and the stoichiometry of the binding (polyanion/tetrameric hemoglobin) is less than unity; sedimentation analysis gives indication for the existence of octomers. The oxygen affinity of hemoglobin is decreased, to the same extent, by both dextran sulphate and heparin. This effect is pH-dependent. In addition the polyanions affect the position and the magnitude of the Bohr effect. In the presence of dextran sulphate the recombination of hemoglobin with carbon monoxide after flash photolysis is biphasic and the fraction of quickly reacting material increases with dilution of the protein.
|
The effect of macromolecular polyanions on the functional properties of human hemoglobin. The binding of dextran sulphate and heparin to human hemoglobin and their effect on the properties of gas transport have been investigated. Both dextran sulphate and heparin are strongly bound by oxy-hemoglobin as well as deoxyhemoglobin and the stoichiometry of the binding (polyanion/tetrameric hemoglobin) is less than unity; sedimentation analysis gives indication for the existence of octomers. The oxygen affinity of hemoglobin is decreased, to the same extent, by both dextran sulphate and heparin. This effect is pH-dependent. In addition the polyanions affect the position and the magnitude of the Bohr effect. In the presence of dextran sulphate the recombination of hemoglobin with carbon monoxide after flash photolysis is biphasic and the fraction of quickly reacting material increases with dilution of the protein.
|
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] |
PMID:19247
|
Nuclear-magnetic-resonance-spectroscopic studies of the amino groups of insulin.
|
The amino groups of insulin have been studied by 1H and 13C nuclear magnetic resonance spectroscopy in insulin, zinc-free insulin and methylated insulin. By difference spectroscopy it is possible to follow the shift with pH of the epsilon-CH2 and delta-CH2 proton resonances of lysine-B29 in insulin. In methylated insulin the dimethyl proton resonances of glycine-A1, phenylalanine-B1 and lysine-B29 can be followed as a function of pH. In native insulin pKapp values of 6.7 and 8.0 are obtained for phenylalanine-B1 and glycine-A1 (the assignment is tentative) and 11.2 for lysine-B29. Separate resonances have been observed from the lysine-B29 Nepsilon-(CH3)2 group for the monomeric and dimeric forms of methylated insulin, which indicates a small change in the environment of lysine-B29 on dimerisation. The nuclear magnetic resonance spectral characteristics of these groups are, in general, consistent with the overall structure of the crystal form of the 2-zinc insulin hexamer.
|
Nuclear-magnetic-resonance-spectroscopic studies of the amino groups of insulin. The amino groups of insulin have been studied by 1H and 13C nuclear magnetic resonance spectroscopy in insulin, zinc-free insulin and methylated insulin. By difference spectroscopy it is possible to follow the shift with pH of the epsilon-CH2 and delta-CH2 proton resonances of lysine-B29 in insulin. In methylated insulin the dimethyl proton resonances of glycine-A1, phenylalanine-B1 and lysine-B29 can be followed as a function of pH. In native insulin pKapp values of 6.7 and 8.0 are obtained for phenylalanine-B1 and glycine-A1 (the assignment is tentative) and 11.2 for lysine-B29. Separate resonances have been observed from the lysine-B29 Nepsilon-(CH3)2 group for the monomeric and dimeric forms of methylated insulin, which indicates a small change in the environment of lysine-B29 on dimerisation. The nuclear magnetic resonance spectral characteristics of these groups are, in general, consistent with the overall structure of the crystal form of the 2-zinc insulin hexamer.
|
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] |
PMID:19249
|
Bacteriorhodopsin-mediated photophosphorylation in Halobacterium halobium.
|
The rate of halobacterial photophosphorylation was found to be a linear function of light intensity over a wide range (between 1 and 20 mW/cm2). At higher light intensities (above 25 mW/cm2) the ATP-synthesizing system itself limits the maximal rate of photophosphorylation. The optimal external pH range for this type of photophosphorylation is between pH 6.2 and 7.2 external. The photophosphorylation rate is directly proportional to the bacteriorhodopsin content of the cells. The quantum requirement for photophosphorylation was found to be 22 +/- 5 photons per ATP molecule synthesized. According to Mitchell's chemiosmotic hypothesis of energy coupling phosphorylation can be driven by a membrane potential or a pH gradient or a combination of both. From the results of experiments with drugs which abolish or reduce either one of the two components we conclude that the major driving force for photophosphorylation above an external pH value of 6.5 is the membrane potential, while at more acidic pH value the pH gradient becomes dominating. We did not observe a correlation between a transient alkalinization of the medium and ATP-synthesis upon illumination under certain conditions.
|
Bacteriorhodopsin-mediated photophosphorylation in Halobacterium halobium. The rate of halobacterial photophosphorylation was found to be a linear function of light intensity over a wide range (between 1 and 20 mW/cm2). At higher light intensities (above 25 mW/cm2) the ATP-synthesizing system itself limits the maximal rate of photophosphorylation. The optimal external pH range for this type of photophosphorylation is between pH 6.2 and 7.2 external. The photophosphorylation rate is directly proportional to the bacteriorhodopsin content of the cells. The quantum requirement for photophosphorylation was found to be 22 +/- 5 photons per ATP molecule synthesized. According to Mitchell's chemiosmotic hypothesis of energy coupling phosphorylation can be driven by a membrane potential or a pH gradient or a combination of both. From the results of experiments with drugs which abolish or reduce either one of the two components we conclude that the major driving force for photophosphorylation above an external pH value of 6.5 is the membrane potential, while at more acidic pH value the pH gradient becomes dominating. We did not observe a correlation between a transient alkalinization of the medium and ATP-synthesis upon illumination under certain conditions.
|
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] |
PMID:19250
|
The effective proton conductance of the inner membrane of mitochondria from brown adipose tissue. Dependency on proton electrochemical potential gradient.
|
The nucleotide-sensitive H+ (OH-) conducting pathway of mitochondria from the brown-adipose tissue of cold-adapted guinea-pigs passes an effective proton current which is directly proportional to the proton electrochemical gradient. At 23 degrees C and pH 7.0 this conductance is 16 nmol H+ - min-1 - mg-1 - mV-1. Addition of 0.2 mM GDP results in a conductance which is linear and low (0.7 nmol H+ - min-1 - mg-1 - mV-1) until deltamicronH+ exceeds 220 mV. At higher values of deltamicronH+, which can be attained by glycerol 3-phosphate oxidation but not palmitoyl-L-carnitine plus malate oxidation, the membrane conductance greatly increases, effectively limiting the maximal deltamicronH+ to 240 mV. High glycerol 3-phosphate concentrations which have the thermodynamic potential to exceed this value of deltamicronH+ instead create a greatly increased rate of controlled respiration. The generality and significance of this device to limit deltamicronH+, and its relation to the nucleotide-sensitive conductance, are discussed.
|
The effective proton conductance of the inner membrane of mitochondria from brown adipose tissue. Dependency on proton electrochemical potential gradient. The nucleotide-sensitive H+ (OH-) conducting pathway of mitochondria from the brown-adipose tissue of cold-adapted guinea-pigs passes an effective proton current which is directly proportional to the proton electrochemical gradient. At 23 degrees C and pH 7.0 this conductance is 16 nmol H+ - min-1 - mg-1 - mV-1. Addition of 0.2 mM GDP results in a conductance which is linear and low (0.7 nmol H+ - min-1 - mg-1 - mV-1) until deltamicronH+ exceeds 220 mV. At higher values of deltamicronH+, which can be attained by glycerol 3-phosphate oxidation but not palmitoyl-L-carnitine plus malate oxidation, the membrane conductance greatly increases, effectively limiting the maximal deltamicronH+ to 240 mV. High glycerol 3-phosphate concentrations which have the thermodynamic potential to exceed this value of deltamicronH+ instead create a greatly increased rate of controlled respiration. The generality and significance of this device to limit deltamicronH+, and its relation to the nucleotide-sensitive conductance, are discussed.
|
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] |
PMID:19253
|
alpha-D-galactosidase from soybeans destroying blood-group B antigens. Purification by affinity chromatography and properties.
|
alpha-D-Galactosidase was isolated from untoasted soybean meal and purified to homogeneity by affinity chromatography on N-epsilon-aminoacaproyl alpha-D-galactopyranosylamine-Sepharose. The purified enzyme destroyed the B-specificity of human ovarian cyst B-glycoprotein with an accompanying increase in H-specificity, and converted human type-B erythrocytes to type O. The enzyme consists primarily of a tetramer, molecular weight 150 000 +/- 5 000 at pH 4.0 and of a monomer, molecular weight 40 000 +/- 3 000 at pH 8.0. Polyacrylamide gel electrophoresis in dodecyl sulfate at pH 7.2 distinguished between two types of monomeric unit of similar molecular weight. N-terminal alanine was identified as the sole N-terminal amino acid residue. The enzyme was shown to be devoid of carbohydrate.
|
alpha-D-galactosidase from soybeans destroying blood-group B antigens. Purification by affinity chromatography and properties. alpha-D-Galactosidase was isolated from untoasted soybean meal and purified to homogeneity by affinity chromatography on N-epsilon-aminoacaproyl alpha-D-galactopyranosylamine-Sepharose. The purified enzyme destroyed the B-specificity of human ovarian cyst B-glycoprotein with an accompanying increase in H-specificity, and converted human type-B erythrocytes to type O. The enzyme consists primarily of a tetramer, molecular weight 150 000 +/- 5 000 at pH 4.0 and of a monomer, molecular weight 40 000 +/- 3 000 at pH 8.0. Polyacrylamide gel electrophoresis in dodecyl sulfate at pH 7.2 distinguished between two types of monomeric unit of similar molecular weight. N-terminal alanine was identified as the sole N-terminal amino acid residue. The enzyme was shown to be devoid of carbohydrate.
|
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] |
PMID:19254
|
Ionization dependence of camphor binding and spin conversion of the complex between cytochrome P-450 and camphor. Kinetic and static studies at sub-zero temperatures.
