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PMID:10388 | Bioanalysis of picomole amounts of acetylcholine by ion-pair partition chromatography applied to rat sciatic nerve. | A method for determination of acetylcholine in small, discrete biological objects by use of ion-pair technique has been developed. Acetylcholine is extracted as an ion pair with 3,5-di-t-butyl-2-hydroxybenzenesulphonate and separated from co-extracted components by ion-pair partition chromatography with picrate as the counter ion and porous cellulose as support. The quantitative evaluation is made from the acetylcholine peak in the chromatogram obtained by ultraviolet detection. Acetylcholine has been analysed in 1 cm large pieces of rat sciatic nerve containing about 60 pmol (10 ng). The overall recovery of the method is 100 +/- 10% at the 120 pmol level of acetylcholine in a sample. | Bioanalysis of picomole amounts of acetylcholine by ion-pair partition chromatography applied to rat sciatic nerve. A method for determination of acetylcholine in small, discrete biological objects by use of ion-pair technique has been developed. Acetylcholine is extracted as an ion pair with 3,5-di-t-butyl-2-hydroxybenzenesulphonate and separated from co-extracted components by ion-pair partition chromatography with picrate as the counter ion and porous cellulose as support. The quantitative evaluation is made from the acetylcholine peak in the chromatogram obtained by ultraviolet detection. Acetylcholine has been analysed in 1 cm large pieces of rat sciatic nerve containing about 60 pmol (10 ng). The overall recovery of the method is 100 +/- 10% at the 120 pmol level of acetylcholine in a sample. | [
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PMID:10389 | The effects of administering N-(2-benzoyloxyethyl) norfenfluramine to rats on the hepatic synthesis of glycerolipids. | N-(2-Benzoyloxyethyl) norfenfluramine (S-780) was administered to rats by stomach tube at a dose of 50 mg kg-1 of body weight. Livers of the rats which were given an acute dose of the drug synthesized more triacylglycerol, phosphatidylcholine and phosphatidylethanolamine from [1,3-3H]glycerol and [14C]palmitate than did those of control rats. The measurements were made by injecting a mixture of the radioactive precursors into the portal veins of anaesthetized rats and freeze clamping a portion of the liver 1 min later. Diffferent results were obtained after treating rats daily with S-780 for 5 days. Liver slices from these rats synthesized less triacylglycerol and relatively more phosphatidylinositol plus phosphatidylserine from [3H]glycerol than did those of control rats. S-780 treatment depressed the hepatic synthesis of phosphatidylcholine and phosphatidylethanolamine as measured in vivo after intrapotal injection of [14C]palmitate and [3H]glycerol. Chronic treatment with S-780 also depressed food intake and lowered liver weight and body weight of rats fed the 41B diet. The results are discussed in relation to the effects of S-780 on the synthesis of glycerolipids. | The effects of administering N-(2-benzoyloxyethyl) norfenfluramine to rats on the hepatic synthesis of glycerolipids. N-(2-Benzoyloxyethyl) norfenfluramine (S-780) was administered to rats by stomach tube at a dose of 50 mg kg-1 of body weight. Livers of the rats which were given an acute dose of the drug synthesized more triacylglycerol, phosphatidylcholine and phosphatidylethanolamine from [1,3-3H]glycerol and [14C]palmitate than did those of control rats. The measurements were made by injecting a mixture of the radioactive precursors into the portal veins of anaesthetized rats and freeze clamping a portion of the liver 1 min later. Diffferent results were obtained after treating rats daily with S-780 for 5 days. Liver slices from these rats synthesized less triacylglycerol and relatively more phosphatidylinositol plus phosphatidylserine from [3H]glycerol than did those of control rats. S-780 treatment depressed the hepatic synthesis of phosphatidylcholine and phosphatidylethanolamine as measured in vivo after intrapotal injection of [14C]palmitate and [3H]glycerol. Chronic treatment with S-780 also depressed food intake and lowered liver weight and body weight of rats fed the 41B diet. The results are discussed in relation to the effects of S-780 on the synthesis of glycerolipids. | [
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PMID:10390 | Influence of various substances on prostaglandin biosynthesis by guinea-pig chopped lung. | Guinea-pig chopped lung tissue was used to investigate the inhibitory effect of various steroidal and non-steroidal anti-inflammatory agents, antipyretics, analgesics, local anaesthetics and psychotropic drugs on mechanically induced release of prostaglandin-like material. Low concentrations of non-steroidal anti-inflammatory agents inhibited synthesis, but other antipyretics and analgesics, and the local anaesthetics had little effect. Thymoleptics, neuroleptics and monoamine oxidase inhibitors except phenelzine exhibited weak activity. It is concluded that the method is a useful pharmacological model to study prostaglandin biosynthesis. The weak effects of the psychotropic drugs suggest that they do not exert their clinical effect by inhibiting PG biosynthesis. | Influence of various substances on prostaglandin biosynthesis by guinea-pig chopped lung. Guinea-pig chopped lung tissue was used to investigate the inhibitory effect of various steroidal and non-steroidal anti-inflammatory agents, antipyretics, analgesics, local anaesthetics and psychotropic drugs on mechanically induced release of prostaglandin-like material. Low concentrations of non-steroidal anti-inflammatory agents inhibited synthesis, but other antipyretics and analgesics, and the local anaesthetics had little effect. Thymoleptics, neuroleptics and monoamine oxidase inhibitors except phenelzine exhibited weak activity. It is concluded that the method is a useful pharmacological model to study prostaglandin biosynthesis. The weak effects of the psychotropic drugs suggest that they do not exert their clinical effect by inhibiting PG biosynthesis. | [
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PMID:10391 | Metabolic N-oxidation of atropine, hyoscine and the corresponding nor-alkaloids by guinea-pig liver microsomal preparations. | Incubation of guinea-pig liver microsomal preparations with atropine or hyoscine resulted in the formation of the corresponding nor-alkaloids and both isomers of atropine N-oxide from atropine and one isomer of hyoscine N-oxide from hyoscine. Separate incubations of guinea-pig liver microsomal preparations with nor-atropine and nor-hyoscine yielded the corresponding hydroxylamines. The N-oxide and hydroxylamine metabolites were identified by comparison of their t.l.c. behavior and m.x. with prepared compounds and also by their reduction to the corresponding tertiary or secondary amines. | Metabolic N-oxidation of atropine, hyoscine and the corresponding nor-alkaloids by guinea-pig liver microsomal preparations. Incubation of guinea-pig liver microsomal preparations with atropine or hyoscine resulted in the formation of the corresponding nor-alkaloids and both isomers of atropine N-oxide from atropine and one isomer of hyoscine N-oxide from hyoscine. Separate incubations of guinea-pig liver microsomal preparations with nor-atropine and nor-hyoscine yielded the corresponding hydroxylamines. The N-oxide and hydroxylamine metabolites were identified by comparison of their t.l.c. behavior and m.x. with prepared compounds and also by their reduction to the corresponding tertiary or secondary amines. | [
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PMID:10392 | The microsomal N-oxidation of phentermine. | The microsomal N-oxidation of phentermine (Ia) to N-hydroxyphentermine (Ib) and to alpha,alpha-dimethyl-alpha-nitroso-beta-phenylethane (Ic was investigated. Maximum activities were obtained with microsomal (9000 g supernatant and microsomes) fractions of rabbit liver in the presence of an NADPH generating system. Incubation of Ia with hepatic washed microsomes from a phenobarbitone pretreated rabbit increased the formation of Ib and decreased that of Ic but the total amount of N-oxidized metabolites (i.e. Ib + Ic) was not affected. The ratio of the metabolically produced Ic to Ib but not the total amount of N-oxygenated metabolites varied greatly depending of the liver microsomal fractions used in the incubation mixtures of Ia; more Ib was produced from Ia using 9000 g supernatant and conversely, more Ic was formed using the washed microsomes of the same liver. The nitroso compound (Ic) was metabolically reduced to Ib and Ib to Ia by the hepatic 9000 g supernatant and soluble fraction; under the same conditions, washed microsomes had only limited reductive properties towards Ic and Ib. N-Hydroxyphentermine (Ib) was not metabolically oxidized to Ic when incubated with washed microsomes from rabbit liver. The use of known carbon-oxidation inhibitors showed that cytochrome P-450 is not involved in the incorporation of oxygen at the nitrogen centre of Ia. The metabolic characteristics and kinetic behavior of the microsomal N-oxidation of Ia supported a recently proposed mechanism explaining the independent formation of Ib and Ic from a common precursor resulting from metabolic N-oxidation of Ia. | The microsomal N-oxidation of phentermine. The microsomal N-oxidation of phentermine (Ia) to N-hydroxyphentermine (Ib) and to alpha,alpha-dimethyl-alpha-nitroso-beta-phenylethane (Ic was investigated. Maximum activities were obtained with microsomal (9000 g supernatant and microsomes) fractions of rabbit liver in the presence of an NADPH generating system. Incubation of Ia with hepatic washed microsomes from a phenobarbitone pretreated rabbit increased the formation of Ib and decreased that of Ic but the total amount of N-oxidized metabolites (i.e. Ib + Ic) was not affected. The ratio of the metabolically produced Ic to Ib but not the total amount of N-oxygenated metabolites varied greatly depending of the liver microsomal fractions used in the incubation mixtures of Ia; more Ib was produced from Ia using 9000 g supernatant and conversely, more Ic was formed using the washed microsomes of the same liver. The nitroso compound (Ic) was metabolically reduced to Ib and Ib to Ia by the hepatic 9000 g supernatant and soluble fraction; under the same conditions, washed microsomes had only limited reductive properties towards Ic and Ib. N-Hydroxyphentermine (Ib) was not metabolically oxidized to Ic when incubated with washed microsomes from rabbit liver. The use of known carbon-oxidation inhibitors showed that cytochrome P-450 is not involved in the incorporation of oxygen at the nitrogen centre of Ia. The metabolic characteristics and kinetic behavior of the microsomal N-oxidation of Ia supported a recently proposed mechanism explaining the independent formation of Ib and Ic from a common precursor resulting from metabolic N-oxidation of Ia. | [
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PMID:10393 | A new class of irreversible muscarinic antagonists: beta-haloethylamine furoates. | The synthesis of a class of ultra-long acting muscarinic antagonists is described. Furthermore, it is noted that the stability of the, in situ, aziridinium ion is sufficiently great so that these agents can be used to study the effect of temperature upon the conformation of the muscarinic cholinoceptor. The inactivation kinetics of these receptors as well as the dissociation constants, Kd, for all probes are presented. | A new class of irreversible muscarinic antagonists: beta-haloethylamine furoates. The synthesis of a class of ultra-long acting muscarinic antagonists is described. Furthermore, it is noted that the stability of the, in situ, aziridinium ion is sufficiently great so that these agents can be used to study the effect of temperature upon the conformation of the muscarinic cholinoceptor. The inactivation kinetics of these receptors as well as the dissociation constants, Kd, for all probes are presented. | [
