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PMID:15102 | Active form of ketamine in squid giant axons. | The active form of ketamine has been studied with internally perfused squid qiant axons. The drug was applied internally, and decreases in peak transient and steady-state conductances as measured by voltage clamp technique were taken as an index of activity. When the total internal ketamine concentration was maintained constant, the suppression peak transient and steady-state conductances decreased with an increase in internal pH from 7.0 TO 8.4. When the concentration of the internally present charged form of ketamine was kept constant, the suppression of the peak transient conductance remained almost constant at internal pH values of 7.0, 7.3 and 7.7, but increased at pH 8.4. However, the suppression of the stead-state conductance became more prominent as the pH was raised from 7.0 to 8.4. With a constant internal concentration of the uncharged form, the suppression of both conductances decreased as the pH was raised from 7.0 to 8.4. Computation of dissociation constants to suppretamine is more potent than the charged form at all internal pH values examined. These data also show that the potency to suppress the peak transient conductance by the charged and uncharged forms of ketamine decreased as the intertance of the charged form increased, and that of the uncharged form decreased considerably with increase in the internal pH. | Active form of ketamine in squid giant axons. The active form of ketamine has been studied with internally perfused squid qiant axons. The drug was applied internally, and decreases in peak transient and steady-state conductances as measured by voltage clamp technique were taken as an index of activity. When the total internal ketamine concentration was maintained constant, the suppression peak transient and steady-state conductances decreased with an increase in internal pH from 7.0 TO 8.4. When the concentration of the internally present charged form of ketamine was kept constant, the suppression of the peak transient conductance remained almost constant at internal pH values of 7.0, 7.3 and 7.7, but increased at pH 8.4. However, the suppression of the stead-state conductance became more prominent as the pH was raised from 7.0 to 8.4. With a constant internal concentration of the uncharged form, the suppression of both conductances decreased as the pH was raised from 7.0 to 8.4. Computation of dissociation constants to suppretamine is more potent than the charged form at all internal pH values examined. These data also show that the potency to suppress the peak transient conductance by the charged and uncharged forms of ketamine decreased as the intertance of the charged form increased, and that of the uncharged form decreased considerably with increase in the internal pH. | [
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PMID:15103 | Uptake of uric acid by separated renal tubules of the rabbit. I. Characteristics of transport. | A rapid filtration procedure was used to determine rates of uric acid uptake by a preparation of separated renal cortical tubules of the rabbit. The rate of uric acid uptake was temperature dependent and showed saturation kinetics with a K of 3.2 mM. The uptake was 50% lower when measured under nitrogen as compared with uptake under oxygen. The uptake rate increased with increasing sodium concentration and decreased with increasing potassium concentration. Uptake was stimulated by citrate, succinate and pyruvate, and inhibited by alpha-ketoglutarate. Parathyroid hormone and 10(-4) M adenosine 3':5'-monophosphate increased the uric acid uptake rate when preincubated with the tubules for 130 minutes before the addition of uric acid. Uric acid uptake in this preparation appears to occur by some form of carrier-mediated active transport. | Uptake of uric acid by separated renal tubules of the rabbit. I. Characteristics of transport. A rapid filtration procedure was used to determine rates of uric acid uptake by a preparation of separated renal cortical tubules of the rabbit. The rate of uric acid uptake was temperature dependent and showed saturation kinetics with a K of 3.2 mM. The uptake was 50% lower when measured under nitrogen as compared with uptake under oxygen. The uptake rate increased with increasing sodium concentration and decreased with increasing potassium concentration. Uptake was stimulated by citrate, succinate and pyruvate, and inhibited by alpha-ketoglutarate. Parathyroid hormone and 10(-4) M adenosine 3':5'-monophosphate increased the uric acid uptake rate when preincubated with the tubules for 130 minutes before the addition of uric acid. Uric acid uptake in this preparation appears to occur by some form of carrier-mediated active transport. | [
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|
PMID:15104 | Further evaluation of the discriminative effects of morphine in the rat. | Rats were trained in a two-choice discrete trial avoidance paradigm to discriminate between saline and 3.0 mg/kg of morphine. Behavior was considered to be under stimulus ocntrol when the rats completed at least 90% of the trials in a 20-trial session on the morphine-appropriate choice lever after receiving morphine and when they completed at least 90% of the trials on the saline-appropriate choice lever after receiving saline. The discriminative effects of morphine, measured by responding on the morphine-appropriate lever, were then evaluated by determining the dose-response characteristics of representative narcotic analgesics, analgesics with mixed agonist and narcotic antagonist properties and nonopioid psychoactive drugs. Eight narcotic analgesics each produced dose-related responding on the morphine-appropriate lever. The relative potency for producing discriminative effects equivalent to those produced by 3.0 mg/kg of morphine ranged form etonitazene = 1000 x morphine to propoxyphene = 0.0175 x morphine. Of the narcotic antagonist analgesics tested, butorphanol and nalmexone produced discriminative effects equivalent to those of the morphine training dose whereas nalorphine, levallorphan, oxilorphan, nalbuphine and ketocyclazocine did not. The nonopioid psychoactive drugs, mescaline, ketamine, physostigmine and scopolamine, also failed to produce discriminative effect equivalent to those produced by 3.0 mg/kg of morphine. These results confirm and extend our previous findings that of those drugs which have also been evaluated in man, discriminative effects equivalent to the training dose of morphine are produced uniquely by narcotic analgesics and narcotic antagonists which produce morphine-like subjective effects. These results are compatible with the hypothesis that the properties of morphine which enable it to function as a discriminative stimulus in the rat are analogous to those responsible for producing subjective effects in man. | Further evaluation of the discriminative effects of morphine in the rat. Rats were trained in a two-choice discrete trial avoidance paradigm to discriminate between saline and 3.0 mg/kg of morphine. Behavior was considered to be under stimulus ocntrol when the rats completed at least 90% of the trials in a 20-trial session on the morphine-appropriate choice lever after receiving morphine and when they completed at least 90% of the trials on the saline-appropriate choice lever after receiving saline. The discriminative effects of morphine, measured by responding on the morphine-appropriate lever, were then evaluated by determining the dose-response characteristics of representative narcotic analgesics, analgesics with mixed agonist and narcotic antagonist properties and nonopioid psychoactive drugs. Eight narcotic analgesics each produced dose-related responding on the morphine-appropriate lever. The relative potency for producing discriminative effects equivalent to those produced by 3.0 mg/kg of morphine ranged form etonitazene = 1000 x morphine to propoxyphene = 0.0175 x morphine. Of the narcotic antagonist analgesics tested, butorphanol and nalmexone produced discriminative effects equivalent to those of the morphine training dose whereas nalorphine, levallorphan, oxilorphan, nalbuphine and ketocyclazocine did not. The nonopioid psychoactive drugs, mescaline, ketamine, physostigmine and scopolamine, also failed to produce discriminative effect equivalent to those produced by 3.0 mg/kg of morphine. These results confirm and extend our previous findings that of those drugs which have also been evaluated in man, discriminative effects equivalent to the training dose of morphine are produced uniquely by narcotic analgesics and narcotic antagonists which produce morphine-like subjective effects. These results are compatible with the hypothesis that the properties of morphine which enable it to function as a discriminative stimulus in the rat are analogous to those responsible for producing subjective effects in man. | [
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|
PMID:15105 | Discriminative effects of morphine in the squirrel monkey. | Squirrel monkeys were trained in a two-choice discrete trial avoidance task to discriminate between intramuscular injections of saline and 3.0 mg/kg of morphine. Morphine (0.1-10 mg/kg) produced a dose-related increase in the number of trials completed on the morphine-appropriate lever. The stimulus control produced by the discriminative effects of morphine met the following criteria for classification as a specific narcotic effect: 1) morphine-like stimulus control was produced by all other narcotic analgesics tested (fentanyl, oxymorphone, levorphanol, methadone and meperidine); 2) in so doing, these drugs spanned a 900-fold potency range relative to morphine; 3) stimulus control was blocked by the specific narcotic antagonist naloxone; and 4) stereospecificity was a requirement for stimulus control--levorphanol produced stimulus control equivalent to 3.0 mg/kg of mrophine but its optical isomer dextrorphan did not. The time course of the stimulus control produced by 3.0 mg/kg of morphine showed that the animals continued to respond on the morphine-appropriate lever up to 14 hours after morphine administration. In contrast, monkeys administered 0.01 mg/kg of fentanyl responded on the morphine lever for only as lone as 1/2 hour after fentanyl administration. Naloxone, d-amphetamine and pentobarbital all failed to substitute for morphine, Thus, this study has extended previous observations of the discriminative properties of morphine in rats by demonstrating that qualitatively similar data are produced in a second species, the squirrel monkey. | Discriminative effects of morphine in the squirrel monkey. Squirrel monkeys were trained in a two-choice discrete trial avoidance task to discriminate between intramuscular injections of saline and 3.0 mg/kg of morphine. Morphine (0.1-10 mg/kg) produced a dose-related increase in the number of trials completed on the morphine-appropriate lever. The stimulus control produced by the discriminative effects of morphine met the following criteria for classification as a specific narcotic effect: 1) morphine-like stimulus control was produced by all other narcotic analgesics tested (fentanyl, oxymorphone, levorphanol, methadone and meperidine); 2) in so doing, these drugs spanned a 900-fold potency range relative to morphine; 3) stimulus control was blocked by the specific narcotic antagonist naloxone; and 4) stereospecificity was a requirement for stimulus control--levorphanol produced stimulus control equivalent to 3.0 mg/kg of mrophine but its optical isomer dextrorphan did not. The time course of the stimulus control produced by 3.0 mg/kg of morphine showed that the animals continued to respond on the morphine-appropriate lever up to 14 hours after morphine administration. In contrast, monkeys administered 0.01 mg/kg of fentanyl responded on the morphine lever for only as lone as 1/2 hour after fentanyl administration. Naloxone, d-amphetamine and pentobarbital all failed to substitute for morphine, Thus, this study has extended previous observations of the discriminative properties of morphine in rats by demonstrating that qualitatively similar data are produced in a second species, the squirrel monkey. | [
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|
PMID:15108 | Control of hepatic and intestinal blood flow: effect of isovolaemic haemodilution on blood flow and oxygen uptake in the intact liver and intestines. | 1. Limited isovolaemic haemodilution was produced in cats by addition of dextran 75-Ringer solution to an extracorporeal blood reservoir connected in series with the cat. Total hepatic venous outflow was neasured using a hepatic venous long-circuit and hepatic arterial flow was measured with an electromagnetic flow probe. Oxygen uptake was monitored in the guts and liver. Na-pentobarbitone anaesthesia was used. 2. Following reduction of the haematocrit (from 31 to 22) the oxygen uptake of the gut segment and liver were maintained. Gut conductance increased to 125% of control while the oxygen extraction ratio increased to only 109%. The hepatic arterial conductance did not change in spite of a greatly reduced (to 68%) oxygen delivery. Hepatic extraction increased to 140% of control. 3. The hepatic artery did not dilate to maintain constant oxygen supply to the liver thus confirming our previous observation that blood flow is not coupled to hepatic metabolism. 4. Oxygen extraction in the gut correlated well with changes in portal blood flow but not with changes in vascular conductance, arterial blood pressure or oxygen delivery. 5. The blood flow of the gut (vascular beds draining into the portal vein in the splenectomized preparation) was controlled in a manner that prevented changes in portal venous PO2 in spite of a reduction in oxygen content. Local PO2 and perhaps pH, are suggested as the factors controlling gut blood flow following haemodilution. 6. Changes in portal blood flow correlated with changes in portal vascular (intrahepatic) conductance such that increased portal flow produced an increased portal conductance thereby maintaining portal venous pressure constant. | Control of hepatic and intestinal blood flow: effect of isovolaemic haemodilution on blood flow and oxygen uptake in the intact liver and intestines. 1. Limited isovolaemic haemodilution was produced in cats by addition of dextran 75-Ringer solution to an extracorporeal blood reservoir connected in series with the cat. Total hepatic venous outflow was neasured using a hepatic venous long-circuit and hepatic arterial flow was measured with an electromagnetic flow probe. Oxygen uptake was monitored in the guts and liver. Na-pentobarbitone anaesthesia was used. 2. Following reduction of the haematocrit (from 31 to 22) the oxygen uptake of the gut segment and liver were maintained. Gut conductance increased to 125% of control while the oxygen extraction ratio increased to only 109%. The hepatic arterial conductance did not change in spite of a greatly reduced (to 68%) oxygen delivery. Hepatic extraction increased to 140% of control. 3. The hepatic artery did not dilate to maintain constant oxygen supply to the liver thus confirming our previous observation that blood flow is not coupled to hepatic metabolism. 4. Oxygen extraction in the gut correlated well with changes in portal blood flow but not with changes in vascular conductance, arterial blood pressure or oxygen delivery. 5. The blood flow of the gut (vascular beds draining into the portal vein in the splenectomized preparation) was controlled in a manner that prevented changes in portal venous PO2 in spite of a reduction in oxygen content. Local PO2 and perhaps pH, are suggested as the factors controlling gut blood flow following haemodilution. 6. Changes in portal blood flow correlated with changes in portal vascular (intrahepatic) conductance such that increased portal flow produced an increased portal conductance thereby maintaining portal venous pressure constant. | [
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|
PMID:15109 | The measurement of distortion: theoretical considerations. | When very find measurements are recorded for the purpose of establishing distortion values, from the standpoint of accuracy, specific procedures must be developed. In the procedural system defined here, a potential major source of error is indicated and the means of overcoming this error established. From the standpoint of modern electronic technology, the use of an analogue-to-digital interface with a digital computer is considered extremely advantageous and allows the utilization of technical help to carry out the preliminary measurements. Under these circumstances, major error sources, such as reader error and calculation error, are eliminated. Further, multiple readings on each point can be made without significant sacrifice of time. This latter statement can help define the reading accuracy on each point and allows an immediate further set of readings on any point that has a large variation in the initially established set of readings. The applicability of this approach to any system in which an accurate set of readings is required to define distortion or some other parameter is limited only by the imagination of the investigator. Measuring points can be established at any desired location, and the number required is a function of the mathematical analysis being attempted. The use of a digital computer to compute the final values allows an immediate remeasurement of the initial values if necessary, since the computer response time is of the order of seconds for the calculations carried out. This is very important if changes in the experimental setup are likely to occur with time. In general, a computerized scheme such as is proposed here allows the investigator to concentrate on the experiment rather than the calculation of results. Also, the use of technical help to carry out delicate measuring procedures can allow the investigator more time to analyze the final results. | The measurement of distortion: theoretical considerations. When very find measurements are recorded for the purpose of establishing distortion values, from the standpoint of accuracy, specific procedures must be developed. In the procedural system defined here, a potential major source of error is indicated and the means of overcoming this error established. From the standpoint of modern electronic technology, the use of an analogue-to-digital interface with a digital computer is considered extremely advantageous and allows the utilization of technical help to carry out the preliminary measurements. Under these circumstances, major error sources, such as reader error and calculation error, are eliminated. Further, multiple readings on each point can be made without significant sacrifice of time. This latter statement can help define the reading accuracy on each point and allows an immediate further set of readings on any point that has a large variation in the initially established set of readings. The applicability of this approach to any system in which an accurate set of readings is required to define distortion or some other parameter is limited only by the imagination of the investigator. Measuring points can be established at any desired location, and the number required is a function of the mathematical analysis being attempted. The use of a digital computer to compute the final values allows an immediate remeasurement of the initial values if necessary, since the computer response time is of the order of seconds for the calculations carried out. This is very important if changes in the experimental setup are likely to occur with time. In general, a computerized scheme such as is proposed here allows the investigator to concentrate on the experiment rather than the calculation of results. Also, the use of technical help to carry out delicate measuring procedures can allow the investigator more time to analyze the final results. | [
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PMID:15110 | Cathepsin D activity in bovine articular cartilage, synovial membrane and fluid: degradation of cartilage proteoglycans from same joint. | Cathepsin D type proteases were extracted from articular cartilage, synovial membrane, and synovial fluid from normal, adult bovine knee joints. A sensitive enzyme assay made it possible to measure protease activity in the different tissues from individual joints. Highest activity was found in the synovial membrane, while cell free synovial fluids contained comparatively low activity. The degrading effect on articular cartilage proteoglycans (PGC and PGS), isolated from the same joints, was demonstrated by gelfiltration on Sepharose columns and by viscometry. Gelfiltration profiles of incubation mixtures indicated a proteolytic effect on PGC and on PGS), at pH 3.5, in concentrations of enzyme and proteoglycans found in cartilage tissue. No effect at neutral pH was obtained despite a 100-fold increase of enzyme concentration. These findings were supported by viscometry data. The degrading effect of enzymes from all sources was completely inhibited by pepstatin. | Cathepsin D activity in bovine articular cartilage, synovial membrane and fluid: degradation of cartilage proteoglycans from same joint. Cathepsin D type proteases were extracted from articular cartilage, synovial membrane, and synovial fluid from normal, adult bovine knee joints. A sensitive enzyme assay made it possible to measure protease activity in the different tissues from individual joints. Highest activity was found in the synovial membrane, while cell free synovial fluids contained comparatively low activity. The degrading effect on articular cartilage proteoglycans (PGC and PGS), isolated from the same joints, was demonstrated by gelfiltration on Sepharose columns and by viscometry. Gelfiltration profiles of incubation mixtures indicated a proteolytic effect on PGC and on PGS), at pH 3.5, in concentrations of enzyme and proteoglycans found in cartilage tissue. No effect at neutral pH was obtained despite a 100-fold increase of enzyme concentration. These findings were supported by viscometry data. The degrading effect of enzymes from all sources was completely inhibited by pepstatin. | [
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|
PMID:15111 | 3-Halo-5,7-dimethylpyrazolo [1,5-a]pyrimidines, a nonbenzodiazepinoid class of antianxiety agents devoid of potentiation of central nervous system depressant effects of ethanol or barbiturates. | Forty derivatives (1-40) of pyrazolo[1,5-a]pyrimidine were synthesized and evaluated for antianxiety properties via gross behavioral observations in rats. Five of these compounds, including 5,7-dimethylpyrazolo[1,5-a]pyrimidine (6) and the 3-fluoro (7), 3-chloro (8), 3-bromo (9), and 3-iodo (10) derivatives, were selected for advanced evaluation. Although 6 and 7 had marginal activity, 8-10 had an anxiolytic effect in animals comparable to the clinically useful benzodiazepines, diazepam, and chlorodiazepoxide. Comparison with chlorpromazine indicated that 6-10 are probably not antipsychotic agents. These compounds also lacked activity in anticonvulsant and analgesic tests. Acute toxicity data (mouse, ip and po) indicated that 8-10 had excellent therapeutic ratios, although 10 was more poorly absorbed than 8 and 9. Further demonstration of anxiolytic efficacy was obtained by comparing the effects of 8 and 9 with the benzodiazepines in modifying provoked aggression in monkeys, rats (muricide), and fighting mice. The most remarkable observation, however, was that 8 and 9 had no effect, at the anxiolytic threshold, in potentiating the CNS depressant effects of ethanol or sodium barbital (po) in treated mice. In contrast, diazepam and chlorodiazepoxide potentiated this drug interaction effect at minimal anxiolytic doses. | 3-Halo-5,7-dimethylpyrazolo [1,5-a]pyrimidines, a nonbenzodiazepinoid class of antianxiety agents devoid of potentiation of central nervous system depressant effects of ethanol or barbiturates. Forty derivatives (1-40) of pyrazolo[1,5-a]pyrimidine were synthesized and evaluated for antianxiety properties via gross behavioral observations in rats. Five of these compounds, including 5,7-dimethylpyrazolo[1,5-a]pyrimidine (6) and the 3-fluoro (7), 3-chloro (8), 3-bromo (9), and 3-iodo (10) derivatives, were selected for advanced evaluation. Although 6 and 7 had marginal activity, 8-10 had an anxiolytic effect in animals comparable to the clinically useful benzodiazepines, diazepam, and chlorodiazepoxide. Comparison with chlorpromazine indicated that 6-10 are probably not antipsychotic agents. These compounds also lacked activity in anticonvulsant and analgesic tests. Acute toxicity data (mouse, ip and po) indicated that 8-10 had excellent therapeutic ratios, although 10 was more poorly absorbed than 8 and 9. Further demonstration of anxiolytic efficacy was obtained by comparing the effects of 8 and 9 with the benzodiazepines in modifying provoked aggression in monkeys, rats (muricide), and fighting mice. The most remarkable observation, however, was that 8 and 9 had no effect, at the anxiolytic threshold, in potentiating the CNS depressant effects of ethanol or sodium barbital (po) in treated mice. In contrast, diazepam and chlorodiazepoxide potentiated this drug interaction effect at minimal anxiolytic doses. | [
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|
PMID:15112 | Synthesis and adrenergic beta-blocking activity of some 1,3-benzodioxole derivatives. | A series of 1,3-benzodioxole derivatives was synthesized. We found four compounds (2,3,10 and 11 in Table IV) to have about the same order of beta-blocking activity as that of sotalol. In addition, it is of interest that some of the compounds (2-4) were found to have hypotensive activites, although they were about one-tenth of that of hydralazine. Sotalol did not produce any change in blood pressure, and propranolol raised the blood pressure. | Synthesis and adrenergic beta-blocking activity of some 1,3-benzodioxole derivatives. A series of 1,3-benzodioxole derivatives was synthesized. We found four compounds (2,3,10 and 11 in Table IV) to have about the same order of beta-blocking activity as that of sotalol. In addition, it is of interest that some of the compounds (2-4) were found to have hypotensive activites, although they were about one-tenth of that of hydralazine. Sotalol did not produce any change in blood pressure, and propranolol raised the blood pressure. | [
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|
PMID:15113 | Beta-adrenoceptor studies. 2. Effects of alkyl substitution on beta-adrenoceptor blocking, antiarrhythmic, and local anesthetic activities of 1,1'-(o-phenylenedioxy)bis(3-isopropylamino-2-propanol). | A series of bis(2-hydroxy-3-isopropylaminopropyl) ethers of nuclear-substituted catechols (1-7) has been synthesized and examined in vitro for beta-adrenoceptor blocking activity, antagonism of ouabain-induced arrhythmias, and local anesthetic activity. Both tracheal and right atrial beta-adrenoceptor blocking activity are markedly decreased by alkyl substitution in position 3 of parent catechol diether 1. Substitution in position 4 still lowers the affinity to cardiac arrhythmias and local anesthetic activity increases with introduction of alkyl substituents in the 3 as well as in the 4 position. In contrast with biological activities, the partition coefficient 1-octanol-phosphate buffer, pH 7.40, of 1 did not change significantly by 3- and 4-methyl substitution. Stepwise multiple regression analyses were performed using log P or pi values in combination with pKa(m), E8, or sigma. With cardiac beta-adrenoceptor blocking activity the optimal equation contained E8 and pi parameters, tracheal activity appeared to depend mainly on the E8 parameter, whereas for antiarrhythmic and local anesthetic activities the lipophilicity of the substituents appeared to be the determinant factor. | Beta-adrenoceptor studies. 2. Effects of alkyl substitution on beta-adrenoceptor blocking, antiarrhythmic, and local anesthetic activities of 1,1'-(o-phenylenedioxy)bis(3-isopropylamino-2-propanol). A series of bis(2-hydroxy-3-isopropylaminopropyl) ethers of nuclear-substituted catechols (1-7) has been synthesized and examined in vitro for beta-adrenoceptor blocking activity, antagonism of ouabain-induced arrhythmias, and local anesthetic activity. Both tracheal and right atrial beta-adrenoceptor blocking activity are markedly decreased by alkyl substitution in position 3 of parent catechol diether 1. Substitution in position 4 still lowers the affinity to cardiac arrhythmias and local anesthetic activity increases with introduction of alkyl substituents in the 3 as well as in the 4 position. In contrast with biological activities, the partition coefficient 1-octanol-phosphate buffer, pH 7.40, of 1 did not change significantly by 3- and 4-methyl substitution. Stepwise multiple regression analyses were performed using log P or pi values in combination with pKa(m), E8, or sigma. With cardiac beta-adrenoceptor blocking activity the optimal equation contained E8 and pi parameters, tracheal activity appeared to depend mainly on the E8 parameter, whereas for antiarrhythmic and local anesthetic activities the lipophilicity of the substituents appeared to be the determinant factor. | [
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|