|
The kinetic rate constants of formation and dissociation of the cytochrome-P-450 - camphor complex (Fe3+-RH) have been obtained by low-temperature (+ 5 degrees C to -20 degrees C) stopped-flow experiments. Simiarly the high-spin/low-spin equilibrium of this complex has been studied as a function of temperature and protonic activity. Both the camphor-binding mechanism and the high-spin/low-spin thermodynamic parameters of Fe3+-RH depend on the protonic activity of the medium in the physiological pH range. The binding rate constants are shown to depend on the ionization of a residue of the protein, probably a histidine. Linear enthalpy-entropy compensation is observed for the camphor binding as well as for the spin-state transition. A camphor-binding-induced change of the electrostatic potential is discussed.
|
Ionization dependence of camphor binding and spin conversion of the complex between cytochrome P-450 and camphor. Kinetic and static studies at sub-zero temperatures. The kinetic rate constants of formation and dissociation of the cytochrome-P-450 - camphor complex (Fe3+-RH) have been obtained by low-temperature (+ 5 degrees C to -20 degrees C) stopped-flow experiments. Simiarly the high-spin/low-spin equilibrium of this complex has been studied as a function of temperature and protonic activity. Both the camphor-binding mechanism and the high-spin/low-spin thermodynamic parameters of Fe3+-RH depend on the protonic activity of the medium in the physiological pH range. The binding rate constants are shown to depend on the ionization of a residue of the protein, probably a histidine. Linear enthalpy-entropy compensation is observed for the camphor binding as well as for the spin-state transition. A camphor-binding-induced change of the electrostatic potential is discussed.
|
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] |
PMID:19257
|
Hemoglobin S Travis: a sickling hemoglobin with two amino acid substitutions [beta6(A3)glutamic acid leads to valine and beta142 (h20) alanine leads to valine).
|
Hb S Travis is a previously undescribed sickling hemoglobin with two amino acid substitutions in the beta chain: beta6 Glu leads to Val and beta142 Ala leads to Val. The beta6 Glu leads to Val mutation imparts to Hb S Travis the characteristic properties of sickling hemoglobin, namely its association with erythrocyte sickling, the insolubility of the hemoglobin in the reduced form, and a minimum gelling concentration value identical to Hb S. Unlike Hb S, Hb S Travis exhibits an increased oxygen affinity and a decreased affinity for 2,3-bisphosphoglycerate and inositol hexakisphosphate. In addition, the variant hemoglobin's tendency to autoxidize and its mechanical precipitability suggest that there are conformational differences between Hb S and Hb S Travis.
|
Hemoglobin S Travis: a sickling hemoglobin with two amino acid substitutions [beta6(A3)glutamic acid leads to valine and beta142 (h20) alanine leads to valine). Hb S Travis is a previously undescribed sickling hemoglobin with two amino acid substitutions in the beta chain: beta6 Glu leads to Val and beta142 Ala leads to Val. The beta6 Glu leads to Val mutation imparts to Hb S Travis the characteristic properties of sickling hemoglobin, namely its association with erythrocyte sickling, the insolubility of the hemoglobin in the reduced form, and a minimum gelling concentration value identical to Hb S. Unlike Hb S, Hb S Travis exhibits an increased oxygen affinity and a decreased affinity for 2,3-bisphosphoglycerate and inositol hexakisphosphate. In addition, the variant hemoglobin's tendency to autoxidize and its mechanical precipitability suggest that there are conformational differences between Hb S and Hb S Travis.
|
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] |
PMID:19260
|
Mechanism of oestrogen and progesterone effects on lipid and carbohydrate metabolism: alteration in the insulin: glucagon molar ratio and hepatic enzyme activity.
|
As in women receiving oestrogens the administration of 17beta-oestradiol to ovariectomized female rats caused a rise in fasting plasma triglycerides and a fall in plasma glucose. Progesterone, on the other hand, had no significant effects. In the oestradiol treated rats, the portal vein basal insulin levels were slightly reduced. Oestradiol, however, had a marked suppressive effect on the alpha cells of the pancreas resulting in a greater reduction in basal glucagon and impaired glucagon response to alanine infusions. The relative insulin to glucagon (I/G) molar concentration ratio in portal vein blood was increased. Oestradiol also produced a dose dependent increase in the activity of the liver lipogenic enzymes, acetyl CoA carboxylase and fatty acid synthetase. On the other hand, the activity of the gluconeogenic rate limiting enzyme phosphoenol-pyruvate carboxykinase (PEPCK) was inhibited. The cross-over pattern of gluconeogenic intermediates confirmed inhibition of gluconeogenesis at this step, an effect which is similar to that induced by relative insulin 'excess'. Progesterone produced an increase in the portal vein insulin concentrations. Both the basal and the alanine-stimulated glucagon levels were also increased. The I/G molar ratio in portal vein blood of progesterone treated rats remained unaltered and the hepatic lipogenic and gluconeogenic enzyme activities were similar to control animals. These data suggest that insulin activity is increased relative to glucagon in the liver of oestradiol-treated rats due to the rise in portal vein I/G ratio. The changes in liver lipogenic and gluconeogenic enzymes and the alterations in fasting plasma triglycerides and glucose in response to oestrogens could be secondary to this effect.
|
Mechanism of oestrogen and progesterone effects on lipid and carbohydrate metabolism: alteration in the insulin: glucagon molar ratio and hepatic enzyme activity. As in women receiving oestrogens the administration of 17beta-oestradiol to ovariectomized female rats caused a rise in fasting plasma triglycerides and a fall in plasma glucose. Progesterone, on the other hand, had no significant effects. In the oestradiol treated rats, the portal vein basal insulin levels were slightly reduced. Oestradiol, however, had a marked suppressive effect on the alpha cells of the pancreas resulting in a greater reduction in basal glucagon and impaired glucagon response to alanine infusions. The relative insulin to glucagon (I/G) molar concentration ratio in portal vein blood was increased. Oestradiol also produced a dose dependent increase in the activity of the liver lipogenic enzymes, acetyl CoA carboxylase and fatty acid synthetase. On the other hand, the activity of the gluconeogenic rate limiting enzyme phosphoenol-pyruvate carboxykinase (PEPCK) was inhibited. The cross-over pattern of gluconeogenic intermediates confirmed inhibition of gluconeogenesis at this step, an effect which is similar to that induced by relative insulin 'excess'. Progesterone produced an increase in the portal vein insulin concentrations. Both the basal and the alanine-stimulated glucagon levels were also increased. The I/G molar ratio in portal vein blood of progesterone treated rats remained unaltered and the hepatic lipogenic and gluconeogenic enzyme activities were similar to control animals. These data suggest that insulin activity is increased relative to glucagon in the liver of oestradiol-treated rats due to the rise in portal vein I/G ratio. The changes in liver lipogenic and gluconeogenic enzymes and the alterations in fasting plasma triglycerides and glucose in response to oestrogens could be secondary to this effect.
|
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] |
PMID:19261
|
Effects of bile and bile acids on cultured human fibroblasts.
|
Impaired healing induced by leakage of bile has been postulated as one factor responsible for complications after reconstructive bile duct surgery. The cytotoxicity of human bile and its major bile acids on cultured human fibroblasts was therefore studied by evaluation of their effects on cell morphology and growth, on synthesis and secretion of 35SO4-mucopolysaccharides and on release of a lysosomal enzyme. Normal human fibroblasts derived from a standard culture strain (MRC-5) were grown to confluence and exposed to: (1) sterile human T-tube bile, (2) a mixture of bile acids resembling that of human bile, or (3) various concentrations of the glycine- and taurine conjugates of cholic, chenodeoxycholic or deoxycholic acid. Medium containing whole bile (total bile acid concentration 0.25, 0.75 or 1.6 mmol/l) exerted time and dose dependent cytotoxic effects on morphology and growth and release of lysosomal enzyme. Synthesis and secretion of 35SO4-mucopolysaccharides were markedly inhibited. The bile acid mixture exhibited the same time and dose dependent effects. The conjugates of deoxycholic acid were found to be the most toxic of the individual bile acids studied.
|
Effects of bile and bile acids on cultured human fibroblasts. Impaired healing induced by leakage of bile has been postulated as one factor responsible for complications after reconstructive bile duct surgery. The cytotoxicity of human bile and its major bile acids on cultured human fibroblasts was therefore studied by evaluation of their effects on cell morphology and growth, on synthesis and secretion of 35SO4-mucopolysaccharides and on release of a lysosomal enzyme. Normal human fibroblasts derived from a standard culture strain (MRC-5) were grown to confluence and exposed to: (1) sterile human T-tube bile, (2) a mixture of bile acids resembling that of human bile, or (3) various concentrations of the glycine- and taurine conjugates of cholic, chenodeoxycholic or deoxycholic acid. Medium containing whole bile (total bile acid concentration 0.25, 0.75 or 1.6 mmol/l) exerted time and dose dependent cytotoxic effects on morphology and growth and release of lysosomal enzyme. Synthesis and secretion of 35SO4-mucopolysaccharides were markedly inhibited. The bile acid mixture exhibited the same time and dose dependent effects. The conjugates of deoxycholic acid were found to be the most toxic of the individual bile acids studied.
|
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] |
PMID:19263
|
Mechanisms of B cell tolerance: I. Dynamic nature of the induction of hapten-specific unresponsiveness by hapten-conjugated pneumococcal polysaccharide.
|
Exposure of 2,4-dinitrophenyl (DNP)-hemocyanin-primed spleen cells to DNP-conjugated pneumococcal polysaccharide (DNP-S3) in vivo or in vitro renders such cells unresponsive to DNP-hemocyanin challenge following adoptive transfer. The unresponsiveness is hapten-specific, independent of the presence of T cells and adherent cells, and not due to either toxic effect of S3 or carry-over of tolerogenic amounts of cell-bound DNP-S3, and thus presumably represents a model of B cell tolerance. The degree of suppression induced depends upon the dose of the tolerogen and most strikingly on the duration of exposure of cells to the tolerogen. Thus 2 to 6 h exposure to DNP-S3 has an insignificant effect on anti-DNP responses, while 24 and 48 h exposures in vitro and in vivo are highly suppressive. Such a dynamic process of tolerance induction suggests a "multiple hit" phenomenon, implying the generation of suppressive signals by repeated cycles of tolerogen-receptor interactions. The process can be interrupted by removal of extracellular DNP-S3, although this does not reverse the unresponsiveness that has already been induced.