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PMID:10394 | The influence of polyvinylpyrrolidone on the solution and bioavailability of hydrochlorothiazide. | The dissolution properties of hydrochlorothiazide-PVP 10 000 mechanical mix and coprecipitate systems were qualitatively similar to those previously reported using hydroflumethiazide. Quantitative differences were dependent on the proportion of PVP present, its molecular weight and method of incorporation. Cumulative urinary excretion data from test capsule preparations showed that bioavailability was enhanced by the presence of PVP. However, the degree of enhancement was less than that expected from constant surface area disc rate studies. Dissolution tests on the capsule formulations, using the U.S.P. basket stirrer assembly, did not correlate with in vivo results. Using the Levy beaker method and a stirring speed of 40 rev min-1, good correlation between amount dissolved in 30 min and amount excreted in urine after 24 h was obtained. The dissolution tests revealed that PVP retards the initial dissolution from capsule dosage forms, probably by retarding deaggregation and dispersion of drug particles. | The influence of polyvinylpyrrolidone on the solution and bioavailability of hydrochlorothiazide. The dissolution properties of hydrochlorothiazide-PVP 10 000 mechanical mix and coprecipitate systems were qualitatively similar to those previously reported using hydroflumethiazide. Quantitative differences were dependent on the proportion of PVP present, its molecular weight and method of incorporation. Cumulative urinary excretion data from test capsule preparations showed that bioavailability was enhanced by the presence of PVP. However, the degree of enhancement was less than that expected from constant surface area disc rate studies. Dissolution tests on the capsule formulations, using the U.S.P. basket stirrer assembly, did not correlate with in vivo results. Using the Levy beaker method and a stirring speed of 40 rev min-1, good correlation between amount dissolved in 30 min and amount excreted in urine after 24 h was obtained. The dissolution tests revealed that PVP retards the initial dissolution from capsule dosage forms, probably by retarding deaggregation and dispersion of drug particles. | [
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PMID:10395 | Data point weighting in pharmacokinetic analysis: intravenous paracetamol in man. | Compartmental analysis of plasma paracetamol concentrations following intravenous injection of 12 mg kg-1 in aqueous solution to normal subjects was performed using analogue and digital computer methods. Using a "simplex" non-linear optimization procedure, the pharmacokinetic parameters were found to be influenced considerably by the choice of the weighting factors (W1) attributed to individual data points. The plasma half-life of paracetamol varied by up to a seven-fold with the weighting factors selected. However, the predicted mean steady-state plasma concentrations were shown to be relatively little affected by the different weighting factors. | Data point weighting in pharmacokinetic analysis: intravenous paracetamol in man. Compartmental analysis of plasma paracetamol concentrations following intravenous injection of 12 mg kg-1 in aqueous solution to normal subjects was performed using analogue and digital computer methods. Using a "simplex" non-linear optimization procedure, the pharmacokinetic parameters were found to be influenced considerably by the choice of the weighting factors (W1) attributed to individual data points. The plasma half-life of paracetamol varied by up to a seven-fold with the weighting factors selected. However, the predicted mean steady-state plasma concentrations were shown to be relatively little affected by the different weighting factors. | [
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PMID:10408 | Electrochemistry of drug action I: electrooreduction of ferredoxins. | Ferredoxin serves as an electron carrier in the oxidation-reduction system in anaerobic microorganisms, transferring electrons from a low potential donor to electron-accepting biochemicals. The anaerobicidal activity of some drugs may be due to their interference with the electron transport function of ferredoxin. Two types of ferredoxins (isolated from Clostridium pasteurianum and spinach) were studied, and their electrochemical reduction and biochemical properties were analyzed using a sensitive ac polarographic technique. The reduction potential of both ferredoxins was linearly related to pH. The mechanisms of electron transport in ferredoxin molecules were found to be related to their sulfur-iron bonds. The dissociation of the sulfur-iron bonds resulted in the formation of a free sulfhydryl group and the interruption of the electroactivity of ferredoxin. This sulfur-iron dissociation process was found to be pH dependent. The electroreduction of ferredoxins was an energy-requiring, pH-dependent process. | Electrochemistry of drug action I: electrooreduction of ferredoxins. Ferredoxin serves as an electron carrier in the oxidation-reduction system in anaerobic microorganisms, transferring electrons from a low potential donor to electron-accepting biochemicals. The anaerobicidal activity of some drugs may be due to their interference with the electron transport function of ferredoxin. Two types of ferredoxins (isolated from Clostridium pasteurianum and spinach) were studied, and their electrochemical reduction and biochemical properties were analyzed using a sensitive ac polarographic technique. The reduction potential of both ferredoxins was linearly related to pH. The mechanisms of electron transport in ferredoxin molecules were found to be related to their sulfur-iron bonds. The dissociation of the sulfur-iron bonds resulted in the formation of a free sulfhydryl group and the interruption of the electroactivity of ferredoxin. This sulfur-iron dissociation process was found to be pH dependent. The electroreduction of ferredoxins was an energy-requiring, pH-dependent process. | [
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PMID:10409 | Liquid chromatography in pharmaceutical analysis V: determination of an isoniazid-pyridoxine hydrochloride mixture. | Operating parameters are described for the qualitative and quantitative analysis of an isoniazid-pyridoxine hydrochloride mixture by high-pressure liquid chromatography. Each compound was chromatographed on an octadecyl column, using absolute methanol-water (60:40) (pH 2.5) containing 0.01 M dioctyl sodium sulfosuccinate. The flow rate was 2.0 ml/min (2500 psig), and the peaks were detected at 293 nm. The analysis was accomplished using ion-pair formation for effecting chromatographic separation. The time required for separation of the drug mixture is approximately 12 min with an accuracy of 1.17-0.30%. | Liquid chromatography in pharmaceutical analysis V: determination of an isoniazid-pyridoxine hydrochloride mixture. Operating parameters are described for the qualitative and quantitative analysis of an isoniazid-pyridoxine hydrochloride mixture by high-pressure liquid chromatography. Each compound was chromatographed on an octadecyl column, using absolute methanol-water (60:40) (pH 2.5) containing 0.01 M dioctyl sodium sulfosuccinate. The flow rate was 2.0 ml/min (2500 psig), and the peaks were detected at 293 nm. The analysis was accomplished using ion-pair formation for effecting chromatographic separation. The time required for separation of the drug mixture is approximately 12 min with an accuracy of 1.17-0.30%. | [
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PMID:10410 | Simple and rapid determination of a new pharmaceutically active amine hydrochloride. | The quantitative analysis of a new pharmaceutically active amine hydrochloride is described. Samples are extracted with chloroform. A yellow amine-dye complex is formed by buffering a sample-bromthymol blue solution at pH 8.5 +/- 0.1 and subsequently extracted with chloroform. The complex is treated with 0.01 N NaOH to convert it back to the sodium salt of bromthymol blue, which is then measured at 615 nm in the aqueous layer. The amount of complex extracted is linearly related to the amount of amine present, from 0.020 to 0.20 mg/ml. Under the selected conditions, the compound can be determined in the presence of degradation products and there is no interference from common pharmaceutical excipients. The method is suitable for stability studies. | Simple and rapid determination of a new pharmaceutically active amine hydrochloride. The quantitative analysis of a new pharmaceutically active amine hydrochloride is described. Samples are extracted with chloroform. A yellow amine-dye complex is formed by buffering a sample-bromthymol blue solution at pH 8.5 +/- 0.1 and subsequently extracted with chloroform. The complex is treated with 0.01 N NaOH to convert it back to the sodium salt of bromthymol blue, which is then measured at 615 nm in the aqueous layer. The amount of complex extracted is linearly related to the amount of amine present, from 0.020 to 0.20 mg/ml. Under the selected conditions, the compound can be determined in the presence of degradation products and there is no interference from common pharmaceutical excipients. The method is suitable for stability studies. | [
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PMID:10411 | Phenylbutazone ionization kinetics. | Phenylbutazone has been associated with bioavailability problems and has shown nonclassical behavior in phase-transport studies. This nonclassical behavior has been attributed, in part, to the fact that phenylbutazone, as a carbon acid, undergoes noninstantaneous ionization kinetics. Instantaneous reaction is an assumption made in many diffusion-limited transport models involving a simultaneous ionization reaction. The ionization kinetics of phenylbutazone were determined at an ionic strength of 0.1 and 25 degrees using a stopped-flow spectrophotometer. A log kobs versus pH profile for the approach to the ionization equilibrium was determined, and a mechanism consistent with the profile was postulated. The percent enol versus the diketo form of phenylbutazone acid as well as pKaenol and pKadiketo was kinetically calculated. The protonation reaction was highly catalyzed by general acids while the deprotonation reaction was highly catalyzed by general bases. The general acid, water, was a poor proton donor to the anionic form (the so-called mesomericanion) of phenylbutazone. | Phenylbutazone ionization kinetics. Phenylbutazone has been associated with bioavailability problems and has shown nonclassical behavior in phase-transport studies. This nonclassical behavior has been attributed, in part, to the fact that phenylbutazone, as a carbon acid, undergoes noninstantaneous ionization kinetics. Instantaneous reaction is an assumption made in many diffusion-limited transport models involving a simultaneous ionization reaction. The ionization kinetics of phenylbutazone were determined at an ionic strength of 0.1 and 25 degrees using a stopped-flow spectrophotometer. A log kobs versus pH profile for the approach to the ionization equilibrium was determined, and a mechanism consistent with the profile was postulated. The percent enol versus the diketo form of phenylbutazone acid as well as pKaenol and pKadiketo was kinetically calculated. The protonation reaction was highly catalyzed by general acids while the deprotonation reaction was highly catalyzed by general bases. The general acid, water, was a poor proton donor to the anionic form (the so-called mesomericanion) of phenylbutazone. | [