PMID:15114 | Synthesis and hypoglycemic activity of S-acyl derivatives of 3-mercaptopicolinic acid. | A series of S-alkanoyl and benzoyl derivatives of 3-mercaptopicolinic acid (3-MPA) was prepared and studied for hypoglycemic activity. Three alkanoyl derivatives (propionyl, pivaloyl, and 1-adamantanecarbonyl, 19-21) were prepared with increasing bulk around the thio ester bond. The benzoyl derivatives contained aromatic substituents chosen from a sigma-pi cluster chart so that the esters prepared had a wide range of electronic and solubility properties. In general, compounds with substituents which increased lipid solubility [p-chlorobenzoyl (4), p-trifluoromethylbenzoyl (6), and pivaloyl (20)] had the greatest potency at a dose of 300 mg/kg. Hydrolysis rates, measured at pH 6 and 8, indicated that in vivo breakdown to 3-MPA probably did not account for the observed hypoglycemic activity of the esters. 4, 6, and 20 were less potent than 3-MPA in comparative dose range studies. | Synthesis and hypoglycemic activity of S-acyl derivatives of 3-mercaptopicolinic acid. A series of S-alkanoyl and benzoyl derivatives of 3-mercaptopicolinic acid (3-MPA) was prepared and studied for hypoglycemic activity. Three alkanoyl derivatives (propionyl, pivaloyl, and 1-adamantanecarbonyl, 19-21) were prepared with increasing bulk around the thio ester bond. The benzoyl derivatives contained aromatic substituents chosen from a sigma-pi cluster chart so that the esters prepared had a wide range of electronic and solubility properties. In general, compounds with substituents which increased lipid solubility [p-chlorobenzoyl (4), p-trifluoromethylbenzoyl (6), and pivaloyl (20)] had the greatest potency at a dose of 300 mg/kg. Hydrolysis rates, measured at pH 6 and 8, indicated that in vivo breakdown to 3-MPA probably did not account for the observed hypoglycemic activity of the esters. 4, 6, and 20 were less potent than 3-MPA in comparative dose range studies. | [
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|
PMID:15115 | Relative concentrations of zwitterionic and uncharged species in catecholamines and the effect of N-substituents. | The relative concentrations of zwitterionic and uncharged species for the series of N-substituted catecholamines (I, R1 = R2 = OH; R = H, Me, Et, i-Pr, t-Bu) are derived from the pKa data published in 1962 by Sinistri and Villa. The concentration ratios, represented by the tautomeric equilibrium constant Kt, show a definite trend and are respectively 1.8, 4.3, 4.7, 4.7, and 7.1. These values suggest that any mechanism of action involving proton transfer, which might transform the zwitterion into the uncharged form, would be most favorable for norepinephrine and least favorable fo the t-Bu derivative. | Relative concentrations of zwitterionic and uncharged species in catecholamines and the effect of N-substituents. The relative concentrations of zwitterionic and uncharged species for the series of N-substituted catecholamines (I, R1 = R2 = OH; R = H, Me, Et, i-Pr, t-Bu) are derived from the pKa data published in 1962 by Sinistri and Villa. The concentration ratios, represented by the tautomeric equilibrium constant Kt, show a definite trend and are respectively 1.8, 4.3, 4.7, 4.7, and 7.1. These values suggest that any mechanism of action involving proton transfer, which might transform the zwitterion into the uncharged form, would be most favorable for norepinephrine and least favorable fo the t-Bu derivative. | [
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|
PMID:15123 | Interviewing skills: a comprehensive approach to teaching and evaluation. | Evaluation procedures utilizing standardized interviews scored by means of a formal objective rating system which make it possible to measure the interviewing skills of health professional students have been developed in recent years. These procedures have been used in the Child Health Associate Program at the University of Colorado Medical School to evaluate the ability of a group of students to achieve the objectives of a practice-oriented interviewing course. Results from the standardized interviews indicate that students taking the course gathered an average of 76 percent of all available data and used 86 percent of the process skills defined as necessary for positive interview interaction. A previous group of child health associate students who did not take the course gathered an average of 47 percent of all available data and used an average of 62 percent of the necessary process skills. | Interviewing skills: a comprehensive approach to teaching and evaluation. Evaluation procedures utilizing standardized interviews scored by means of a formal objective rating system which make it possible to measure the interviewing skills of health professional students have been developed in recent years. These procedures have been used in the Child Health Associate Program at the University of Colorado Medical School to evaluate the ability of a group of students to achieve the objectives of a practice-oriented interviewing course. Results from the standardized interviews indicate that students taking the course gathered an average of 76 percent of all available data and used 86 percent of the process skills defined as necessary for positive interview interaction. A previous group of child health associate students who did not take the course gathered an average of 47 percent of all available data and used an average of 62 percent of the necessary process skills. | [
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|
PMID:15124 | Evaluation of a child health associate program. | The staff of the University of Colorado Child Health Associate Program critically reviewed the effectiveness of the program's structure and content during an intensive two-day seminar. The review was conducted through workshops involving participants representing students, graduates, faculty, employers, funding agencies, university administrators, and educational consultants. The agenda for the evaluation included workshops on specific topics such as basic and clinical sciences, psychosocial skills, and proficiency testing. Information obtained provided extremely valuable data which were used to improve the program. | Evaluation of a child health associate program. The staff of the University of Colorado Child Health Associate Program critically reviewed the effectiveness of the program's structure and content during an intensive two-day seminar. The review was conducted through workshops involving participants representing students, graduates, faculty, employers, funding agencies, university administrators, and educational consultants. The agenda for the evaluation included workshops on specific topics such as basic and clinical sciences, psychosocial skills, and proficiency testing. Information obtained provided extremely valuable data which were used to improve the program. | [
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|
PMID:15125 | A protonmotive force as the source of energy for galactoside transport in energy depleted Escherichia coli. | An artificially produced electrochemical potential difference for protons (portonmotive force) provided the energy for the transport of galactosides in Escherichia coli cells which were depleted of their endogenous energy reserves. The driving force for the entry of protons was provided by either a transmembrane pH gradient or a membrane potential. The pH gradient across the membrane was created by acidifying the external medium. The membrane potential (inside negative) was established by the outward diffusion of potassium (in the presence of valinomycin) or by the inward diffusion of the permeant thiocyanate ion. The magnitude of the electrochemical potential difference for protons agreed well with magnitude of the chemical potential difference of the lactose analog, thiomethylgalactoside. The observations are consistent with the view that the carrier-mediated entry of each galactoside molecule is accompanied by the entry of one proton. | A protonmotive force as the source of energy for galactoside transport in energy depleted Escherichia coli. An artificially produced electrochemical potential difference for protons (portonmotive force) provided the energy for the transport of galactosides in Escherichia coli cells which were depleted of their endogenous energy reserves. The driving force for the entry of protons was provided by either a transmembrane pH gradient or a membrane potential. The pH gradient across the membrane was created by acidifying the external medium. The membrane potential (inside negative) was established by the outward diffusion of potassium (in the presence of valinomycin) or by the inward diffusion of the permeant thiocyanate ion. The magnitude of the electrochemical potential difference for protons agreed well with magnitude of the chemical potential difference of the lactose analog, thiomethylgalactoside. The observations are consistent with the view that the carrier-mediated entry of each galactoside molecule is accompanied by the entry of one proton. | [
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PMID:15126 | Studies on the cation permeability of human red cell ghosts. Characterization and biological significance of two membrane sites with high affinities for Ca. | Net K movements in reconstituted human red cell ghosts and the resealing of ghosts to cations after osmotic hemolysis of red cells have been studied as functions of the free Ca ion concentration. The Ca-dependent specific increase in K permeability was shown to be mediated by a site close to the internal surface of the membrane with an apparent dissociation constant ap pH 7.2 for Ca (K'p1) of 3-5 X 10(-7) M, for Sr of 7 X 10(-6) M. Ba and Mg did not increase the K-permeability of the membrane but inhibited the Ca-mediated permeability changes. K'D1 decreased in a nonlinear fashion when the pH was increased from 6.0 to 8.5. Two different pK' values of this membrane site were found at pH 8.3 and 6.3. The Ca-activated net K efflux into a K-free medium was almost completely inhibited by an increase in intracellular Na from 4 to 70mM. Extracellular K antagonized this Na effect. Changes in the extracellular Na (0.1-140 mM) or K (0.1-6 mM) concentrations had little effect and did not change K'p1. The Ca-stimulated recovery of a low cation permeability in ghost cells appeared to be mediated by a second membrane site which was accessible to divalent cations only during the process of hemolysis in media of low ionic strength. The apparent dissociation constant for Ca at this site (K'p2) varied between 6 X 10(-7) and 4 X 10(-6) M at pH 7.2 Mg, Sr, and Ba could replace Ca functionally. The selectivity sequence was Ca greater than Sr greater than Ba greater than Mg. K'p2 was independent on the pH value in the range between 6.0 and 8.0 Hill coefficients of 2 were observed for the interaction of Ca with both membrane sites suggesting that more than one Ca ion is bound per site. The Hill cofficients were affected neither by the ion composition nor by the Ph values of the intra-and extracellular media. It is concluded that two different pathways for the permeation of cations across the membrane are controlled by membrane sites with high affinities for Ca: One specific for K, one unspecific with respect to cations. The K-specific "channel" has properties similar to the K channel in excitable tissues. | Studies on the cation permeability of human red cell ghosts. Characterization and biological significance of two membrane sites with high affinities for Ca. Net K movements in reconstituted human red cell ghosts and the resealing of ghosts to cations after osmotic hemolysis of red cells have been studied as functions of the free Ca ion concentration. The Ca-dependent specific increase in K permeability was shown to be mediated by a site close to the internal surface of the membrane with an apparent dissociation constant ap pH 7.2 for Ca (K'p1) of 3-5 X 10(-7) M, for Sr of 7 X 10(-6) M. Ba and Mg did not increase the K-permeability of the membrane but inhibited the Ca-mediated permeability changes. K'D1 decreased in a nonlinear fashion when the pH was increased from 6.0 to 8.5. Two different pK' values of this membrane site were found at pH 8.3 and 6.3. The Ca-activated net K efflux into a K-free medium was almost completely inhibited by an increase in intracellular Na from 4 to 70mM. Extracellular K antagonized this Na effect. Changes in the extracellular Na (0.1-140 mM) or K (0.1-6 mM) concentrations had little effect and did not change K'p1. The Ca-stimulated recovery of a low cation permeability in ghost cells appeared to be mediated by a second membrane site which was accessible to divalent cations only during the process of hemolysis in media of low ionic strength. The apparent dissociation constant for Ca at this site (K'p2) varied between 6 X 10(-7) and 4 X 10(-6) M at pH 7.2 Mg, Sr, and Ba could replace Ca functionally. The selectivity sequence was Ca greater than Sr greater than Ba greater than Mg. K'p2 was independent on the pH value in the range between 6.0 and 8.0 Hill coefficients of 2 were observed for the interaction of Ca with both membrane sites suggesting that more than one Ca ion is bound per site. The Hill cofficients were affected neither by the ion composition nor by the Ph values of the intra-and extracellular media. It is concluded that two different pathways for the permeation of cations across the membrane are controlled by membrane sites with high affinities for Ca: One specific for K, one unspecific with respect to cations. The K-specific "channel" has properties similar to the K channel in excitable tissues. | [
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PMID:15129 | Enhancement of 5-iododeoxyuridine-induced endogenous C-type virus activation by polycyclic hydrocarbons: apparent lack of parallelism between enhancement and carcinogenicity. | When mouse MLg cells were treated with 3-methylcholanthrene or 7,12-dimethylbenz[alpha]anthracene in the presence of microsomal enzymes and NADPH after 5-iododeoxyuridine (IUDR) treatment, the induction rate of the endogenous C-type virus was increased fivefold to sixfold in comparison with the culture treated with IUDR only. In this reaction, both the microsomal enzymes and NADPH were indispensable. 7,8-Benzoflavone, an inhibitor of the metabolism of hydrocarbons in hamster embryo cultures, inhibited the reaction. For detecting the enhancing activity, the concentration of IUDR for the pretreatment, the concentration of the test products, and the duration of the treatment with the products were important factors. In screening 30 polycyclic hydrocarbons, we were unable to detect a correlation between the in vivo carcinogenicity in the skin and the enhancing activity in the conditions tested. | Enhancement of 5-iododeoxyuridine-induced endogenous C-type virus activation by polycyclic hydrocarbons: apparent lack of parallelism between enhancement and carcinogenicity. When mouse MLg cells were treated with 3-methylcholanthrene or 7,12-dimethylbenz[alpha]anthracene in the presence of microsomal enzymes and NADPH after 5-iododeoxyuridine (IUDR) treatment, the induction rate of the endogenous C-type virus was increased fivefold to sixfold in comparison with the culture treated with IUDR only. In this reaction, both the microsomal enzymes and NADPH were indispensable. 7,8-Benzoflavone, an inhibitor of the metabolism of hydrocarbons in hamster embryo cultures, inhibited the reaction. For detecting the enhancing activity, the concentration of IUDR for the pretreatment, the concentration of the test products, and the duration of the treatment with the products were important factors. In screening 30 polycyclic hydrocarbons, we were unable to detect a correlation between the in vivo carcinogenicity in the skin and the enhancing activity in the conditions tested. | [
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|
PMID:15130 | Lack of influence of hypophysectomy on estrogen-induced DNA synthesis in Leydig cells of BALB/c mice. | In mice of strains susceptible to Leydig cell tumor induction, treatment with estrogens induced a "spurt" of DNA synthesis within the first few days. This synthetic activity generally subsided, until areas of Leydig cell hyperplasia developed several months later. Autoradiographic and quantitative biochemical studies indicated that in BALB/c mice this initial DNA synthetic activity occurred in the absence of the hypophysis and apparently was the result of effects of estrogen directly on Leydig cells. Although hypophysectomy inhibited sperm maturation, [3H]thymidine incorporation into spermatogonia was reduced only slightly 2 weeks after surgery, as was the induced DNA spurt in the interstitial tissues. | Lack of influence of hypophysectomy on estrogen-induced DNA synthesis in Leydig cells of BALB/c mice. In mice of strains susceptible to Leydig cell tumor induction, treatment with estrogens induced a "spurt" of DNA synthesis within the first few days. This synthetic activity generally subsided, until areas of Leydig cell hyperplasia developed several months later. Autoradiographic and quantitative biochemical studies indicated that in BALB/c mice this initial DNA synthetic activity occurred in the absence of the hypophysis and apparently was the result of effects of estrogen directly on Leydig cells. Although hypophysectomy inhibited sperm maturation, [3H]thymidine incorporation into spermatogonia was reduced only slightly 2 weeks after surgery, as was the induced DNA spurt in the interstitial tissues. | [
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|
PMID:15131 | Effect of hyperthermia and environmental acidity on the proteolytic activity in murine ascites tumor cells. | The influence of hyperthermia and environmental pH on proteolytic activity was studied in murine ascites tumor cells in vitro. PNJ ascites tumor cells were incubated with [125I]cytochrome c at 42.5 or 37 degrees C in a modified Krebs-Ringer buffer adjusted to pH 7.2 or 6.4. Incubation at normal temperature at pH 6.4 and 7.2 or at 42.5 degrees C and pH 7.2 resulted in identical protein digestion. However, hyperthermic incubation at pH 6.4 resulted in a significant increased activity. This was also observed after only 1 hour of hyperthermic incubation followed by subsequent incubation in an acidic environment at normal temperature. The increased proteolytic activity following hyperthermic treatment under acidic conditions may support the hypothesis that increased lysosomal activity is of primary importance in the hyperthermic tumor-cell destruction in vivo. | Effect of hyperthermia and environmental acidity on the proteolytic activity in murine ascites tumor cells. The influence of hyperthermia and environmental pH on proteolytic activity was studied in murine ascites tumor cells in vitro. PNJ ascites tumor cells were incubated with [125I]cytochrome c at 42.5 or 37 degrees C in a modified Krebs-Ringer buffer adjusted to pH 7.2 or 6.4. Incubation at normal temperature at pH 6.4 and 7.2 or at 42.5 degrees C and pH 7.2 resulted in identical protein digestion. However, hyperthermic incubation at pH 6.4 resulted in a significant increased activity. This was also observed after only 1 hour of hyperthermic incubation followed by subsequent incubation in an acidic environment at normal temperature. The increased proteolytic activity following hyperthermic treatment under acidic conditions may support the hypothesis that increased lysosomal activity is of primary importance in the hyperthermic tumor-cell destruction in vivo. | [
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|
PMID:15132 | Nature of the delayed graft-versus-host reactivity of fetal liver cell transplants in mice. | Experiments were designed to determine which actual differences in the cellular composition between fetal liver and bone marrow account for the distinct types of graft-versus-host (GvH) disease. The assay of reactive lymphocytes (by in vitro mitogenic stimulation) in fetal liver transplants in mice, the purification of hemopoietic stem cells (HSC) of the transplants, and the quantitation of HSC numbers in the grafts traced the basis for the distinctly weak type of GvH disease after fetal liver cell grafts. It was found that transplantation of purified HSC concentrates did not modify the severity of GvH mortality. The moderate character of the delayed GvH disease was shown to depend on the presence of an HSC population in fetal liver with different qualities and not on numerical differences between the HSC in fetal liver and bone marrow. Data collected also demonstrated that when GvH disease occurred in the recipients of transplants of fetal liver, it shared the characteristic histologic features of the bone marrow GvH syndrome. The recovery of mitogen responsiveness of spleen cells may have been delayed in fetal liver allotransplantation as compared to syngeneic grafting. By supportive infusion of lymphoid cells, it was suggested that the immunodeficiency coinciding with GvH disease represented a secondary manifestation of the disease rather than a primary impairment in lymphoid differentiation. | Nature of the delayed graft-versus-host reactivity of fetal liver cell transplants in mice. Experiments were designed to determine which actual differences in the cellular composition between fetal liver and bone marrow account for the distinct types of graft-versus-host (GvH) disease. The assay of reactive lymphocytes (by in vitro mitogenic stimulation) in fetal liver transplants in mice, the purification of hemopoietic stem cells (HSC) of the transplants, and the quantitation of HSC numbers in the grafts traced the basis for the distinctly weak type of GvH disease after fetal liver cell grafts. It was found that transplantation of purified HSC concentrates did not modify the severity of GvH mortality. The moderate character of the delayed GvH disease was shown to depend on the presence of an HSC population in fetal liver with different qualities and not on numerical differences between the HSC in fetal liver and bone marrow. Data collected also demonstrated that when GvH disease occurred in the recipients of transplants of fetal liver, it shared the characteristic histologic features of the bone marrow GvH syndrome. The recovery of mitogen responsiveness of spleen cells may have been delayed in fetal liver allotransplantation as compared to syngeneic grafting. By supportive infusion of lymphoid cells, it was suggested that the immunodeficiency coinciding with GvH disease represented a secondary manifestation of the disease rather than a primary impairment in lymphoid differentiation. | [
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|
PMID:15133 | Characterization of group H streptococcal temperate bacteriophage phi 227. | phi 227, a temperate phage from a group H streptococcus (Streptococcus sanguis), was propagated vegetatively in group H strain Wicky 4-EryR, and its characteristics were determined. A procedure dependent on multiplicity of infection, incubation time, and treatment of crude lysates with diatomaceous earth was found to optimize phage yield, resulting in titers of 1 X 10(10) to 2 X 10(10) PFU/ml. Without prior treatment with diatomaceous earth, subsequent purification procedures (methanol, ammonium sulfate, polyethylene glycol) gave recoveries of less than 1% of crude lysate titers. Adsorption of phi227 to host cells was relatively unaffected by the medium, but calcium (not substituted by magnesium) was required for formation of infectious centers. The phage receptor was present on purified cell walls, resisted trypsin and heat, and was removed ty hydrochloric acid, trichloracetic acid, and hot formamide: however, formamide-extracted material failed to inactivate phage, and the nature of the receptor is unknown. Single-step growth experiments showed a latent period of 39 min and a burst size of 100 PFU/infectious center; results were unaffected by omission of supplemental Ca2+, by supplementation with Mg2, addition of glucose, or changes of pH between 6.35 and 8.0; but increased temperature (40 to 43 degrees C) shortened the latent period and decreased the burst size. The latent period was prolonged in genetically competent host cells and in chemically defined medium; and in the latter, the burst size was smaller. Phage replication was sensitive to those metabolic inhibitors which inhibited the host streptococcus: these included rifampin, fluorodeoxyuridine, hydroxyurea, dihydrostreptomycin, and 6-P-hydroxyphenylazouracil. The data suggest that phi227 does not code for a rifampin-resistant RNA polymerase. However, in a rifampin-resistant host strain, phage replication and lysogen formation were both decreased suggesting that altered host core polymerase had less affinity for (some) promotors on the phi227 template. In transfection, a Ca2+-dependent stabilization step that was inhibited by Mg2+ was demonstrated; transformation was not affected by either Ca2+ or Mg2+, and the site and nature of the stabilization are unknown. More than one molecule of DNA was required for plaque formation. Biophysical characterization showed a type B phage of buoyant density (CsCl) 1.50, containing five proteins and 54.8% DNA. The duplex linear DNA had a molecular weight (calculated from contour length) of 23.2 X 10(6) and a guanine plus cytosine content (calculated from melting point) of 42.3 mol%. Similar characterizations of streptococcal phages, including biophysical data, have not been previously available. | Characterization of group H streptococcal temperate bacteriophage phi 227. phi 227, a temperate phage from a group H streptococcus (Streptococcus sanguis), was propagated vegetatively in group H strain Wicky 4-EryR, and its characteristics were determined. A procedure dependent on multiplicity of infection, incubation time, and treatment of crude lysates with diatomaceous earth was found to optimize phage yield, resulting in titers of 1 X 10(10) to 2 X 10(10) PFU/ml. Without prior treatment with diatomaceous earth, subsequent purification procedures (methanol, ammonium sulfate, polyethylene glycol) gave recoveries of less than 1% of crude lysate titers. Adsorption of phi227 to host cells was relatively unaffected by the medium, but calcium (not substituted by magnesium) was required for formation of infectious centers. The phage receptor was present on purified cell walls, resisted trypsin and heat, and was removed ty hydrochloric acid, trichloracetic acid, and hot formamide: however, formamide-extracted material failed to inactivate phage, and the nature of the receptor is unknown. Single-step growth experiments showed a latent period of 39 min and a burst size of 100 PFU/infectious center; results were unaffected by omission of supplemental Ca2+, by supplementation with Mg2, addition of glucose, or changes of pH between 6.35 and 8.0; but increased temperature (40 to 43 degrees C) shortened the latent period and decreased the burst size. The latent period was prolonged in genetically competent host cells and in chemically defined medium; and in the latter, the burst size was smaller. Phage replication was sensitive to those metabolic inhibitors which inhibited the host streptococcus: these included rifampin, fluorodeoxyuridine, hydroxyurea, dihydrostreptomycin, and 6-P-hydroxyphenylazouracil. The data suggest that phi227 does not code for a rifampin-resistant RNA polymerase. However, in a rifampin-resistant host strain, phage replication and lysogen formation were both decreased suggesting that altered host core polymerase had less affinity for (some) promotors on the phi227 template. In transfection, a Ca2+-dependent stabilization step that was inhibited by Mg2+ was demonstrated; transformation was not affected by either Ca2+ or Mg2+, and the site and nature of the stabilization are unknown. More than one molecule of DNA was required for plaque formation. Biophysical characterization showed a type B phage of buoyant density (CsCl) 1.50, containing five proteins and 54.8% DNA. The duplex linear DNA had a molecular weight (calculated from contour length) of 23.2 X 10(6) and a guanine plus cytosine content (calculated from melting point) of 42.3 mol%. Similar characterizations of streptococcal phages, including biophysical data, have not been previously available. | [
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PMID:15134 | Characterization of components released by alkali disruption of simian virus 40. | Treatment of simian virus 40 (SV40) particles at pH 9.8 in the presence of 1 mM dithiothreitol for 5 min at 37 degrees C disrupted the virions into a 60S DNA-protein complex and DNA-free 7S protein particles. The DNA-protein complex contained approximately equal amounts of DNA and protein, and appeared by electron microscopy to be relaxed circular structures with an average of 21 beads joined by short, thin bridges. The major protein components in the complex were host cell histones, but SV40-specific proteins VP3 and VP2 were also present. The 7S protein particles were almost exclusively VP1 and, in negatively stained samples, resembled the capsomer structures of intact virions. | Characterization of components released by alkali disruption of simian virus 40. Treatment of simian virus 40 (SV40) particles at pH 9.8 in the presence of 1 mM dithiothreitol for 5 min at 37 degrees C disrupted the virions into a 60S DNA-protein complex and DNA-free 7S protein particles. The DNA-protein complex contained approximately equal amounts of DNA and protein, and appeared by electron microscopy to be relaxed circular structures with an average of 21 beads joined by short, thin bridges. The major protein components in the complex were host cell histones, but SV40-specific proteins VP3 and VP2 were also present. The 7S protein particles were almost exclusively VP1 and, in negatively stained samples, resembled the capsomer structures of intact virions. | [