|
Mechanisms of B cell tolerance: I. Dynamic nature of the induction of hapten-specific unresponsiveness by hapten-conjugated pneumococcal polysaccharide. Exposure of 2,4-dinitrophenyl (DNP)-hemocyanin-primed spleen cells to DNP-conjugated pneumococcal polysaccharide (DNP-S3) in vivo or in vitro renders such cells unresponsive to DNP-hemocyanin challenge following adoptive transfer. The unresponsiveness is hapten-specific, independent of the presence of T cells and adherent cells, and not due to either toxic effect of S3 or carry-over of tolerogenic amounts of cell-bound DNP-S3, and thus presumably represents a model of B cell tolerance. The degree of suppression induced depends upon the dose of the tolerogen and most strikingly on the duration of exposure of cells to the tolerogen. Thus 2 to 6 h exposure to DNP-S3 has an insignificant effect on anti-DNP responses, while 24 and 48 h exposures in vitro and in vivo are highly suppressive. Such a dynamic process of tolerance induction suggests a "multiple hit" phenomenon, implying the generation of suppressive signals by repeated cycles of tolerogen-receptor interactions. The process can be interrupted by removal of extracellular DNP-S3, although this does not reverse the unresponsiveness that has already been induced.
|
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] |
PMID:19265
|
pD2-, pA2- and pD2'-values of a series of compounds in a histaminic and a cholinergic system.
|
(1) Affinity and intrinsic activity values of 75 compounds for a histaminergic and a cholinergic system are presented. The quantitative correlations between the affinity values of 35 compounds and some physicochemical constants (Van der Waals volume, lipophilicity, number of hydrogen atoms on the protonated amine) are discussed. (2) Absence of systematic differences between pD2 and pA2 of partial agonists supports the assumption that these values are equivalent expressions of the same affinity. (3) The "mimetic moiety" in a number of the antihistaminic test compounds hardly contributes to their affinity. The affinity mainly depends on an interaction tendency with additional receptor areas. (4) The correlation between pA2 and pD2' of the whole series of compounds in the histaminergic system is artificial. The method only allows determination of both values if their ratio lies between certain limits. (5) The correlation between pA2 and pD2' for 16 closely related compounds in the guinea pig ileum and for nearly all compounds in the rat intestine has to be explained by an influence of the structural differences on drug transference and/or the less specific binding forces. (6) The metactoid receptors in the two systems are different structures. (7) Possible molecular modifications to maximize the separation of antihistaminic form cholinergic affinity are suggested.
|
pD2-, pA2- and pD2'-values of a series of compounds in a histaminic and a cholinergic system. (1) Affinity and intrinsic activity values of 75 compounds for a histaminergic and a cholinergic system are presented. The quantitative correlations between the affinity values of 35 compounds and some physicochemical constants (Van der Waals volume, lipophilicity, number of hydrogen atoms on the protonated amine) are discussed. (2) Absence of systematic differences between pD2 and pA2 of partial agonists supports the assumption that these values are equivalent expressions of the same affinity. (3) The "mimetic moiety" in a number of the antihistaminic test compounds hardly contributes to their affinity. The affinity mainly depends on an interaction tendency with additional receptor areas. (4) The correlation between pA2 and pD2' of the whole series of compounds in the histaminergic system is artificial. The method only allows determination of both values if their ratio lies between certain limits. (5) The correlation between pA2 and pD2' for 16 closely related compounds in the guinea pig ileum and for nearly all compounds in the rat intestine has to be explained by an influence of the structural differences on drug transference and/or the less specific binding forces. (6) The metactoid receptors in the two systems are different structures. (7) Possible molecular modifications to maximize the separation of antihistaminic form cholinergic affinity are suggested.
|
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] |
PMID:19266
|
Regional effects of neuroleptics on dopamine metabolism and dopamine-sensitive adenylate cyclase activity.
|
The effect of haloperidol, chlorpromazine, thioridazine and sulpride on the levels of DOPAC and HVA, as an index of DA turnover, and on the activity of DA-stimulated adenylate cyclase was investigated inthe striatum, the nucleus accumbens and the tuberculum olfactorium of the rat brain. Haloperidol, chlorpromazine and thioridazine caused a more marked increase in DA turnover in the striatum than in the mesolimbic areas, while the reverse was true for sulpiride. In contrast, although the relative potency of these compounds varied greatly, the Ki of each drug for the DA-sensitive adenylate cyclase was similar in three structures of rat brain. The results indicate that in the three brain structures investigated there was no correlation between the differential effects of neuroleptics on dopamine turnover in vivo and the blockade by these drug of the DA-sensitive adenylate cyclase activity in vitro.
|
Regional effects of neuroleptics on dopamine metabolism and dopamine-sensitive adenylate cyclase activity. The effect of haloperidol, chlorpromazine, thioridazine and sulpride on the levels of DOPAC and HVA, as an index of DA turnover, and on the activity of DA-stimulated adenylate cyclase was investigated inthe striatum, the nucleus accumbens and the tuberculum olfactorium of the rat brain. Haloperidol, chlorpromazine and thioridazine caused a more marked increase in DA turnover in the striatum than in the mesolimbic areas, while the reverse was true for sulpiride. In contrast, although the relative potency of these compounds varied greatly, the Ki of each drug for the DA-sensitive adenylate cyclase was similar in three structures of rat brain. The results indicate that in the three brain structures investigated there was no correlation between the differential effects of neuroleptics on dopamine turnover in vivo and the blockade by these drug of the DA-sensitive adenylate cyclase activity in vitro.
|
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0.009530415758490562,
-0.028901781886816025,
-0.015763837844133377,
-0.0444280169904232,
0.010579120367765427,
0.052762821316719055,
-0.010139014571905136,
-0.04171978682279587,
0.01537611149251461,
0.021289147436618805,
0.0464051179587841,
0.0695374608039856
] |
PMID:19267
|
The effect of antipsychotic drugs and their clinically inactive analogs on dopamine metabolism.
|
Changes in dopamine metabolite levels in the rat striatum and tuberculum olfactorium, following the administration of three non-antipsychotic butyrophenones (AL-499, AHR-1900 and U-25,927) and a non-antipsychotic benzazepine (SCH-12,679), were compared to the effects seen following the antipsychotics haloperidol, chlorpromazine and clozapine. The non-antipsychotics, although clinically ineffective, were reported as active in a variety of animal screening tests. Haloperidol, chlorpromazine and clozapine produced a dose-dependent increase in 3,4-dihydroxyphenylacetic acid (DOPAC) levels in both regions. Of the non-antipsychotic drugs only AHR-1900 significantly elevated the level of DOPAC, however, the slope of its dose-response curve was atypically flat in comparison to the dose-response curves of drugs with known antipsychotic efficacy. Moreover, the maximal effect of AHR-1900 observed at a dose of 40 mg/kg was less than the ED50 effect of haloperidol which occurs at a 250 fold lower dose. It is concluded that the dose-dependent elevation of DOPAC in the striatum and tuberculum olfactorium of the rat is a good predictor of antipsychotic efficacy.
|
The effect of antipsychotic drugs and their clinically inactive analogs on dopamine metabolism. Changes in dopamine metabolite levels in the rat striatum and tuberculum olfactorium, following the administration of three non-antipsychotic butyrophenones (AL-499, AHR-1900 and U-25,927) and a non-antipsychotic benzazepine (SCH-12,679), were compared to the effects seen following the antipsychotics haloperidol, chlorpromazine and clozapine. The non-antipsychotics, although clinically ineffective, were reported as active in a variety of animal screening tests. Haloperidol, chlorpromazine and clozapine produced a dose-dependent increase in 3,4-dihydroxyphenylacetic acid (DOPAC) levels in both regions. Of the non-antipsychotic drugs only AHR-1900 significantly elevated the level of DOPAC, however, the slope of its dose-response curve was atypically flat in comparison to the dose-response curves of drugs with known antipsychotic efficacy. Moreover, the maximal effect of AHR-1900 observed at a dose of 40 mg/kg was less than the ED50 effect of haloperidol which occurs at a 250 fold lower dose. It is concluded that the dose-dependent elevation of DOPAC in the striatum and tuberculum olfactorium of the rat is a good predictor of antipsychotic efficacy.
|
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0.042972784489393234
] |
PMID:19268
|
The anticonvulsant action of methazolamide in mice: antagonism by various inhibitors of dopamine beta-hydroxylase.
|
The dopamine beta-hydroxylase inhibitors, FLA-63 and picolinic acid, antagonized the anticonvulsant action of methazolamine in mice; disulfiram and pyrimidinethiol were inactive. FLA-63 and picolinic acid, but not disulfiram or pyrimidinethiol prevented pheniprazine restoration of the anticonvulsant action of methazolamide in reserpinized mice. The present findings clearly demonstrate that differences exist among inhibitors of dopamine beta-hydroxylase regarding their ability to antagonize the anticonvulsant action of methazolamide under various test conditions.
|
The anticonvulsant action of methazolamide in mice: antagonism by various inhibitors of dopamine beta-hydroxylase. The dopamine beta-hydroxylase inhibitors, FLA-63 and picolinic acid, antagonized the anticonvulsant action of methazolamine in mice; disulfiram and pyrimidinethiol were inactive. FLA-63 and picolinic acid, but not disulfiram or pyrimidinethiol prevented pheniprazine restoration of the anticonvulsant action of methazolamide in reserpinized mice. The present findings clearly demonstrate that differences exist among inhibitors of dopamine beta-hydroxylase regarding their ability to antagonize the anticonvulsant action of methazolamide under various test conditions.
|
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] |
PMID:19269
|
Acetylcholine-norepinephrine interactions on potassium movements in the sinus node.