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PMID:10412 | Conversion of cefamandole nafate to cefamandole sodium. | The rate of hydrolysis of the formyl moiety of cefamandole nafate was determined as a function of pH, temperature, and concentration of added sodium carbonate or tromethamine. The reaction rate was sensitive to hydroxide ion in the pH 5.5-8.0 range with half-life values of hours to minutes. Hydrolysis was rapid upon the addition of sodium carbonate or tromethamine. Chirality in the 7-D-mandelamido side chain was unaffected by hydrolysis. | Conversion of cefamandole nafate to cefamandole sodium. The rate of hydrolysis of the formyl moiety of cefamandole nafate was determined as a function of pH, temperature, and concentration of added sodium carbonate or tromethamine. The reaction rate was sensitive to hydroxide ion in the pH 5.5-8.0 range with half-life values of hours to minutes. Hydrolysis was rapid upon the addition of sodium carbonate or tromethamine. Chirality in the 7-D-mandelamido side chain was unaffected by hydrolysis. | [
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PMID:10413 | Structure of aluminum hydroxide gel I: initial precipitate. | The initial aluminum hydroxide gel precipitate resulting from the reaction of aluminum chloride or aluminum sulfate with ammonium hydroxide is shown by potentiometric titration, chemical analysis, and the ratio of bound hydroxide to aluminum to fit a polymer model described previously. The formation of polynuclear hydroxyaluminum particles is treated as a stepwise process involving a deprotonation-dehydration mechanism, which results in the formation of six-membered rings; these rings may further coalesce by the same mechanism. The aluminum hydroxide gel precipitated from aluminum chloride can be represented by the formula Al(OH)2.55(Cl)0.45 and probably exists as a polymer of 10 fused six-membered rings. The aluminum hydroxide gel precipitated from aluminum sulfate can be represented by the formula Al(OH)2.30(SO4)0.35. This species probably exists as a polymer of three fused six-membered rings. | Structure of aluminum hydroxide gel I: initial precipitate. The initial aluminum hydroxide gel precipitate resulting from the reaction of aluminum chloride or aluminum sulfate with ammonium hydroxide is shown by potentiometric titration, chemical analysis, and the ratio of bound hydroxide to aluminum to fit a polymer model described previously. The formation of polynuclear hydroxyaluminum particles is treated as a stepwise process involving a deprotonation-dehydration mechanism, which results in the formation of six-membered rings; these rings may further coalesce by the same mechanism. The aluminum hydroxide gel precipitated from aluminum chloride can be represented by the formula Al(OH)2.55(Cl)0.45 and probably exists as a polymer of 10 fused six-membered rings. The aluminum hydroxide gel precipitated from aluminum sulfate can be represented by the formula Al(OH)2.30(SO4)0.35. This species probably exists as a polymer of three fused six-membered rings. | [
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PMID:10414 | Structure of aluminum hydroxide gel II: aging mechanism. | The aging of aluminum hydroxide gel prepared by the reaction of aluminum chloride and ammonium hydroxide was studied by measurement of pH, acid-consuming capacity, hydroxide to aluminum ratio, chloride activity, and X-ray line broadening. The results were consistent with a polymer model involving particle growth by a deprotonation-dehydration mechanism. Anions inhibit this reaction by binding to the positively charged edges of the hydroxyaluminum polymers. | Structure of aluminum hydroxide gel II: aging mechanism. The aging of aluminum hydroxide gel prepared by the reaction of aluminum chloride and ammonium hydroxide was studied by measurement of pH, acid-consuming capacity, hydroxide to aluminum ratio, chloride activity, and X-ray line broadening. The results were consistent with a polymer model involving particle growth by a deprotonation-dehydration mechanism. Anions inhibit this reaction by binding to the positively charged edges of the hydroxyaluminum polymers. | [
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] |
PMID:10415 | Structure of aluminum hydroxide gel III: mechanism of stabilization by sorbitol. | The effect of sorbitol on the aging of aluminum hydroxide gel, prepared by the reaction of aluminum chloride solution with strong ammonia solution to a final pH of 7.0, was studied by potentiometric titration, acid-consuming capacity, pH, hydroxide to aluminum ratio, chloride activity, X-ray diffraction, and IR spectroscopy. Gels containing sorbitol lost less than 10% of their acid-consuming capacity during a 6-month aging period compared with a loss of more than 60% for an identical gel without sorbitol. The mechanism by which sorbitol stabilizes the gel appears to be inhibition of the secondary polymerization reaction which takes place upon aging. Another polyhydroxy compound, quercetin, also stabilizes aluminum hydroxide gel. | Structure of aluminum hydroxide gel III: mechanism of stabilization by sorbitol. The effect of sorbitol on the aging of aluminum hydroxide gel, prepared by the reaction of aluminum chloride solution with strong ammonia solution to a final pH of 7.0, was studied by potentiometric titration, acid-consuming capacity, pH, hydroxide to aluminum ratio, chloride activity, X-ray diffraction, and IR spectroscopy. Gels containing sorbitol lost less than 10% of their acid-consuming capacity during a 6-month aging period compared with a loss of more than 60% for an identical gel without sorbitol. The mechanism by which sorbitol stabilizes the gel appears to be inhibition of the secondary polymerization reaction which takes place upon aging. Another polyhydroxy compound, quercetin, also stabilizes aluminum hydroxide gel. | [
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PMID:10416 | Kinetics and mechanisms of hydrolysis of 1,4-benzodiazepines I: chlordiazepoxide and demoxepam. | Differential absorbance spectroscopy was successfully used to follow the hydrolysis kinetics of chlordiazepoxide and demoxepam from pH 1 to 11. Loss of the methylamino group from chlordiazepoxide produced demoxepam. Demoxepam degraded by a parallel consecutive reaction to 2-amino-5-chlorobenzophenone and a glycine derivative. Two intermediates were observed by TLC for demoxepam hydrolysis. One was assigned the open-ring structure resulting from amide hydrolysis, which kinetically appears to be the major mechanistic route leading to the benzophenone product. The other intermediate, representing an alternative but minor pathway, presumably results from initial scission of the azomethine linkage. Protonation of the N-oxide slightly alters the importance of these two pathways. Recyclization of the carboxylic acid intermediate was facile at pH values below the pKa of this intermediate. The stability parameters involving buffer catalysis, ionic strength effects, and temperature dependence of rate constants are reported. | Kinetics and mechanisms of hydrolysis of 1,4-benzodiazepines I: chlordiazepoxide and demoxepam. Differential absorbance spectroscopy was successfully used to follow the hydrolysis kinetics of chlordiazepoxide and demoxepam from pH 1 to 11. Loss of the methylamino group from chlordiazepoxide produced demoxepam. Demoxepam degraded by a parallel consecutive reaction to 2-amino-5-chlorobenzophenone and a glycine derivative. Two intermediates were observed by TLC for demoxepam hydrolysis. One was assigned the open-ring structure resulting from amide hydrolysis, which kinetically appears to be the major mechanistic route leading to the benzophenone product. The other intermediate, representing an alternative but minor pathway, presumably results from initial scission of the azomethine linkage. Protonation of the N-oxide slightly alters the importance of these two pathways. Recyclization of the carboxylic acid intermediate was facile at pH values below the pKa of this intermediate. The stability parameters involving buffer catalysis, ionic strength effects, and temperature dependence of rate constants are reported. | [
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PMID:10417 | Spectrofluorometric determination of hydroflumethiazide in plasma and urine. | A rapid, accurate, sensitive, and reproducible assay for hydroflumethiazide in plasma and urine was developed after studies of its UV and fluorescence spectral properties and partitioning behavior. The assay is based on initial extraction from acidified plasma or urine into ether, back-extraction into basic solution followed by acidification to about pH 1, and measurement of the fluorescence derived from the unionized molecule. Analysis of variance indicated no significant differences in assays performed on the same day. The mean recovery was 98.8 +/- 7.4% for plasma over a concentration range of 0.2-2.0 mug/ml. The method is convenient for routine clinical use and has sufficient sensitivity to quantify hydroflumethiazide levels after administration of therapeutic doses. | Spectrofluorometric determination of hydroflumethiazide in plasma and urine. A rapid, accurate, sensitive, and reproducible assay for hydroflumethiazide in plasma and urine was developed after studies of its UV and fluorescence spectral properties and partitioning behavior. The assay is based on initial extraction from acidified plasma or urine into ether, back-extraction into basic solution followed by acidification to about pH 1, and measurement of the fluorescence derived from the unionized molecule. Analysis of variance indicated no significant differences in assays performed on the same day. The mean recovery was 98.8 +/- 7.4% for plasma over a concentration range of 0.2-2.0 mug/ml. The method is convenient for routine clinical use and has sufficient sensitivity to quantify hydroflumethiazide levels after administration of therapeutic doses. | [
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PMID:10418 | Spectrofluorometric determination of bufuralol in blood and urine. | The benzofuran analog bufuralol, a beta-adrenergic blocker, was determined in blood and urine by a specific and sensitive spectrofluorometric assay. The compound was extracted into ether from blood or urine adjusted to pH 10. The ether extract was separated by TLC to resolve the parent drug from any basic metabolites present, and the spots were eluted off the silica gel and quantitated fluorometrically in 0.1 N HCl. The overall recovery of the assay was 85 +/- 3.0%; the sensitivity limit was 2-4 ng/ml of blood or urine, using a 2.5-ml specimen/analysis. The method was applied to the determination of blood levels in a dog following a single 10-mg/kg oral dose and in two human subjects administered a single 20-mg oral dose. | Spectrofluorometric determination of bufuralol in blood and urine. The benzofuran analog bufuralol, a beta-adrenergic blocker, was determined in blood and urine by a specific and sensitive spectrofluorometric assay. The compound was extracted into ether from blood or urine adjusted to pH 10. The ether extract was separated by TLC to resolve the parent drug from any basic metabolites present, and the spots were eluted off the silica gel and quantitated fluorometrically in 0.1 N HCl. The overall recovery of the assay was 85 +/- 3.0%; the sensitivity limit was 2-4 ng/ml of blood or urine, using a 2.5-ml specimen/analysis. The method was applied to the determination of blood levels in a dog following a single 10-mg/kg oral dose and in two human subjects administered a single 20-mg oral dose. | [