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PMID:15135 | Temperature-sensitive defect of vesicular stomatitis virus in complementation group II. | The prototype member of the complementation group II temperature-sensitive (ts) mutants of vesicular stomatitis virus, ts II 052, has been investigated. In ts II 052-infected HeLa cells at the restrictive temperature (39.5 degrees C), reduced viral RNA synthesis was observed by comparison with infections conducted at the permissive temperature (30 degrees C). It was found that for an infection conducted at 39.5 degrees C, no 38S RNA or intracytoplasmic nucleocapsids were present. For nucleocapsids isolated from ts II 052 purified virions or from ts II 052-infected cells at 30 degrees C, the RNA was sensitive to pancreatic RNase after an exposure at 39.5 degrees C in contrast to the resistance observed for wild-type virus. The nucleocapsid stability of wild-type virus when heated to 63 degrees C or submitted to varying pH was not found in nucleocapsids extracted from ts II 052 purified virions. The data suggest that for ts II 052 there is an altered relationship between the viral 38S RNA and the nucleocapsid protein(s) by comparison with wild-type virus. Such results argue for the complementation group II gene product being N protein, so that the ts defect in ts II 052 represents an altered N protein. | Temperature-sensitive defect of vesicular stomatitis virus in complementation group II. The prototype member of the complementation group II temperature-sensitive (ts) mutants of vesicular stomatitis virus, ts II 052, has been investigated. In ts II 052-infected HeLa cells at the restrictive temperature (39.5 degrees C), reduced viral RNA synthesis was observed by comparison with infections conducted at the permissive temperature (30 degrees C). It was found that for an infection conducted at 39.5 degrees C, no 38S RNA or intracytoplasmic nucleocapsids were present. For nucleocapsids isolated from ts II 052 purified virions or from ts II 052-infected cells at 30 degrees C, the RNA was sensitive to pancreatic RNase after an exposure at 39.5 degrees C in contrast to the resistance observed for wild-type virus. The nucleocapsid stability of wild-type virus when heated to 63 degrees C or submitted to varying pH was not found in nucleocapsids extracted from ts II 052 purified virions. The data suggest that for ts II 052 there is an altered relationship between the viral 38S RNA and the nucleocapsid protein(s) by comparison with wild-type virus. Such results argue for the complementation group II gene product being N protein, so that the ts defect in ts II 052 represents an altered N protein. | [
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|
PMID:15136 | Receptor interaction between eastern equine encephalitis virus and chicken embryo fibroblasts. | The attachment of eastern equine encephalitis virus to chicken embryo fibroblasts was studied at 0 degrees C. The binding specifically responsible for initiating infection was studied in the initial experiments by employing plaque-forming ability as the measured response. Results from these initial studies were closely paralleled in studies of binding of radiolabeled virus under the same conditions. Binding that had occurred at the pH optimum, pH 6.5, could be reversed only at higher pH. The observed pH dependence of virus attachment suggested the interaction of at least two ionizable species in the initial binding of virus to cell, and that one to three attachments must occur between virus and cell prior to infection. | Receptor interaction between eastern equine encephalitis virus and chicken embryo fibroblasts. The attachment of eastern equine encephalitis virus to chicken embryo fibroblasts was studied at 0 degrees C. The binding specifically responsible for initiating infection was studied in the initial experiments by employing plaque-forming ability as the measured response. Results from these initial studies were closely paralleled in studies of binding of radiolabeled virus under the same conditions. Binding that had occurred at the pH optimum, pH 6.5, could be reversed only at higher pH. The observed pH dependence of virus attachment suggested the interaction of at least two ionizable species in the initial binding of virus to cell, and that one to three attachments must occur between virus and cell prior to infection. | [
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|
PMID:15137 | Priapism: evolution of management in 48 patients in a 22-year series. | The choice of an effective method to treat priapism is challenging because precise causes in the majority of patients have not been well defined. A review of 48 patients treated during a 22-year period shows evolution of a regimen of management that has yielded a high percentage of success. Idiopathic priapism and sickle cell disease accounted for 81 per cent of the subjects. An evaluation should include a medication history, a search for specific diseases, as well as a thorough physical examination to detect possible etiologic factors. The explanation for the frequent association of fever deserves further investigation. Initial therapy consisting of aspiration and irrigation, and intermittent pneumatic cuff compression should be undertaken for a trial period of 12 to 36 hours, repeating the aspiration 2 or 3 times if necessary. The failure of priapism to resolve after such treatment is an indication for a shunt operation. Patients with known etiology should be treated specifically for the primary disease and usually more conservatively for priapism. Resolution occurred in all patients and approximately 50 per cent regained sexual potency. | Priapism: evolution of management in 48 patients in a 22-year series. The choice of an effective method to treat priapism is challenging because precise causes in the majority of patients have not been well defined. A review of 48 patients treated during a 22-year period shows evolution of a regimen of management that has yielded a high percentage of success. Idiopathic priapism and sickle cell disease accounted for 81 per cent of the subjects. An evaluation should include a medication history, a search for specific diseases, as well as a thorough physical examination to detect possible etiologic factors. The explanation for the frequent association of fever deserves further investigation. Initial therapy consisting of aspiration and irrigation, and intermittent pneumatic cuff compression should be undertaken for a trial period of 12 to 36 hours, repeating the aspiration 2 or 3 times if necessary. The failure of priapism to resolve after such treatment is an indication for a shunt operation. Patients with known etiology should be treated specifically for the primary disease and usually more conservatively for priapism. Resolution occurred in all patients and approximately 50 per cent regained sexual potency. | [
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|
PMID:15139 | Venographic localization of the non-palpable undescended testis in children. | In cases of bilateral non-palpable undescended testes in which human chorionic gonadotropin stimulation has shown the presence of testicular tissue and in cases of unilateral non-palpable undescended testes selective transfemoral gonadal venography with a modified Seldinger technique has been used for the preoperative localization of the non-palpable testis. since the undescended testis may be located anywhere along the course of its embryologic descent, that is from the level of the renal fossa to its exit from the inguinal canal, preoperative localization will aid in the surgical management. Gonadal venography has proved to be accurate and safe, and has aided in the determination of the extent of surgical exploration in 9 children with 12 non-palpable undescended testes (6 right and 6 left). | Venographic localization of the non-palpable undescended testis in children. In cases of bilateral non-palpable undescended testes in which human chorionic gonadotropin stimulation has shown the presence of testicular tissue and in cases of unilateral non-palpable undescended testes selective transfemoral gonadal venography with a modified Seldinger technique has been used for the preoperative localization of the non-palpable testis. since the undescended testis may be located anywhere along the course of its embryologic descent, that is from the level of the renal fossa to its exit from the inguinal canal, preoperative localization will aid in the surgical management. Gonadal venography has proved to be accurate and safe, and has aided in the determination of the extent of surgical exploration in 9 children with 12 non-palpable undescended testes (6 right and 6 left). | [
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|
PMID:15141 | The effects of glucose infusion on myocardial performance during acute hypoxia. | The effects of hypoxia with or without glucose infusion on the cardiac contractility, blood pressure, electrocardiogram, blood electrolytes (sodium and potassium), glucose, pH, Po2, and Pco2 in anesthetized dogs were studied. Hypoxia was induced by ventilating the dogs with reduced oxygen (10%) in the inspired air. Hypoxia produced a decrease in the cardiac contractility and blood pressure, and an increase in the heart rate and central venous pressure. It produced a decrease in the blood pH, Po2, and Pco2, and an increase in the blood glucose and potassium. Glucose infusion during hypoxia delayed the rate of decrease in the contractility and blood pressure significantly. The time for decrease in the contracility to 45 to 50% was increased by 67%. Glucose infusion prevented the loss of potassium from the cell. Glucose infusion however, was unable to correct acidosis. These results indicate that glucose infusion during hypoxia might prevent or delay the deterioration of myocardial function. | The effects of glucose infusion on myocardial performance during acute hypoxia. The effects of hypoxia with or without glucose infusion on the cardiac contractility, blood pressure, electrocardiogram, blood electrolytes (sodium and potassium), glucose, pH, Po2, and Pco2 in anesthetized dogs were studied. Hypoxia was induced by ventilating the dogs with reduced oxygen (10%) in the inspired air. Hypoxia produced a decrease in the cardiac contractility and blood pressure, and an increase in the heart rate and central venous pressure. It produced a decrease in the blood pH, Po2, and Pco2, and an increase in the blood glucose and potassium. Glucose infusion during hypoxia delayed the rate of decrease in the contractility and blood pressure significantly. The time for decrease in the contracility to 45 to 50% was increased by 67%. Glucose infusion prevented the loss of potassium from the cell. Glucose infusion however, was unable to correct acidosis. These results indicate that glucose infusion during hypoxia might prevent or delay the deterioration of myocardial function. | [
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|
PMID:15142 | Effects of beta-adrenergic blocking agents on the blood pressure, plasma renin activity and hemodynamics of hypertensive patients. | Changes in blood pressure, plasma renin activity, and hemodynamic components were studied in 23 patients with essential hypertension treated with oral pindolol or propranolol. These beta-adrenergic blocking agents effectively lowered the blood pressure in the majority of the patients. Although plasma renin activity was not significantly changed, the higher was the pretreatment level, the more it tended to be decreased. Systemic vascular resistence was significantly decreased, while changes in cardiac index and circulating blood volume were variable. Pindolol showed less effect in reducing the heart rate than propranolol. The antihypertensive effect of these drugs had no correlation with the change in plasma renin activity or in any one of hemodynamic components. | Effects of beta-adrenergic blocking agents on the blood pressure, plasma renin activity and hemodynamics of hypertensive patients. Changes in blood pressure, plasma renin activity, and hemodynamic components were studied in 23 patients with essential hypertension treated with oral pindolol or propranolol. These beta-adrenergic blocking agents effectively lowered the blood pressure in the majority of the patients. Although plasma renin activity was not significantly changed, the higher was the pretreatment level, the more it tended to be decreased. Systemic vascular resistence was significantly decreased, while changes in cardiac index and circulating blood volume were variable. Pindolol showed less effect in reducing the heart rate than propranolol. The antihypertensive effect of these drugs had no correlation with the change in plasma renin activity or in any one of hemodynamic components. | [
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|
PMID:15147 | [Metabolism of tetrachloroethylene in guinea pigs (author's transl)]. | Tetrachloroethylene oxide was chemically prepared from tetrachloroethylene, and the metabolites of the oxide in guinea pigs were analyzed by gaschromatography and Fujiwara reaction. The results obtained are as follows: 1) Trichloroacetic acid appeared in gaschromatogram after injection of tetrachloroethylene oxide, but trichloroethanol did not. 2) The metabolites analyzed by Fujiwara reaction after injection of tetrachloroethylene oxide were composed of large proportion of trichloroacetic acid and small proportion of trichloroethanol. 3) The ratio of trichloroacetic acid to trichloroethanol in the urine in case of tetrachloroethylene oxide was relatively similar to that of tetrachloroethylene. 4) The effects of pH (2.0 and 10.0) and temperature (4 degrees C and 37 degrees C) on the urinary metabolites suggest that the substance equivalent to trichloroethanol by Fujiwara reaction in metabolites may not be a real one. 5) Toxicity of tetrachloroethylene oxide seems to be much higher as compared with that of tetrachloroethylene in consideration of the maximum allowable dose in guinea pigs. 6) It is supposed from our experiments that tetrachloroethylene oxide is an intermediary metabolite of tetrachloroethylene. | [Metabolism of tetrachloroethylene in guinea pigs (author's transl)]. Tetrachloroethylene oxide was chemically prepared from tetrachloroethylene, and the metabolites of the oxide in guinea pigs were analyzed by gaschromatography and Fujiwara reaction. The results obtained are as follows: 1) Trichloroacetic acid appeared in gaschromatogram after injection of tetrachloroethylene oxide, but trichloroethanol did not. 2) The metabolites analyzed by Fujiwara reaction after injection of tetrachloroethylene oxide were composed of large proportion of trichloroacetic acid and small proportion of trichloroethanol. 3) The ratio of trichloroacetic acid to trichloroethanol in the urine in case of tetrachloroethylene oxide was relatively similar to that of tetrachloroethylene. 4) The effects of pH (2.0 and 10.0) and temperature (4 degrees C and 37 degrees C) on the urinary metabolites suggest that the substance equivalent to trichloroethanol by Fujiwara reaction in metabolites may not be a real one. 5) Toxicity of tetrachloroethylene oxide seems to be much higher as compared with that of tetrachloroethylene in consideration of the maximum allowable dose in guinea pigs. 6) It is supposed from our experiments that tetrachloroethylene oxide is an intermediary metabolite of tetrachloroethylene. | [
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|
PMID:15148 | Serum gamma-glutamyl transpeptidase as a diagnostic aid in the periodic health examination. | Serum gamma-glutamyl transpeptidase is one of the enzymes in diagnosis of liver diseases, since a new colorimetric method was devised by Orlowski, M. et al. Forty mU/ml of serum gamma-glutamyl transpeptidase activity is said to be the upper limit at clinical level. When this value is set up as a screening level in the periodic health examination, about 35% of the subjects including daily drinkers can be evaluated as abnormal. In the present study, the upper limits of serum gamma-glutamyl transpeptidase activity were 102 mU/ml for 147 normal subjects including daily drinkers and 49 mU/ml in 70 non-drinkers selected from the subjects. Therefore, we propose that the standards for screening the abnormal from the normal in the periodic health examination should be 50 mU/ml for non-drinkers and 100 mU/ml for drinkers. | Serum gamma-glutamyl transpeptidase as a diagnostic aid in the periodic health examination. Serum gamma-glutamyl transpeptidase is one of the enzymes in diagnosis of liver diseases, since a new colorimetric method was devised by Orlowski, M. et al. Forty mU/ml of serum gamma-glutamyl transpeptidase activity is said to be the upper limit at clinical level. When this value is set up as a screening level in the periodic health examination, about 35% of the subjects including daily drinkers can be evaluated as abnormal. In the present study, the upper limits of serum gamma-glutamyl transpeptidase activity were 102 mU/ml for 147 normal subjects including daily drinkers and 49 mU/ml in 70 non-drinkers selected from the subjects. Therefore, we propose that the standards for screening the abnormal from the normal in the periodic health examination should be 50 mU/ml for non-drinkers and 100 mU/ml for drinkers. | [
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|
PMID:15149 | [Studies on the adsorption removal of ammonia gas. 1) Adsorption of ammonia gas on several kinds of activated carbons (author's transl)]. | The present study was designed to secure some fundamental informations on the adsorption removal of ammonia gas by the static method. In order to find out the suitable activated carbons for the adsorption removal of ammonia gas, amounts of ammonia gas adsorbed on twelve kinds of activated carbons were measured at 30 degrees C and up to 70,000 ppm of ammonia gas. The relations between the amounts of ammonia gas adsorbed on the activated carbons and the physical properties of them were discussed through the results of specific surface area, pore volume, mean pore radius, scanning electron micrograph, pH, and amount of base. The results were as follows: 1) Among the twelve kinds of activated carbons, the activated carbons No. 2, No. 3, and No. 6 adsorbed larger amount of ammonia gas than the others. 2) Adsorption of ammonia gas on the activated carbons seemed to be mainly physical as judged from the values of heat of adsorption. 3) The adsorption capacity of the used activated carbons can be recovered to the original capacity by some treatment. 4) The period to reach adsorption equilibrium was about 5 minutes. 5) It may be concluded that adsorption of ammonia gas on the activated carbons was decided mainly by the surface properties (pH and amount of base) of the activated carbons rather than their porous structures. | [Studies on the adsorption removal of ammonia gas. 1) Adsorption of ammonia gas on several kinds of activated carbons (author's transl)]. The present study was designed to secure some fundamental informations on the adsorption removal of ammonia gas by the static method. In order to find out the suitable activated carbons for the adsorption removal of ammonia gas, amounts of ammonia gas adsorbed on twelve kinds of activated carbons were measured at 30 degrees C and up to 70,000 ppm of ammonia gas. The relations between the amounts of ammonia gas adsorbed on the activated carbons and the physical properties of them were discussed through the results of specific surface area, pore volume, mean pore radius, scanning electron micrograph, pH, and amount of base. The results were as follows: 1) Among the twelve kinds of activated carbons, the activated carbons No. 2, No. 3, and No. 6 adsorbed larger amount of ammonia gas than the others. 2) Adsorption of ammonia gas on the activated carbons seemed to be mainly physical as judged from the values of heat of adsorption. 3) The adsorption capacity of the used activated carbons can be recovered to the original capacity by some treatment. 4) The period to reach adsorption equilibrium was about 5 minutes. 5) It may be concluded that adsorption of ammonia gas on the activated carbons was decided mainly by the surface properties (pH and amount of base) of the activated carbons rather than their porous structures. | [
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|
PMID:15153 | Effects of bufetolol and propranolol on active and passive membrane properties of dog papillary muscle. | Effects of bufetolol and propranolol, adrenergic beta-receptor blocking and anti-arrhythmic drugs, on active and passive membrane properties of the dog papillary muscle were investigated with microelectrode and sucrose-gap methods. Bufetolol (10(-5) to 10(-4) g/ml) and propranolol (10(-6) g/ml) significantly decreased the maximum rate of rise of the action potential. The maximum responsive frequency to driving stimulus was decreased in the presence of bufetolol (3 X 10(-5) g/ml) and propranolol (10(-5 g/ml), whereas the effective refractory period was not affected. The critical threshold potential was shifted to more positive potential in the presence of the drugs. The passive membrane property, the space constant (lambda), the time constant (tau) and the current-voltage relations of the muscle membrane were not significantly altered by the drugs. It is concluded that bufetolol and propranolol suppress the excitability of the muscle membrane and this action may be ascribed to the decrease in the sodium conductance (gNa) and to the rise of gNa onset potential without alteration in the passive membrane property. | Effects of bufetolol and propranolol on active and passive membrane properties of dog papillary muscle. Effects of bufetolol and propranolol, adrenergic beta-receptor blocking and anti-arrhythmic drugs, on active and passive membrane properties of the dog papillary muscle were investigated with microelectrode and sucrose-gap methods. Bufetolol (10(-5) to 10(-4) g/ml) and propranolol (10(-6) g/ml) significantly decreased the maximum rate of rise of the action potential. The maximum responsive frequency to driving stimulus was decreased in the presence of bufetolol (3 X 10(-5) g/ml) and propranolol (10(-5 g/ml), whereas the effective refractory period was not affected. The critical threshold potential was shifted to more positive potential in the presence of the drugs. The passive membrane property, the space constant (lambda), the time constant (tau) and the current-voltage relations of the muscle membrane were not significantly altered by the drugs. It is concluded that bufetolol and propranolol suppress the excitability of the muscle membrane and this action may be ascribed to the decrease in the sodium conductance (gNa) and to the rise of gNa onset potential without alteration in the passive membrane property. | [
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|
PMID:15154 | Effects of L-glutamine on acetylsalycylic acid induced gastric lesions and acid back diffusion in dogs. | Effects of L-glutamine on acetylsalicylic acid (ASA)-induced gastric mucosal lesions were studied in mongrel dogs. It was confirmed that when oral ASA at 1.0 or 2.0 g per dog is given in two divided doses, there is severe and consistent dose-dependent mucosal damage in the glandular portion of the stomach in fasted dogs. However, when L-glutamine 2.0 or 4.0 g per dog in two divided doses is given concomitantly with ASA 2.0 g per dog orally, the gastric irritation is significantly inhibited. Instillation of 20 mM of ASA in 100 mM HCl solution into the Heidenhain pouch of Beagle dogs produced a significant loss of H+ from the pouch and a gain of Na+ in the lumen compared with ASA-free controls. When L-glutamine (100 mM) was given concomitantly with ASA (20 mM) into the pouch, changes of electrolyte fluxes in response to ASA alone were significantly suppressed. However, 50 mM of L-glutamine had no appreciable effect on acid back diffusion caused by ASA 20 mM. The amino acid itself had little effect on the ionic movement in the pouch. Gross bleeding from the pouch treated with ASA was never observed with the concomitant dosing of ASA and L-glutamine 50 or 100 mM. | Effects of L-glutamine on acetylsalycylic acid induced gastric lesions and acid back diffusion in dogs. Effects of L-glutamine on acetylsalicylic acid (ASA)-induced gastric mucosal lesions were studied in mongrel dogs. It was confirmed that when oral ASA at 1.0 or 2.0 g per dog is given in two divided doses, there is severe and consistent dose-dependent mucosal damage in the glandular portion of the stomach in fasted dogs. However, when L-glutamine 2.0 or 4.0 g per dog in two divided doses is given concomitantly with ASA 2.0 g per dog orally, the gastric irritation is significantly inhibited. Instillation of 20 mM of ASA in 100 mM HCl solution into the Heidenhain pouch of Beagle dogs produced a significant loss of H+ from the pouch and a gain of Na+ in the lumen compared with ASA-free controls. When L-glutamine (100 mM) was given concomitantly with ASA (20 mM) into the pouch, changes of electrolyte fluxes in response to ASA alone were significantly suppressed. However, 50 mM of L-glutamine had no appreciable effect on acid back diffusion caused by ASA 20 mM. The amino acid itself had little effect on the ionic movement in the pouch. Gross bleeding from the pouch treated with ASA was never observed with the concomitant dosing of ASA and L-glutamine 50 or 100 mM. | [
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|
PMID:15157 | Complement-independent nephrotoxic nephritis in the guinea pig. | Immunologic mechanisms of proteinuria were investigated in guinea pigs (GP) injected with sheep antiserum (NTS) to GP glomerular basement membrane (GBM). Linear deposition of sheep gamma 1 and gamma 2 IgG led to a prompt but transient (36 hr) increase in albumin excretion from control values of 0.026 +/- 0.013 mg/hr to maximal values of 26+/-12.1 mg/rh at six hours without detectable histologic or electron microscopic changes except for decreased staining for glomerular polyanion and epithelial cell foot process fusion. GBM permeability to anionic ferritin was not increased during proteinuria. Anti-GBM antibody deposits did not fix GP C3 or C4 in vivo or in vitro. NTS-induced proteinuria was the same in guinea pigs that were normal, greater than 95% depleted of C3 through C9, genetically deficient in C4, and depleted of circulating polymorphonuclear leukocytes (PMN). Prior administration of antihistamines, steroids, azathioprine, colchicine, indomethacin, heparin, aprotinin (Trasylol), and niridazole also failed to reduced proteinuria. Initial proteinuria subsided by 36 hr, did not recur despite linear deposition of GP gemma 1 and gemma 2 after day seven, and antibody to GMB-bound sheep globlin. In the GP nephrotoxic nephritis model, anti-GBM antibody deposits apparently mediate increased permeability to albumin by a currently undefined mechanism which is independent of complement, PMN, and other know mediators of inflammation. | Complement-independent nephrotoxic nephritis in the guinea pig. Immunologic mechanisms of proteinuria were investigated in guinea pigs (GP) injected with sheep antiserum (NTS) to GP glomerular basement membrane (GBM). Linear deposition of sheep gamma 1 and gamma 2 IgG led to a prompt but transient (36 hr) increase in albumin excretion from control values of 0.026 +/- 0.013 mg/hr to maximal values of 26+/-12.1 mg/rh at six hours without detectable histologic or electron microscopic changes except for decreased staining for glomerular polyanion and epithelial cell foot process fusion. GBM permeability to anionic ferritin was not increased during proteinuria. Anti-GBM antibody deposits did not fix GP C3 or C4 in vivo or in vitro. NTS-induced proteinuria was the same in guinea pigs that were normal, greater than 95% depleted of C3 through C9, genetically deficient in C4, and depleted of circulating polymorphonuclear leukocytes (PMN). Prior administration of antihistamines, steroids, azathioprine, colchicine, indomethacin, heparin, aprotinin (Trasylol), and niridazole also failed to reduced proteinuria. Initial proteinuria subsided by 36 hr, did not recur despite linear deposition of GP gemma 1 and gemma 2 after day seven, and antibody to GMB-bound sheep globlin. In the GP nephrotoxic nephritis model, anti-GBM antibody deposits apparently mediate increased permeability to albumin by a currently undefined mechanism which is independent of complement, PMN, and other know mediators of inflammation. | [
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|
PMID:15159 | [Chloroquine keratopathy as an example of drug-induced phospholipidosis (contribution to the pathogenesis of cornea verticillata) (author's transl)]. | Chronic treatment with certain drugs induces morphological alterations in the eye which are histologically and electron microscopically identical with those found in hereditary lipidoses (cornea verticillata, e.g. in M. Fabry). Hereditary storage diseases are the consequence of enzyme defects, the mechanism underlying the side effect of certain drugs, however, is quite different. "Amphiphilic" drugs from completely different pharmacological groups, like chloroquine, amiodarone, chlorpromazine form complexes with cellular phospholipids which cannot be metabolised by lysosomal phospholipases. Thus in all tissues with high phospholipid content or turnover typical intracellular deposites with lamellary or crystalloid structure may occur (myelin figures). Such deposites were observed in different parts of the eye and are known e.g. from the cornea as "cornea verticillata". | [Chloroquine keratopathy as an example of drug-induced phospholipidosis (contribution to the pathogenesis of cornea verticillata) (author's transl)]. Chronic treatment with certain drugs induces morphological alterations in the eye which are histologically and electron microscopically identical with those found in hereditary lipidoses (cornea verticillata, e.g. in M. Fabry). Hereditary storage diseases are the consequence of enzyme defects, the mechanism underlying the side effect of certain drugs, however, is quite different. "Amphiphilic" drugs from completely different pharmacological groups, like chloroquine, amiodarone, chlorpromazine form complexes with cellular phospholipids which cannot be metabolised by lysosomal phospholipases. Thus in all tissues with high phospholipid content or turnover typical intracellular deposites with lamellary or crystalloid structure may occur (myelin figures). Such deposites were observed in different parts of the eye and are known e.g. from the cornea as "cornea verticillata". | [
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|