|
Possible adrenergic mechanisms involved in acetylcholine (ACh) induced potassium movements in the sinus node have been investigated using a tracer potassium (42K) and a microelectrode technique. The ACh-induced increase in 42K uptake was enhanced by propranolol and was unaffected by phentolamine. Reserpinization neither prevented the ACh-induced increase in 42K uptake nor the enhanced effect in the presence of propranolol. In reserpinized preparations, ACh-induced 42K uptake was the same before and after norepinephrine (NE) administration, but was greater after acute reserpinization. NE alone induced an increase in 42K uptake but the simultaneous administration of ACh and NE provoked an increase in 42K uptake similar to that obtained with ACh alone. When both neuromediators and atropine were given simultaneously, an increase in 42K uptake did not occur. Thus, the ACh induces an increase in 42K uptake independently of NE. ACh antagonizes the NE effect on 42K uptake independently of a muscarinic interaction.
|
Acetylcholine-norepinephrine interactions on potassium movements in the sinus node. Possible adrenergic mechanisms involved in acetylcholine (ACh) induced potassium movements in the sinus node have been investigated using a tracer potassium (42K) and a microelectrode technique. The ACh-induced increase in 42K uptake was enhanced by propranolol and was unaffected by phentolamine. Reserpinization neither prevented the ACh-induced increase in 42K uptake nor the enhanced effect in the presence of propranolol. In reserpinized preparations, ACh-induced 42K uptake was the same before and after norepinephrine (NE) administration, but was greater after acute reserpinization. NE alone induced an increase in 42K uptake but the simultaneous administration of ACh and NE provoked an increase in 42K uptake similar to that obtained with ACh alone. When both neuromediators and atropine were given simultaneously, an increase in 42K uptake did not occur. Thus, the ACh induces an increase in 42K uptake independently of NE. ACh antagonizes the NE effect on 42K uptake independently of a muscarinic interaction.
|
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0.043514855206012726,
0.10133227705955505
] |
PMID:19277
|
Modification of the action of pentagastrin on acid secretion by botulinum toxin.
|
I.v botulinum toxin after 60-90 min abolished the dose-response relationship between pentagastrin and gastric acid secretion in anesthetized rats and guinea-pigs. The toxin reduced but did not abolish the acid stimulatory effect of histamine. As expected, the acid response to vagal stimulation was abolished and that to methacholine in rats was unaltered by the toxin.
|
Modification of the action of pentagastrin on acid secretion by botulinum toxin. I.v botulinum toxin after 60-90 min abolished the dose-response relationship between pentagastrin and gastric acid secretion in anesthetized rats and guinea-pigs. The toxin reduced but did not abolish the acid stimulatory effect of histamine. As expected, the acid response to vagal stimulation was abolished and that to methacholine in rats was unaltered by the toxin.
|
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] |
PMID:19279
|
The effect of ion pair formation on the antimuscarinic activity of methantheline.
|
The quaternary ammonium compound, methantheline, was found to antagonize acetylcholine induced contractions in isolated guinea pig ileum by a mechanism which did not conform to the criteria for either competitive or non-competitive inhibition. Enhancement of the lipid solubility of methantheline by formation of an ion pair with trichloracetate failed to influence its cholinergic inhibitory activity. The results suggest that in the guinea pig ileum a) an intracellular site of action does not exist for methantheline and b) the membrane receptors for methantheline most likely are located in an aqueous environment.
|
The effect of ion pair formation on the antimuscarinic activity of methantheline. The quaternary ammonium compound, methantheline, was found to antagonize acetylcholine induced contractions in isolated guinea pig ileum by a mechanism which did not conform to the criteria for either competitive or non-competitive inhibition. Enhancement of the lipid solubility of methantheline by formation of an ion pair with trichloracetate failed to influence its cholinergic inhibitory activity. The results suggest that in the guinea pig ileum a) an intracellular site of action does not exist for methantheline and b) the membrane receptors for methantheline most likely are located in an aqueous environment.
|
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] |
PMID:19281
|
Interaction of extrinsic fluorescent probes with E. coli glutamine synthetase.
|
Binding of 2-p-toluidinylnaphthalene-6-sulfonate (TNS) to adenylylated (E--11) glutamine synthetase is cooperative and time-dependent, with 3 dye sites per subunit. In fluorescence polarization experiments TNS and pyrene butyrate give normalized Perrin plots that indicate a symmetrical arrangement of dye excited state dipoles, relative to the rotational axis of the oblate ellipsoid of the dodecameric native enzyme.
|
Interaction of extrinsic fluorescent probes with E. coli glutamine synthetase. Binding of 2-p-toluidinylnaphthalene-6-sulfonate (TNS) to adenylylated (E--11) glutamine synthetase is cooperative and time-dependent, with 3 dye sites per subunit. In fluorescence polarization experiments TNS and pyrene butyrate give normalized Perrin plots that indicate a symmetrical arrangement of dye excited state dipoles, relative to the rotational axis of the oblate ellipsoid of the dodecameric native enzyme.
|
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] |
PMID:19282
|
Effects of alpha- and beta-adrenergic blockers on the actions of noradrenaline on body temperature in the newborn guinea-pig.
|
The effect of NA injected into the lateral cerebral ventricle on Tc was blocked by alpha-adrenergic receptor blockers, but not by beta-receptor blockers, whereas the effect of systemically administered NA was blocked by i.p. administered beta-receptor blockers, but not by alpha-blockers.
|
Effects of alpha- and beta-adrenergic blockers on the actions of noradrenaline on body temperature in the newborn guinea-pig. The effect of NA injected into the lateral cerebral ventricle on Tc was blocked by alpha-adrenergic receptor blockers, but not by beta-receptor blockers, whereas the effect of systemically administered NA was blocked by i.p. administered beta-receptor blockers, but not by alpha-blockers.
|
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] |
PMID:19283
|
Chemical studies of marine invertebrates. XXVII. On the absolute configuration of aromadendrane sesquiterpenes from the soft coral Cespitularia aff. subviridis.
|
Allo-aromadendrene, (-)viridiflorol and (+)ledol have been identified as minor constituents of the soft coral Cespitularia aff. subviridis.
|
Chemical studies of marine invertebrates. XXVII. On the absolute configuration of aromadendrane sesquiterpenes from the soft coral Cespitularia aff. subviridis. Allo-aromadendrene, (-)viridiflorol and (+)ledol have been identified as minor constituents of the soft coral Cespitularia aff. subviridis.
|
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] |
PMID:19285
|
Inhibitory effect of fentonium on gastric acid secretion in different laboratory animals.
|
The influence of fentonium, a new derivative of hyoscyamine, on the gastric secretion has been evaluated in rats, cats and dogs. The compound appeared to be able to markedly reduce the gastric hypersecretion produced by pentagastrin, caerulein and bethanechol; its antagonizing activity against histamine stimulation was poor.
|
Inhibitory effect of fentonium on gastric acid secretion in different laboratory animals. The influence of fentonium, a new derivative of hyoscyamine, on the gastric secretion has been evaluated in rats, cats and dogs. The compound appeared to be able to markedly reduce the gastric hypersecretion produced by pentagastrin, caerulein and bethanechol; its antagonizing activity against histamine stimulation was poor.
|
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] |
PMID:19307
|
Hydrogen ion and carbon dioxide content of the oviductal fluid of the rhesus monkey (Macaca mulatta).
|
The pH in the oviduct lumen of adult female rhesus monkey (Macaca mulatta) during the menstrual cycle was measured by means of miniaturized pH electrodes. Two types were used: one was a flexible electrode 1.4 mm in diameter; the other was a rigid electrode 0.8 mm in diameter. The same results were obtained with both types. For PCO2 measurements, the pH electrodes was converted to a Severinghaus tyep electrode covered with a Teflon membrane. During the follicular phase, the pH remains constant in the range 7.1 to 7.3. In association with ovulation, there is a sudden increase in the pH to the range 7.5 to 7.8. The pH remains high through the luteal phase. The PCO2 values obtained showed no evident changes with day of cycle. The range of values estimated was wide because of drift in electrode calibration, with the average level about 89 Torr, compared with 40 Torr for blood. At the average pH of 7.2 in the follicular phase, this value of PCO2 corresponds to 35 mM HCO3-; at the average pH of 7.6 of the secretory phase, this value corresponds to 90 mM HCO3-. This increase in pH and HCO3- after ovulation would be expected to enhance dispersal of corona cells during fertilization and to provide a favorable environment for the developing embryo.
|
Hydrogen ion and carbon dioxide content of the oviductal fluid of the rhesus monkey (Macaca mulatta). The pH in the oviduct lumen of adult female rhesus monkey (Macaca mulatta) during the menstrual cycle was measured by means of miniaturized pH electrodes. Two types were used: one was a flexible electrode 1.4 mm in diameter; the other was a rigid electrode 0.8 mm in diameter. The same results were obtained with both types. For PCO2 measurements, the pH electrodes was converted to a Severinghaus tyep electrode covered with a Teflon membrane. During the follicular phase, the pH remains constant in the range 7.1 to 7.3. In association with ovulation, there is a sudden increase in the pH to the range 7.5 to 7.8. The pH remains high through the luteal phase. The PCO2 values obtained showed no evident changes with day of cycle. The range of values estimated was wide because of drift in electrode calibration, with the average level about 89 Torr, compared with 40 Torr for blood. At the average pH of 7.2 in the follicular phase, this value of PCO2 corresponds to 35 mM HCO3-; at the average pH of 7.6 of the secretory phase, this value corresponds to 90 mM HCO3-. This increase in pH and HCO3- after ovulation would be expected to enhance dispersal of corona cells during fertilization and to provide a favorable environment for the developing embryo.
|
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] |
PMID:19327
|
Determining optimum dose and acute tolerance of triazolam.
|
The present two-phase study was designed to determine the optimum hynotic dose of triazolam and to assess any evidence of acute tolerance. Subjective and objective data were used to assess hypnotic efficacy. In the dose-titration phase, the dose of triazolam was increased in 0-5 mg increments to the highest level tolerated by each individual patient. Maximum tolerable doses ranged between 0-5 and 3-0 mg; however, all subjects experienced maximum therapeutic effect at 1-5 mg triazolam. During the 4-night acute tolerance portion of the study no clear evidence of tolerance was found.
|
Determining optimum dose and acute tolerance of triazolam. The present two-phase study was designed to determine the optimum hynotic dose of triazolam and to assess any evidence of acute tolerance. Subjective and objective data were used to assess hypnotic efficacy. In the dose-titration phase, the dose of triazolam was increased in 0-5 mg increments to the highest level tolerated by each individual patient. Maximum tolerable doses ranged between 0-5 and 3-0 mg; however, all subjects experienced maximum therapeutic effect at 1-5 mg triazolam. During the 4-night acute tolerance portion of the study no clear evidence of tolerance was found.