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PMID:10419 | Kinetics of acid neutralization by aluminum hydroxide gel. | The rate of acid neutralization by an aluminum hydroxide gel prepared by the reaction of aluminum chloride solution and strong ammonia solution was studied. The decrease in acid-consuming capacity during aging as measured by the USP test is due to a decrease in the rate of reaction rather than to a decrease in equilibrium reactivity. The reactivity profile has three phases, which are shown to be related to the structure of the gel. The rate of loss of reactivity is directly related to the extent of washing. | Kinetics of acid neutralization by aluminum hydroxide gel. The rate of acid neutralization by an aluminum hydroxide gel prepared by the reaction of aluminum chloride solution and strong ammonia solution was studied. The decrease in acid-consuming capacity during aging as measured by the USP test is due to a decrease in the rate of reaction rather than to a decrease in equilibrium reactivity. The reactivity profile has three phases, which are shown to be related to the structure of the gel. The rate of loss of reactivity is directly related to the extent of washing. | [
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PMID:10420 | Effect of histamine and antihistamines on renal hemodynamics and functions in the isolated perfused canine kidney. | Histamine was infused intra-arterially (10-40 mug/min) into isolated blood perfused canine kidneys while functional and hemodynamic parameters were monitored. When perfusion pressure (PP) was kept constant during the infusion of histamine, renal blood flow (RBF) increased from 137 +/- 9 to 181 +/- 9 ml/min (P less than .001). A greater increase in RBF occurred to the inner renal cortex than the outer renal cortex as measured by radioactive microspheres. The fractional outer/inner cortical blood flow changed from 79:21 before histamine to 74:26 during its infusion (P less than .001). Histamine did not alter creatinine clearance (Ccr), urine volume (V), sodium excretion (UNaV) or the fractional excretion of sodium (FENa) or water (FEH20) under these conditions. When renal blood flow was held constant during the infusion of histamine, PP decreased from 126/106 +/- 2 to 100/81 +/- 2 mm Hg (P less than .001). This resulted in a reduction of absolute outer cortical (outer/inner) changed from 77:23 before histamine to 72:28 during its infusion (P less than .001). In contrast to the effects of histamine at constant PP, CCr, V, UNaV, FENa, and FEH20 were decreased when histamine infusion reduced the PP. Administration of the H1 receptor antagonist, diphenhydramine, blocked the hemodynamic effects of histamine whereas the administration of an H2 receptor antagonist, metiamide, did not alter the histamine response. Similar vasodilatory responses to histamine were observed in isolated blood perfused dog and cat kidneys. In contrast, vasoconstrictor responses to histamine occurred in isolated dog and cat kidneys perfused with Krebs' solution and in isolated rabbit kidneys whether perfused with blood or Krebs' solution. | Effect of histamine and antihistamines on renal hemodynamics and functions in the isolated perfused canine kidney. Histamine was infused intra-arterially (10-40 mug/min) into isolated blood perfused canine kidneys while functional and hemodynamic parameters were monitored. When perfusion pressure (PP) was kept constant during the infusion of histamine, renal blood flow (RBF) increased from 137 +/- 9 to 181 +/- 9 ml/min (P less than .001). A greater increase in RBF occurred to the inner renal cortex than the outer renal cortex as measured by radioactive microspheres. The fractional outer/inner cortical blood flow changed from 79:21 before histamine to 74:26 during its infusion (P less than .001). Histamine did not alter creatinine clearance (Ccr), urine volume (V), sodium excretion (UNaV) or the fractional excretion of sodium (FENa) or water (FEH20) under these conditions. When renal blood flow was held constant during the infusion of histamine, PP decreased from 126/106 +/- 2 to 100/81 +/- 2 mm Hg (P less than .001). This resulted in a reduction of absolute outer cortical (outer/inner) changed from 77:23 before histamine to 72:28 during its infusion (P less than .001). In contrast to the effects of histamine at constant PP, CCr, V, UNaV, FENa, and FEH20 were decreased when histamine infusion reduced the PP. Administration of the H1 receptor antagonist, diphenhydramine, blocked the hemodynamic effects of histamine whereas the administration of an H2 receptor antagonist, metiamide, did not alter the histamine response. Similar vasodilatory responses to histamine were observed in isolated blood perfused dog and cat kidneys. In contrast, vasoconstrictor responses to histamine occurred in isolated dog and cat kidneys perfused with Krebs' solution and in isolated rabbit kidneys whether perfused with blood or Krebs' solution. | [
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PMID:10421 | Phenacetin: renal tubular transport and intrarenal distribution in the dog. | In a total of 22 studies in the anesthetized dog, the renal clearnace of phenacetin was measured over a range of plasma concentrations and at different urinary pH and rates of urine flow. Phenacetin is reabsorbed by passive diffusion. The urine/plasma ratio is essentially unity under all conditions. As measured by ultraviolet-spectrophotometric and colorometric methods and by high-pressure liquid chromatography, a labile metabolite appears in urine and renal papilla which upon hydrolysis gives rise to falsely high values for phenacetin. Renal cortical and papillary concentrations were determined in six dogs during hydropenia or diuresis. All tissue/plasma concentration ratios for phenacetin were essentially unity. The results indicate that the papillary localization of the earliest lesions of analgesic nephropathy cannot be attributed to high phenacetin concentrations within the papilla. This is in contrast to acetaminophen, the major metabolite of phenacetin. | Phenacetin: renal tubular transport and intrarenal distribution in the dog. In a total of 22 studies in the anesthetized dog, the renal clearnace of phenacetin was measured over a range of plasma concentrations and at different urinary pH and rates of urine flow. Phenacetin is reabsorbed by passive diffusion. The urine/plasma ratio is essentially unity under all conditions. As measured by ultraviolet-spectrophotometric and colorometric methods and by high-pressure liquid chromatography, a labile metabolite appears in urine and renal papilla which upon hydrolysis gives rise to falsely high values for phenacetin. Renal cortical and papillary concentrations were determined in six dogs during hydropenia or diuresis. All tissue/plasma concentration ratios for phenacetin were essentially unity. The results indicate that the papillary localization of the earliest lesions of analgesic nephropathy cannot be attributed to high phenacetin concentrations within the papilla. This is in contrast to acetaminophen, the major metabolite of phenacetin. | [
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PMID:10422 | Loperamide binding to opiate receptor sites of brain and myenteric plexus. | Loperamide, a new antidiarrheal agent, was tested to determine whether its biological activity involves binding to opiate receptor sites. Loperamide and morphine competitively inhibited 3H-naloxone binding to homogenates a guinea-pig brain and myenteric plexus. The Kp values obtain in the presence of Na+ were: morphine, 9.60-10(-9) M (brain), 1.66-10(-7) M (myenteric plexus); loperamide, 7.20-10(-9) M (brain), 1.33-10(-7) M (myenteric plexus); naloxone, 4.78-10(-10) M (brain), 1.27-10(-9) M (myenteric plexus. In the absence of Na+, binding a loperamide and morphine to brain homogenate was enhanced while the binding of naloxone was reduced. Morphine (IC50 = 7.5-10(-8) M) and loperamide (IC50 = 6.9-10(-9) M) inhibited the electrically induced contractions of longitudinal muscle from guinea-pig ileum, and naloxone competitively antagonized these effects. The Kd value calculated for the interaction of naloxone with binding sites associated with the contracting muscle was between 0.98-10(-9) M and 1.85-10(-9) M. In the mouse hot plate test, subcutaneous administration of morphine (minimal effective dose = 6.6 mugmol/kg) and loperamide (minimal effective dose = 78 mugmol/kg) delayed the response to heat stimuli and this effect was completely blocked by prior administration of naloxone. In the anesthetixed dog, intravenous administration of morphine (100 mug/kg) and loperamide (100 mug/kg) enhanced the contractile activity of circular muscle in proximal and distal duodenum, distal ileum and proximal colon but duodenal longitudinal muscle was relaxed; these effects were completely reversed by subsequent administration of naloxone. It is concluded that loperamide binds to opiate receptor sites and possesses opiate agonist activity both in vivo and in vitro. | Loperamide binding to opiate receptor sites of brain and myenteric plexus. Loperamide, a new antidiarrheal agent, was tested to determine whether its biological activity involves binding to opiate receptor sites. Loperamide and morphine competitively inhibited 3H-naloxone binding to homogenates a guinea-pig brain and myenteric plexus. The Kp values obtain in the presence of Na+ were: morphine, 9.60-10(-9) M (brain), 1.66-10(-7) M (myenteric plexus); loperamide, 7.20-10(-9) M (brain), 1.33-10(-7) M (myenteric plexus); naloxone, 4.78-10(-10) M (brain), 1.27-10(-9) M (myenteric plexus. In the absence of Na+, binding a loperamide and morphine to brain homogenate was enhanced while the binding of naloxone was reduced. Morphine (IC50 = 7.5-10(-8) M) and loperamide (IC50 = 6.9-10(-9) M) inhibited the electrically induced contractions of longitudinal muscle from guinea-pig ileum, and naloxone competitively antagonized these effects. The Kd value calculated for the interaction of naloxone with binding sites associated with the contracting muscle was between 0.98-10(-9) M and 1.85-10(-9) M. In the mouse hot plate test, subcutaneous administration of morphine (minimal effective dose = 6.6 mugmol/kg) and loperamide (minimal effective dose = 78 mugmol/kg) delayed the response to heat stimuli and this effect was completely blocked by prior administration of naloxone. In the anesthetixed dog, intravenous administration of morphine (100 mug/kg) and loperamide (100 mug/kg) enhanced the contractile activity of circular muscle in proximal and distal duodenum, distal ileum and proximal colon but duodenal longitudinal muscle was relaxed; these effects were completely reversed by subsequent administration of naloxone. It is concluded that loperamide binds to opiate receptor sites and possesses opiate agonist activity both in vivo and in vitro. | [