PMID:15163 | Carbon tetrachloride-induced changes in mixed function oxidases and microsomal cytochromes in the rat lung. | The effects of a single exposure, by gastric intubation or inhalation, to carbon tetrachloride (CCL4) on rat lungs were assessed. By 1 to 7 days, focal areas of alveolar collapse, septal edema, and modification of type II pneumonocytes were observed. By 24 hours after exposure to the toxin, there were no identifiable changes in surfactant levels or distribution. Microsomes obtained from the lungs and prepared for analysis revealed marked decreases in cytochrome P-450 content and P-450-related N-demethylation of dimethylaniline. Only a transient reduction of cytochrome b5 occurred, with a rebound exceeding control values during the period of pulmonary healing. Whether the lung acted as an excretory route (following intubation) or as an absorption path (after inhalation) made little difference. Carbon tetrachloride had no effect on in vitro microsome composition and function unless supplemented with a reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) generating system. Under these circumstances, there was a reduction in both cytochromes b5 and P-450. Our data indicate that a considerable chemical modification of the pulmonary tissues had taken place, with no accompanying easily recognized changes in cellular structure. Furthermore, evidence for the in vitro destruction of pulmonary microsomal cytochromes P-450 and b5, unrelated to peroxidation, is indicated by these findings. | Carbon tetrachloride-induced changes in mixed function oxidases and microsomal cytochromes in the rat lung. The effects of a single exposure, by gastric intubation or inhalation, to carbon tetrachloride (CCL4) on rat lungs were assessed. By 1 to 7 days, focal areas of alveolar collapse, septal edema, and modification of type II pneumonocytes were observed. By 24 hours after exposure to the toxin, there were no identifiable changes in surfactant levels or distribution. Microsomes obtained from the lungs and prepared for analysis revealed marked decreases in cytochrome P-450 content and P-450-related N-demethylation of dimethylaniline. Only a transient reduction of cytochrome b5 occurred, with a rebound exceeding control values during the period of pulmonary healing. Whether the lung acted as an excretory route (following intubation) or as an absorption path (after inhalation) made little difference. Carbon tetrachloride had no effect on in vitro microsome composition and function unless supplemented with a reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) generating system. Under these circumstances, there was a reduction in both cytochromes b5 and P-450. Our data indicate that a considerable chemical modification of the pulmonary tissues had taken place, with no accompanying easily recognized changes in cellular structure. Furthermore, evidence for the in vitro destruction of pulmonary microsomal cytochromes P-450 and b5, unrelated to peroxidation, is indicated by these findings. | [
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|
PMID:15164 | Acid hydrolases. Assay of activity and latency in the varied mixed cell populations of canine gastric mucosa. | The specific enzymic properties, membrane or particle binding capacities, and the total activities of certain acid hydrolases, including cathepsin D, acid phosphatase, arylsulfatase, and five acid glycosidases have been compared in normal canine antral and fundic mucosae and in liver. The two major regions of the gastric mucosa, whose cell populations are comparable in type but have very distinct functions, also differ in many properties of their lysosomal enzymes. These differences necessitate several major modification in their method of assay. Using optimal conditions, the activities of most of these enzymes were found to differ: levels in the antrum, in spite of its high water and mucin-glycoprotein content, were significantly greater, suggesting that the high lysosomal hydrolytic activity may be associated with the rapid autophagic processes of normal turnover of its surface epithelial and mucous neck cells. Lysosomal membrane stability or latency is also greater in the antrum; this may account, in part at least, for antral resistance to erosions brought about by stress. | Acid hydrolases. Assay of activity and latency in the varied mixed cell populations of canine gastric mucosa. The specific enzymic properties, membrane or particle binding capacities, and the total activities of certain acid hydrolases, including cathepsin D, acid phosphatase, arylsulfatase, and five acid glycosidases have been compared in normal canine antral and fundic mucosae and in liver. The two major regions of the gastric mucosa, whose cell populations are comparable in type but have very distinct functions, also differ in many properties of their lysosomal enzymes. These differences necessitate several major modification in their method of assay. Using optimal conditions, the activities of most of these enzymes were found to differ: levels in the antrum, in spite of its high water and mucin-glycoprotein content, were significantly greater, suggesting that the high lysosomal hydrolytic activity may be associated with the rapid autophagic processes of normal turnover of its surface epithelial and mucous neck cells. Lysosomal membrane stability or latency is also greater in the antrum; this may account, in part at least, for antral resistance to erosions brought about by stress. | [
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|
PMID:15165 | An evaluation of the immune state in leprosy. | An evaluation of the immune state in leprosy was done by the application of a system of graft-versus-host reaction. Peripheral blood lymphocytes obtained from patients with different forms of leprosy and from normal healthy individuals were injected intravenously into the irradiated mice. The rate of blast transformation of the donor cells was measured by the radio-active thymidine uptake. The number of cells labelled with tritiated-thymidine was much higher in the normal individuals and patients with tuberculoid leprosy than in the patients with lepromatous leprosy with the borderline group placed in between the two. However, following successful treatment with DDS, an increased responsiveness and active DNA synthesis could be observed in the previously less responsive lepromatous lymphocytes. | An evaluation of the immune state in leprosy. An evaluation of the immune state in leprosy was done by the application of a system of graft-versus-host reaction. Peripheral blood lymphocytes obtained from patients with different forms of leprosy and from normal healthy individuals were injected intravenously into the irradiated mice. The rate of blast transformation of the donor cells was measured by the radio-active thymidine uptake. The number of cells labelled with tritiated-thymidine was much higher in the normal individuals and patients with tuberculoid leprosy than in the patients with lepromatous leprosy with the borderline group placed in between the two. However, following successful treatment with DDS, an increased responsiveness and active DNA synthesis could be observed in the previously less responsive lepromatous lymphocytes. | [
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|
PMID:15161 | [Intestinal autoflora of the test subjects in a 6-month biological engineering experiment]. | During a 6-month bioengineering experiment the intestinal microflora of four test subjects was examined. Changes in the composition of different groups of intestinal microflora (bifidobacteria, lactic-acid bacteria, sporogenous anaerobes, proteus, etc) were found. In spite of the unstable pattern of the intestinal microflora and its tendency for simplification, the total number of microorganisms in 1 g of feces remained relatively unchanged in all the test subjects. | [Intestinal autoflora of the test subjects in a 6-month biological engineering experiment]. During a 6-month bioengineering experiment the intestinal microflora of four test subjects was examined. Changes in the composition of different groups of intestinal microflora (bifidobacteria, lactic-acid bacteria, sporogenous anaerobes, proteus, etc) were found. In spite of the unstable pattern of the intestinal microflora and its tendency for simplification, the total number of microorganisms in 1 g of feces remained relatively unchanged in all the test subjects. | [
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PMID:15162 | [Sensitivity of animals to central nervous system stimulants in hypokinesia]. | The experiments were carried out on 1150 non-inbred white male rats weighing 200+/-50 g. The animals were housed in small cages for 1, 5, 10, 15, 30, 45 and 60 days. Control rats remained normally active. The experimental and control animals were given a typical diet. On the above days the rats were injected intraperitoneally with central nervous stimulants--caffeine, phenamine and strychnine. Changes in the animal sensitivity to the stimulants were measured with respect to the alterating of LD16, LD50 and LD54 in test animals as compared with the controls and in regards to the emergence and duration of behavioural reactions: adynamics (caffeine), stereotype behavior (phenamine) and convulsions (strychnine). The greatest changes in the animal sensitivity were noted in response to phenamine. A significant increase in the sensitivity to caffeine was found on the 5, 15 and 45th experimental days and to strychnine only on the 5 and 45th days. Convulsions in response to strychnine were recorded in experimental animals earlier than in the controls and their duration was dependent on the doses injected. Adynamics in response to caffeine developed in experimental rats later than in the controls (on the 15th day) and its duration changed cyclically. Stereotype behavior in response to phenamine showed cyclic pattern and its duration in experimental rats was shorter than in the controls. | [Sensitivity of animals to central nervous system stimulants in hypokinesia]. The experiments were carried out on 1150 non-inbred white male rats weighing 200+/-50 g. The animals were housed in small cages for 1, 5, 10, 15, 30, 45 and 60 days. Control rats remained normally active. The experimental and control animals were given a typical diet. On the above days the rats were injected intraperitoneally with central nervous stimulants--caffeine, phenamine and strychnine. Changes in the animal sensitivity to the stimulants were measured with respect to the alterating of LD16, LD50 and LD54 in test animals as compared with the controls and in regards to the emergence and duration of behavioural reactions: adynamics (caffeine), stereotype behavior (phenamine) and convulsions (strychnine). The greatest changes in the animal sensitivity were noted in response to phenamine. A significant increase in the sensitivity to caffeine was found on the 5, 15 and 45th experimental days and to strychnine only on the 5 and 45th days. Convulsions in response to strychnine were recorded in experimental animals earlier than in the controls and their duration was dependent on the doses injected. Adynamics in response to caffeine developed in experimental rats later than in the controls (on the 15th day) and its duration changed cyclically. Stereotype behavior in response to phenamine showed cyclic pattern and its duration in experimental rats was shorter than in the controls. | [
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|
PMID:15169 | Immunohistochemical studies on the localization and distribution of monoamine neuron systems in the rat brain II. Tyrosine hydroxylase in the telencephalon. | Extensive plexuses of TH-positive nerve terminals were found in many parts of the telencephalon, mainly confined to the subcortical and limbic cortical structures. Of special interest were the distinct networks of varying densities in the amygdaloid cortex, the entorhinal cortex, the prepiriform cortex, the anterior cingulate cortex and the (pre-)frontal cortex. Their distribution is identical with the patterns observed in recent studies on cortical dopamine nerve terminals using certain modifications of the Falck-Hillarp technique. The extremely dense TH innervations patterns of the caudate nucleus, nucleus accumbens, tuberculum olfactorium and the less dense basket-like innervation of the lateral septal nuclei could also be demonstrated. TH-positive cell bodies in a periglomerular position could be observed in the olfactory bulb. A few TH-positive cell bodies were observed in the area around the anterior commissure and in the cingulate cortex. In one area, the hippocampal formation, TH-positive dotlike structures were located in the position of the mossy fibres. In all probability they do not belong to monoamine neurons but may contain a cross-reacting protein. In general, the distribution and density of TH-positive terminals agrees well with extensive regional, biochemical studies on TH activity performed by other groups. Minor discrepancies are discussed. As stated in a parallel study on the distribution of TH in the mes- and diencephalon these findings indicate that TH activity is closely related to the amount of enzyme protein. The TH enzyme levels seem to be much higher in the DA than in the NA nerve terminals of the forebrain which would explain the preferential demonstration of DA terminals in the forebrain using TH antiserum and the high and low TH enzyme activity in DA and NA rich regions, respectively. | Immunohistochemical studies on the localization and distribution of monoamine neuron systems in the rat brain II. Tyrosine hydroxylase in the telencephalon. Extensive plexuses of TH-positive nerve terminals were found in many parts of the telencephalon, mainly confined to the subcortical and limbic cortical structures. Of special interest were the distinct networks of varying densities in the amygdaloid cortex, the entorhinal cortex, the prepiriform cortex, the anterior cingulate cortex and the (pre-)frontal cortex. Their distribution is identical with the patterns observed in recent studies on cortical dopamine nerve terminals using certain modifications of the Falck-Hillarp technique. The extremely dense TH innervations patterns of the caudate nucleus, nucleus accumbens, tuberculum olfactorium and the less dense basket-like innervation of the lateral septal nuclei could also be demonstrated. TH-positive cell bodies in a periglomerular position could be observed in the olfactory bulb. A few TH-positive cell bodies were observed in the area around the anterior commissure and in the cingulate cortex. In one area, the hippocampal formation, TH-positive dotlike structures were located in the position of the mossy fibres. In all probability they do not belong to monoamine neurons but may contain a cross-reacting protein. In general, the distribution and density of TH-positive terminals agrees well with extensive regional, biochemical studies on TH activity performed by other groups. Minor discrepancies are discussed. As stated in a parallel study on the distribution of TH in the mes- and diencephalon these findings indicate that TH activity is closely related to the amount of enzyme protein. The TH enzyme levels seem to be much higher in the DA than in the NA nerve terminals of the forebrain which would explain the preferential demonstration of DA terminals in the forebrain using TH antiserum and the high and low TH enzyme activity in DA and NA rich regions, respectively. | [
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|
PMID:15170 | pH-dependency in diffusion of some weakly acidic drugs, warfarin, sulfaethidole, and barbital, into organic phases. | The diffusion of three weakly acidic drugs (warfarin, sulfaethidole, and barbital) in cyclohexane-buffer and n-octanol-buffer systems was studied by two different methods; shaking and model cell type of experimentation. At low pH values the drugs moved generally more readily into the organic phases than at higher pH. Moreover, when the shaking method was used the drugs moved readily to the n-octanol phase even at higher pH levels. Lecithin improved the diffusion at low more than at high pH levels. Thus, the results varied according to the organic phase selected and the method used. Poor correlation was found between some results of this study and previously reported results of in situ absorption from the rat gastrointestinal tract of the same drugs. The results do not support the assumption that phospholipids may have a role in the absorption of ionized moieties. | pH-dependency in diffusion of some weakly acidic drugs, warfarin, sulfaethidole, and barbital, into organic phases. The diffusion of three weakly acidic drugs (warfarin, sulfaethidole, and barbital) in cyclohexane-buffer and n-octanol-buffer systems was studied by two different methods; shaking and model cell type of experimentation. At low pH values the drugs moved generally more readily into the organic phases than at higher pH. Moreover, when the shaking method was used the drugs moved readily to the n-octanol phase even at higher pH levels. Lecithin improved the diffusion at low more than at high pH levels. Thus, the results varied according to the organic phase selected and the method used. Poor correlation was found between some results of this study and previously reported results of in situ absorption from the rat gastrointestinal tract of the same drugs. The results do not support the assumption that phospholipids may have a role in the absorption of ionized moieties. | [
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|
PMID:15177 | Involvement of cytochrome b5 in the oxidative desaturation of linoleic acid to gamma-linolenic acid in rat liver microsomes. | The effects of antibodies against microsomal electron-transport components on the in vitro activity of delta6-desaturation of linoleic acid to gamma-linolenic acid have been studied in intact microsomal membranes of rat liver. Reduced nicotinamide adenine dinucleotide (NADH) or reduced nicotinamide adenine dinucleotide phosphate (NADPH) (0.87 mM) served as electron donors, and effectively prompted the delta6-desaturase activities with yields of about 1.1 to 1.3 nmol per mg of protein in 10 min. Of the two antibodies studied under the same in vitro conditions, i.e., rabbit antisera preparations against rat liver microsomal hydrophilic parts of cytochrome b5 and NADPH-cytochrome c reductase, only the antibody against cytochrome b5 demonstrated a marked ability to inhibit the delta6-desaturase activity. This evidence supports a participation of cytochrome b5 in the delta6-desaturation of linoleic acid and suggests a pathway analogous to the delta9-desaturation of stearyl-CoA. | Involvement of cytochrome b5 in the oxidative desaturation of linoleic acid to gamma-linolenic acid in rat liver microsomes. The effects of antibodies against microsomal electron-transport components on the in vitro activity of delta6-desaturation of linoleic acid to gamma-linolenic acid have been studied in intact microsomal membranes of rat liver. Reduced nicotinamide adenine dinucleotide (NADH) or reduced nicotinamide adenine dinucleotide phosphate (NADPH) (0.87 mM) served as electron donors, and effectively prompted the delta6-desaturase activities with yields of about 1.1 to 1.3 nmol per mg of protein in 10 min. Of the two antibodies studied under the same in vitro conditions, i.e., rabbit antisera preparations against rat liver microsomal hydrophilic parts of cytochrome b5 and NADPH-cytochrome c reductase, only the antibody against cytochrome b5 demonstrated a marked ability to inhibit the delta6-desaturase activity. This evidence supports a participation of cytochrome b5 in the delta6-desaturation of linoleic acid and suggests a pathway analogous to the delta9-desaturation of stearyl-CoA. | [
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|
PMID:15178 | Glycerokinase in human adipose tissue. | The presence of glycerokinase has been demonstrated in human omental and subcutaneous adipose tissue. The enzyme reaction showed a linear time course for 5 min at 30 C and pH optima at pH 7.6 and 9.0. Saturation of the enzyme was observed at 1.8 mM adenosine triphosphate (ATP) and the double reciprocal plot of activity vs. ATP concentration was nonlinear giving two apparent Km values of 0.094 and 0.518 mM. The apparent Km for glycerol, 0.112 mM, was obtained from a linear double reciprocal plot, and the enzyme was saturated at about 0.4 mM glycerol. The activity of glycerokinase in human adipose tissue excised under general anaesthesia was low and was unrelated to adipose cell size or the degree of obesity of the subject from whom the fat was obtained. | Glycerokinase in human adipose tissue. The presence of glycerokinase has been demonstrated in human omental and subcutaneous adipose tissue. The enzyme reaction showed a linear time course for 5 min at 30 C and pH optima at pH 7.6 and 9.0. Saturation of the enzyme was observed at 1.8 mM adenosine triphosphate (ATP) and the double reciprocal plot of activity vs. ATP concentration was nonlinear giving two apparent Km values of 0.094 and 0.518 mM. The apparent Km for glycerol, 0.112 mM, was obtained from a linear double reciprocal plot, and the enzyme was saturated at about 0.4 mM glycerol. The activity of glycerokinase in human adipose tissue excised under general anaesthesia was low and was unrelated to adipose cell size or the degree of obesity of the subject from whom the fat was obtained. | [
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|
PMID:15181 | Some respiratory and metabolic effects of exercise in moderately obese men. | The effects of varying levels of exercise on oxygen uptake, CO2 production, blood pressure, arterial blood gasses, and arterial concentrations of glucose, insulin, and growth hormone were examined in ten normal weight and ten moderately overweight young men. At comparable external work loads with a bicycle ergometer, the lean men required less oxygen than the obese men. When oxygen uptakes were matched during exercise on a treadmill, the lean men were walking on a steeper grade or at a higher rate than the obese men. The efficiency of exercise as assessed by the relation between oxygen uptake and work did not differ between the two groups. Blood pressure rose more in the obese during exercise than in the lean. The fall in lactate and rise in bicarbonate was of greater magnitude during cycle ergometry than during treadmill exercise. Obese and lean men, however, showed similar changes. With each level of exercise, there was a fall in arterial insulin levels, but the concentrations in the blood of overweight men always remained significantly above that of the normal men. Growth hormones tended to be higher in the normal weight men, but the differences were usually not significant, and there was no significant rise with exercise in either group until the highest levels of work were achieved. Glucose concentrations tended to be higher in the obese men, but fell to constant levels in both groups during exercise. Blood pressure rose to a greater extent in the overweight men during exercise. | Some respiratory and metabolic effects of exercise in moderately obese men. The effects of varying levels of exercise on oxygen uptake, CO2 production, blood pressure, arterial blood gasses, and arterial concentrations of glucose, insulin, and growth hormone were examined in ten normal weight and ten moderately overweight young men. At comparable external work loads with a bicycle ergometer, the lean men required less oxygen than the obese men. When oxygen uptakes were matched during exercise on a treadmill, the lean men were walking on a steeper grade or at a higher rate than the obese men. The efficiency of exercise as assessed by the relation between oxygen uptake and work did not differ between the two groups. Blood pressure rose more in the obese during exercise than in the lean. The fall in lactate and rise in bicarbonate was of greater magnitude during cycle ergometry than during treadmill exercise. Obese and lean men, however, showed similar changes. With each level of exercise, there was a fall in arterial insulin levels, but the concentrations in the blood of overweight men always remained significantly above that of the normal men. Growth hormones tended to be higher in the normal weight men, but the differences were usually not significant, and there was no significant rise with exercise in either group until the highest levels of work were achieved. Glucose concentrations tended to be higher in the obese men, but fell to constant levels in both groups during exercise. Blood pressure rose to a greater extent in the overweight men during exercise. | [
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|
PMID:15198 | Paradoxical reaction to a new minor tranquilizer. | This case report demonstrates that paradoxical reactions to the benzodiazepine class of minor tranquilizer can occur, and that lorazepam, a new derivative, is not free from this effect. It also lends support to the hypothesis that such reactions may be precipitated by a frustrating stimulus, and this has clinical relevance in that such precipitants may well be similar to those situations producing anxiety for which the drug had been prescribed. | Paradoxical reaction to a new minor tranquilizer. This case report demonstrates that paradoxical reactions to the benzodiazepine class of minor tranquilizer can occur, and that lorazepam, a new derivative, is not free from this effect. It also lends support to the hypothesis that such reactions may be precipitated by a frustrating stimulus, and this has clinical relevance in that such precipitants may well be similar to those situations producing anxiety for which the drug had been prescribed. | [
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|
PMID:15207 | Enzyme transformations. | The paper briefly discusses the current theories on the processes underlying the transformation of the catalytic activity of a number of enzymes. The phenomenon of transformation, which is induced by mild chemical effects acting on a protein, is of interest both from the point of view of the investigation of its molecular mechanisms and from the point of view of the study of several pathological states under which this phenomenon is observed. Gorkin's own data are presented. | Enzyme transformations. The paper briefly discusses the current theories on the processes underlying the transformation of the catalytic activity of a number of enzymes. The phenomenon of transformation, which is induced by mild chemical effects acting on a protein, is of interest both from the point of view of the investigation of its molecular mechanisms and from the point of view of the study of several pathological states under which this phenomenon is observed. Gorkin's own data are presented. | [
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|
PMID:15209 | Interaction of transfer RNA with 50S ribosomal subunits of Escherichia coli in absence of templates. | The equilibrium constant of a complex of tRNA with the 50S ribosomal subunit was measured in the absence of a template. It was shown that the stability of the complex increases with an increase in the concentration of Mg2+, it decreases with an increase in the concentration of univalent ions, and does not depend on the pH of the medium in the range of 7.0-8.2. Removal of the 3'-terminal nucleoside of tRNA weakens the association approximately 40-fold; the subsequent successive splitting off of another three nucleotides has little effect on the association constant. In 90% 2H2O the stability of the complex increases approximately four-fold, which points to the large contribution of the hydrogen bonds to the free energy of the interaction. The tetranucleotide TphiCG competes slightly with tRNA for sites on the 50S subparticles; this means that the TphiC segment of tRNA does not play an important role in the formation of the complex under investigation. | Interaction of transfer RNA with 50S ribosomal subunits of Escherichia coli in absence of templates. The equilibrium constant of a complex of tRNA with the 50S ribosomal subunit was measured in the absence of a template. It was shown that the stability of the complex increases with an increase in the concentration of Mg2+, it decreases with an increase in the concentration of univalent ions, and does not depend on the pH of the medium in the range of 7.0-8.2. Removal of the 3'-terminal nucleoside of tRNA weakens the association approximately 40-fold; the subsequent successive splitting off of another three nucleotides has little effect on the association constant. In 90% 2H2O the stability of the complex increases approximately four-fold, which points to the large contribution of the hydrogen bonds to the free energy of the interaction. The tetranucleotide TphiCG competes slightly with tRNA for sites on the 50S subparticles; this means that the TphiC segment of tRNA does not play an important role in the formation of the complex under investigation. | [
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|
PMID:15208 | Presence of a proteinase in polyribosomes of rat liver. | The presence of a proteinase in the composition of the ribosomal proteins of rat liver is demonstrated. The enzyme possesses optimal activity in the zone of pH 7.0-7.2. Soybean trypsin inhibitor and 1-chloro-4-phenyl-3-tosylamido-2-butanone inhibit the enzyme by 50-60%. | Presence of a proteinase in polyribosomes of rat liver. The presence of a proteinase in the composition of the ribosomal proteins of rat liver is demonstrated. The enzyme possesses optimal activity in the zone of pH 7.0-7.2. Soybean trypsin inhibitor and 1-chloro-4-phenyl-3-tosylamido-2-butanone inhibit the enzyme by 50-60%. | [
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|