|
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] |
PMID:19328
|
Antihypertensive effect and tolerability of metoprolol during long-term treatment: a multicentre study.
|
One hundred and forty-seven patients with essential hypertension participated in this multicentre study. The results indicate that metoprolol in a dosage of 150-450 mg daily is an effective and well tolerated therapy both in patients previously untreated and in patients unsatisfactorily treated with other antihypertensive agents.
|
Antihypertensive effect and tolerability of metoprolol during long-term treatment: a multicentre study. One hundred and forty-seven patients with essential hypertension participated in this multicentre study. The results indicate that metoprolol in a dosage of 150-450 mg daily is an effective and well tolerated therapy both in patients previously untreated and in patients unsatisfactorily treated with other antihypertensive agents.
|
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] |
PMID:19330
|
6-Phosphogluconate/glucose-6-phosphate ratio in rat pancreatic islets during inhibition of insulin release by exogenous insulin.
|
Inhibition of glucose-stimulated insulin release by exogenous insulin has been demonstrated in pancreatic islets to be associated with a decrease of the NADPH/NADP ratio and the pentose-phosphate cycle activity. Batches of five islets were incubated for 15 and 90 minutes in 1 ml. of KRB buffer with 2 per cent albumin containing 3 mg./ml. glucose and 0, 200, 400, or 800 microU./ml. of rat insulin, and the glucose-6-phosphate (G6P) and 6-phosphogluconate (6PG) contents were determined by enzymatic cycling. In response to a rise in the concentration of insulin, the 6PG/G6P ratio decreased. A close relationship was observed between this decrease of 6PG/G6P ratio and the net insulin release, the absolute rate of glucose oxidation via the pentose phosphate cycle, and the NADPH/NADP ratios measured under similar conditions. The results suggest that exogenous insulin, directly or indirectly, regulates the pentose cycle activity in the pancreatic islets at the G6P dehydrogenase step.
|
6-Phosphogluconate/glucose-6-phosphate ratio in rat pancreatic islets during inhibition of insulin release by exogenous insulin. Inhibition of glucose-stimulated insulin release by exogenous insulin has been demonstrated in pancreatic islets to be associated with a decrease of the NADPH/NADP ratio and the pentose-phosphate cycle activity. Batches of five islets were incubated for 15 and 90 minutes in 1 ml. of KRB buffer with 2 per cent albumin containing 3 mg./ml. glucose and 0, 200, 400, or 800 microU./ml. of rat insulin, and the glucose-6-phosphate (G6P) and 6-phosphogluconate (6PG) contents were determined by enzymatic cycling. In response to a rise in the concentration of insulin, the 6PG/G6P ratio decreased. A close relationship was observed between this decrease of 6PG/G6P ratio and the net insulin release, the absolute rate of glucose oxidation via the pentose phosphate cycle, and the NADPH/NADP ratios measured under similar conditions. The results suggest that exogenous insulin, directly or indirectly, regulates the pentose cycle activity in the pancreatic islets at the G6P dehydrogenase step.
|
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] |
PMID:19331
|
Effect of caerulein on gamma-glutamyl transpeptidase and alkaline phosphatase in the duodenal juice in chronic pancreatitis.
|
During infusion of caerulein, the output of gamma-glutamyl transpeptidase and biliary alkaline phosphatase is significantly more increased in chronic pancreatitis patients than in normal subjects. The presence of the gall-bladder is necessary to observe this effect of caerulein. It is not related to the discharge of large amounts of both enzymes by the diseases pancreas.
|
Effect of caerulein on gamma-glutamyl transpeptidase and alkaline phosphatase in the duodenal juice in chronic pancreatitis. During infusion of caerulein, the output of gamma-glutamyl transpeptidase and biliary alkaline phosphatase is significantly more increased in chronic pancreatitis patients than in normal subjects. The presence of the gall-bladder is necessary to observe this effect of caerulein. It is not related to the discharge of large amounts of both enzymes by the diseases pancreas.
|
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] |
PMID:19333
|
Inhibitory effect of sulpiride on cinchophen gastric ulceration in dogs.
|
An intraventricular administration of sulpiride suppressed the experimental cinchophen ulcer, thereby the effect of repeated administration of the material on cinchophen ulcerogenesis is evident, whereas intermittent administration lacked this effect. Such an effect as reducing experimental cinchophen ulceration, which is due to central neurohumoral mechanism, following intraventricular infusion of sulpiride appears to be ascribable to a chemical blockage of the hypothalamic structure. It can thus be concluded that sulpiride acts principally at the level of hypothalamus. Its inhibitory action on cinchophen gastric ulcer development is probably exerted via the hypothalomo-pituitary-adrenal axis, which is essential to the occurrence of an ulcer, and may be regarded as favoring the central neurohumoral theory of cinchophen ulcerogenesis.
|
Inhibitory effect of sulpiride on cinchophen gastric ulceration in dogs. An intraventricular administration of sulpiride suppressed the experimental cinchophen ulcer, thereby the effect of repeated administration of the material on cinchophen ulcerogenesis is evident, whereas intermittent administration lacked this effect. Such an effect as reducing experimental cinchophen ulceration, which is due to central neurohumoral mechanism, following intraventricular infusion of sulpiride appears to be ascribable to a chemical blockage of the hypothalamic structure. It can thus be concluded that sulpiride acts principally at the level of hypothalamus. Its inhibitory action on cinchophen gastric ulcer development is probably exerted via the hypothalomo-pituitary-adrenal axis, which is essential to the occurrence of an ulcer, and may be regarded as favoring the central neurohumoral theory of cinchophen ulcerogenesis.
|
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] |
PMID:19334
|
Role of intragastric pressure, pH, and pepsin in gastric ulceration in the rat.
|
A method of continuous gastric perfusion with "artificial gastric juice" was used in a study of individual factors (intragastric pressure, pH, and pepsin) known to participate in the pathogenesis of peptic ulcers. This method allowed change of only one factor at a time, while the other two remained constant. The gastric mucosa of normal rats, fasted for 48 hr, was found to be resistant to the ulcerogenic effects of artificial gastric juice perfused through the stomach for 6 hr without increasing the intragastric pressure. Perfusion of hydrochloric acid (pH 1.3) under increasing pressure produced ulceration of the corpus as well as forestomach portion of the stomach. The degree of gastric ulceration paralleled increases in intragastric pressure, acidity, and pepsin proteolytic activity. Inhibition of pepsin activity by a pepsin inhibitor protected the gastric mucosa even at the very low pH of 1.3. These results demonstrate that under the experimental conditions used, hydrochloric acid alone in the absence of pepsin does not produce ulceration of the rat stomach.
|
Role of intragastric pressure, pH, and pepsin in gastric ulceration in the rat. A method of continuous gastric perfusion with "artificial gastric juice" was used in a study of individual factors (intragastric pressure, pH, and pepsin) known to participate in the pathogenesis of peptic ulcers. This method allowed change of only one factor at a time, while the other two remained constant. The gastric mucosa of normal rats, fasted for 48 hr, was found to be resistant to the ulcerogenic effects of artificial gastric juice perfused through the stomach for 6 hr without increasing the intragastric pressure. Perfusion of hydrochloric acid (pH 1.3) under increasing pressure produced ulceration of the corpus as well as forestomach portion of the stomach. The degree of gastric ulceration paralleled increases in intragastric pressure, acidity, and pepsin proteolytic activity. Inhibition of pepsin activity by a pepsin inhibitor protected the gastric mucosa even at the very low pH of 1.3. These results demonstrate that under the experimental conditions used, hydrochloric acid alone in the absence of pepsin does not produce ulceration of the rat stomach.
|
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] |
PMID:19335
|
Binding properties in vitro of antacids for conjugated bile acids.
|
The binding properties of various commercial antacids for pure conjugated bile acids in an aqueous solution or the bile acids present in human gallbladder bile were characterized. Aluminum hydroxide was a potent binding agent, similar in affinity and capacity to cholestyramine. With aluminum hydroxide, dihydroxy bile acid conjugates were bound more strongly than trihydroxy bile acid conjugates, and glycine conjugates were bound more strongly than taurine conjugates; pH had no effect per se on binding. Magnesium hydroxide and aluminum phosphate bound bile acids much more weakly. Aluminum hydroxide warrants exploration in therapeutic situations in which bile acid binding is sought.
|
Binding properties in vitro of antacids for conjugated bile acids. The binding properties of various commercial antacids for pure conjugated bile acids in an aqueous solution or the bile acids present in human gallbladder bile were characterized. Aluminum hydroxide was a potent binding agent, similar in affinity and capacity to cholestyramine. With aluminum hydroxide, dihydroxy bile acid conjugates were bound more strongly than trihydroxy bile acid conjugates, and glycine conjugates were bound more strongly than taurine conjugates; pH had no effect per se on binding. Magnesium hydroxide and aluminum phosphate bound bile acids much more weakly. Aluminum hydroxide warrants exploration in therapeutic situations in which bile acid binding is sought.
|
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] |
PMID:19336
|
Obstructive jaundice after bone marrow transplantation.
|
Jaundice after bone marrow transplantation is usually a consequence of graft versus host disease. Reported is a patient who presented with obstructive jaundice several months after a successful marrow allograft. Despite a benign bone marrow examination, abdominal ultrasound, upper gastrointestinal series, and endoscopic biopsy were utilized to diagnose recurrent leukemia at the pancreatic head and descending duodenum. The entities of graft versus host disease as related to jaundice, and gastrointestinal leukemia, in the presence of a "remission" bone marrow, are reviewed.
|
Obstructive jaundice after bone marrow transplantation. Jaundice after bone marrow transplantation is usually a consequence of graft versus host disease. Reported is a patient who presented with obstructive jaundice several months after a successful marrow allograft. Despite a benign bone marrow examination, abdominal ultrasound, upper gastrointestinal series, and endoscopic biopsy were utilized to diagnose recurrent leukemia at the pancreatic head and descending duodenum. The entities of graft versus host disease as related to jaundice, and gastrointestinal leukemia, in the presence of a "remission" bone marrow, are reviewed.