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PMID:10423 | Cellular mechanisms of renal tubular transport of I-dopa and its derivatives in the rat: microperfusion studies. | Proximal tubular transport of L-3,4-dihydroxyphenylalanine (L-dopa) and its derivatives were studied in the rat kidney by microperfusion and capillary perfusion techniques in situ. With the use of microperfusion techniques alone, it was found that L-dopa, L-3-methoxy-4-hydroxyphenylalanine, L-tyrosine and L-phenylalanine rapidly disappeared from the perfusate. The rates of reabsorption were calculated to be 2.05 X 10(-12), 2.13 X 10(-12), 6.35 X 10(-12) and 7.14 X 10(-12) mol/cm/sec, respectively. In contrast, L-alpha-methyldopa and dopamine were only slightly reabsorbed. The permeability coefficients were calculated to be 0.35 X 10(-4) and 0.21 x 10(-4) and 0.21 x 10(-4) cm/sec, respectively. The rate of reabsorption of L-dopa was greatly reduced in the presence of L-phenylalanine in the perfusate, but was not affected by D-dopa. With the stop-flow microperfusion technique with simultaneous capillary perfusion, the zero net flux transtubular concentration difference (deltaC) of labeled dopa was measured. When the initial concentration of 2 mM labeled L-dopa was perfused, the luminal concentration fell to a plateau level of 1.5 mM after a contact time of 20 seconds; i.e., deltaC was 0.5 mM. The net flux was much less than the efflux, suggesting the presence of a secretory or passive back flux of L-dopa. The deltaC of L-dopa was not influenced by the decarboxylase inhibitor, MK-486, and monoamine oxidase inhibitor, pheniprazine, but was significantly reduced by NaCN. Thus the reabsorption of L-dopa in the proximal convoluted tubule is an active process with great structural specificity. | Cellular mechanisms of renal tubular transport of I-dopa and its derivatives in the rat: microperfusion studies. Proximal tubular transport of L-3,4-dihydroxyphenylalanine (L-dopa) and its derivatives were studied in the rat kidney by microperfusion and capillary perfusion techniques in situ. With the use of microperfusion techniques alone, it was found that L-dopa, L-3-methoxy-4-hydroxyphenylalanine, L-tyrosine and L-phenylalanine rapidly disappeared from the perfusate. The rates of reabsorption were calculated to be 2.05 X 10(-12), 2.13 X 10(-12), 6.35 X 10(-12) and 7.14 X 10(-12) mol/cm/sec, respectively. In contrast, L-alpha-methyldopa and dopamine were only slightly reabsorbed. The permeability coefficients were calculated to be 0.35 X 10(-4) and 0.21 x 10(-4) and 0.21 x 10(-4) cm/sec, respectively. The rate of reabsorption of L-dopa was greatly reduced in the presence of L-phenylalanine in the perfusate, but was not affected by D-dopa. With the stop-flow microperfusion technique with simultaneous capillary perfusion, the zero net flux transtubular concentration difference (deltaC) of labeled dopa was measured. When the initial concentration of 2 mM labeled L-dopa was perfused, the luminal concentration fell to a plateau level of 1.5 mM after a contact time of 20 seconds; i.e., deltaC was 0.5 mM. The net flux was much less than the efflux, suggesting the presence of a secretory or passive back flux of L-dopa. The deltaC of L-dopa was not influenced by the decarboxylase inhibitor, MK-486, and monoamine oxidase inhibitor, pheniprazine, but was significantly reduced by NaCN. Thus the reabsorption of L-dopa in the proximal convoluted tubule is an active process with great structural specificity. | [
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PMID:10424 | Biosynthesis and metabolism of endogenous tyramine and its normal presence in sympathetic nerves. | By use of a sensitive and specific enzymatic isotopic method for the determination of tyramine, the small quantities of this amine which are present endogenously in rat tissues, including brain, heart, kidney and salivary gland, have been quantitated. The levels of tyramine in brain were increased to a similar extent by injecting animals with a monoamine oxidase inhibitor, pargyline, and a dopamine beta-hydroxylase inhibitor, FLA-63; in contrast, pretreatment of animals with alpha-methyl-para-tyrosine, a tyrosine hydroxylase inhibitor, did not lead to an increase in tyramine levels in brain. Pretreatment of rats with 6-hydroxydopamine resulted in a marked diminution in the tyramine content of rat atria and salivary gland. Denervation of the salivary gland decreased the endogenous level of tyramine approximately 50% in denervated glands compared to undenervated glands. These results suggest that tyramine exists at least partly in sympathetic nerves in many tissues. | Biosynthesis and metabolism of endogenous tyramine and its normal presence in sympathetic nerves. By use of a sensitive and specific enzymatic isotopic method for the determination of tyramine, the small quantities of this amine which are present endogenously in rat tissues, including brain, heart, kidney and salivary gland, have been quantitated. The levels of tyramine in brain were increased to a similar extent by injecting animals with a monoamine oxidase inhibitor, pargyline, and a dopamine beta-hydroxylase inhibitor, FLA-63; in contrast, pretreatment of animals with alpha-methyl-para-tyrosine, a tyrosine hydroxylase inhibitor, did not lead to an increase in tyramine levels in brain. Pretreatment of rats with 6-hydroxydopamine resulted in a marked diminution in the tyramine content of rat atria and salivary gland. Denervation of the salivary gland decreased the endogenous level of tyramine approximately 50% in denervated glands compared to undenervated glands. These results suggest that tyramine exists at least partly in sympathetic nerves in many tissues. | [
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PMID:10425 | The beta adrenergic receptors of chromatophores of the frog, Rana pipiens. | The isolated skin of Rana pipiens was found to be a suitable model for the quantitative study of chromatophore beta adrenergic receptors uninfluenced by prejunctional phenomena. Cumulative concentration-response curves for adrenergic agonists were obtained in preparations in which effective alpha adrenergic blockade had been produced with phenoxybenzamine. The beta adrenergic agonists darkened the preparation, as did melanocyte-stimulating hormone, but the maximum effects differed. The maximum of the l-isoproterenol cumulative concentration-response curve was approximately 50% less than that of melanocyte-stimulating hormone, while the maxima for l-epinephrine and l-norepinephrine were significantly less than that for isoproterenol. Microscopic examination revealed a qualitative difference: while maximal darkening produced by melanocyte-stimulating hormone was associated with maximal changes in both interspot melanophores and iridophores, maximal adrenergic-induced darkening was associated with maximal iridophore granule concentration only. No qualitative differences could be observed in the darkening caused by the three adrenergic agonists. The beta adrenergic potencies of l-norepinephrine and l-isoproterenol relative to l-epinephrine were determined by four-point bioassay. Isoproterenol was found to be 138 times as potent as epinephrine, while norepinephrine was 4 times as potent. Similarly, antagonism of isoproterenol-induced darkening of phenoxybenzamine-pretreated skin samples by the beta adrenergic blocking agents dl-propranolol, dl-sotalol, dl-practolol, l-butoxamine and d-butoxamine was studied, and their KB and pA2 values, respectively, were found to be: dl-propranolol (1.44 X 10(-8)M, 7.81); dl-sotalol (7.25 X 10(-8)M, 7.23); l-butoxamine (6.92 X 10(-6)M, 5.10); dl-practolol (1.91 X 10(-5)M, 4.96); d-butoxamine (no activity). Comparison of the potency ratios and pA2 values cited above with similar parameters obtained by other investigators in several mammalian tissues suggests that there is wide variation among beta adrenergic receptors. | The beta adrenergic receptors of chromatophores of the frog, Rana pipiens. The isolated skin of Rana pipiens was found to be a suitable model for the quantitative study of chromatophore beta adrenergic receptors uninfluenced by prejunctional phenomena. Cumulative concentration-response curves for adrenergic agonists were obtained in preparations in which effective alpha adrenergic blockade had been produced with phenoxybenzamine. The beta adrenergic agonists darkened the preparation, as did melanocyte-stimulating hormone, but the maximum effects differed. The maximum of the l-isoproterenol cumulative concentration-response curve was approximately 50% less than that of melanocyte-stimulating hormone, while the maxima for l-epinephrine and l-norepinephrine were significantly less than that for isoproterenol. Microscopic examination revealed a qualitative difference: while maximal darkening produced by melanocyte-stimulating hormone was associated with maximal changes in both interspot melanophores and iridophores, maximal adrenergic-induced darkening was associated with maximal iridophore granule concentration only. No qualitative differences could be observed in the darkening caused by the three adrenergic agonists. The beta adrenergic potencies of l-norepinephrine and l-isoproterenol relative to l-epinephrine were determined by four-point bioassay. Isoproterenol was found to be 138 times as potent as epinephrine, while norepinephrine was 4 times as potent. Similarly, antagonism of isoproterenol-induced darkening of phenoxybenzamine-pretreated skin samples by the beta adrenergic blocking agents dl-propranolol, dl-sotalol, dl-practolol, l-butoxamine and d-butoxamine was studied, and their KB and pA2 values, respectively, were found to be: dl-propranolol (1.44 X 10(-8)M, 7.81); dl-sotalol (7.25 X 10(-8)M, 7.23); l-butoxamine (6.92 X 10(-6)M, 5.10); dl-practolol (1.91 X 10(-5)M, 4.96); d-butoxamine (no activity). Comparison of the potency ratios and pA2 values cited above with similar parameters obtained by other investigators in several mammalian tissues suggests that there is wide variation among beta adrenergic receptors. | [
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PMID:10426 | A polypeptide (AP-A) from sea anemone (Anthopleura xanthogrammica) with potent positive inotropic action. | Anthopleurin-A (AP-A), a polypeptide with MW ca. 5500 (53 amino acids), isolated from the sea anemone, Anthopleura xanthogrammica (Brandt), elicited a potent positive inotropic effect but without an accompanying chronotropic effect on the isolated cardiac muscles of rat, rabbit, guinea pig and cat. Similarly in dogs and cats in situ, i.p. injections of AP-A increased the contractile force without effect on heart rate or blood pressure. The cardiotonic potency for AP-A was equivalent to that of isoproterenol but much greater than that for ouabain or glucagon on the isolated cardiac muscle. AP-A increased the contractile force (cardiac output) and decreased atrial pressure in dog heart during pentobarbital-induced failure. This inotropic effect was not inhibited by propranolol pretreatment. The Ca++ requirement to restore the contractile force was less in AP-A-treated than in ouabain or isoproterenol-treated tissues. After AP-A treatment, the cardiac contractility was more resistant to hypoxia and to low or high temperature stress than ouabain-treated or control preparations. AP-A at 5 10(-9) M increased the duration of the action potential, its mean rate of rise and conduction in the guinea-pig atria and ventricles. At the maximum effective concentration, AP-A did not inhibit Na+, K+-activated adenosine triphosphatase, phosphodiesterase (high Km and low Km) and cyclic 3',5'-adenosine monophosphate content of guinea-pig heart. AP-A (5 X 10(-8) to 5 X 10(-7) M) neither contracted nor relaxed the isolated vascular smooth muscle. The results suggest that AP-A may be useful in the clinical management of cardiac failure and as an experimental tool to study the pharmacology and physiology of cardiac muscle. | A polypeptide (AP-A) from sea anemone (Anthopleura xanthogrammica) with potent positive inotropic action. Anthopleurin-A (AP-A), a polypeptide with MW ca. 5500 (53 amino acids), isolated from the sea anemone, Anthopleura xanthogrammica (Brandt), elicited a potent positive inotropic effect but without an accompanying chronotropic effect on the isolated cardiac muscles of rat, rabbit, guinea pig and cat. Similarly in dogs and cats in situ, i.p. injections of AP-A increased the contractile force without effect on heart rate or blood pressure. The cardiotonic potency for AP-A was equivalent to that of isoproterenol but much greater than that for ouabain or glucagon on the isolated cardiac muscle. AP-A increased the contractile force (cardiac output) and decreased atrial pressure in dog heart during pentobarbital-induced failure. This inotropic effect was not inhibited by propranolol pretreatment. The Ca++ requirement to restore the contractile force was less in AP-A-treated than in ouabain or isoproterenol-treated tissues. After AP-A treatment, the cardiac contractility was more resistant to hypoxia and to low or high temperature stress than ouabain-treated or control preparations. AP-A at 5 10(-9) M increased the duration of the action potential, its mean rate of rise and conduction in the guinea-pig atria and ventricles. At the maximum effective concentration, AP-A did not inhibit Na+, K+-activated adenosine triphosphatase, phosphodiesterase (high Km and low Km) and cyclic 3',5'-adenosine monophosphate content of guinea-pig heart. AP-A (5 X 10(-8) to 5 X 10(-7) M) neither contracted nor relaxed the isolated vascular smooth muscle. The results suggest that AP-A may be useful in the clinical management of cardiac failure and as an experimental tool to study the pharmacology and physiology of cardiac muscle. | [