PMID:15211 | Ribonuclease and deoxyribonuclease activity of exonuclease A5. | A comparative study was made of the specificity and some properties of the RNase and DNase activities of exonuclease A5. The results obtained indicate that RNA and denatured DNA are hydrolyzed in the same active center of the enzyme and, consequently, exonuclease A5 is an enzyme which is not specific for the sugar of nucleic acids. The data reported are important when exonuclease A5 is used as a specific reagent in investigations of nucleic acids. | Ribonuclease and deoxyribonuclease activity of exonuclease A5. A comparative study was made of the specificity and some properties of the RNase and DNase activities of exonuclease A5. The results obtained indicate that RNA and denatured DNA are hydrolyzed in the same active center of the enzyme and, consequently, exonuclease A5 is an enzyme which is not specific for the sugar of nucleic acids. The data reported are important when exonuclease A5 is used as a specific reagent in investigations of nucleic acids. | [
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|
PMID:15215 | The influence of pH on the effects of 2,4-D (2,4-dichlorophenoxyacetic acid, Na salt) on Saccharomyces cerevisiae and Salmonella typhimurium. | The genetic effects of 2,4-D (2,4-dichlorophenoxyacetic acid, Na salt) have been investigated in cells of the yeast Saccharomyces cerevisiae and of the bacterium Salmonella typhimurium in experiments in vitro and in vivo. Experiments in vitro showed that the killing of both yeast and bacteria is dependent on the pH in the treatment solution of 2,4-D. A dose-dependent increase of the frequency of mitotic gene conversion and mitotic recombination in yeast was observed at pH 4.50 and 4.30. In experiments in vitro with two strains of Salmonella no significant increase of the number of revertants to prototrophy was obtained. The positive correlation between survival of cells and dissociation of 2,4-D in the pH region 2.8-5.0 indicates that the cells are unable to take up dissociated 2,4-D. Therefore the survival is high at a high pH when most 2,4-D is in dissociated form, and the survival is low at a relatively low pH when more of the 2,4-D is in its undissociated form. No genetic effects were induced by oral administration of tolerable doses of 2,4-D in host-mediated assays using mice as hosts and yeast or Salmonella as indicator cells. | The influence of pH on the effects of 2,4-D (2,4-dichlorophenoxyacetic acid, Na salt) on Saccharomyces cerevisiae and Salmonella typhimurium. The genetic effects of 2,4-D (2,4-dichlorophenoxyacetic acid, Na salt) have been investigated in cells of the yeast Saccharomyces cerevisiae and of the bacterium Salmonella typhimurium in experiments in vitro and in vivo. Experiments in vitro showed that the killing of both yeast and bacteria is dependent on the pH in the treatment solution of 2,4-D. A dose-dependent increase of the frequency of mitotic gene conversion and mitotic recombination in yeast was observed at pH 4.50 and 4.30. In experiments in vitro with two strains of Salmonella no significant increase of the number of revertants to prototrophy was obtained. The positive correlation between survival of cells and dissociation of 2,4-D in the pH region 2.8-5.0 indicates that the cells are unable to take up dissociated 2,4-D. Therefore the survival is high at a high pH when most 2,4-D is in dissociated form, and the survival is low at a relatively low pH when more of the 2,4-D is in its undissociated form. No genetic effects were induced by oral administration of tolerable doses of 2,4-D in host-mediated assays using mice as hosts and yeast or Salmonella as indicator cells. | [
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|
PMID:15210 | Physicochemical properties of Sendai virus RNA. II. Effect of ionic strength on thermostability of RNA. | The dependence of the melting point (Tm) and width of the melting range (deltaTm) on the ionic strength and pH of the medium was investigated for the double-stranded RNA formed through self-hybridization during the isolation of RNA from Sendai virus. It was shown that Tm is a linear function of the logarithm of the sodium ion concentration in the range of concentrations from 10(-1) to 10(-4) M, with a slope of 11.5 degrees toward the abscissa for each order of magnitude. The width of the melting range increased slightly with a decrease in the ionic strength. A change in the pH of the solutions from 5 to 8 had almost no effect on the melting point or the width of the melting range. The degree of purification of the preparations of RNA and the presence of EDTA in the solutions affected the form of the dependence of the mp on the logarithm of the sodium ion concentration very strongly, especially in the region of low ionic strengths. | Physicochemical properties of Sendai virus RNA. II. Effect of ionic strength on thermostability of RNA. The dependence of the melting point (Tm) and width of the melting range (deltaTm) on the ionic strength and pH of the medium was investigated for the double-stranded RNA formed through self-hybridization during the isolation of RNA from Sendai virus. It was shown that Tm is a linear function of the logarithm of the sodium ion concentration in the range of concentrations from 10(-1) to 10(-4) M, with a slope of 11.5 degrees toward the abscissa for each order of magnitude. The width of the melting range increased slightly with a decrease in the ionic strength. A change in the pH of the solutions from 5 to 8 had almost no effect on the melting point or the width of the melting range. The degree of purification of the preparations of RNA and the presence of EDTA in the solutions affected the form of the dependence of the mp on the logarithm of the sodium ion concentration very strongly, especially in the region of low ionic strengths. | [
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PMID:15216 | Induction of DNA single-strand breaks in barley by sodium azide applied at pH 3. | Sodium azide (1 to 50 mM), adjusted to pH 3 and applied for 2 h to presoaked barley seeds, induced a dose-dependent frequency of single-strand breaks in DNA of non-germinating embryos. This was demonstrated by sedimentation analyses of isolated DNA samples in alkaline sucrose gradients and in neutral sucrose gradients with 80% formamide. The doses applied also inhibited dose dependently the root length, seed germination and partially the seedling height. Only the sub-lethal doses (10 and 12.5 mM) induced a low frequency of chromatid breaks and translocations in the root tip metaphases. The sedimentation rate (in alkaline sucrose gradients) of calf thymus DNA treated with sodium azide at pH 3, was similar to that of the control DNA treated with buffer (pH 3) alone. | Induction of DNA single-strand breaks in barley by sodium azide applied at pH 3. Sodium azide (1 to 50 mM), adjusted to pH 3 and applied for 2 h to presoaked barley seeds, induced a dose-dependent frequency of single-strand breaks in DNA of non-germinating embryos. This was demonstrated by sedimentation analyses of isolated DNA samples in alkaline sucrose gradients and in neutral sucrose gradients with 80% formamide. The doses applied also inhibited dose dependently the root length, seed germination and partially the seedling height. Only the sub-lethal doses (10 and 12.5 mM) induced a low frequency of chromatid breaks and translocations in the root tip metaphases. The sedimentation rate (in alkaline sucrose gradients) of calf thymus DNA treated with sodium azide at pH 3, was similar to that of the control DNA treated with buffer (pH 3) alone. | [
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|
PMID:15212 | Acceptor activity of tRNAPhe from yeasts under special conditions of aminoacylation. | The reaction of aminoacylation of tRNAPhe from yeasts and the erroneous acylation of total tRNA from E. coli by yeast phenylalanyl-tRNA synthetase under special conditions was studied. It was shown that the decrease in the degree of acylation of tRNAPhe and the increase in the degree of erroneous acylation of the total tRNA from E. coli are associated with the influence of these conditions on the structure of tRNA, and not on the structure or specificity of the enzyme. It was found that under special conditions of acylation, tRNAPhe exists in two conformations: acylatable and nonacylatable. The ability for complete acylation is restored after the transfer of tRNAPhe under classical conditions of acylation. The results are discussed from the standpoint of possible mechanisms of the recognition of tRNA by aminoacyl-tRNA synthetases. | Acceptor activity of tRNAPhe from yeasts under special conditions of aminoacylation. The reaction of aminoacylation of tRNAPhe from yeasts and the erroneous acylation of total tRNA from E. coli by yeast phenylalanyl-tRNA synthetase under special conditions was studied. It was shown that the decrease in the degree of acylation of tRNAPhe and the increase in the degree of erroneous acylation of the total tRNA from E. coli are associated with the influence of these conditions on the structure of tRNA, and not on the structure or specificity of the enzyme. It was found that under special conditions of acylation, tRNAPhe exists in two conformations: acylatable and nonacylatable. The ability for complete acylation is restored after the transfer of tRNAPhe under classical conditions of acylation. The results are discussed from the standpoint of possible mechanisms of the recognition of tRNA by aminoacyl-tRNA synthetases. | [
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|
PMID:15214 | Thermodynamic parameters of helix-random coil transitions in polypeptide chains. IV. Random copolymers of L-alanine with L-glutamic acid. | pH-Induced helix-random coil transitions in random copolymers of Ala with Glu have been investigated in order to determine the effect of Ala on the stability of the helical state of polyglutamic acid. The free energies for the transfer of one uncharged Glu residue from a random coil to a helix (deltaGo) have been determined from potentiometric titration curves by the method of Zimm and Rice. It has been shown that the introduction of Ala hampers the transfer of a Glu residue from a random coil to a helix (reduces -deltaGo), although Ala itself is a helix-forming residue, i.e., its free energy decreases during helix formation. This has suggested that its introduction weakens the helix-stabilizing interactions between the uncharged Glu residues (apparently hydrogen bonds). The evaluation of the intrinsic helix-random coil equilibrium constant s for uncharged Glu residues with consideration of this situation yields a value which is smaller than the value of s for (Glu)n and in good agreement with the theoretical values. | Thermodynamic parameters of helix-random coil transitions in polypeptide chains. IV. Random copolymers of L-alanine with L-glutamic acid. pH-Induced helix-random coil transitions in random copolymers of Ala with Glu have been investigated in order to determine the effect of Ala on the stability of the helical state of polyglutamic acid. The free energies for the transfer of one uncharged Glu residue from a random coil to a helix (deltaGo) have been determined from potentiometric titration curves by the method of Zimm and Rice. It has been shown that the introduction of Ala hampers the transfer of a Glu residue from a random coil to a helix (reduces -deltaGo), although Ala itself is a helix-forming residue, i.e., its free energy decreases during helix formation. This has suggested that its introduction weakens the helix-stabilizing interactions between the uncharged Glu residues (apparently hydrogen bonds). The evaluation of the intrinsic helix-random coil equilibrium constant s for uncharged Glu residues with consideration of this situation yields a value which is smaller than the value of s for (Glu)n and in good agreement with the theoretical values. | [
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|
PMID:15219 | Use of amylum derivatives for isolation of amylolytic enzymes. | In a leading article literature is reviewed concerning isolation of amylolytic enzymes by adsorption on differently modified starches, resp. on other adsorbents. DEAE amylum, DEAHP amylum and DEAE-Sephadex A 25 were found to be most suitable adsorbents. The other adsorbents examined did not reach claimed parameters. | Use of amylum derivatives for isolation of amylolytic enzymes. In a leading article literature is reviewed concerning isolation of amylolytic enzymes by adsorption on differently modified starches, resp. on other adsorbents. DEAE amylum, DEAHP amylum and DEAE-Sephadex A 25 were found to be most suitable adsorbents. The other adsorbents examined did not reach claimed parameters. | [
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PMID:15213 | Labilization of the phosphoester linkage in enzyme-inhibitor complexes of aspartate aminotransferase. | Individual enzyme-inhibitor complexes with characteristic absorption spectra have been obtained as a result of the reaction of the apoenzyme of aspartate aminotransferase with Nalpha-(5'-phosphopyridoxyl)-L-glutamic acid, Nalpha-(5'-phosphopyridoxyl)-D-glutamic acid, and Nalpha-(5'-phosphopyridoxyl)-L-pyroglutamic acid. The stability of the enzyme-inhibitor complexes has been investigated under various conditions, viz., reactivation by the coenzyme, denaturation by urea, variations in the pH. It has been shown that the complexes formed by the last two inhibitors are reactivated by pyridoxal-5'-phosphate and that the inhibitor can be released under mild conditions. The enzyme-inhibitor complex formed by Nalpha-(5'-phosphopyridoxyl)-L-glutamic acid, on the other hand, was not reactivated by the coenzyme. Pyridoxylglutamic acid has been isolate in attempts to release the inhibitor. The dephosphorylation of the inhibitor has been associated both with the hydrolysis of a phosphate bond involving the enzyme and with the phosphorylation of aspartate aminotransferase. A 32P peptide containing 13 amino acids has been isolated from the tryptic hydrolysate of the enzyme-inhibitor complex (formed by a 32P inhibitor). The data obtained have been interpreted on the basis of an assumption that the phosphate group of the coenzyme has an active role in the enzymatic transamination reaction. | Labilization of the phosphoester linkage in enzyme-inhibitor complexes of aspartate aminotransferase. Individual enzyme-inhibitor complexes with characteristic absorption spectra have been obtained as a result of the reaction of the apoenzyme of aspartate aminotransferase with Nalpha-(5'-phosphopyridoxyl)-L-glutamic acid, Nalpha-(5'-phosphopyridoxyl)-D-glutamic acid, and Nalpha-(5'-phosphopyridoxyl)-L-pyroglutamic acid. The stability of the enzyme-inhibitor complexes has been investigated under various conditions, viz., reactivation by the coenzyme, denaturation by urea, variations in the pH. It has been shown that the complexes formed by the last two inhibitors are reactivated by pyridoxal-5'-phosphate and that the inhibitor can be released under mild conditions. The enzyme-inhibitor complex formed by Nalpha-(5'-phosphopyridoxyl)-L-glutamic acid, on the other hand, was not reactivated by the coenzyme. Pyridoxylglutamic acid has been isolate in attempts to release the inhibitor. The dephosphorylation of the inhibitor has been associated both with the hydrolysis of a phosphate bond involving the enzyme and with the phosphorylation of aspartate aminotransferase. A 32P peptide containing 13 amino acids has been isolated from the tryptic hydrolysate of the enzyme-inhibitor complex (formed by a 32P inhibitor). The data obtained have been interpreted on the basis of an assumption that the phosphate group of the coenzyme has an active role in the enzymatic transamination reaction. | [
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PMID:15220 | Isolation of amylolytic system of Aspergillus oryzae by sorption on DEAHP amylum. | Conditions of effective sorption of amylolytic enzyme from a solution or from fermentative liquid on DEAHP amylum were studied. Isolating action is in a direct dependence on the relation between activity and amount of DEAHP amylum, the curve of this dependence was illustrated. The enzyme can be released by elution or adsorbate can be used in a pulverised from. In the conclusion of the work laboratory isolation technique is described. | Isolation of amylolytic system of Aspergillus oryzae by sorption on DEAHP amylum. Conditions of effective sorption of amylolytic enzyme from a solution or from fermentative liquid on DEAHP amylum were studied. Isolating action is in a direct dependence on the relation between activity and amount of DEAHP amylum, the curve of this dependence was illustrated. The enzyme can be released by elution or adsorbate can be used in a pulverised from. In the conclusion of the work laboratory isolation technique is described. | [
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|
PMID:15232 | [Fauna and ecology of bloodsucking mosquitoes of Evenkia]. | This paper presents data on the specific composition, number, hatching sites, seasonal and daily activity of mosquitoes attacking man and reindeer and the influence of weather factors on the attacking activity of mosquitoes in Central Siberia. 23 species of 3 genera are reported from Central Siberia as follows: Anopheles maculipennis, Culiseta alaskaensis, Aedes beklemishevi, A. cantans, A. caspius dorsalis, A. cataphylla, A. communis, A. cyprius, A. diantaeus, A. excrucians, A. fitchii, A. flavescens, A. hexodontus, A. impiger, A. intrudens, A. leucomelas, A. nigripes, A. pionips, A. pullatus, A. punctor, A. rempeli, A. stricticus, A. cinereus. | [Fauna and ecology of bloodsucking mosquitoes of Evenkia]. This paper presents data on the specific composition, number, hatching sites, seasonal and daily activity of mosquitoes attacking man and reindeer and the influence of weather factors on the attacking activity of mosquitoes in Central Siberia. 23 species of 3 genera are reported from Central Siberia as follows: Anopheles maculipennis, Culiseta alaskaensis, Aedes beklemishevi, A. cantans, A. caspius dorsalis, A. cataphylla, A. communis, A. cyprius, A. diantaeus, A. excrucians, A. fitchii, A. flavescens, A. hexodontus, A. impiger, A. intrudens, A. leucomelas, A. nigripes, A. pionips, A. pullatus, A. punctor, A. rempeli, A. stricticus, A. cinereus. | [
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|
PMID:15242 | [Nature of the enzymes participating in the transformation of proinsulin into insulin]. | A study was made of the enzymes of the islands of Langerhans which could participate in the transformation of proinsulin into insulin. The homogenate of the islands of Langerhans of rat and man catalized the hyppuril-L-arginine splitting at pH 5.4-5.8 and 6.8-7.2 which was completely blocked with N-ethyl maleimide. The enzyme of the islands with the optimum action pH of 5.4-5.8 was similar to the enzyme of the exocrine tissue and was possibly a catheptic carboxypeptidase. The second enzyme of the islands differing from the exocrine carboxypeptidase could apparently participate in the insulin formation from the intermediate forms of proinsulin. In the formation of these proinsulin fomrs the participation of the enzyme of the endopeptidase character with a more acid optimum of the action pH is supposed. | [Nature of the enzymes participating in the transformation of proinsulin into insulin]. A study was made of the enzymes of the islands of Langerhans which could participate in the transformation of proinsulin into insulin. The homogenate of the islands of Langerhans of rat and man catalized the hyppuril-L-arginine splitting at pH 5.4-5.8 and 6.8-7.2 which was completely blocked with N-ethyl maleimide. The enzyme of the islands with the optimum action pH of 5.4-5.8 was similar to the enzyme of the exocrine tissue and was possibly a catheptic carboxypeptidase. The second enzyme of the islands differing from the exocrine carboxypeptidase could apparently participate in the insulin formation from the intermediate forms of proinsulin. In the formation of these proinsulin fomrs the participation of the enzyme of the endopeptidase character with a more acid optimum of the action pH is supposed. | [
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|
PMID:15243 | [Laparoscopic diagnosis of cryptorchism]. | On the basis of observations of 28 patients suffering from cryptorchism in which laparoscopy was conducted for the purpose of diagnosis the authors came to the conclusion on the expediency of using this method in the practice of endocrinologist's work. | [Laparoscopic diagnosis of cryptorchism]. On the basis of observations of 28 patients suffering from cryptorchism in which laparoscopy was conducted for the purpose of diagnosis the authors came to the conclusion on the expediency of using this method in the practice of endocrinologist's work. | [
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|
PMID:15244 | [Change in the activity of tyrosine aminotransferase in the tissues of rabbits with various levels of corticosteroids in the body]. | Experiments were conducted on rabbits. As revealed, hypocoriticism was accompanied by a reduction of the tyrosineaminotranspherase in the liver, muscles and blood plasma; the enzyme activity was unchanged in the brain and the spleen. An increase of the corticosteroid level in the organism after the administration of hydrocortisone to the intact and adrenalectomized animals led to increase of the enzyme activity in the tissues under study; the effect of hydrocortisone action depended on the initial hormonal background in the organism and the duration of the hormone administration. A single ACTH administration to the intact rabbits was accompanied by increase in the enzyme activity in the liver and the spleen, whereas to the adrenalectomized animals--in the brain, muscles and the blood plasma. A repeated administration of both hormones decreased the enzyme activity in the brain and the liver. | [Change in the activity of tyrosine aminotransferase in the tissues of rabbits with various levels of corticosteroids in the body]. Experiments were conducted on rabbits. As revealed, hypocoriticism was accompanied by a reduction of the tyrosineaminotranspherase in the liver, muscles and blood plasma; the enzyme activity was unchanged in the brain and the spleen. An increase of the corticosteroid level in the organism after the administration of hydrocortisone to the intact and adrenalectomized animals led to increase of the enzyme activity in the tissues under study; the effect of hydrocortisone action depended on the initial hormonal background in the organism and the duration of the hormone administration. A single ACTH administration to the intact rabbits was accompanied by increase in the enzyme activity in the liver and the spleen, whereas to the adrenalectomized animals--in the brain, muscles and the blood plasma. A repeated administration of both hormones decreased the enzyme activity in the brain and the liver. | [
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|
PMID:15247 | A monomeric form of pyruvate kinase in human pyruvate kinase deficiency. | A mutant pyruvate kinase (ATP:pyruvate 2-O-phosphotransferase, EC 2.7.1.40) from human erythrocytes which is easily separated into monomers and dimers by gel chromatography is described. Tht mutant enzyme shows almost the same pH optimum and thermostability as normal enzyme, but has a decreased stability on shaking with air, a decreased Km for phosphoenolpyruvate and a loss of allosteric properties. The apparent Km values for phosphoenolpyruvate of tetramers and monomers were the same. The tetrameric enzyme was slightly activated by fructose-1,6-diphosphate but the monomeric form was not. The tetrameric enzyme was found to dissociate spontaneously to dimeric and monomeric forms. | A monomeric form of pyruvate kinase in human pyruvate kinase deficiency. A mutant pyruvate kinase (ATP:pyruvate 2-O-phosphotransferase, EC 2.7.1.40) from human erythrocytes which is easily separated into monomers and dimers by gel chromatography is described. Tht mutant enzyme shows almost the same pH optimum and thermostability as normal enzyme, but has a decreased stability on shaking with air, a decreased Km for phosphoenolpyruvate and a loss of allosteric properties. The apparent Km values for phosphoenolpyruvate of tetramers and monomers were the same. The tetrameric enzyme was slightly activated by fructose-1,6-diphosphate but the monomeric form was not. The tetrameric enzyme was found to dissociate spontaneously to dimeric and monomeric forms. | [
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|
PMID:15248 | Membrane-associated assembly of M13 phage in extracts of virus-infected Escherichia coli. | Assembly of coliphage M13 is known to occur as the viral DNA crosses the cytoplasmic membrane, shedding its virus-coded DNA unwinding protein and acquiring from the membrane approximately 2400 copies of the major coat protein. Conditions are described in which extracts of M13-infected E. coli and membranes prepared from such extracts will support virus assembly at a rate equivalent to that of intact cells. Extracts prepared from cells infected with temperature-sensitive M13 mutants in genes 1, 3, 4, or 5 are temperature-sensitive in this cell-free assembly reaction. Phage assembly in vitro requires magnesium and as yet an unidentified heat-stable cofactor of low molecular weight. The rate of virus assembly is approximately linear with respect to extract concentration over a 10(4)-fold range, consistent with the observation that the entire M13 assembly activity copurifies with the cell membrane fraction. | Membrane-associated assembly of M13 phage in extracts of virus-infected Escherichia coli. Assembly of coliphage M13 is known to occur as the viral DNA crosses the cytoplasmic membrane, shedding its virus-coded DNA unwinding protein and acquiring from the membrane approximately 2400 copies of the major coat protein. Conditions are described in which extracts of M13-infected E. coli and membranes prepared from such extracts will support virus assembly at a rate equivalent to that of intact cells. Extracts prepared from cells infected with temperature-sensitive M13 mutants in genes 1, 3, 4, or 5 are temperature-sensitive in this cell-free assembly reaction. Phage assembly in vitro requires magnesium and as yet an unidentified heat-stable cofactor of low molecular weight. The rate of virus assembly is approximately linear with respect to extract concentration over a 10(4)-fold range, consistent with the observation that the entire M13 assembly activity copurifies with the cell membrane fraction. | [
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|
PMID:15249 | Catecholamine binding to the beta-adrenergic receptor. | The adenylate cyclase-coupled beta-adrenergic receptors of frog erythrocyte membranes have been identified by direct radioligand binding techniques using the potent catecholamine agonist (+/-)[3H]hydroxybenzylisproterenol (2-[3, 4-dihydroxyphenyl]-2-hydroxy-1', 1'-dimethyl-2'-[4-hydroxyphenyl]-diethylamine). The successful experimental conditions included the use of (i) high concentrations of catechol and ascorbic acid to suppress nonreceptor binding, (ii) a very potent radiolabeled catecholamine (10 times more potent than isoproterenol), and (iii) membranes rich in binding sites for beta-adrenergic receptors. Thus, previous problems in accomplishing successful catecholamine binding to the beta-receptors have been overcome. The binding sites identified with (+/-)[3H]hydroxybenzylisoproterenol in the erythrocyte membranes have all the characteristics expected of true beta-adrenergic receptors. These include rapidity of binding, saturability, specificity for beta-agonists and antagonists, and stereospecificity [(-)isomers more potent than (+)isomers]. Physiologically inactive compounds containing a catechol moiety do not compete for occupancy of these binding sites. Dissociation of the radiolabeled agonist from the receptors is slow and incomplete in the absence of guanine nucleotides. In the presence of nucleotide, however, dissociation is rapid and complete. beta-Adrenergic agonists and antagonists compete for the (+/-)[3H]hydroxybenzylisoproterenol binding sites in a fashion parallel to their competition for the receptors, as previously delineated with the beta-adrenergic antagonist (-)[3H]dihydroalprenolol. | Catecholamine binding to the beta-adrenergic receptor. The adenylate cyclase-coupled beta-adrenergic receptors of frog erythrocyte membranes have been identified by direct radioligand binding techniques using the potent catecholamine agonist (+/-)[3H]hydroxybenzylisproterenol (2-[3, 4-dihydroxyphenyl]-2-hydroxy-1', 1'-dimethyl-2'-[4-hydroxyphenyl]-diethylamine). The successful experimental conditions included the use of (i) high concentrations of catechol and ascorbic acid to suppress nonreceptor binding, (ii) a very potent radiolabeled catecholamine (10 times more potent than isoproterenol), and (iii) membranes rich in binding sites for beta-adrenergic receptors. Thus, previous problems in accomplishing successful catecholamine binding to the beta-receptors have been overcome. The binding sites identified with (+/-)[3H]hydroxybenzylisoproterenol in the erythrocyte membranes have all the characteristics expected of true beta-adrenergic receptors. These include rapidity of binding, saturability, specificity for beta-agonists and antagonists, and stereospecificity [(-)isomers more potent than (+)isomers]. Physiologically inactive compounds containing a catechol moiety do not compete for occupancy of these binding sites. Dissociation of the radiolabeled agonist from the receptors is slow and incomplete in the absence of guanine nucleotides. In the presence of nucleotide, however, dissociation is rapid and complete. beta-Adrenergic agonists and antagonists compete for the (+/-)[3H]hydroxybenzylisoproterenol binding sites in a fashion parallel to their competition for the receptors, as previously delineated with the beta-adrenergic antagonist (-)[3H]dihydroalprenolol. | [
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|