|
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] |
PMID:19337
|
Peptic ulcer disease--a heterogeneous group of disorders?
|
The familial aggregation of peptic ulcer disease has been well established, as has its association with such clear-cut genetic factors as blood group O and nonsecretor status. However, the genetics of this disorder, or group of disorders, is still in question. Polygenic inheritance is the prevailing hypothesis that has been proposed for peptic ulcer. This hypothesis was based primarily on the exclusion of a simple mode of inheritance for all ulcer disease. Genetic heterogeneity is an alternative hypothesis that can explain both the familial aggregation of peptic ulcer disease and the lack of a simple Mendelian pattern of inheritance. The evidence for genetic heterogeneity in peptic ulcer disease is reviewed, and studies are proposed to test this hypothesis.
|
Peptic ulcer disease--a heterogeneous group of disorders? The familial aggregation of peptic ulcer disease has been well established, as has its association with such clear-cut genetic factors as blood group O and nonsecretor status. However, the genetics of this disorder, or group of disorders, is still in question. Polygenic inheritance is the prevailing hypothesis that has been proposed for peptic ulcer. This hypothesis was based primarily on the exclusion of a simple mode of inheritance for all ulcer disease. Genetic heterogeneity is an alternative hypothesis that can explain both the familial aggregation of peptic ulcer disease and the lack of a simple Mendelian pattern of inheritance. The evidence for genetic heterogeneity in peptic ulcer disease is reviewed, and studies are proposed to test this hypothesis.
|
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] |
PMID:19338
|
Effect of beta adrenergic blocking drugs on the renin-aldosterone system, sodium excretion, and renal hemodynamics in cirrhosis with ascites.
|
As a result of effective beta adrenergic blockade with either propranolol or practolol, plasma renin activity was suppressed in all of 11 patients with cirrhosis and ascites. In contrast, the effect on the rate of renal excretion of aldosterone was variable, suggesting that factors other than the renin-angiotension system are responsible for the control of aldosterone secretion in cirrhosis. The changes in aldosterone could not be explained on the basis of changes in the plasma concentrations of potassium or sodium. The renal sodium excretion was inversely related to the values for aldosterone both before and after beta adrenergic blockade, indicating a major role for aldosterone in regulating sodium excretion. A number of patients had an abnormal intrarenal distribution of plasma flow with a relative hypoperfusion of the renin-secreting outer cortical nephrons. Plasma renin activity was inversely related to outer cortical plasma flow, suggesting that the reduced outer cortical flow may be a stimulus to increased renin secretion. Because the abnormal intrarenal hemodynamic pattern was not corrected by suppression of plasma renin activity, and presumably angiotension II concentrations, it is unlikely that it is attributable to the known renal vasonconstrictor effects of angiotensin II.
|
Effect of beta adrenergic blocking drugs on the renin-aldosterone system, sodium excretion, and renal hemodynamics in cirrhosis with ascites. As a result of effective beta adrenergic blockade with either propranolol or practolol, plasma renin activity was suppressed in all of 11 patients with cirrhosis and ascites. In contrast, the effect on the rate of renal excretion of aldosterone was variable, suggesting that factors other than the renin-angiotension system are responsible for the control of aldosterone secretion in cirrhosis. The changes in aldosterone could not be explained on the basis of changes in the plasma concentrations of potassium or sodium. The renal sodium excretion was inversely related to the values for aldosterone both before and after beta adrenergic blockade, indicating a major role for aldosterone in regulating sodium excretion. A number of patients had an abnormal intrarenal distribution of plasma flow with a relative hypoperfusion of the renin-secreting outer cortical nephrons. Plasma renin activity was inversely related to outer cortical plasma flow, suggesting that the reduced outer cortical flow may be a stimulus to increased renin secretion. Because the abnormal intrarenal hemodynamic pattern was not corrected by suppression of plasma renin activity, and presumably angiotension II concentrations, it is unlikely that it is attributable to the known renal vasonconstrictor effects of angiotensin II.
|
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-0.054341837763786316,
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0.019967176020145416,
0.04202459752559662
] |
PMID:19340
|
Crohn's disease involving the colon: an audit of clinical management.
|
This paper describes the clinical management of 204 patients with Crohn's disease involving the colon who have been reviewed regularly for many years. The results have been analyzed to define the morbidity, mortality, and long term prognosis of the disorder. Despite a high morbidity, the long term prognosis is good. Although the mean interval since the onset of symptoms is more than 13 years, 77% of the patients are symptom-free and 60% have normal laboratory indices. This analysis suggests that until the etiology of Crohn's disease is established and a specific therapy is developed, good results can be obtained with regular medical review and appropriate surgical treatment with the limited use of immunosuppressive agents or total parenteral nutrition.
|
Crohn's disease involving the colon: an audit of clinical management. This paper describes the clinical management of 204 patients with Crohn's disease involving the colon who have been reviewed regularly for many years. The results have been analyzed to define the morbidity, mortality, and long term prognosis of the disorder. Despite a high morbidity, the long term prognosis is good. Although the mean interval since the onset of symptoms is more than 13 years, 77% of the patients are symptom-free and 60% have normal laboratory indices. This analysis suggests that until the etiology of Crohn's disease is established and a specific therapy is developed, good results can be obtained with regular medical review and appropriate surgical treatment with the limited use of immunosuppressive agents or total parenteral nutrition.
|
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] |
PMID:19341
|
Effect of acetylsalicylic acid on the amphibian gastric mucosa. I. Electrophysiological and permeability changes.
|
The effects of acetylsalicylic acid on the in vitro bullfrog gastric mucosa were defined. After mucosal exposure of 10 or 20 mM acetylsalicylic acid, a characteristic series of changes evolved. The potential difference and resistance rose initially. After 15 to 30 min the potential difference declined to zero. Transmucosal resistance remained elevated considerably longer but began to decline toward zero after 30 to 60 min of exposure. Mucosal permeability to mannitol increased as transmucosal resistance declined, but was not markedly elevated until the 2nd hr of exposure to acetylsalicylic acid. The initial rise of potential difference was associated with inhibition of H+ secretion whereas the later decline of potential difference was due to inhibition of Cl- transport. The abolition of short circuit current could not be attributed to mucosal to serosal flux of acetylsalicylate.
|
Effect of acetylsalicylic acid on the amphibian gastric mucosa. I. Electrophysiological and permeability changes. The effects of acetylsalicylic acid on the in vitro bullfrog gastric mucosa were defined. After mucosal exposure of 10 or 20 mM acetylsalicylic acid, a characteristic series of changes evolved. The potential difference and resistance rose initially. After 15 to 30 min the potential difference declined to zero. Transmucosal resistance remained elevated considerably longer but began to decline toward zero after 30 to 60 min of exposure. Mucosal permeability to mannitol increased as transmucosal resistance declined, but was not markedly elevated until the 2nd hr of exposure to acetylsalicylic acid. The initial rise of potential difference was associated with inhibition of H+ secretion whereas the later decline of potential difference was due to inhibition of Cl- transport. The abolition of short circuit current could not be attributed to mucosal to serosal flux of acetylsalicylate.
|
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] |
PMID:19344
|
Characterization of Salmonella typhimurium mutants with altered glutamine synthetase activity.
|
A number of glutamine auxotrophs of Salmonella typhimurium were isolated and characterized genetically. Three of the mutations appear to be closely linked and are complemented by episomes carrying the glnA region of Escherichia coli. The lesions in these strains are approximately 20% linked by P1 transduction with a mutation in the rha gene, but are unlinked to ilv. Another mutation causing glutamine auxotrophy in strain JB674 is genetically distinct from the others. Strain JB674 grown in glucose medium containing ammonia as the nitrogen source has reduced levels of glutamine synthetase that is more adenylylated than in the parent strain, suggesting that the enzyme can not be deadenylylated normally. The lesion causing glutamine auxotrophy in JB674 lies in the region corresponding to the glnB and glnE genes affecting glutamine synthetase modification in Klebsiella areogenes. Four Gln+ revertants of JB674 have glutamine synthetase activities 4 to 6 fold higher than normal. One mutation causing this increased enzyme synthesis has been shown by three-factor crosses with the glnA mutations to lie near or within the glnA gene.
|
Characterization of Salmonella typhimurium mutants with altered glutamine synthetase activity. A number of glutamine auxotrophs of Salmonella typhimurium were isolated and characterized genetically. Three of the mutations appear to be closely linked and are complemented by episomes carrying the glnA region of Escherichia coli. The lesions in these strains are approximately 20% linked by P1 transduction with a mutation in the rha gene, but are unlinked to ilv. Another mutation causing glutamine auxotrophy in strain JB674 is genetically distinct from the others. Strain JB674 grown in glucose medium containing ammonia as the nitrogen source has reduced levels of glutamine synthetase that is more adenylylated than in the parent strain, suggesting that the enzyme can not be deadenylylated normally. The lesion causing glutamine auxotrophy in JB674 lies in the region corresponding to the glnB and glnE genes affecting glutamine synthetase modification in Klebsiella areogenes. Four Gln+ revertants of JB674 have glutamine synthetase activities 4 to 6 fold higher than normal. One mutation causing this increased enzyme synthesis has been shown by three-factor crosses with the glnA mutations to lie near or within the glnA gene.
|
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] |
PMID:19345
|
[Bacterial recombination].
|
The most perspective trends in studying the mechanisms of genetic exchanges in bacteria are discussed. They are the following. 1. Comparison of the recombination stages in various biological phenomena such as transformation, transduction (generalized), conjugation (common and single-stranded) and transfection. 2. Characterization of genetic determinants and basic enzymes of the recombination process in Rec-systems of different bacteria. 3. Determination of some peculiar features of the intragenic recombination and finding a possible approach to their research. 4. Analysis of some plastic properties of the DNA structure and their application to the modern ideas in the formulation of a molecular mechanism of genetic recombination.