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PMID:10427 | Isoproterenol antagonism of cardioselective beta adrenergic receptor blocking agents: a comparative study of human and guinea-pig cardiac and bronchial beta adrenergic receptors. | pA2 values against isoproterenol were determined for a number of cardioselective and noncardioselective beta adrenergic receptor blocking agents using human and guinea-pig isolated atrial and bronchial or tracheal preparations to study possible species differences. No significant differences in pA2 values for propranolol, pindolol, Ro 3-4787, acebutolol, atenolol, practolol, metoprolol, H 87/07 and tolamolol on bronchial or tracheal beta adrenergic receptors of both species were found. With respect to atrial beta adrenergic receptors, significantly lower pA2 values for human preparations, as compared to guinea-pig preparations, were found for tolamolol and CI 775. These are the only two agents in the series that derive their cardioselectivities from specific nitrogen substitutents. The different potencies of only these two compounds in antagonizing isoproterenol on atrial beta adrenergic receptors of both species suggest a difference in an accessory receptor area close to the site that interacts with the nitrogen atom of beta adrenergic agents. | Isoproterenol antagonism of cardioselective beta adrenergic receptor blocking agents: a comparative study of human and guinea-pig cardiac and bronchial beta adrenergic receptors. pA2 values against isoproterenol were determined for a number of cardioselective and noncardioselective beta adrenergic receptor blocking agents using human and guinea-pig isolated atrial and bronchial or tracheal preparations to study possible species differences. No significant differences in pA2 values for propranolol, pindolol, Ro 3-4787, acebutolol, atenolol, practolol, metoprolol, H 87/07 and tolamolol on bronchial or tracheal beta adrenergic receptors of both species were found. With respect to atrial beta adrenergic receptors, significantly lower pA2 values for human preparations, as compared to guinea-pig preparations, were found for tolamolol and CI 775. These are the only two agents in the series that derive their cardioselectivities from specific nitrogen substitutents. The different potencies of only these two compounds in antagonizing isoproterenol on atrial beta adrenergic receptors of both species suggest a difference in an accessory receptor area close to the site that interacts with the nitrogen atom of beta adrenergic agents. | [
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PMID:10428 | Interactions between narcotic analgesics and benzodiazepine derivatives on behavior in the mouse. | Interactions between the benzodiazepine derivatives, diazepam and oxazepam, and the narcotic analgesics, morphine and methadone, were evaluated on locomotor activity and in the tail-flick and hot-plate tests for analgesia in the mouse. The dose-related stimulation of locomotor activity by morphine was reduced by diazepam and oxazepam at doses which alone had no effect on locomotor activity. However, only oxazepam reduced the dose-related stimulation of locomotor activity by methadone. The observed decreases produced by diazepam and oxazepam were comparable in magnitude to those produced by naloxone. Stimulation of locomotor activity by d-amphetamine was not affected by either diazepam or oxazepam. Dose-response curves for locomotor activity were also determined with morphine and methadone administered intraventricularly. As before, diazepam and naloxone given i.p. decreased the stimulation of locomotor activity produced by morphine, but only naloxone affected methadone-stimulated locomotor activity. Neither diazepam, oxazepam nor naloxone reduced the brain or plasma levels of 3H-morphine or 3H-methadone. In contrast to the results on locomotor activity, neither of these benzodiazepines significantly modified the dose-response curves of morphine or methadone in either test for analgesia. The mechanisms involved in the observed interactions on locomotor activity may be related to the influences of benzodiazepines and narcotic analgesics on motor efferent pathways summating in such a manner as to interfere with the ability of the mice to locomote. The present results demonstrate that prominent interactions occur between members of the benzodiazepine and narcotic analgesic classes; these interactions are dependent upon both the specific combination of drugs administered and upon the test procedure. | Interactions between narcotic analgesics and benzodiazepine derivatives on behavior in the mouse. Interactions between the benzodiazepine derivatives, diazepam and oxazepam, and the narcotic analgesics, morphine and methadone, were evaluated on locomotor activity and in the tail-flick and hot-plate tests for analgesia in the mouse. The dose-related stimulation of locomotor activity by morphine was reduced by diazepam and oxazepam at doses which alone had no effect on locomotor activity. However, only oxazepam reduced the dose-related stimulation of locomotor activity by methadone. The observed decreases produced by diazepam and oxazepam were comparable in magnitude to those produced by naloxone. Stimulation of locomotor activity by d-amphetamine was not affected by either diazepam or oxazepam. Dose-response curves for locomotor activity were also determined with morphine and methadone administered intraventricularly. As before, diazepam and naloxone given i.p. decreased the stimulation of locomotor activity produced by morphine, but only naloxone affected methadone-stimulated locomotor activity. Neither diazepam, oxazepam nor naloxone reduced the brain or plasma levels of 3H-morphine or 3H-methadone. In contrast to the results on locomotor activity, neither of these benzodiazepines significantly modified the dose-response curves of morphine or methadone in either test for analgesia. The mechanisms involved in the observed interactions on locomotor activity may be related to the influences of benzodiazepines and narcotic analgesics on motor efferent pathways summating in such a manner as to interfere with the ability of the mice to locomote. The present results demonstrate that prominent interactions occur between members of the benzodiazepine and narcotic analgesic classes; these interactions are dependent upon both the specific combination of drugs administered and upon the test procedure. | [
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PMID:10429 | Precipitation of abstinence-like syndrome in morphine-dependent mice by pargyline. | In mice rendered morphine-dependent by pellet implantation for 3 days, the administration of pargyline 6 hours after pellet removal intensified narcotic abstinence behavior, particularly the narcotic withdrawal jumping response. Pargyline, 75 mg/kg i.p., caused a 6- to 9-fold increase in the incidence of jumping in mice withdrawing from morphine 6 hours after removal of the pellet, whereas this effect was not observed: 1) 1 hour after the injection of pargyline or 2) in animals still implanted with the morphine pellet. The median effective dose (ED50) of pargyline required to elicit withdrawal jumping in mice implanted with morphine decreased with increasing physical dependence. The ED50 for 72 hours was about one-sixth that after 24 hours of implantation. Additionally, pargyline potentiated naloxone-precipitated withdrawal jumping as evidenced by a reduction of the naloxone ED50 by approximately one-half. Administration of other monoamine oxidase inhibitors such as pheniprazine, iproiazid or tranylcypromine failed to alter the indicence of jumping in dependent mice undergoind abrupt morphine with drawal. Further, dopamine receptor stimulation by amphetamine, pheniprazine or amantadine antagonized the pargyline-induced jumping response. These data suggest that the increased incidence of withdrawal jumping observed after pargyline in morphine-dependent mice is not related to monoamine oxidase inhibition but rather to a possible pargyline-induced decrease in dopaminergic activity. | Precipitation of abstinence-like syndrome in morphine-dependent mice by pargyline. In mice rendered morphine-dependent by pellet implantation for 3 days, the administration of pargyline 6 hours after pellet removal intensified narcotic abstinence behavior, particularly the narcotic withdrawal jumping response. Pargyline, 75 mg/kg i.p., caused a 6- to 9-fold increase in the incidence of jumping in mice withdrawing from morphine 6 hours after removal of the pellet, whereas this effect was not observed: 1) 1 hour after the injection of pargyline or 2) in animals still implanted with the morphine pellet. The median effective dose (ED50) of pargyline required to elicit withdrawal jumping in mice implanted with morphine decreased with increasing physical dependence. The ED50 for 72 hours was about one-sixth that after 24 hours of implantation. Additionally, pargyline potentiated naloxone-precipitated withdrawal jumping as evidenced by a reduction of the naloxone ED50 by approximately one-half. Administration of other monoamine oxidase inhibitors such as pheniprazine, iproiazid or tranylcypromine failed to alter the indicence of jumping in dependent mice undergoind abrupt morphine with drawal. Further, dopamine receptor stimulation by amphetamine, pheniprazine or amantadine antagonized the pargyline-induced jumping response. These data suggest that the increased incidence of withdrawal jumping observed after pargyline in morphine-dependent mice is not related to monoamine oxidase inhibition but rather to a possible pargyline-induced decrease in dopaminergic activity. | [