PMID:15250 | Trans-synaptic induction of adrenomedullary tyrosine hydroxylase activity by choline: evidence that choline administration can increase cholinergic transmission. | Twenty-four hours after rats receive choline chloride (20 mmol/kg, by stomach tube) the activity of tyrosine hydroxylase [tyrosine 3-monooxygenase; L-tyrosine, tetrahydropteridine:oxygen oxidoreductase (3-hydroxylating), EC 1.14.16.2] increases by 31% within adrenomedullary chromaffin cells. This treatment also causes major elevations in the levels of choline and acetylcholine within the adrenal gland; however, acetylcholine levels return to normal by 16 hr after the choline is given. The daily administration of 10 or 20 mmol/kg of choline for 4 days elevates adrenal tyrosine hydroxylase activity by 29% or 51%, respectively. Such increases in tyrosine hydroxylase activity are not observed in animals given ammonium chloride, another basic chloride-containing compound, by stomach tube or in animals treated with cycloheximide, an inhibitor of adrenal protein synthesis. They are also absent in denervated adrenals. These observations demonstrate that the increase in presynaptic acetylcholine levels produced by giving animals the neurotransmitter's precursor (choline) can be associated with parallel changes in the transmission of signals across cholinergic synapses, probably because more of the transmitter is released per nerve impulse. | Trans-synaptic induction of adrenomedullary tyrosine hydroxylase activity by choline: evidence that choline administration can increase cholinergic transmission. Twenty-four hours after rats receive choline chloride (20 mmol/kg, by stomach tube) the activity of tyrosine hydroxylase [tyrosine 3-monooxygenase; L-tyrosine, tetrahydropteridine:oxygen oxidoreductase (3-hydroxylating), EC 1.14.16.2] increases by 31% within adrenomedullary chromaffin cells. This treatment also causes major elevations in the levels of choline and acetylcholine within the adrenal gland; however, acetylcholine levels return to normal by 16 hr after the choline is given. The daily administration of 10 or 20 mmol/kg of choline for 4 days elevates adrenal tyrosine hydroxylase activity by 29% or 51%, respectively. Such increases in tyrosine hydroxylase activity are not observed in animals given ammonium chloride, another basic chloride-containing compound, by stomach tube or in animals treated with cycloheximide, an inhibitor of adrenal protein synthesis. They are also absent in denervated adrenals. These observations demonstrate that the increase in presynaptic acetylcholine levels produced by giving animals the neurotransmitter's precursor (choline) can be associated with parallel changes in the transmission of signals across cholinergic synapses, probably because more of the transmitter is released per nerve impulse. | [
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|
PMID:15251 | Ionic regulation in genetic translation systems. | The polyelectrolyte theory can provide an interpretation of the interdependence of pH, ionic strength, and polyamines one observes in the activity of ribonuclease acting on RNA. According to this theory: (i) A nucleic acid-enzyme complex and the suspending medium may be considered as two phases in equilibrium, even though within limits, the complex is soluble in water. (ii) The enzymatic catalysis is under tight control of the electrostatic potential generated by the system. Consequently, modification in electrostatic potential will induce a concomitant change in activity. (iii) The electrostatic potential can be modified through action on the system of "modulators", either "external" (ionic strength, pH, temperature, etc.) or "internal" (specific ligands, substrates, protein factors, etc.). Similarities between the reaction of ribonuclease (ribonuclease 3'-pyrimidino-oligonucleotidohydrolase; EC 3.1.4.22) and RNA and those observed with highly organized systems catalyzing DNA, RNA, and protein synthesis suggest that the electrostatic potential also provides an important regulatory mechanism in genetic translation. In this view, an essential function of nucleic acids is to provide their enzyme partners with polyanionic microenvironments within which their catalytic activities are controlled by variation in physicochemical parameters, including the proton concentration induced through "modulation" of the local electrostatic potential. | Ionic regulation in genetic translation systems. The polyelectrolyte theory can provide an interpretation of the interdependence of pH, ionic strength, and polyamines one observes in the activity of ribonuclease acting on RNA. According to this theory: (i) A nucleic acid-enzyme complex and the suspending medium may be considered as two phases in equilibrium, even though within limits, the complex is soluble in water. (ii) The enzymatic catalysis is under tight control of the electrostatic potential generated by the system. Consequently, modification in electrostatic potential will induce a concomitant change in activity. (iii) The electrostatic potential can be modified through action on the system of "modulators", either "external" (ionic strength, pH, temperature, etc.) or "internal" (specific ligands, substrates, protein factors, etc.). Similarities between the reaction of ribonuclease (ribonuclease 3'-pyrimidino-oligonucleotidohydrolase; EC 3.1.4.22) and RNA and those observed with highly organized systems catalyzing DNA, RNA, and protein synthesis suggest that the electrostatic potential also provides an important regulatory mechanism in genetic translation. In this view, an essential function of nucleic acids is to provide their enzyme partners with polyanionic microenvironments within which their catalytic activities are controlled by variation in physicochemical parameters, including the proton concentration induced through "modulation" of the local electrostatic potential. | [
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|
PMID:15252 | Catecholamine hormone receptor differences identified on 3T3 and simian virus-transformed 3T3 cells. | Identification and characterization of hormone receptors on the cell surface is an effective tool for studying the plasma membrane. Using the direct binding of a radiolabeled antagonist, (-)[3H]alprenolol, to crude membrane preparations, and a physiological response (cellular cyclic AMP levels), I demonstrated a catecholamine (beta-adrenergic) hormone receptor site coupled to a catecholamine responsive adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] on 3T3 and simian virus 40 (SV40)-transformed 3T3 cells. At a concentration of 1 muM, epinephrine and isoproterenol elevate cellular cyclic AMP levels 8- and 12-fold, respectively, in both cell lines. Norepinephrine was also a potent agonist on 3T3 cells (8-fold stimulation), but SV3T3 cells showed a lesser (2-fold) response to this hormone. The specificity of the physiological response (as well as the direct binding studies using the alprenolol radiolabel) is indicated by the increased effectiveness of (-) compared to (+) stereoisomers, rapid and reversible kinetics (steady state within 2 min), high affinity (Kd approximately 30 nM) and saturability (indicating a finite number of hormone receptors). These hormone receptor studies indicate the 3T3 cells have a beta1 adrenergic receptor while the SV3T3 cells have a receptor with beta2 qualities. In addition, the number of beta-adrenergic hormone receptors appear to be increased in the normal 3T3 cells by approximately 2-fold over the SV3T3 cells (300 versus versus 120 femtomol/mg of protein). | Catecholamine hormone receptor differences identified on 3T3 and simian virus-transformed 3T3 cells. Identification and characterization of hormone receptors on the cell surface is an effective tool for studying the plasma membrane. Using the direct binding of a radiolabeled antagonist, (-)[3H]alprenolol, to crude membrane preparations, and a physiological response (cellular cyclic AMP levels), I demonstrated a catecholamine (beta-adrenergic) hormone receptor site coupled to a catecholamine responsive adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] on 3T3 and simian virus 40 (SV40)-transformed 3T3 cells. At a concentration of 1 muM, epinephrine and isoproterenol elevate cellular cyclic AMP levels 8- and 12-fold, respectively, in both cell lines. Norepinephrine was also a potent agonist on 3T3 cells (8-fold stimulation), but SV3T3 cells showed a lesser (2-fold) response to this hormone. The specificity of the physiological response (as well as the direct binding studies using the alprenolol radiolabel) is indicated by the increased effectiveness of (-) compared to (+) stereoisomers, rapid and reversible kinetics (steady state within 2 min), high affinity (Kd approximately 30 nM) and saturability (indicating a finite number of hormone receptors). These hormone receptor studies indicate the 3T3 cells have a beta1 adrenergic receptor while the SV3T3 cells have a receptor with beta2 qualities. In addition, the number of beta-adrenergic hormone receptors appear to be increased in the normal 3T3 cells by approximately 2-fold over the SV3T3 cells (300 versus versus 120 femtomol/mg of protein). | [
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PMID:15253 | Genetic control of renin activity in the submaxillary gland of the mouse. | Administration of androgen to female mice is known to increase the level of several proteins in the submaxillary gland, including nerve growth factor, epidermal growth factor, esteroproteolytic activity, and renin. In the present study, renin activity has been assessed in extracts of submaxillary gland of female mice from two inbred strains (SWR/J and C57BL/10J), from F1 and F2 hybrids, and from backcrosses between F1 and parental strains. In both uninduced and induced mice, renin activity of submaxillary gland was more than 100-fold greater in SWR/J than in C57BL/10J mice as measured by either an enzymatic assay or an immunodiffusion method. This difference was not due to differences in plasma testosterone levels between the strains, and the enzymes from the two strains had similar pH optima, substrate specificities, heat stabilities, and apparent Michaelis constants. In the submaxillary gland the difference was relatively specific for renin because increases in esteroproteolytic activity, nerve growth factor, and epidermal growth factor after androgen treatment appeared to be similar in both strains. Studies with the various hybrids indicate that the difference in renin activity between the two strains is apparently due to a single regulatory gene. | Genetic control of renin activity in the submaxillary gland of the mouse. Administration of androgen to female mice is known to increase the level of several proteins in the submaxillary gland, including nerve growth factor, epidermal growth factor, esteroproteolytic activity, and renin. In the present study, renin activity has been assessed in extracts of submaxillary gland of female mice from two inbred strains (SWR/J and C57BL/10J), from F1 and F2 hybrids, and from backcrosses between F1 and parental strains. In both uninduced and induced mice, renin activity of submaxillary gland was more than 100-fold greater in SWR/J than in C57BL/10J mice as measured by either an enzymatic assay or an immunodiffusion method. This difference was not due to differences in plasma testosterone levels between the strains, and the enzymes from the two strains had similar pH optima, substrate specificities, heat stabilities, and apparent Michaelis constants. In the submaxillary gland the difference was relatively specific for renin because increases in esteroproteolytic activity, nerve growth factor, and epidermal growth factor after androgen treatment appeared to be similar in both strains. Studies with the various hybrids indicate that the difference in renin activity between the two strains is apparently due to a single regulatory gene. | [
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PMID:15254 | Alterations of spore coat processing and protein turnover in a Bacillus cereus mutant with a defective postexponential intracellular protease. | A mutant with an alteration in the major intracellular serine protease produced by postexponential Bacillus cereus was isolated by screening mutants defective in spore germination. The purified enzyme from the mutant is more labile to heat and alkaline pH than the protease from the wild type. Protease activity appears at the same time as in the wild type but only reaches 50% of the specific activity and decays more rapidly during sporulation. Coincident with the decay is a decrease in the rate of protein turnover. Generation of amino acids by turnover seems to be important for sporulation because the number of spores produced by the mutant is increased 4- to 10-fold by addition of casamino acids. As anticipated, the mutant produces spores that germinate poorly but, surprisingly, these spores are very deficient in coat protein. Coat antigen is present in cell extracts of mutant and wild type, however, both as large molecules not found on mature spores and as spore coat protein monomers. The large molecules rapidly disappear in a pulse chase experiment in the wild type with some increase in the coat monomers. In mutant extracts, however, this large coat antigen is slowly and improperly processed. | Alterations of spore coat processing and protein turnover in a Bacillus cereus mutant with a defective postexponential intracellular protease. A mutant with an alteration in the major intracellular serine protease produced by postexponential Bacillus cereus was isolated by screening mutants defective in spore germination. The purified enzyme from the mutant is more labile to heat and alkaline pH than the protease from the wild type. Protease activity appears at the same time as in the wild type but only reaches 50% of the specific activity and decays more rapidly during sporulation. Coincident with the decay is a decrease in the rate of protein turnover. Generation of amino acids by turnover seems to be important for sporulation because the number of spores produced by the mutant is increased 4- to 10-fold by addition of casamino acids. As anticipated, the mutant produces spores that germinate poorly but, surprisingly, these spores are very deficient in coat protein. Coat antigen is present in cell extracts of mutant and wild type, however, both as large molecules not found on mature spores and as spore coat protein monomers. The large molecules rapidly disappear in a pulse chase experiment in the wild type with some increase in the coat monomers. In mutant extracts, however, this large coat antigen is slowly and improperly processed. | [
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PMID:15255 | Dehydrogenase and transhydrogenase properties of the soluble NADH dehydrogenase of bovine heart mitochondria. | The soluble NADH dehydrogenase of low molecular weight, isolated from complex I (NADH:ubiquinone oxidoreductase, EC 1.6.5.3) of the respiratory chain, has been shown to have NADPH dehydrogenase and NADPH leads to NAD transhydrogenase activities. Both activities are greatly increased in the presence of added guanidine-HCl and at pH values less than 6.5. The chromophores of the soluble enzyme (flavin and iron--sulfur centers) are reduced by NADH and NADPH to the same extent. The latter reduction is extremely slow, and is considerably stimulated in the presence of guanidine-HCl. The soluble dehydrogenase has little or no NADH leads to NADP and NADPH leads to NADP transhydrogenase activity. The former reaction is known to be energy-linked in submitochondrial particles; the latter was shown in the present studies also to be energy-linked. In view of the above and earlier results, possible mechanisms for dehydrogenation and transhydrogenation (nonenergy-linked and energy-linked) involving reduced and oxidized NAD and NADP are proposed. | Dehydrogenase and transhydrogenase properties of the soluble NADH dehydrogenase of bovine heart mitochondria. The soluble NADH dehydrogenase of low molecular weight, isolated from complex I (NADH:ubiquinone oxidoreductase, EC 1.6.5.3) of the respiratory chain, has been shown to have NADPH dehydrogenase and NADPH leads to NAD transhydrogenase activities. Both activities are greatly increased in the presence of added guanidine-HCl and at pH values less than 6.5. The chromophores of the soluble enzyme (flavin and iron--sulfur centers) are reduced by NADH and NADPH to the same extent. The latter reduction is extremely slow, and is considerably stimulated in the presence of guanidine-HCl. The soluble dehydrogenase has little or no NADH leads to NADP and NADPH leads to NADP transhydrogenase activity. The former reaction is known to be energy-linked in submitochondrial particles; the latter was shown in the present studies also to be energy-linked. In view of the above and earlier results, possible mechanisms for dehydrogenation and transhydrogenation (nonenergy-linked and energy-linked) involving reduced and oxidized NAD and NADP are proposed. | [
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|
PMID:15256 | 31P nuclear magnetic resonance studies of glycogen phosphorylase from rabbit skeletal muscle: ionization states of pyridoxal 5'-phosphate. | 31P nuclear magnetic resonance (NMR) at 72.8 MHZ has been used to study glycogen phosphorylase from rabbit muscle (1,4-alpha-D-glucan:orthophosphate alpha-glucosyltransferase, EC 2.4.1.1) at concentrations as low as 25 mg/ml, using a WH-180 wide-bore superconducting spectrometer. The use of a thio analogue for 5'-AMP and arsenate for inorganic phosphate allowed the observation of three distinct forms of enzyme-bound pyridoxal 5'-phosphate at --0.2 ppm (Form I), --2 to --3 ppm (Form II), and --3.5 ppm (Form III) relative to triethylphosphate. Conversion of I to III occurs by activation of phosphorylase either by formation of a ternary complex of phosphorylase b with effector and arsenate or, more efficiently, by direct phosphorylation to give the a form of the enzyme. The ionization state and exposure to solvent of each of the three forms is inferred from the 31P NMR data. | 31P nuclear magnetic resonance studies of glycogen phosphorylase from rabbit skeletal muscle: ionization states of pyridoxal 5'-phosphate. 31P nuclear magnetic resonance (NMR) at 72.8 MHZ has been used to study glycogen phosphorylase from rabbit muscle (1,4-alpha-D-glucan:orthophosphate alpha-glucosyltransferase, EC 2.4.1.1) at concentrations as low as 25 mg/ml, using a WH-180 wide-bore superconducting spectrometer. The use of a thio analogue for 5'-AMP and arsenate for inorganic phosphate allowed the observation of three distinct forms of enzyme-bound pyridoxal 5'-phosphate at --0.2 ppm (Form I), --2 to --3 ppm (Form II), and --3.5 ppm (Form III) relative to triethylphosphate. Conversion of I to III occurs by activation of phosphorylase either by formation of a ternary complex of phosphorylase b with effector and arsenate or, more efficiently, by direct phosphorylation to give the a form of the enzyme. The ionization state and exposure to solvent of each of the three forms is inferred from the 31P NMR data. | [
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|
PMID:15257 | High-resolution 31P nuclear magnetic resonance studies of metabolism in aerobic Escherichia coli cells. | 31P nuclear magnetic resonance spectra at 145.7 MHZ were obtained of concentrated suspensions of E. coli cells. The position of the Pi resonance was used to determine the pH, and in most experiments it was possible to distinguish the intracellular (pHin) and extracellular (pHex) values. During respiration pHin approached 7.55, while pHex varied from 6.0 to 8.0. With succinate as a carbon source and in a N2 environment, pHin - pHex. Upon addition of glucose, pHin greater than pHex. In the presence of an ATPase (adenosinetriphosphatase; ATP phosphohydrolase; EC 3.6.1.3) inhibitor dicyclohexylcarbodiimide, pHin remained equal to pHex even in the presence of glucose. In other experiments, oxygenation brought pHin above pHex even in the presence of dicyclohexylcarbodiimide. These experiments are consistent with Mitchell's hypothesis that, first, delta pH can be created by the reversal of the ATPase reaction and, second, that protons are pumped outward during respiration. In addition to Pi, about 10 more resonances were resolved, several of which were assigned to different phosphate metabolites. | High-resolution 31P nuclear magnetic resonance studies of metabolism in aerobic Escherichia coli cells. 31P nuclear magnetic resonance spectra at 145.7 MHZ were obtained of concentrated suspensions of E. coli cells. The position of the Pi resonance was used to determine the pH, and in most experiments it was possible to distinguish the intracellular (pHin) and extracellular (pHex) values. During respiration pHin approached 7.55, while pHex varied from 6.0 to 8.0. With succinate as a carbon source and in a N2 environment, pHin - pHex. Upon addition of glucose, pHin greater than pHex. In the presence of an ATPase (adenosinetriphosphatase; ATP phosphohydrolase; EC 3.6.1.3) inhibitor dicyclohexylcarbodiimide, pHin remained equal to pHex even in the presence of glucose. In other experiments, oxygenation brought pHin above pHex even in the presence of dicyclohexylcarbodiimide. These experiments are consistent with Mitchell's hypothesis that, first, delta pH can be created by the reversal of the ATPase reaction and, second, that protons are pumped outward during respiration. In addition to Pi, about 10 more resonances were resolved, several of which were assigned to different phosphate metabolites. | [
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|
PMID:15258 | Intracellular binding of radioactive hydroxocobalamin to cobalamin-dependent apoenzymes in rat liver. | We identified previously an intracellular cobalamin (Cbl) binding protein(s) in cultured human fibroblasts, distinct from known Cbl "R" binders and absent from mutant cells deficient in the synthesis of the two Cbl coenzymes. In order to further characterize this binding activity, we have investigated its homologue in rat liver. After being transported to the liver by the serum protein transcobalamin II, [57Co]Cbl was bound by at least two distinct proteins, one cytosolic, the other mitochondrial. Labeled Cbl bound to cytosolic protein faster than or prior to the mitochondrial protein. With time there was a decline in radioactivity associated with the cytosolic binder and a coordinate increase in that associated with the mitochondrial binder. Although both proteins cochromatographed on Sephadex G-150 and had apparent molecular weights of 120,000, they were separated into two discrete components by polyacrylamide gel electrophoresis and by DEAE-cellulose chromatography. The cytosolic binder cochromatographed with N5-methyltetrahydrofolate:homocysteine methyltransferase activity (5-methyltetrahydropteroyl-L-glutamate:L-homocysteine S-methyltransferase, EC 2.1.1.13); the mitochondrial one with methylmalonyl CoA mutase activity (methylmalonyl-CoA CoA-carbonylmutase, EC 5.4.99.2). These proteins were distinguished further by the chemical forms of [57Co]Cbl found with them, hydroxocobalamin and methylcobalamin with the cytosolic protein and adenosylcobalamin with the mitochondrial one. These results suggest that intracellular Cbl binding activity in rat liver can be accounted for by attachment of Cbl to the two known Cbl-dependent apoenzymes, methylmalonyl CoA mutase and methyltetrahydrofolate methyltransferase. The mechanism and significance of the observered binding protein deficiency in mutant human fibroblasts must, therefore, be re-evaluated. | Intracellular binding of radioactive hydroxocobalamin to cobalamin-dependent apoenzymes in rat liver. We identified previously an intracellular cobalamin (Cbl) binding protein(s) in cultured human fibroblasts, distinct from known Cbl "R" binders and absent from mutant cells deficient in the synthesis of the two Cbl coenzymes. In order to further characterize this binding activity, we have investigated its homologue in rat liver. After being transported to the liver by the serum protein transcobalamin II, [57Co]Cbl was bound by at least two distinct proteins, one cytosolic, the other mitochondrial. Labeled Cbl bound to cytosolic protein faster than or prior to the mitochondrial protein. With time there was a decline in radioactivity associated with the cytosolic binder and a coordinate increase in that associated with the mitochondrial binder. Although both proteins cochromatographed on Sephadex G-150 and had apparent molecular weights of 120,000, they were separated into two discrete components by polyacrylamide gel electrophoresis and by DEAE-cellulose chromatography. The cytosolic binder cochromatographed with N5-methyltetrahydrofolate:homocysteine methyltransferase activity (5-methyltetrahydropteroyl-L-glutamate:L-homocysteine S-methyltransferase, EC 2.1.1.13); the mitochondrial one with methylmalonyl CoA mutase activity (methylmalonyl-CoA CoA-carbonylmutase, EC 5.4.99.2). These proteins were distinguished further by the chemical forms of [57Co]Cbl found with them, hydroxocobalamin and methylcobalamin with the cytosolic protein and adenosylcobalamin with the mitochondrial one. These results suggest that intracellular Cbl binding activity in rat liver can be accounted for by attachment of Cbl to the two known Cbl-dependent apoenzymes, methylmalonyl CoA mutase and methyltetrahydrofolate methyltransferase. The mechanism and significance of the observered binding protein deficiency in mutant human fibroblasts must, therefore, be re-evaluated. | [
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|
PMID:15259 | Recognition of two intracellular cobalamin binding proteins and their identification as methylmalonyl-CoA mutase and methionine synthetase. | The granulocyte R-type cobalamin binding protein delivers cobalamin (Cbl) exclusively to hepatocytes, and transcobalamin II delivers Cbl to various mammalian cells. Both protein-Cbl complexes enter cells by pinocytosis, and the protein moieties are rapidly degraded in lysosomes. The liberated Cbl is subsequently bound to a high-molecular-weight intracellular cobalamin binding protein (ICB). The nature of ICB-Cbl is unknown but appears important because ICB-[57Co]Cbl is missing from cultured fibroblasts of a group of patients whose cells take up CN-[57Co]Cbl normally but do not convert it to either of its coenzyme forms. We have examined supernatants of sonicated rabbit livers and have found that 65% of the total endogenous Cbl elutes from Sephadex G-150 as ICB-Cbl and that this fraction also contains the two mammalian Cbl-dependent enzymes, methylmalonyl-CoA mutase (methylmalonyl-CoA CoA-carbonylmutase;EC 5.4.99.2) and methionine synthetase (tetrahydropteroylglutamate methyltransferase; 5-methyltetrahydropteroyl-L-glutamate:L-homocysteine-S-methyltransferase; EC 2.1.1.13). Gradient elution from DEAE-Sephadex reveals that 90--95% of the ICB--Cbl elutes with methylmalonyl-CoA mutase and 5--10% elutes with methionine synthetase. ICB--[57Co]Cbl first appears 2 hr after the intravenous injection of CN[57Co]Cbl bound to granulocyte R-type protein. This ICB-[57Co]Cbl is associated with either methylmalonyl-CoA mutase or methionine synthetase although the latter appears to be formed at a relatively faster rate. Our studies indicate that mammalian cells contain two ICBs, that these proteins are methylmalonyl-CoA mutase and methionine synthetase, and that the primary abnormality in the group of patients mentioned above lies at a step that is common to the formation of both Cbl coenzymes and that precedes the stable binding of Cbl to both methylmalonyl-CoA mutase and methionine synthetase. | Recognition of two intracellular cobalamin binding proteins and their identification as methylmalonyl-CoA mutase and methionine synthetase. The granulocyte R-type cobalamin binding protein delivers cobalamin (Cbl) exclusively to hepatocytes, and transcobalamin II delivers Cbl to various mammalian cells. Both protein-Cbl complexes enter cells by pinocytosis, and the protein moieties are rapidly degraded in lysosomes. The liberated Cbl is subsequently bound to a high-molecular-weight intracellular cobalamin binding protein (ICB). The nature of ICB-Cbl is unknown but appears important because ICB-[57Co]Cbl is missing from cultured fibroblasts of a group of patients whose cells take up CN-[57Co]Cbl normally but do not convert it to either of its coenzyme forms. We have examined supernatants of sonicated rabbit livers and have found that 65% of the total endogenous Cbl elutes from Sephadex G-150 as ICB-Cbl and that this fraction also contains the two mammalian Cbl-dependent enzymes, methylmalonyl-CoA mutase (methylmalonyl-CoA CoA-carbonylmutase;EC 5.4.99.2) and methionine synthetase (tetrahydropteroylglutamate methyltransferase; 5-methyltetrahydropteroyl-L-glutamate:L-homocysteine-S-methyltransferase; EC 2.1.1.13). Gradient elution from DEAE-Sephadex reveals that 90--95% of the ICB--Cbl elutes with methylmalonyl-CoA mutase and 5--10% elutes with methionine synthetase. ICB--[57Co]Cbl first appears 2 hr after the intravenous injection of CN[57Co]Cbl bound to granulocyte R-type protein. This ICB-[57Co]Cbl is associated with either methylmalonyl-CoA mutase or methionine synthetase although the latter appears to be formed at a relatively faster rate. Our studies indicate that mammalian cells contain two ICBs, that these proteins are methylmalonyl-CoA mutase and methionine synthetase, and that the primary abnormality in the group of patients mentioned above lies at a step that is common to the formation of both Cbl coenzymes and that precedes the stable binding of Cbl to both methylmalonyl-CoA mutase and methionine synthetase. | [
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|