|
[Bacterial recombination]. The most perspective trends in studying the mechanisms of genetic exchanges in bacteria are discussed. They are the following. 1. Comparison of the recombination stages in various biological phenomena such as transformation, transduction (generalized), conjugation (common and single-stranded) and transfection. 2. Characterization of genetic determinants and basic enzymes of the recombination process in Rec-systems of different bacteria. 3. Determination of some peculiar features of the intragenic recombination and finding a possible approach to their research. 4. Analysis of some plastic properties of the DNA structure and their application to the modern ideas in the formulation of a molecular mechanism of genetic recombination.
|
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] |
PMID:19346
|
Cimetidine and the gastric mucosal barrier.
|
The histamine H2-receptor antagonist, cimetidine, when given topically or parenterally, did not affect net luminal Na+ gain or net luminal H+ loss in vagally-denervated pouches of antrectomised dogs. Cimetidine did not affect disruption of the mucosal barrier by 5 mmol/l or 20 mmol/l sodium taurocholate.
|
Cimetidine and the gastric mucosal barrier. The histamine H2-receptor antagonist, cimetidine, when given topically or parenterally, did not affect net luminal Na+ gain or net luminal H+ loss in vagally-denervated pouches of antrectomised dogs. Cimetidine did not affect disruption of the mucosal barrier by 5 mmol/l or 20 mmol/l sodium taurocholate.
|
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] |
PMID:19347
|
[Treatment of peptic ulcer with histamine H2 receptor antagonists].
|
Histamine H2-receptor antagonists (Burimamide, Metiamide and Cimetidine as the most recent generation) may drastically inhibit gastric acid secretion stimulated by histamine, pentagastrin, insulin, 2-deoxyglucose or an intragastrically instilled meal, respectively. This inhibitory action may explain the beneficial effects of H2-antagonists in the treatment of active peptic ulceration. On Cimetidine administered at a usual dosage over a 4--6 week period, serious side-effects must not be expected. At present studies aim to establish a Cimetidine dosage which, on long-term treatment, may reduce ulcer recurrency.
|
[Treatment of peptic ulcer with histamine H2 receptor antagonists]. Histamine H2-receptor antagonists (Burimamide, Metiamide and Cimetidine as the most recent generation) may drastically inhibit gastric acid secretion stimulated by histamine, pentagastrin, insulin, 2-deoxyglucose or an intragastrically instilled meal, respectively. This inhibitory action may explain the beneficial effects of H2-antagonists in the treatment of active peptic ulceration. On Cimetidine administered at a usual dosage over a 4--6 week period, serious side-effects must not be expected. At present studies aim to establish a Cimetidine dosage which, on long-term treatment, may reduce ulcer recurrency.
|
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] |
PMID:19348
|
[Antacids and gastric mucosa. Protective effect of an antacid on acetylsalicylic acid induced functional lesions in the human gastric mucosa].
|
Salicylates do induce functional and morphological changes of the gastric mucosa: increased H+-back-diffusion, increased Na+-influx, and increased blood loss. Several functional parameters of the gastric mucosal barrier correlate with the transmural electrical potential difference (PD) which can be used as a sensitive, but unspecific parameter to detect functional changes of the gastric mucosal barrier. The effect of a salicylate alone and in presence of a liquid antiacid (Gastropulgit Gel) on transmural PD was measured in healthy volunteers. The presence of an antacid prevented the salicylate-induced drop of transmural PD. The tested antacid seems to exert a protective effect towards acute damage of the gastric mucosa induced by salicylates.
|
[Antacids and gastric mucosa. Protective effect of an antacid on acetylsalicylic acid induced functional lesions in the human gastric mucosa]. Salicylates do induce functional and morphological changes of the gastric mucosa: increased H+-back-diffusion, increased Na+-influx, and increased blood loss. Several functional parameters of the gastric mucosal barrier correlate with the transmural electrical potential difference (PD) which can be used as a sensitive, but unspecific parameter to detect functional changes of the gastric mucosal barrier. The effect of a salicylate alone and in presence of a liquid antiacid (Gastropulgit Gel) on transmural PD was measured in healthy volunteers. The presence of an antacid prevented the salicylate-induced drop of transmural PD. The tested antacid seems to exert a protective effect towards acute damage of the gastric mucosa induced by salicylates.
|
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] |
PMID:19350
|
Interaction of colchicine with DNA molecules.
|
Colchicine (7.5 X 10-6M or 3.3 X 10-5M) was incubated at pH 7.0, 10.0, or 12.0 with high-molecular DNA from salmon sperm (7 X 10-5M or 3.5 X 10-5M DNA-P) at 40 degrees C. Interaction was monitored by UV spectrophotometry; Amax/Amin ratios, difference and additive spectra, and the quantity of colchicine bound to DNA were presented as functions of time, ionic strength and DNA concentration. Using NMR techniques, delta deltav 1/2/deltac values, chemical shifts and half-line widths of colchicine protons were analyzed as a function of the DNA/colchicine ratio, thus proving the interaction between alkaloid and DNA. An intercalation of the tropolone moiety of colchicine between the nitrogenous bases of DNA is suggested.
|
Interaction of colchicine with DNA molecules. Colchicine (7.5 X 10-6M or 3.3 X 10-5M) was incubated at pH 7.0, 10.0, or 12.0 with high-molecular DNA from salmon sperm (7 X 10-5M or 3.5 X 10-5M DNA-P) at 40 degrees C. Interaction was monitored by UV spectrophotometry; Amax/Amin ratios, difference and additive spectra, and the quantity of colchicine bound to DNA were presented as functions of time, ionic strength and DNA concentration. Using NMR techniques, delta deltav 1/2/deltac values, chemical shifts and half-line widths of colchicine protons were analyzed as a function of the DNA/colchicine ratio, thus proving the interaction between alkaloid and DNA. An intercalation of the tropolone moiety of colchicine between the nitrogenous bases of DNA is suggested.
|
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] |
PMID:19351
|
Glutamine synthetase from pig brain: binding of adenosine triphosphate.
|
Glutamine synthetase was purified to homogeneity from fresh brain homogenates by a procedure that makes use of the affinity of this ATP requiring ligase to Cibacron 3G-A Blue-Sephadex G-75. It is shown that the interaction of pig brain enzyme with the dye does not concern the active centre but a non-catalytic although specific ATP binding site. Both sides contain a cysteine residue which may react with N-ethylmaleimide. The unprotected succinimidated enzyme is inactive, and aggregates. ATP alone protects only against aggregation, not against inactivation; the ATP/Mg2 complex also hinders inactivation. A tryptic heptadecapeptide isolated from the derivatized enzyme representing the carboxy terminal seems to belong to the active centre; its amino acid sequence was partially determined.
|
Glutamine synthetase from pig brain: binding of adenosine triphosphate. Glutamine synthetase was purified to homogeneity from fresh brain homogenates by a procedure that makes use of the affinity of this ATP requiring ligase to Cibacron 3G-A Blue-Sephadex G-75. It is shown that the interaction of pig brain enzyme with the dye does not concern the active centre but a non-catalytic although specific ATP binding site. Both sides contain a cysteine residue which may react with N-ethylmaleimide. The unprotected succinimidated enzyme is inactive, and aggregates. ATP alone protects only against aggregation, not against inactivation; the ATP/Mg2 complex also hinders inactivation. A tryptic heptadecapeptide isolated from the derivatized enzyme representing the carboxy terminal seems to belong to the active centre; its amino acid sequence was partially determined.
|
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] |
PMID:19354
|
Enzymatically active peptide from the adenosine diphosphate-ribosylating toxin of Pseudomonas aeruginosa.
|
A nontoxic peptide (molecular weight, 26,000), which is active in catalyzing the adenosine diphosphate (ADP)-ribosylation of elongation factor 2, has been isolated from the culture supernatant of Pseudomonas aeruginosa strain 103 in stationary phase. Like fragment A from diphtheria toxin, the active peptide catalyzed the hydrolysis of nicotinamide adenine dinucleotide as well as the ADP-ribosylation of elongation factor 2 and showed similarities to fragment A in specific activity, kinetic constants, pH optimum, and ionic sensitivity. These results provide strong evidence for a high degree of homology in the structures of their active sites. That the peptide is not identical to fragment A is shown by the fact that it was not neutralized by fragment A-specific antiserum and was different in amino acid composition and pH and thermal labilities. Although definitive evidence is lacking, there are data suggesting that this peptide is a proteolytic fragment from the ADP-ribosylating toxin (exotoxin A; molecular weight, 66,000) produced by the same strain of P. aeruginosa.
|
Enzymatically active peptide from the adenosine diphosphate-ribosylating toxin of Pseudomonas aeruginosa. A nontoxic peptide (molecular weight, 26,000), which is active in catalyzing the adenosine diphosphate (ADP)-ribosylation of elongation factor 2, has been isolated from the culture supernatant of Pseudomonas aeruginosa strain 103 in stationary phase. Like fragment A from diphtheria toxin, the active peptide catalyzed the hydrolysis of nicotinamide adenine dinucleotide as well as the ADP-ribosylation of elongation factor 2 and showed similarities to fragment A in specific activity, kinetic constants, pH optimum, and ionic sensitivity. These results provide strong evidence for a high degree of homology in the structures of their active sites. That the peptide is not identical to fragment A is shown by the fact that it was not neutralized by fragment A-specific antiserum and was different in amino acid composition and pH and thermal labilities. Although definitive evidence is lacking, there are data suggesting that this peptide is a proteolytic fragment from the ADP-ribosylating toxin (exotoxin A; molecular weight, 66,000) produced by the same strain of P. aeruginosa.
|
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] |
PMID:19355
|
Correlation between oral toxicity and in vitro stability of Clostridium botulinum type A and B toxins of different molecular sizes.
|
The in vitro sensitivity to acid and pepsin differed markedly among Clostridium botulinum type A and B toxins of different molecular sizes. The larger the molecular size of the toxin, the higher the resistance to these agents. Tye B derivative toxin was rapidly inactivated, but the progenitor toxins resisted in vitro exposure to rat intestinal juice. The molecular dissociation of the progenitor toxins did not occur in rat intestinal juice of pH 7.0, but did occur in a buffer solution of the same pH. The oral toxicity may depend mostly on the stability of toxin molecules in the stomach and, to a less extent, in the intestine. The present results seem to justify the conclusion that C. botulinum type A and B progenitor toxins with molecular sizes larger than 16S are more potent oral toxins than 12S progenitor toxins.