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PMID:10430 | Effect of sodium para-aminosalicylate on oxygen affinity in normal, sickle and fetal human blood. | Sodium para-aminosalicylate (sodium salt of 2-hydroxy-4-aminobenzoic acid, Na-PAS) lowers the oxygen affinity of normal adult human placental, heterozygous and homozygous sickle cell anemic whole blood at 37 degrees C. The reduction of oxygen affinity is related to the type of hemoglobin in the blood. The mean P50 +/- S.E. at pH 7.40 for normal, placental, heterozygoud and homozygous sickle cell anemic blood in 26.2 +/- 0.1, 20.8 +/- 0.3, 26.8 +/- 0.3 and 31.0 +/- 0.5 mm Hg; in the presence of 5.7 mmol of Na-PAS per liter of blood the P50 values are increased to 28.0 +/- 0.3, 22.9 +/- 0.8, 30.5 +/- 0.6 and 33.9 +/- 0.3 mm Hg, respectively. The Bohr effect in normal and placental blood at this Na-PAS concentration is essentially unchanged: in heterozygous and homozygous sickle cell anemic blood, the Bohr factor (deta log P50/deta pH) is reduced from -0.48 +/- 0.02 to -0.41 +/- 0.01 and from -0.53 +/- 0.03 to -0.48 +/- 0.01. The Hill constants (n) of normal and placental blood are not affected by Na-PAS. In homozygous and heterozygous sickle blood, high concentrations of Na-PAS (22.9 mmol/l) decrease the Hill constant from 2.55 to 2.35 and from 2.56 to 2.28, respectively. Na-PAS is more firmly bound to red blood cells than to plasma. The binding of Na-PAS is probably primarily ionic in nature since the drug can be almost completely removed from blood components by dialysis. The changes in oxygen affinity caused by Na-PAS are consistent with conformational changes (R leads to T) which enhance the presence of deoxyhemoglobin. | Effect of sodium para-aminosalicylate on oxygen affinity in normal, sickle and fetal human blood. Sodium para-aminosalicylate (sodium salt of 2-hydroxy-4-aminobenzoic acid, Na-PAS) lowers the oxygen affinity of normal adult human placental, heterozygous and homozygous sickle cell anemic whole blood at 37 degrees C. The reduction of oxygen affinity is related to the type of hemoglobin in the blood. The mean P50 +/- S.E. at pH 7.40 for normal, placental, heterozygoud and homozygous sickle cell anemic blood in 26.2 +/- 0.1, 20.8 +/- 0.3, 26.8 +/- 0.3 and 31.0 +/- 0.5 mm Hg; in the presence of 5.7 mmol of Na-PAS per liter of blood the P50 values are increased to 28.0 +/- 0.3, 22.9 +/- 0.8, 30.5 +/- 0.6 and 33.9 +/- 0.3 mm Hg, respectively. The Bohr effect in normal and placental blood at this Na-PAS concentration is essentially unchanged: in heterozygous and homozygous sickle cell anemic blood, the Bohr factor (deta log P50/deta pH) is reduced from -0.48 +/- 0.02 to -0.41 +/- 0.01 and from -0.53 +/- 0.03 to -0.48 +/- 0.01. The Hill constants (n) of normal and placental blood are not affected by Na-PAS. In homozygous and heterozygous sickle blood, high concentrations of Na-PAS (22.9 mmol/l) decrease the Hill constant from 2.55 to 2.35 and from 2.56 to 2.28, respectively. Na-PAS is more firmly bound to red blood cells than to plasma. The binding of Na-PAS is probably primarily ionic in nature since the drug can be almost completely removed from blood components by dialysis. The changes in oxygen affinity caused by Na-PAS are consistent with conformational changes (R leads to T) which enhance the presence of deoxyhemoglobin. | [
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PMID:10432 | Drug blockade of open end-plate channels. | 1. The actions of amylobarbitone, thiopentone, methohexitone and methyprylone at voltage-clamped frog end-plates were studied. 2. In the presence of barbiturates the conductance change evoked by an iontophoretic carbachol application was reduced by a prepulse of carbachol. The extra inhibition evoked by a prepulse disappeared exponentially with a time constant of 150-200 ms. 3. Barbiturates produce an increased rate of decay of nerve evoked endplate currents. Tne concentration and voltage dependence of the barbtiruate e.p.c. decay rates tally with the hypothesis that the increased rate of decay is due to block of active receptor-channel complexes by barbiturates with a rate constant of 10(6) M-1S-1. 4. Conductance changes produced by bath applied agonists were depressed by thiopentone, the effect becoming greater the higher the agonist concentration. This effect, and also the observation that the concentration of thiopentone required to depress the bath agonist response is much greater than the apparent dissociation constant for binding to active receptor-channel complexes calculated from kinetic measurements, suggest that the selectivity for binding to open receptor-channel complexes is very high. 5. Methyprylone, which is structurally similar to the barbiturates, is only a weak antagonist and shows no interpulse interaction. It was predicted that methyprylone should produce fast and slow components in the e.p.c. decay, and this prediction was verified. 6. In the presence of barbiturates large iontophoretic carbachol applications produce conductance changes which show fast and slow components. Under these conditions the effects of carbachol prepulses become complex. However the effects are qualitatively consistent with the notion that different components of the response are contributed by channels located at various distances from the iontophoretic pipette tip. 7. All the data agree with a model in which the channel has three stages: closed, open and blocked. Only open channels can block, and blocked channels can only open. | Drug blockade of open end-plate channels. 1. The actions of amylobarbitone, thiopentone, methohexitone and methyprylone at voltage-clamped frog end-plates were studied. 2. In the presence of barbiturates the conductance change evoked by an iontophoretic carbachol application was reduced by a prepulse of carbachol. The extra inhibition evoked by a prepulse disappeared exponentially with a time constant of 150-200 ms. 3. Barbiturates produce an increased rate of decay of nerve evoked endplate currents. Tne concentration and voltage dependence of the barbtiruate e.p.c. decay rates tally with the hypothesis that the increased rate of decay is due to block of active receptor-channel complexes by barbiturates with a rate constant of 10(6) M-1S-1. 4. Conductance changes produced by bath applied agonists were depressed by thiopentone, the effect becoming greater the higher the agonist concentration. This effect, and also the observation that the concentration of thiopentone required to depress the bath agonist response is much greater than the apparent dissociation constant for binding to active receptor-channel complexes calculated from kinetic measurements, suggest that the selectivity for binding to open receptor-channel complexes is very high. 5. Methyprylone, which is structurally similar to the barbiturates, is only a weak antagonist and shows no interpulse interaction. It was predicted that methyprylone should produce fast and slow components in the e.p.c. decay, and this prediction was verified. 6. In the presence of barbiturates large iontophoretic carbachol applications produce conductance changes which show fast and slow components. Under these conditions the effects of carbachol prepulses become complex. However the effects are qualitatively consistent with the notion that different components of the response are contributed by channels located at various distances from the iontophoretic pipette tip. 7. All the data agree with a model in which the channel has three stages: closed, open and blocked. Only open channels can block, and blocked channels can only open. | [
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PMID:10433 | Conductances, diffusion and streaming potentials in the rat proximal tubule. | 1. Transtubular potential differences and specific resistances were measured in rat proximal tubules by means of single and double barrelled glass micro-electrodes. 2. Tip localization was made by observation of effective resistance changes measured with double barrelled micro-electrodes upon passage of oil droplets, and by perfusion with choline C1. 3. Mean early proximal p.d.s. of the order of -1 to -2 mV, and late values of +0-5 to +1mV were found. Mean specific resistances ranged from 12 to 15 omega cm2. 4. Diffusion potentials and single ion relative conductances were evaluated, perfusing the lumen with solutions differing only with respect to one salt concentration. Na and K conductances were similar and greater than those of C1. 5. Luminal and peritubular perfusions with hypotonic solutions showed the occurrence of streaming potentials in this structure suggesting the existence of pores lined with negative charges. The effective diameter of these pores appeared to be reduced by hypotonic perfusion, as evidenced by a significant increase in resistance, indicating that the main ion path across this structure is represented by intercellular spaces. | Conductances, diffusion and streaming potentials in the rat proximal tubule. 1. Transtubular potential differences and specific resistances were measured in rat proximal tubules by means of single and double barrelled glass micro-electrodes. 2. Tip localization was made by observation of effective resistance changes measured with double barrelled micro-electrodes upon passage of oil droplets, and by perfusion with choline C1. 3. Mean early proximal p.d.s. of the order of -1 to -2 mV, and late values of +0-5 to +1mV were found. Mean specific resistances ranged from 12 to 15 omega cm2. 4. Diffusion potentials and single ion relative conductances were evaluated, perfusing the lumen with solutions differing only with respect to one salt concentration. Na and K conductances were similar and greater than those of C1. 5. Luminal and peritubular perfusions with hypotonic solutions showed the occurrence of streaming potentials in this structure suggesting the existence of pores lined with negative charges. The effective diameter of these pores appeared to be reduced by hypotonic perfusion, as evidenced by a significant increase in resistance, indicating that the main ion path across this structure is represented by intercellular spaces. | [
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PMID:10434 | Some aspects of foetal and uteroplacental metabolism in cows with indwelling umbilical and uterine vascular catheters. | 1. The experiments were carried out on conscious pregnant Jersey cows with intravascular catheters implanted during late gestation in umbilical and uterine vessels. All but three of fifteen animals delivered live healthy calves. 2. Rountine daily analyses were made of blood gas tensions, pH and packed cell volume in foetal and maternal blood; plasma concentrations of glucose, fructose, lactate and urea were also determined. Measurements of plasma free fatty acids and blood acetate concentrations were made less frequently. Foetal heart rate and arterial blood pressure were recorded in animals with an umbilical arterial catheter. 3. The concentration differences between foetal and maternal blood or plasma in glucose, urea and acetate were measured in fifteen animals. The maternal-to-foetal glucose and acetate gradients across the placenta were high while the foetal-to-maternal plasma urea differences were small. 4. In those animals with patent arterial and venous catheters, uterine and umbilical blood flows were measured together with the arteriovenous differences in 02, glucose, acetate and lactate so that rates of foetal and uterine consumption could be estimated. The rates of utilization of O2, glucose and acetate by the foetus were lower than the values for the whole uterus, while the uteroplacental metabolism of these substrates was very high. 5. Significant amounts of lactate, which appeared to be produced by the uteroplacental tissue, were utilized by the foetus; the remainder passed into the uterine venous blood. 6. The total substrate/O2 quotient for the foetus, calculated from the utilization of known metabolites, appeared to be greater than 1. Thus, in the calf some carbon accumulation from sources other than amino acids, the uptake of which was not measured, would seem to occur. These results and the metabolic activity of the uterine tissues are discussed in relation to comparable findings in the sheep. | Some aspects of foetal and uteroplacental metabolism in cows with indwelling umbilical and uterine vascular catheters. 