PMID:15260 | Affinity labeling of gamma-glutamyl transpeptidase and location of the gamma-glutamyl binding site on the light subunit. | Gamma-Glutamyl transpeptidase, which consists of two nonidentical subunits, is rapidly inactivated with respect to its transpeptidase and hydrolase activities by the gamma-glutamyl analogs 6-diazo-5-oxo-L-norleucine and L-azaserine. Inactivation, which is prevented by gamma-glutamyl substrates (but not by acceptor substrates), is accelerated by maleate, which was previously shown to enhance utilization of glutamine by transpeptidase. 6-Diazo-5-oxo--norleucine reacts specifically, covalently, and stoichiometrically at the gamma-glutamyl site of the enzyme, which was localized through studies with 6-diazo-5-OXO-[14C]norleucine to the light subunits of both the transpeptidase of rat kidney (which has subunits of molecular weights 22,000 and 46,000) and the transpeptidase of human kidney (which has subunits of molecular weights 22,000 and 62,000). The findings, which indicate that these enzymes have similar gamma-glutamyl binding subunits, are relevant to the structure-function relationships of this membrane-bound enzyme and its physiological role. | Affinity labeling of gamma-glutamyl transpeptidase and location of the gamma-glutamyl binding site on the light subunit. Gamma-Glutamyl transpeptidase, which consists of two nonidentical subunits, is rapidly inactivated with respect to its transpeptidase and hydrolase activities by the gamma-glutamyl analogs 6-diazo-5-oxo-L-norleucine and L-azaserine. Inactivation, which is prevented by gamma-glutamyl substrates (but not by acceptor substrates), is accelerated by maleate, which was previously shown to enhance utilization of glutamine by transpeptidase. 6-Diazo-5-oxo--norleucine reacts specifically, covalently, and stoichiometrically at the gamma-glutamyl site of the enzyme, which was localized through studies with 6-diazo-5-OXO-[14C]norleucine to the light subunits of both the transpeptidase of rat kidney (which has subunits of molecular weights 22,000 and 46,000) and the transpeptidase of human kidney (which has subunits of molecular weights 22,000 and 62,000). The findings, which indicate that these enzymes have similar gamma-glutamyl binding subunits, are relevant to the structure-function relationships of this membrane-bound enzyme and its physiological role. | [
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|
PMID:15261 | Detection in bovine adrenal cortex of a lipoidal substance that yields pregnenolone upon treatment with alkali. | Bovine adrenal cortical tissue contains a lipoidal derivative of pregnenolone (3beta-hydroxy-pregn-5-en-20-one) from which the free steroid can be liberated by treatment with alkali. Evidence for the presence of such an entity comes from examination of a nonpolar extract of tissue from which pregnenolone and its sulfate had been removed by chromatography. Treatment of the nonpolar fraction with alkali followed by exhaustive chromatographic analysis led to the detection of pregnenolone. The steroid was identified by both gas chromatography/mass spectrometry and double isotope procedures. Quantitative analysis indicated that the three forms of pregnenolone are present in bovine adrenal cortical tissue in the following amounts (mug/kg): lipoidal derivative, 290; free steroid, 435; and sulfate, 65. Because the only known metabolic function of pregnenolone is to serve as a precursor of the steroid hormones, these findings have far-reaching implications for steroid hormone biochemistry. | Detection in bovine adrenal cortex of a lipoidal substance that yields pregnenolone upon treatment with alkali. Bovine adrenal cortical tissue contains a lipoidal derivative of pregnenolone (3beta-hydroxy-pregn-5-en-20-one) from which the free steroid can be liberated by treatment with alkali. Evidence for the presence of such an entity comes from examination of a nonpolar extract of tissue from which pregnenolone and its sulfate had been removed by chromatography. Treatment of the nonpolar fraction with alkali followed by exhaustive chromatographic analysis led to the detection of pregnenolone. The steroid was identified by both gas chromatography/mass spectrometry and double isotope procedures. Quantitative analysis indicated that the three forms of pregnenolone are present in bovine adrenal cortical tissue in the following amounts (mug/kg): lipoidal derivative, 290; free steroid, 435; and sulfate, 65. Because the only known metabolic function of pregnenolone is to serve as a precursor of the steroid hormones, these findings have far-reaching implications for steroid hormone biochemistry. | [
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|
PMID:15262 | Regulation of hepatic nuclear guanylate cyclase. | In immunohistochemical studies of rat liver tissue slices and purified nuclei, adenosine 3':5'-cyclic monophosphate (cAMP) and guanosine 3':5'-cyclic monophosphate (cGMP) immunofluorescence on the nuclear membrane are sequentially increased after glucagon administration. An explanation for the increased cGMP immunofluorescence was sought in experiments in which guanylate cyclase [GTP pyrophosphate-lyase (cyclizing), EC 4.6.1.2]activity of hepatic subcellular fractions was determined. The results showed that a nuclear guanylate cyclase exists which can be distinguished from the soluble and crude particulate guanylate cyclases. The activity of the nuclear enzyme was increased by 35% in nuclei isolated from rats 30 min after glucagon injection, the time at which maximal nuclear membrane cGMP immunofluorescence is observed. Because glucagon altered both cAMP location and levels prior to the observed changes in nuclear cGMP metabolism, the hypothesis that cAMP acted as the second messenger was tested. In vitro incubation of nuclei isolated from control rats with 10(-5) M cAMP produced a 25% increase in nuclear guanylate cyclase activity. With nuclei isolated from glucagon-treated rats, no significant increase in enzyme activity was observed; this indicates that maximal stimulation of nuclear guanylate cyclase by cAMP occurred at levels that are obtained in vivo after glucagon administration. These findings suggest that hepatic nuclear cGMP content may be regulated by a specific organelle guanylate cyclase and that cAMP may be one of the determinants of this enzyme's activity. | Regulation of hepatic nuclear guanylate cyclase. In immunohistochemical studies of rat liver tissue slices and purified nuclei, adenosine 3':5'-cyclic monophosphate (cAMP) and guanosine 3':5'-cyclic monophosphate (cGMP) immunofluorescence on the nuclear membrane are sequentially increased after glucagon administration. An explanation for the increased cGMP immunofluorescence was sought in experiments in which guanylate cyclase [GTP pyrophosphate-lyase (cyclizing), EC 4.6.1.2]activity of hepatic subcellular fractions was determined. The results showed that a nuclear guanylate cyclase exists which can be distinguished from the soluble and crude particulate guanylate cyclases. The activity of the nuclear enzyme was increased by 35% in nuclei isolated from rats 30 min after glucagon injection, the time at which maximal nuclear membrane cGMP immunofluorescence is observed. Because glucagon altered both cAMP location and levels prior to the observed changes in nuclear cGMP metabolism, the hypothesis that cAMP acted as the second messenger was tested. In vitro incubation of nuclei isolated from control rats with 10(-5) M cAMP produced a 25% increase in nuclear guanylate cyclase activity. With nuclei isolated from glucagon-treated rats, no significant increase in enzyme activity was observed; this indicates that maximal stimulation of nuclear guanylate cyclase by cAMP occurred at levels that are obtained in vivo after glucagon administration. These findings suggest that hepatic nuclear cGMP content may be regulated by a specific organelle guanylate cyclase and that cAMP may be one of the determinants of this enzyme's activity. | [
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PMID:15263 | Effect of batrachotoxin on the electroplax of electric eel: evidence for voltage-dependent interaction with sodium channels. | Batrachotoxin under certain conditions has a strong depolarizing effect on the innervated membrane of the monocellular electroplax preparation from the electric eel, El-ectrophorus electricus. No effect is observed when the toxin (50-200 nM) is applied to the resting membrane for periods up to 1 hr. However, if the membrane is exposed to batrachotoxin and the cell is subjected to stimulation at a stimulus voltage slightly above the threshold for action potential firing, a progressive prolongation of the action potential and concomitant progressive depolarization of the innervated membrane is observed. When the membrane is depolarized by 15-20 mV, a further abrupt all-or-none depolarization occurs, and the potential attains a steady-state value between 0 and -10 mV. Brief stimulation of a cell in the presence of batrachotoxin is sufficient to define a batrachotoxin-treated cell, even though negligible depolarization occurs. If depolarizing agents such as carbamoylcholine or potassium chloride are introduced to such a cell in concentrations that normally produce a 20-30 mV depolarization, the abrupt all-or-none depolarization immediately occurs. All-or-none depolarizations arising from either electrical stimulation or depolarizing agents are unaffected by d-tubocurarine but are completely reversed by tetrodotoxin. Batrachotoxin thus appears to activate only the action potential sodium channels. In the batrachotoxin-treated membrane, these channels can attain stable steady states in either a closed configuration at the normal resting potential or in an open configuration after complete depolarization. A striking hysteresis cycle thus can be generated, which is strongly indicative of a voltage-dependent interaction of the toxin with the action potential sodium channels. | Effect of batrachotoxin on the electroplax of electric eel: evidence for voltage-dependent interaction with sodium channels. Batrachotoxin under certain conditions has a strong depolarizing effect on the innervated membrane of the monocellular electroplax preparation from the electric eel, El-ectrophorus electricus. No effect is observed when the toxin (50-200 nM) is applied to the resting membrane for periods up to 1 hr. However, if the membrane is exposed to batrachotoxin and the cell is subjected to stimulation at a stimulus voltage slightly above the threshold for action potential firing, a progressive prolongation of the action potential and concomitant progressive depolarization of the innervated membrane is observed. When the membrane is depolarized by 15-20 mV, a further abrupt all-or-none depolarization occurs, and the potential attains a steady-state value between 0 and -10 mV. Brief stimulation of a cell in the presence of batrachotoxin is sufficient to define a batrachotoxin-treated cell, even though negligible depolarization occurs. If depolarizing agents such as carbamoylcholine or potassium chloride are introduced to such a cell in concentrations that normally produce a 20-30 mV depolarization, the abrupt all-or-none depolarization immediately occurs. All-or-none depolarizations arising from either electrical stimulation or depolarizing agents are unaffected by d-tubocurarine but are completely reversed by tetrodotoxin. Batrachotoxin thus appears to activate only the action potential sodium channels. In the batrachotoxin-treated membrane, these channels can attain stable steady states in either a closed configuration at the normal resting potential or in an open configuration after complete depolarization. A striking hysteresis cycle thus can be generated, which is strongly indicative of a voltage-dependent interaction of the toxin with the action potential sodium channels. | [
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|
PMID:15284 | Phencyclidine-induced rotational behavior in rats with nigrostriatal lesions and its modulation by dopaminergic and cholinergic agents. | The peripheral administration of the psychotomimetic drug phencyclidine (1-(phenylcyclohexyl) piperidine hydrochloride) (PCP) induces a dose-related ipsilateral rotation in unilateral substantia nigra electrolytically-lesioned rats. The intensity of this rotation can be modulated by administration of various dopaminergic and cholinergic agents. Injection of alpha-methylparatyrosine methylester (125 mg/kg) or haloperidol (1 mg/kg) inhibited the ipsilateral circling behavior. Pimozide (1 mg/kg) also inhibitied the rotation, but to a lesser extent. The injection of the anticholinergic agent trihexyphenidyl (5 mg/kg) potentiated, and the cholinomimetic drug arecoline (5 mg/kg), depressed the rotation induced by PCP (7.5 mg/kg), It is probable that PCP possesses significant dopaminergic and anticholinergic properties. The capacity of PCP to induce rotation in this model may be related to its effects on dopaminergic and cholingergic neurons in the rat striatum. Thus, PCP may induce rotational behavior by potentiating dopaminergic transmission, by blocking cholinergic activity, or both; both of these effects have been demonstrated to be important in the generation of circling behavior in rats with nigrostriatal lesions. | Phencyclidine-induced rotational behavior in rats with nigrostriatal lesions and its modulation by dopaminergic and cholinergic agents. The peripheral administration of the psychotomimetic drug phencyclidine (1-(phenylcyclohexyl) piperidine hydrochloride) (PCP) induces a dose-related ipsilateral rotation in unilateral substantia nigra electrolytically-lesioned rats. The intensity of this rotation can be modulated by administration of various dopaminergic and cholinergic agents. Injection of alpha-methylparatyrosine methylester (125 mg/kg) or haloperidol (1 mg/kg) inhibited the ipsilateral circling behavior. Pimozide (1 mg/kg) also inhibitied the rotation, but to a lesser extent. The injection of the anticholinergic agent trihexyphenidyl (5 mg/kg) potentiated, and the cholinomimetic drug arecoline (5 mg/kg), depressed the rotation induced by PCP (7.5 mg/kg), It is probable that PCP possesses significant dopaminergic and anticholinergic properties. The capacity of PCP to induce rotation in this model may be related to its effects on dopaminergic and cholingergic neurons in the rat striatum. Thus, PCP may induce rotational behavior by potentiating dopaminergic transmission, by blocking cholinergic activity, or both; both of these effects have been demonstrated to be important in the generation of circling behavior in rats with nigrostriatal lesions. | [
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|
PMID:15285 | Differential motor effects of intraventricular infusion of morphine and etonitazene. | The motor effects produced by intraventricular infusions of morphine were compared to the effects of etonitazene. Despite the similarity in the peripheral actions of these drugs, motor effects of central infusions differed dramatically. Intraventricular morphine infusions resulted in explosive motor behavior whereas etonitazene produced extreme muscular rigidity. The periaqueductal grey (PAG) has been proposed as the substrate of morphine-induced explosive motor behavior. However, considerations of the dose of morphine and the mobility of this drug in tissue suggests that sites other than the PAG may also be involved in explosive motor behavior. | Differential motor effects of intraventricular infusion of morphine and etonitazene. The motor effects produced by intraventricular infusions of morphine were compared to the effects of etonitazene. Despite the similarity in the peripheral actions of these drugs, motor effects of central infusions differed dramatically. Intraventricular morphine infusions resulted in explosive motor behavior whereas etonitazene produced extreme muscular rigidity. The periaqueductal grey (PAG) has been proposed as the substrate of morphine-induced explosive motor behavior. However, considerations of the dose of morphine and the mobility of this drug in tissue suggests that sites other than the PAG may also be involved in explosive motor behavior. | [
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|
PMID:15288 | The different influence of predominantly antiadrenergic and antidopaminergic neuroleptics on the estrous cycle in rats. | In experiments performed on rats, single subcutaneous doses of predominantly antidopaminergic neuroleptics (fluspirilene, pimozide, and thioproperazine in large doses) elicited persistent diestrus, i.e. pseudopregnancy. Neuroleptics with predominantly antiadrenergic action (levomepromazine, thioridazine, chlorpromazine) as well as phenoxybenzamine injected intraventricularly induced mainly prolonged estrus, i.e. blocking of ovulation. Since pseudopregnancy implies a rise of prolactin secretion, our results indirectly indicate that PIF secretion is stimulated by dopamine. The antiovulatory effect of the antiadrenergic agents, which involves an inhibition of the proestral LH surge, suggests that the discharge of LRF is mediated by an adrenergic mechanism. | The different influence of predominantly antiadrenergic and antidopaminergic neuroleptics on the estrous cycle in rats. In experiments performed on rats, single subcutaneous doses of predominantly antidopaminergic neuroleptics (fluspirilene, pimozide, and thioproperazine in large doses) elicited persistent diestrus, i.e. pseudopregnancy. Neuroleptics with predominantly antiadrenergic action (levomepromazine, thioridazine, chlorpromazine) as well as phenoxybenzamine injected intraventricularly induced mainly prolonged estrus, i.e. blocking of ovulation. Since pseudopregnancy implies a rise of prolactin secretion, our results indirectly indicate that PIF secretion is stimulated by dopamine. The antiovulatory effect of the antiadrenergic agents, which involves an inhibition of the proestral LH surge, suggests that the discharge of LRF is mediated by an adrenergic mechanism. | [
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|
PMID:15290 | The functional anatomy of the mantle complex and columellar muscle of tectibranch molluscs (Gastropoda: Opisthobranchia), and its bearing on the evolution of opisthobranch organization. | An account is given of the anatomy of a series of opisthobranch molluscs principally to assess the change in importance and functioning of the mantle cavity and columellar muscle throughout the transition from prosobranch to opisthobranch organization. Intermediate steps are represented by living tectibranchs, of which Philine and Scaphander are investigated in detail, Acteon, Bulla, Haminoea, Akera, Aglaja and Gastropteron more briefly. Though an opisthobranch, Acteon has an organization typical of a monotocardian prosobranch; the remainder show trends affecting the shell and visceral mass, mantle cavity and head-foot, which resulted finally in the production of nudibranch types. It is confirmed that the adaptations exhibited by primitive tectibranchs relate to the assumption of a burrowing mode of life. Initial changes were the reduction of the nuchal area and sealing of the mantle cavity anteriorly so that it opened on the right, where it became restricted, the first perhaps prompting the sealing. A broadening and an anterior elongation of the head-foot produced a wedge to facilitate burrowing. Change in disposition of the mantle edge, incurred by differential growth, produced an involute shell with a large body whorl, alignment changing from erect to horizontal. The resultant streamlining eased infaunal progression; no vertical insinking of the viscera was involved. Subsequently the shell became reduced and finally lost. A section of the mantle edge enlarged to produce a posterior mantle lobe upon which sit both the shell and viscera, and which later became redundant as posterior elongation of the head-foot produced a slug-like form, the viscera being incorporated within the head-foot. As the nuchal area became reduced, mechanical needs prompted alteration to both the form and attachment of the columellar muscle. In Acteon the muscle is like that of a prosobranch, but the proximal region has broadened, a change of proportion required by primitive tectibranchs in order to support the floor of the mantle cavity formed from the section of mantle skirt which in prosobranchs lies on the right. This was followed by reduction and re-alignment of the entire muscle along an anteroposterior axis as emphasis changed from the muscle effecting retraction into a shell to producing contorsions of the head-foot. The shell, similarly reduced, instead of providing anchorage, became itself anchored by additional anterior and posterior attachment zones with, in more advanced forms, dorsoventral muscles of the body wall rather than longitudinal muscles fastening to the former. Importance was placed on the mutual stabilization of constituent parts of the posterior body region. Re-alignments of the muscle induced breaking up of the longitudinal muscle sheet of the head-foot to produce muscle tracts, best exhibited in those tectibranchs which swim; they are derived from both the columellar muscle and intrinsic body wall muscles... | The functional anatomy of the mantle complex and columellar muscle of tectibranch molluscs (Gastropoda: Opisthobranchia), and its bearing on the evolution of opisthobranch organization. An account is given of the anatomy of a series of opisthobranch molluscs principally to assess the change in importance and functioning of the mantle cavity and columellar muscle throughout the transition from prosobranch to opisthobranch organization. Intermediate steps are represented by living tectibranchs, of which Philine and Scaphander are investigated in detail, Acteon, Bulla, Haminoea, Akera, Aglaja and Gastropteron more briefly. Though an opisthobranch, Acteon has an organization typical of a monotocardian prosobranch; the remainder show trends affecting the shell and visceral mass, mantle cavity and head-foot, which resulted finally in the production of nudibranch types. It is confirmed that the adaptations exhibited by primitive tectibranchs relate to the assumption of a burrowing mode of life. Initial changes were the reduction of the nuchal area and sealing of the mantle cavity anteriorly so that it opened on the right, where it became restricted, the first perhaps prompting the sealing. A broadening and an anterior elongation of the head-foot produced a wedge to facilitate burrowing. Change in disposition of the mantle edge, incurred by differential growth, produced an involute shell with a large body whorl, alignment changing from erect to horizontal. The resultant streamlining eased infaunal progression; no vertical insinking of the viscera was involved. Subsequently the shell became reduced and finally lost. A section of the mantle edge enlarged to produce a posterior mantle lobe upon which sit both the shell and viscera, and which later became redundant as posterior elongation of the head-foot produced a slug-like form, the viscera being incorporated within the head-foot. As the nuchal area became reduced, mechanical needs prompted alteration to both the form and attachment of the columellar muscle. In Acteon the muscle is like that of a prosobranch, but the proximal region has broadened, a change of proportion required by primitive tectibranchs in order to support the floor of the mantle cavity formed from the section of mantle skirt which in prosobranchs lies on the right. This was followed by reduction and re-alignment of the entire muscle along an anteroposterior axis as emphasis changed from the muscle effecting retraction into a shell to producing contorsions of the head-foot. The shell, similarly reduced, instead of providing anchorage, became itself anchored by additional anterior and posterior attachment zones with, in more advanced forms, dorsoventral muscles of the body wall rather than longitudinal muscles fastening to the former. Importance was placed on the mutual stabilization of constituent parts of the posterior body region. Re-alignments of the muscle induced breaking up of the longitudinal muscle sheet of the head-foot to produce muscle tracts, best exhibited in those tectibranchs which swim; they are derived from both the columellar muscle and intrinsic body wall muscles... | [
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|
PMID:15294 | [Electrolyte-water balance and acid-base equilibrium in brain tumor patients]. | Electrolyte variations, water-balance disturbances, and acid-base equilibrium disorders observed in patients with brain tumors are due, in the majority of cases, to increases in intracranial pressure and, in a relatively small number of cases, to the particular location of the tumor. Severe pathological pictures are not, in general, observed until the ailment has advanced to a critical state. The authors, after describing the clinical pictures of the various forms of acid-base equilibrium disorders, also discuss methods of treatment. Disturbances of water balance are closely associated with the electrolyte metabolism. Consequently, it is necessary that, if a dehydrating form of therapy is used, careful attention should be given to the corresponding parameters. Disturbances of iatrogenic origin tend to produce particularly adverse effects in brain tumor patients. | [Electrolyte-water balance and acid-base equilibrium in brain tumor patients]. Electrolyte variations, water-balance disturbances, and acid-base equilibrium disorders observed in patients with brain tumors are due, in the majority of cases, to increases in intracranial pressure and, in a relatively small number of cases, to the particular location of the tumor. Severe pathological pictures are not, in general, observed until the ailment has advanced to a critical state. The authors, after describing the clinical pictures of the various forms of acid-base equilibrium disorders, also discuss methods of treatment. Disturbances of water balance are closely associated with the electrolyte metabolism. Consequently, it is necessary that, if a dehydrating form of therapy is used, careful attention should be given to the corresponding parameters. Disturbances of iatrogenic origin tend to produce particularly adverse effects in brain tumor patients. | [
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|
PMID:15295 | Medazepam and the driving ability of anxious patients. | A double-blind crossover trial of Medazepam was carried out in 14 anxious hospital patients. The mean self-adjusted dosage was 16.5 mg daily. The active drug was no more effective than placebo in relieving anxiety, which was rated both clinically and by the Middlesex Health Questionnaire (M.H.Q.) (Crown and Crisp, 1970). This may have been because the dose was relatively low for chronically anxious hospital patients. Even this dosage caused significantly higher scores on the M.H.Q. scale for depression. Braking and driving simulator tests were not adversely affected by Medazepam. In real driving conditions those taking the drug made significantly more technical, but not dangerous, errors. Pulse and blood pressure also were not affected. | Medazepam and the driving ability of anxious patients. A double-blind crossover trial of Medazepam was carried out in 14 anxious hospital patients. The mean self-adjusted dosage was 16.5 mg daily. The active drug was no more effective than placebo in relieving anxiety, which was rated both clinically and by the Middlesex Health Questionnaire (M.H.Q.) (Crown and Crisp, 1970). This may have been because the dose was relatively low for chronically anxious hospital patients. Even this dosage caused significantly higher scores on the M.H.Q. scale for depression. Braking and driving simulator tests were not adversely affected by Medazepam. In real driving conditions those taking the drug made significantly more technical, but not dangerous, errors. Pulse and blood pressure also were not affected. | [
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|
PMID:15296 | Effects of oxazolam, cloxazolam, and CS-386, new anti-anxiety drugs, on socially induced suppression and aggression in pairs of monkeys. | This experiment was conducted to examine effects of oxazolam, cloxazolam, CS-386, and reference drugs on socially induced suppression and aggression in pairs of monkeys. Oxazolam, cloxazolam, and CS-386, as well as other benzodiazepines, at both ataxic and non-ataxic doses, attenuated the socially induced suppression, but failed to show inhibitory effect on the on the socially induced aggression. Chlorpromazine, at both slight-sedative and non-sedative doses, reduced neither socially induced suppression nor aggression. Imipramine did not produce any significant effect in this study. | Effects of oxazolam, cloxazolam, and CS-386, new anti-anxiety drugs, on socially induced suppression and aggression in pairs of monkeys. This experiment was conducted to examine effects of oxazolam, cloxazolam, CS-386, and reference drugs on socially induced suppression and aggression in pairs of monkeys. Oxazolam, cloxazolam, and CS-386, as well as other benzodiazepines, at both ataxic and non-ataxic doses, attenuated the socially induced suppression, but failed to show inhibitory effect on the on the socially induced aggression. Chlorpromazine, at both slight-sedative and non-sedative doses, reduced neither socially induced suppression nor aggression. Imipramine did not produce any significant effect in this study. | [
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|