|
Correlation between oral toxicity and in vitro stability of Clostridium botulinum type A and B toxins of different molecular sizes. The in vitro sensitivity to acid and pepsin differed markedly among Clostridium botulinum type A and B toxins of different molecular sizes. The larger the molecular size of the toxin, the higher the resistance to these agents. Tye B derivative toxin was rapidly inactivated, but the progenitor toxins resisted in vitro exposure to rat intestinal juice. The molecular dissociation of the progenitor toxins did not occur in rat intestinal juice of pH 7.0, but did occur in a buffer solution of the same pH. The oral toxicity may depend mostly on the stability of toxin molecules in the stomach and, to a less extent, in the intestine. The present results seem to justify the conclusion that C. botulinum type A and B progenitor toxins with molecular sizes larger than 16S are more potent oral toxins than 12S progenitor toxins.
|
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] |
PMID:19356
|
Effect of temperature on bacterial killing by serum and by polymorphonuclear leukocytes.
|
Bacterial killing by serum alone and by polymorphonuclear )PMN) leukocytes was studied at 37 degrees C and compared with killing at 39 and 41 degrees C. The test organisms for serum killing were Staphylococcus aureus 502A (serum resistant) and Escherichia coli O14 (serum sensitive). The organisms used in PMN killing tests were Streptococcus pneumoniae type 29 and E. coli O86.S aureus was not killed by serum alone at any temperature. Changes in temperature did not affect the rate of serum killing of E. coli O14 for the first 60 min, but by 90 and 120 min there was a discrepancy with continued killing at 37 degrees C, but no further killing at 39 and 41 degrees C. PMN phagocytic killing of the pneumococcus was enhanced at 39 degrees C compared with 37 degrees C, and phagocytic killing of E. coli O86 was decreased at 41 degrees C when compared with 37 degrees C. Therefore, it appears that under certain circumstances fever may aid the host PMNs in destroying organisms, whereas under other circumstances it may interfere with such destruction.
|
Effect of temperature on bacterial killing by serum and by polymorphonuclear leukocytes. Bacterial killing by serum alone and by polymorphonuclear )PMN) leukocytes was studied at 37 degrees C and compared with killing at 39 and 41 degrees C. The test organisms for serum killing were Staphylococcus aureus 502A (serum resistant) and Escherichia coli O14 (serum sensitive). The organisms used in PMN killing tests were Streptococcus pneumoniae type 29 and E. coli O86.S aureus was not killed by serum alone at any temperature. Changes in temperature did not affect the rate of serum killing of E. coli O14 for the first 60 min, but by 90 and 120 min there was a discrepancy with continued killing at 37 degrees C, but no further killing at 39 and 41 degrees C. PMN phagocytic killing of the pneumococcus was enhanced at 39 degrees C compared with 37 degrees C, and phagocytic killing of E. coli O86 was decreased at 41 degrees C when compared with 37 degrees C. Therefore, it appears that under certain circumstances fever may aid the host PMNs in destroying organisms, whereas under other circumstances it may interfere with such destruction.
|
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] |
PMID:19357
|
Classical and alternative complement pathway activation by pneumococci.
|
Sixty-two strains of Streptococcus pneumoniae were studied for their abilities to consume selected components of classical and alternative complement pathways in human sera. The classical pathway was blocked by chelating calcium with ethyleneglycol-bios (beta-aminoethyl ether)-N,N-tetraacetic acid and by removing C4. The alternative pathway was blocked by removing factor B. Each strain's activation of the two pathways was compared with its nonimmune reactivity with the Fc region of immunoglobulin G (IgG). Activation of the classical complement pathway appeared to be independent of such Fc reactivity. Highly Fc-reactive strains, however, were shown to activate the alternative pathway more effectively than did less Fc-reactive strains. Since pneumococcal activation of the alternative pathway requires non-immunospecific IgG, these findings suggest that nonimmune binding of IgG on the pneumococcal surface endows it with complement-activating properties.
|
Classical and alternative complement pathway activation by pneumococci. Sixty-two strains of Streptococcus pneumoniae were studied for their abilities to consume selected components of classical and alternative complement pathways in human sera. The classical pathway was blocked by chelating calcium with ethyleneglycol-bios (beta-aminoethyl ether)-N,N-tetraacetic acid and by removing C4. The alternative pathway was blocked by removing factor B. Each strain's activation of the two pathways was compared with its nonimmune reactivity with the Fc region of immunoglobulin G (IgG). Activation of the classical complement pathway appeared to be independent of such Fc reactivity. Highly Fc-reactive strains, however, were shown to activate the alternative pathway more effectively than did less Fc-reactive strains. Since pneumococcal activation of the alternative pathway requires non-immunospecific IgG, these findings suggest that nonimmune binding of IgG on the pneumococcal surface endows it with complement-activating properties.
|
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] |
PMID:19358
|
Inhibitory action of fatty acids on the growth of Neisseria gonorrhoeae.
|
Fatty acids of various chain lengths (C(1) to C(24)) were examined for their effects on growth, oxygen consumption, and in vitro reduced nicotinamide adenine dinucleotide oxidase activity of Neisseria gonorrhoeae CS-7. The growth inhibition caused by saturated fatty acids increased with increasing chain length to a maximum with palmitic acid (C(16)). Stearic acid (C(18)) and longer saturated fatty acids showed little inhibition of growth. However, unsaturated fatty acids of chain length C(16) to C(20) were inhibitory. Similar inhibition was observed with Bacillus subtilis and a deep rough mutant of Salmonella typhimurium. Wildtype S. typhimurium and Pseudomonas aeruginosa were more resistant to medium-chain (C(7) to C(10)) fatty acids and completely resistant to long-chain (C(12) to C(18)) fatty acids. Thus, sensitivity of N. gonorrhoeae to long-chain fatty acids appears to be related to the permeability of the outer membrane. Growth inhibition by short-chain (C(1) to C(6)) fatty acids was pH dependent; inhibition of growth increased with decreasing pH. Saturated fatty acids inhibited oxygen consumption by log-phase cells of N. gonorrhoeae. This inhibition increased with increasing chain length to a maximum observed with myristic acid (C(14)). Whereas stearic acid (C(18)) had little effect upon oxygen consumption, unsaturated C(18) fatty acids were inhibitory. An in vitro inhibition of reduced nicotinamide adenine dinucleotide oxidase activity by saturated (C(1) to C(12)) and unsaturated (C(16) to C(20)) fatty acids was also observed. Although the inhibitory concentrations were generally higher than those required to inhibit growth or oxygen consumption, an inhibition of electron transport may be partially responsible for the observed growth inhibition.
|
Inhibitory action of fatty acids on the growth of Neisseria gonorrhoeae. Fatty acids of various chain lengths (C(1) to C(24)) were examined for their effects on growth, oxygen consumption, and in vitro reduced nicotinamide adenine dinucleotide oxidase activity of Neisseria gonorrhoeae CS-7. The growth inhibition caused by saturated fatty acids increased with increasing chain length to a maximum with palmitic acid (C(16)). Stearic acid (C(18)) and longer saturated fatty acids showed little inhibition of growth. However, unsaturated fatty acids of chain length C(16) to C(20) were inhibitory. Similar inhibition was observed with Bacillus subtilis and a deep rough mutant of Salmonella typhimurium. Wildtype S. typhimurium and Pseudomonas aeruginosa were more resistant to medium-chain (C(7) to C(10)) fatty acids and completely resistant to long-chain (C(12) to C(18)) fatty acids. Thus, sensitivity of N. gonorrhoeae to long-chain fatty acids appears to be related to the permeability of the outer membrane. Growth inhibition by short-chain (C(1) to C(6)) fatty acids was pH dependent; inhibition of growth increased with decreasing pH. Saturated fatty acids inhibited oxygen consumption by log-phase cells of N. gonorrhoeae. This inhibition increased with increasing chain length to a maximum observed with myristic acid (C(14)). Whereas stearic acid (C(18)) had little effect upon oxygen consumption, unsaturated C(18) fatty acids were inhibitory. An in vitro inhibition of reduced nicotinamide adenine dinucleotide oxidase activity by saturated (C(1) to C(12)) and unsaturated (C(16) to C(20)) fatty acids was also observed. Although the inhibitory concentrations were generally higher than those required to inhibit growth or oxygen consumption, an inhibition of electron transport may be partially responsible for the observed growth inhibition.
|
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] |
PMID:19359
|
Fungicidal properties of a chymotrypsin-like cationic protein from human neutrophils: adsorption to Candida parapsilosis.
|
Human neutrophils contain a chymotrypsin-like cationic protein (CLCP) that binds to the surface of Candida parapsilosis and is preferentially adsorbed by yeasts mixed with unfractionated extracts of neutrophil granules. Adsorption of CLCP to opsonized or nonopsonized yeasts was rapid at pH 4 through 8. Irreversible inhibition of the enzymatic site of CLCP by phenylmethylsulfonylfluoride or N-acetyl-Ala-Ala-Phe-chloromethyl ketone did not affect its adsorption by yeast. Adsorption, highly sensitive to ionic strength, was abrogated by 0.15 M KCl. The number of CLCP molecules adsorbed per yeast cell and the loss of colony-forming units are described by an exponential relationship.
|
Fungicidal properties of a chymotrypsin-like cationic protein from human neutrophils: adsorption to Candida parapsilosis. Human neutrophils contain a chymotrypsin-like cationic protein (CLCP) that binds to the surface of Candida parapsilosis and is preferentially adsorbed by yeasts mixed with unfractionated extracts of neutrophil granules. Adsorption of CLCP to opsonized or nonopsonized yeasts was rapid at pH 4 through 8. Irreversible inhibition of the enzymatic site of CLCP by phenylmethylsulfonylfluoride or N-acetyl-Ala-Ala-Phe-chloromethyl ketone did not affect its adsorption by yeast. Adsorption, highly sensitive to ionic strength, was abrogated by 0.15 M KCl. The number of CLCP molecules adsorbed per yeast cell and the loss of colony-forming units are described by an exponential relationship.
|
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] |
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