PMID:15297 | Intensive design in evaluating anxiolytic agents. | The purposes of this study were: (1) to test the usefulness of intensive design in detecting the effects of an established antianxiety agent in a single patient studied for a period as brief as 8 weeks and (2) to explore the usefulness of combining intensive and extensive designs by jointly analyzing the results from several similarly treated patients. Fifteen primarily anxious, psychoneurotic patients aged 21-50 and scoring 17 or more on the Taylor Manifest Anxiety Scale were admitted to the study; and 11 completed the full treatment program. Medications were diazepam 5 mg t.i.d. and a matching placebo, administered under double-blind conditions. Patients were treated for 8 weeks, divided into 42-week blocks. In each block, the patient received diazepam 1 week and placebo the other, with the order in each block determined at random. The patient came weekly for evaluation, including, self-ratings on the Hopkins Symptom Checklist (SCL), global status, global change; reports of occupational and social function; resting pulse; reaction time; psychiatrist's ratings on the Hamilton Anxiety Scale, global status and global change. The patient also reported daily his mood on the Profile of Mood States (POMS). Mean deviations from the general trend for post-diazepam and postplacebo scores on each criterion were compared within patients. Diazepam-placebo differences on each criterion were analyzed between patients. Criteria that clearly recorded the anti-anxiety effect of diazepam as compared to placebo included the Hamilton Anxiety Scale, the psychiatrist's global status and global change ratings, the SCL Anxiety and Somatization Scales, and the POMS Anxiety Scale. Other criteria that showed a reliable diazepam effect included SCL Depression (decrease), POMS Vigor (increase), POMS Fatigue (decrease), SCL Anger (increase), and reaction time (increase). The most sensitive criteria distinguished diazepam from placebo even when results were considered only from the first 6 patients during their first 4 weeks of treatment- a total of 24 patient weeks of treatment. The factors contributing to the sensitivity of this design were investigated and discussed. | Intensive design in evaluating anxiolytic agents. The purposes of this study were: (1) to test the usefulness of intensive design in detecting the effects of an established antianxiety agent in a single patient studied for a period as brief as 8 weeks and (2) to explore the usefulness of combining intensive and extensive designs by jointly analyzing the results from several similarly treated patients. Fifteen primarily anxious, psychoneurotic patients aged 21-50 and scoring 17 or more on the Taylor Manifest Anxiety Scale were admitted to the study; and 11 completed the full treatment program. Medications were diazepam 5 mg t.i.d. and a matching placebo, administered under double-blind conditions. Patients were treated for 8 weeks, divided into 42-week blocks. In each block, the patient received diazepam 1 week and placebo the other, with the order in each block determined at random. The patient came weekly for evaluation, including, self-ratings on the Hopkins Symptom Checklist (SCL), global status, global change; reports of occupational and social function; resting pulse; reaction time; psychiatrist's ratings on the Hamilton Anxiety Scale, global status and global change. The patient also reported daily his mood on the Profile of Mood States (POMS). Mean deviations from the general trend for post-diazepam and postplacebo scores on each criterion were compared within patients. Diazepam-placebo differences on each criterion were analyzed between patients. Criteria that clearly recorded the anti-anxiety effect of diazepam as compared to placebo included the Hamilton Anxiety Scale, the psychiatrist's global status and global change ratings, the SCL Anxiety and Somatization Scales, and the POMS Anxiety Scale. Other criteria that showed a reliable diazepam effect included SCL Depression (decrease), POMS Vigor (increase), POMS Fatigue (decrease), SCL Anger (increase), and reaction time (increase). The most sensitive criteria distinguished diazepam from placebo even when results were considered only from the first 6 patients during their first 4 weeks of treatment- a total of 24 patient weeks of treatment. The factors contributing to the sensitivity of this design were investigated and discussed. | [
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|
PMID:15299 | Spectrum of angiographically demonstrable renal pathology in young hypertensive patients. | The renal diseases which cause systemic hypertension in the first two decades of life differ from the adult in their incidence and etiology. Seventeen patients (11 days to 20 years old), studied angiographically, demonstrated a wide spectrum of renal pathology including arterial thrombosis, fibromuscular hyperplasia, vasculitis, neurofibramatosis, cystic disease, pyelonephritis, Page kidney, and congenital anomalies. | Spectrum of angiographically demonstrable renal pathology in young hypertensive patients. The renal diseases which cause systemic hypertension in the first two decades of life differ from the adult in their incidence and etiology. Seventeen patients (11 days to 20 years old), studied angiographically, demonstrated a wide spectrum of renal pathology including arterial thrombosis, fibromuscular hyperplasia, vasculitis, neurofibramatosis, cystic disease, pyelonephritis, Page kidney, and congenital anomalies. | [
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|
PMID:15300 | The influence of hypoxia and acidity on the hyperthermic response of malignant cells in vitro. | Colony formation of JB-1-E tumor cells was studied after hyperthermic treatment (42.5 degrees C) at a pH of 6.4 or 7.2 under hypoxic and euoxic conditions. At a pH of 7.2 and normal oxygen tension, there was a moderate decrease in colony formation with increasing duration of hyperthermic treatment (To = 65 min.). This effect was slightly enhanced under hypoxic conditions (To = 36 min.). The hyperthermic effect was enhanced to a considerably greater degree when treatment was performed at a pH of 6.4 (To = 19 min.), with no observable difference between hypoxia and euoxia. These findings indicate that environmental acidity is a determining factor in the hyperthermic effect. The hypoxic effect at a pH of 7.2 is probably due to a slight decrease in the intracellular pH caused by increased production of lactic acid. | The influence of hypoxia and acidity on the hyperthermic response of malignant cells in vitro. Colony formation of JB-1-E tumor cells was studied after hyperthermic treatment (42.5 degrees C) at a pH of 6.4 or 7.2 under hypoxic and euoxic conditions. At a pH of 7.2 and normal oxygen tension, there was a moderate decrease in colony formation with increasing duration of hyperthermic treatment (To = 65 min.). This effect was slightly enhanced under hypoxic conditions (To = 36 min.). The hyperthermic effect was enhanced to a considerably greater degree when treatment was performed at a pH of 6.4 (To = 19 min.), with no observable difference between hypoxia and euoxia. These findings indicate that environmental acidity is a determining factor in the hyperthermic effect. The hypoxic effect at a pH of 7.2 is probably due to a slight decrease in the intracellular pH caused by increased production of lactic acid. | [
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|
PMID:15301 | Evidence against the release of prostaglandin-like material from isolated intestinal tissue by pure cholera toxin. | Prostaglandin-like material was released from finely cut guinea-pig ileum or human intestinal mucosa during incubation with Krebs solution. The tissue inactivated some significant change in release of prostaglandin-like material when pure cholera toxin was incubated with guinea-pig ileum or human intestinal mucosa. The work is discussed in relation to the action of cholera toxin in vivo. | Evidence against the release of prostaglandin-like material from isolated intestinal tissue by pure cholera toxin. Prostaglandin-like material was released from finely cut guinea-pig ileum or human intestinal mucosa during incubation with Krebs solution. The tissue inactivated some significant change in release of prostaglandin-like material when pure cholera toxin was incubated with guinea-pig ileum or human intestinal mucosa. The work is discussed in relation to the action of cholera toxin in vivo. | [
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|
PMID:15302 | Multiple molecular forms of prostaglandin 15-hydroxydehydrogenase and 9-ketoreductase in chicken kidney. | Prostaglandin-15-hydroxydehydrogenase and prostaglandin-9-keto-reductase were purified from chicken kidney. Both enzymes exist in multiple forms as determined by isoelectric focusing. The dehydrogenases catalyze the transformation of the functional group at C-15 but not the functional group at C-9. The preferred cofactors in these reactions are NAD+ or NADH. The 9-ketoreductases catalyze the reversible transformation of the functional group at C-9 and also the oxidation or reduction of the C-15 functional group. The preferred cofactors are NADP+ or NADPH. Bradykinin does not affect the activities of any of the three prostaglandin 9-ketoreductases. Flavin mononucleotide and the flavonoid, quercetin, as well as indomethacin, ethacrynic acid, and furosemide, inhibit all three 9-ketoreductases. An inhibitor of 9-ketoreductase isolated from chicken breast muscle also inhibits the three separable reductases, but the pattern of inhibition of the reductase that focuses at pH 5.7 differs from that of the reductases focusing at pH 7.8 and 8.2. | Multiple molecular forms of prostaglandin 15-hydroxydehydrogenase and 9-ketoreductase in chicken kidney. Prostaglandin-15-hydroxydehydrogenase and prostaglandin-9-keto-reductase were purified from chicken kidney. Both enzymes exist in multiple forms as determined by isoelectric focusing. The dehydrogenases catalyze the transformation of the functional group at C-15 but not the functional group at C-9. The preferred cofactors in these reactions are NAD+ or NADH. The 9-ketoreductases catalyze the reversible transformation of the functional group at C-9 and also the oxidation or reduction of the C-15 functional group. The preferred cofactors are NADP+ or NADPH. Bradykinin does not affect the activities of any of the three prostaglandin 9-ketoreductases. Flavin mononucleotide and the flavonoid, quercetin, as well as indomethacin, ethacrynic acid, and furosemide, inhibit all three 9-ketoreductases. An inhibitor of 9-ketoreductase isolated from chicken breast muscle also inhibits the three separable reductases, but the pattern of inhibition of the reductase that focuses at pH 5.7 differs from that of the reductases focusing at pH 7.8 and 8.2. | [
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|
PMID:15304 | Studies on membrane receptor sites for serotonin in the brain. | The competitive effect of 5,6-dihydroxytryptamine, morphine and chlorpromazine on the binding of serotonin (5-HT) to rat brain slices was investigated. Ths busynaptosomal localization of the binding of morphine in bovine midbrain preparations was compared to that of 5-HT and found to be considerably higher. The condensation of 5-HT and tryptamine receptor carbonyl groups in brain with phenylisopropylhydrazine was shown in vitro and vivo. Membrane particles labeled with [14C] tryptamine or 5-HT in presence or absence of sodium borohydride (NaBH4) were extracted with chloroform-methanol (C-M) 2:1. The labeled proteolipid precipitated by ether from these extracts showed on electropherograms one single radioautographic spot which was more intense with samples treated with sodium borohydride. In column chromatography, the bound radioactivity peak eluted with the gel void volume, was associated with a protein peak. The eluted, lyophilized material of this fraction was extracted by chloroform methanol (2:1) thus suggesting its proteo-lipid nature. | Studies on membrane receptor sites for serotonin in the brain. The competitive effect of 5,6-dihydroxytryptamine, morphine and chlorpromazine on the binding of serotonin (5-HT) to rat brain slices was investigated. Ths busynaptosomal localization of the binding of morphine in bovine midbrain preparations was compared to that of 5-HT and found to be considerably higher. The condensation of 5-HT and tryptamine receptor carbonyl groups in brain with phenylisopropylhydrazine was shown in vitro and vivo. Membrane particles labeled with [14C] tryptamine or 5-HT in presence or absence of sodium borohydride (NaBH4) were extracted with chloroform-methanol (C-M) 2:1. The labeled proteolipid precipitated by ether from these extracts showed on electropherograms one single radioautographic spot which was more intense with samples treated with sodium borohydride. In column chromatography, the bound radioactivity peak eluted with the gel void volume, was associated with a protein peak. The eluted, lyophilized material of this fraction was extracted by chloroform methanol (2:1) thus suggesting its proteo-lipid nature. | [
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|
PMID:15305 | Action of glucagon and aspirin on ionic flux, mucosal blood flow and bleeding in the fundic pouch of dogs. | Into vagally denervated (Heidenhain) pouches of 4 dogs 25 ml of 0.1 M HCl was instilled and removed at 30 min intervals for 6 hours. During the 4th, 5th, and 6th 30 min periods the acid instillate contained 5 mg/ml of aspirin. Aspirin significantly increased gastric-mucosal clearance of aminopyrine (mucosal blood flow), outputs of Na+, Ca++, Mg++, hemoglobin, and plasma transferrin-Cr51 into the pouch contents, and disappearance of H+ from lumen to mucosa. Glucagon, 50 mug/kg subcutaneously was given during irrigation with aspirin and again 1 hour later. Glucagon did not significantly affect loss of acid from lumen to mucosa or the increase in Na+, K+, Ca++, and Mg++ effluxes caused by aspirin. Glucagon significantly decreased mucosal blood flow and the hemorrhage and loss of plasma protein into the instillate induced by aspirin. | Action of glucagon and aspirin on ionic flux, mucosal blood flow and bleeding in the fundic pouch of dogs. Into vagally denervated (Heidenhain) pouches of 4 dogs 25 ml of 0.1 M HCl was instilled and removed at 30 min intervals for 6 hours. During the 4th, 5th, and 6th 30 min periods the acid instillate contained 5 mg/ml of aspirin. Aspirin significantly increased gastric-mucosal clearance of aminopyrine (mucosal blood flow), outputs of Na+, Ca++, Mg++, hemoglobin, and plasma transferrin-Cr51 into the pouch contents, and disappearance of H+ from lumen to mucosa. Glucagon, 50 mug/kg subcutaneously was given during irrigation with aspirin and again 1 hour later. Glucagon did not significantly affect loss of acid from lumen to mucosa or the increase in Na+, K+, Ca++, and Mg++ effluxes caused by aspirin. Glucagon significantly decreased mucosal blood flow and the hemorrhage and loss of plasma protein into the instillate induced by aspirin. | [
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|
PMID:15306 | The Wien effect in compensated metabolic acidosis. | CSF and blood acid-base values were measured in compensated metabolic acidosis, induced in anesthetized dogs by infusion of dilute HCl during hyperventilation. We observed cerebral blood flow-related steady-state differences in PCO2 between CSF and arterial venous blood from the brain. The steady-state values of CSF-blood PCO2 differences and blood H+ activity were intermediate to those observed by us previously during uncompensated acidosis and alkalosis (J. Appl. Physiol. 34: 249-254, 1973). These results are consistent with predictions of the "charged membrane hypothesis" proposed by us to explain CSF and blood acid-base relationship during uncompensated acid-base derangements (J. Appl. Physiol. 34: 243-248, 1973). | The Wien effect in compensated metabolic acidosis. CSF and blood acid-base values were measured in compensated metabolic acidosis, induced in anesthetized dogs by infusion of dilute HCl during hyperventilation. We observed cerebral blood flow-related steady-state differences in PCO2 between CSF and arterial venous blood from the brain. The steady-state values of CSF-blood PCO2 differences and blood H+ activity were intermediate to those observed by us previously during uncompensated acidosis and alkalosis (J. Appl. Physiol. 34: 249-254, 1973). These results are consistent with predictions of the "charged membrane hypothesis" proposed by us to explain CSF and blood acid-base relationship during uncompensated acid-base derangements (J. Appl. Physiol. 34: 243-248, 1973). | [
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|
PMID:15303 | [An unusual case of small intestine infarction of venous origin]. | The evolution is presented, of a partial infarction of the small bowel, of venous origin, without apparent cause. In the 10-th day after onset, following a segmentary resection of the jejunum and termino-terminal anastomosis, the patient had another segmentar infarction of the ileon, also of venous origin, that was cured after resection. After recovery the hematological investigations evidenced a coagulation disturbance that necessitated longterm heparin therapy, a treatment that is continued 3 years after surgery. Although the repeated infarction of the small bowel of venous origin is theoretically possible the case presented appears to be unique by its evolution and the result of the medico-surgical treatment. | [An unusual case of small intestine infarction of venous origin]. The evolution is presented, of a partial infarction of the small bowel, of venous origin, without apparent cause. In the 10-th day after onset, following a segmentary resection of the jejunum and termino-terminal anastomosis, the patient had another segmentar infarction of the ileon, also of venous origin, that was cured after resection. After recovery the hematological investigations evidenced a coagulation disturbance that necessitated longterm heparin therapy, a treatment that is continued 3 years after surgery. Although the repeated infarction of the small bowel of venous origin is theoretically possible the case presented appears to be unique by its evolution and the result of the medico-surgical treatment. | [
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|
PMID:15307 | Postnatal changes in blood respiratory characteristics in an American opossum (Didelphis virginiana). | The oxygen affinity of opossum blood changes in the postnatal period. The youngest opossums studied were 53 days of age (dated from the day of transit from vagina to pouch) and still confined to the maternal pouch: they had a blood P50 of 32.6 +/- 2.9 mm Hg. The blood P50 value then increased significantly, in two steps. First, while the young were still in the pouch, blood P50 rose secondary to a rise in the concentration of 2,3-diphosphoglycerate (DPG). Blood P50 and DPG concentration then remained stable until the animals left the maternal pouch, when DPG rose again, causing a further increment in the blood P50 which peaked at 48.7 +/- 4.5 mm Hg in animals 148 days old. Blood P50 subsequently decreased significantly, reaching the adult value (42.0 +/- 1.9 mm Hg) at about 250 days. | Postnatal changes in blood respiratory characteristics in an American opossum (Didelphis virginiana). The oxygen affinity of opossum blood changes in the postnatal period. The youngest opossums studied were 53 days of age (dated from the day of transit from vagina to pouch) and still confined to the maternal pouch: they had a blood P50 of 32.6 +/- 2.9 mm Hg. The blood P50 value then increased significantly, in two steps. First, while the young were still in the pouch, blood P50 rose secondary to a rise in the concentration of 2,3-diphosphoglycerate (DPG). Blood P50 and DPG concentration then remained stable until the animals left the maternal pouch, when DPG rose again, causing a further increment in the blood P50 which peaked at 48.7 +/- 4.5 mm Hg in animals 148 days old. Blood P50 subsequently decreased significantly, reaching the adult value (42.0 +/- 1.9 mm Hg) at about 250 days. | [
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|
PMID:15309 | Hemoglobin function in stored blood, XVII. Maintenance of red cell 2,3 DPG (function) and ATP (viability) for six weeks in ACD or CPD-adenine-inosine-methylene blue. | Blood preservatives containing adenine for six week storage have been prepared with inosine and methylene blue at various pH levels in order to maintain, 23-DPG levels for immediate oxygen transport upon transfusion. In one experiment, the adverse effect of a high pH on ATP maintenance was demonstrated in the presence of methylene blue and inosine. In this and other experiments it was clear that ATP was better maintained in low pH preservatives and DPG better maintained in higher pH preservatives. However, 2,3-DPG levels were kept from falling with CPD-adenine-inosine over a wide range of pH values. A CPD-adenine-inosine preservative at a pH 5.8 maintained normal DPG levels for three weeks of storage. A similar preservative but with a pH of 6.6 maintained normal DPG levels for 35 days of storage. It is suggested that if all blood bank units are going to have normal DPG levels for optimal oxygen transport at the time of transfusion then a CPD preservative with a higher pH and/or metabolic nutrients and regulators such as inosine or methylene blue would be required. | Hemoglobin function in stored blood, XVII. Maintenance of red cell 2,3 DPG (function) and ATP (viability) for six weeks in ACD or CPD-adenine-inosine-methylene blue. Blood preservatives containing adenine for six week storage have been prepared with inosine and methylene blue at various pH levels in order to maintain, 23-DPG levels for immediate oxygen transport upon transfusion. In one experiment, the adverse effect of a high pH on ATP maintenance was demonstrated in the presence of methylene blue and inosine. In this and other experiments it was clear that ATP was better maintained in low pH preservatives and DPG better maintained in higher pH preservatives. However, 2,3-DPG levels were kept from falling with CPD-adenine-inosine over a wide range of pH values. A CPD-adenine-inosine preservative at a pH 5.8 maintained normal DPG levels for three weeks of storage. A similar preservative but with a pH of 6.6 maintained normal DPG levels for 35 days of storage. It is suggested that if all blood bank units are going to have normal DPG levels for optimal oxygen transport at the time of transfusion then a CPD preservative with a higher pH and/or metabolic nutrients and regulators such as inosine or methylene blue would be required. | [
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|
PMID:15311 | The effect of intragastric pH-variations on the gastric acid response to insulin hypoglycaemia in healthy subjects and duodenal ulcer patients. | The gastric acid response to i.v. injection of 0.15 U of soluble insulin/kg b.w. was determined in healthy subjects and duodenal ulcer patients during intragastric perfusion with water, 0.1 M HC1, and alkaline buffer (pH 8.3). Perfusion with hydrochloric acid significantly reduced the peak gastric acid output following insulin in 6 healthy subjects (reduction 45%, p less than 0.05) but had no significant effect on the peak gastric acid response to insulin in 7 DU patients (reduction 16%, p greater than 0.05). The 2.5-hour gastric acid response to insulin was, however, significantly reduced in both groups (56% and 35%, respectively) by exogenous acidification of the stomach. The gastric acid response to insulin hypoglycaemia in 3 DU patients was the same with intragastric water and alkaline buffer perfusion. The reduction of the gastric acid response to insulin hypoglycaemia by intragastric acidification corresponded to a reduced volume secretion and could not be ascribed to increased back diffusion of hydrogen ions or duodenal inhibition. These findings suggest that the gastric acid response to insulin hypoglycaemia is inhibited by a low intragastric pH in man, and that DU patients are less sensitive to the inhibitory mechanism than healthy subjects. | The effect of intragastric pH-variations on the gastric acid response to insulin hypoglycaemia in healthy subjects and duodenal ulcer patients. The gastric acid response to i.v. injection of 0.15 U of soluble insulin/kg b.w. was determined in healthy subjects and duodenal ulcer patients during intragastric perfusion with water, 0.1 M HC1, and alkaline buffer (pH 8.3). Perfusion with hydrochloric acid significantly reduced the peak gastric acid output following insulin in 6 healthy subjects (reduction 45%, p less than 0.05) but had no significant effect on the peak gastric acid response to insulin in 7 DU patients (reduction 16%, p greater than 0.05). The 2.5-hour gastric acid response to insulin was, however, significantly reduced in both groups (56% and 35%, respectively) by exogenous acidification of the stomach. The gastric acid response to insulin hypoglycaemia in 3 DU patients was the same with intragastric water and alkaline buffer perfusion. The reduction of the gastric acid response to insulin hypoglycaemia by intragastric acidification corresponded to a reduced volume secretion and could not be ascribed to increased back diffusion of hydrogen ions or duodenal inhibition. These findings suggest that the gastric acid response to insulin hypoglycaemia is inhibited by a low intragastric pH in man, and that DU patients are less sensitive to the inhibitory mechanism than healthy subjects. | [
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|
PMID:15312 | Pancreatic extract and the intestinal uptake of vitamin B12. III. Stimulatory effect in the presence of a non-intrinsic factor vitamin B12 binder. | To determine the mechanism by which pancreatic extract (PE) corrects the malabsorption of vitamin B12 in chronic pancreatic insufficiency (CPI), the following hypotheses were investigated: Firstly, PE might stimulate the absorption of vitamin B12 by changing the intestinal pH, secondly PE might stimulate the intestinal uptake of unbound vitamin B12, thirdly PE might abolish the inhibitory effect of vitamin B12 binders on the intestinal uptake of vitamin B12 bound to intrinsic factor (IF). PE had no effect on the pH in the small intestine and did not stimulate the uptake of unbound 57CoB12 by perfused rat intestinal segments. Preincubation of 57CoB12-IF with a non-IF B12-binder from human saliva (R-binder) reduced the uptake of 57CoB12 from 18.5 pg per cm intestine +/- 3.4 S.E.M. to 7.8 +/- 1.6 (p less than 0.02). PE abolished this inhibitory effect (p less than 0.05). The results indicate that PE corrects the malabsorption of vitamin B12 in CPI by an effect on non-IF B12- binders. | Pancreatic extract and the intestinal uptake of vitamin B12. III. Stimulatory effect in the presence of a non-intrinsic factor vitamin B12 binder. To determine the mechanism by which pancreatic extract (PE) corrects the malabsorption of vitamin B12 in chronic pancreatic insufficiency (CPI), the following hypotheses were investigated: Firstly, PE might stimulate the absorption of vitamin B12 by changing the intestinal pH, secondly PE might stimulate the intestinal uptake of unbound vitamin B12, thirdly PE might abolish the inhibitory effect of vitamin B12 binders on the intestinal uptake of vitamin B12 bound to intrinsic factor (IF). PE had no effect on the pH in the small intestine and did not stimulate the uptake of unbound 57CoB12 by perfused rat intestinal segments. Preincubation of 57CoB12-IF with a non-IF B12-binder from human saliva (R-binder) reduced the uptake of 57CoB12 from 18.5 pg per cm intestine +/- 3.4 S.E.M. to 7.8 +/- 1.6 (p less than 0.02). PE abolished this inhibitory effect (p less than 0.05). The results indicate that PE corrects the malabsorption of vitamin B12 in CPI by an effect on non-IF B12- binders. | [
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|
PMID:15313 | Effect of changes in the intragastric milieu on competence of the gastro-oesophageal region. A study in normal subjects. | Acid-reflux studies were carried out in 10 healthy subjects in the basal state, during continuous infusion of pentagastrin (0.015 mug/kg body-weight) after bolus injection of insulin (0.2 IU/kg bodyweight), and after intragastric instillation of 200 ml hydrochloric acid. Basal gastro-oesophageal sphincter pressure and rise in intragastric pressure on leg raising were measured by means of perfused catheters. The increase of intragastric acidity during infusion of pentagastrin and during insulin-induced hypoglycaemia was not accompanied by changes in the competence of the gastro-oesophageal region. Instillation of hydrochloric acid was followed by a significant enhancement of the reflux tendency. Changes in intragastric pressure-rise were not demonstrated in any of the series of investigation. Gastric acid secretion and its significance at the evaluation of the results of reflux studies by means of pH-measuring equipment has not been clarified in patients. It can therefore reasonably be demanded of future acid reflux studies that details have to be stated with regard to acid secretion, and that these should be taken into account at the assessment of the results of the study. | Effect of changes in the intragastric milieu on competence of the gastro-oesophageal region. A study in normal subjects. Acid-reflux studies were carried out in 10 healthy subjects in the basal state, during continuous infusion of pentagastrin (0.015 mug/kg body-weight) after bolus injection of insulin (0.2 IU/kg bodyweight), and after intragastric instillation of 200 ml hydrochloric acid. Basal gastro-oesophageal sphincter pressure and rise in intragastric pressure on leg raising were measured by means of perfused catheters. The increase of intragastric acidity during infusion of pentagastrin and during insulin-induced hypoglycaemia was not accompanied by changes in the competence of the gastro-oesophageal region. Instillation of hydrochloric acid was followed by a significant enhancement of the reflux tendency. Changes in intragastric pressure-rise were not demonstrated in any of the series of investigation. Gastric acid secretion and its significance at the evaluation of the results of reflux studies by means of pH-measuring equipment has not been clarified in patients. It can therefore reasonably be demanded of future acid reflux studies that details have to be stated with regard to acid secretion, and that these should be taken into account at the assessment of the results of the study. | [
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|
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