1. The experiments were carried out on conscious pregnant Jersey cows with intravascular catheters implanted during late gestation in umbilical and uterine vessels. All but three of fifteen animals delivered live healthy calves. 2. Rountine daily analyses were made of blood gas tensions, pH and packed cell volume in foetal and maternal blood; plasma concentrations of glucose, fructose, lactate and urea were also determined. Measurements of plasma free fatty acids and blood acetate concentrations were made less frequently. Foetal heart rate and arterial blood pressure were recorded in animals with an umbilical arterial catheter. 3. The concentration differences between foetal and maternal blood or plasma in glucose, urea and acetate were measured in fifteen animals. The maternal-to-foetal glucose and acetate gradients across the placenta were high while the foetal-to-maternal plasma urea differences were small. 4. In those animals with patent arterial and venous catheters, uterine and umbilical blood flows were measured together with the arteriovenous differences in 02, glucose, acetate and lactate so that rates of foetal and uterine consumption could be estimated. The rates of utilization of O2, glucose and acetate by the foetus were lower than the values for the whole uterus, while the uteroplacental metabolism of these substrates was very high. 5. Significant amounts of lactate, which appeared to be produced by the uteroplacental tissue, were utilized by the foetus; the remainder passed into the uterine venous blood. 6. The total substrate/O2 quotient for the foetus, calculated from the utilization of known metabolites, appeared to be greater than 1. Thus, in the calf some carbon accumulation from sources other than amino acids, the uptake of which was not measured, would seem to occur. These results and the metabolic activity of the uterine tissues are discussed in relation to comparable findings in the sheep. | [
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PMID:10435 | Stimulation-induced factors which affect augmentation and potentiation of trasmitter release at the neuromuscular junction. | 1. End-plate potentials (e.p.p.s) were recorded from frog sartorius neuromuscular junctions under conditions of decreased transmitter release to study the effect of repetitive stimulation on augmentation and potentiation of transmitter release. 2. The magnitudes and time constants of decay of augmentation and potentiation were determined both following a primary conditioning train and following an identical secondary conditioning train applied from 30 to 170 sec after the primary conditioning train. 3. The magnitude of augmentation following the secondary conditioning trains was increased over that following the primary conditioning trains even though augmentation, with a time constant of decay of about 7 sec, had decayed to insignificant levels before the onset of the secondary trains. This increase in augmentation was not due to a change in its rate of decay during the secondary trains. 4. The increased magnitude of augmentation can be described as arising from an expression factor which, for conditioning trains of 200 impulses at 20/sec, has an initial magnitude of 1-6 +/- 1-2 (S.D. of observation) (the magnitude of augmentation is increased 2-6 times) and decays approximately exponentially with a time constant of 90 +/- 50 (S.D. of observation) sec. The expression factor thus decays about ten times slower than augmentation. 5. Doubling the number of impulses in the primary conditioning train from 100 to 200 led to a 2-8 +/- 1-0 (S.D. of observation) times increase in the magnitude of the expression factor, estimated by placing a 200 impulse secondary conditioning train 40 sec after the primary conditioning train. 6. The expression factor, while increasing the magnitude of augmentation, had little or no effect on the magnitude of potentiation or on trasmitter release in the absence of augmentation. The expression factor decayed about twice as slowly as potentiation. 7. The time constants characterizing the decay of potentiation were greater following the secondary conditioning trains than following the primary conditioning trains. 8. The increased time constant for the decay of potentiation can be described as arising from a time constant factor which, for conditioning trains of 200 impulses at 20/sec, has an initial magnitude of 1-2 +/- 0-7 (S.D. of observation) (the time constant of potentiation is increased 2-2 times) and decays approximately exponentially with a time constant of 130 +/- 45 (S.D. of observation) sec. The time constant factor decayed about three times slower than potentiation. 9. Doubling the number of impulses in the primary conditioning train from 100 to 200 led to a 1-6 +/- 0-8 (S.D. of observation) times increase in the magnitude of the time constant factor, estimated by placing a 200 impulse secondary conditioning train 40 sec after the primary conditioning train. 10... | Stimulation-induced factors which affect augmentation and potentiation of trasmitter release at the neuromuscular junction. 1. End-plate potentials (e.p.p.s) were recorded from frog sartorius neuromuscular junctions under conditions of decreased transmitter release to study the effect of repetitive stimulation on augmentation and potentiation of transmitter release. 2. The magnitudes and time constants of decay of augmentation and potentiation were determined both following a primary conditioning train and following an identical secondary conditioning train applied from 30 to 170 sec after the primary conditioning train. 3. The magnitude of augmentation following the secondary conditioning trains was increased over that following the primary conditioning trains even though augmentation, with a time constant of decay of about 7 sec, had decayed to insignificant levels before the onset of the secondary trains. This increase in augmentation was not due to a change in its rate of decay during the secondary trains. 4. The increased magnitude of augmentation can be described as arising from an expression factor which, for conditioning trains of 200 impulses at 20/sec, has an initial magnitude of 1-6 +/- 1-2 (S.D. of observation) (the magnitude of augmentation is increased 2-6 times) and decays approximately exponentially with a time constant of 90 +/- 50 (S.D. of observation) sec. The expression factor thus decays about ten times slower than augmentation. 5. Doubling the number of impulses in the primary conditioning train from 100 to 200 led to a 2-8 +/- 1-0 (S.D. of observation) times increase in the magnitude of the expression factor, estimated by placing a 200 impulse secondary conditioning train 40 sec after the primary conditioning train. 6. The expression factor, while increasing the magnitude of augmentation, had little or no effect on the magnitude of potentiation or on trasmitter release in the absence of augmentation. The expression factor decayed about twice as slowly as potentiation. 7. The time constants characterizing the decay of potentiation were greater following the secondary conditioning trains than following the primary conditioning trains. 8. The increased time constant for the decay of potentiation can be described as arising from a time constant factor which, for conditioning trains of 200 impulses at 20/sec, has an initial magnitude of 1-2 +/- 0-7 (S.D. of observation) (the time constant of potentiation is increased 2-2 times) and decays approximately exponentially with a time constant of 130 +/- 45 (S.D. of observation) sec. The time constant factor decayed about three times slower than potentiation. 9. Doubling the number of impulses in the primary conditioning train from 100 to 200 led to a 1-6 +/- 0-8 (S.D. of observation) times increase in the magnitude of the time constant factor, estimated by placing a 200 impulse secondary conditioning train 40 sec after the primary conditioning train. 10... | [
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PMID:10436 | Gastric emptying of organic acids in the dog. | Test meals of 300 ml. of six different organic acids were instilled into the stomach of six healthy mongrel dogs. Citric, acetic, propionic, lactic, tartaric and succinic acid were given in 50, 100, 150, and 200 mN concentrations. 2. During the emptying process, the gastric contents were aspirated and immediately re-instilled at 10 min intervals, and the following parameters were recorded: volume, concentration of the organic anion, pH, hydrogen ion concentration and osmolarity. 3. By multiple stepwise regression analysis, the combination of parameters which most effectively determines gastric emptying rate was found to be: concentration of the organic anion, followed by intragastric volume and number of previous test meals given on the same day. These three parameters appear in the equation for gastric emptying rate in which the individual characteristic of each acid is expressed by a constant. 4. Among the various acids, inhibition of emptying rate increases with rising number of carboxylic groups of the acid and its molecular weight. 5. After proximal gastric vagotomy, emptying rate of organic acids is independent of volume, and emptying approaches an exponential pattern. 6. A model for gastric emptying of organic acids with at least three different receptors is proposed: one for the structure of the organic acid, one for concentration and one for intragastric volume. | Gastric emptying of organic acids in the dog. Test meals of 300 ml. of six different organic acids were instilled into the stomach of six healthy mongrel dogs. Citric, acetic, propionic, lactic, tartaric and succinic acid were given in 50, 100, 150, and 200 mN concentrations. 2. During the emptying process, the gastric contents were aspirated and immediately re-instilled at 10 min intervals, and the following parameters were recorded: volume, concentration of the organic anion, pH, hydrogen ion concentration and osmolarity. 3. By multiple stepwise regression analysis, the combination of parameters which most effectively determines gastric emptying rate was found to be: concentration of the organic anion, followed by intragastric volume and number of previous test meals given on the same day. These three parameters appear in the equation for gastric emptying rate in which the individual characteristic of each acid is expressed by a constant. 4. Among the various acids, inhibition of emptying rate increases with rising number of carboxylic groups of the acid and its molecular weight. 5. After proximal gastric vagotomy, emptying rate of organic acids is independent of volume, and emptying approaches an exponential pattern. 6. A model for gastric emptying of organic acids with at least three different receptors is proposed: one for the structure of the organic acid, one for concentration and one for intragastric volume. | [
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PMID:10439 | Evaluation of analgesic action and efficacy of antirheumatic drugs. Study of 10 drugs in 684 patients with rheumatoid arthritis. | A single-blind non-crossover method for assessing the potential effectiveness of antirheumatic drugs has been described. The method employs entirely subjective indices and incorporates a daily pain chart for measuring the pain response over the duration of the trial. In addition, the mean number of days withdrawn and patients' satisfaction rating are measured. The statistical method can correct for initial imbalances between groups and allows for the valid comparison of drugs from separate trials. Ten antirheumatic medications were evaluated using this technique in 684 patients with rheumatoid arthritis, and the results are in agreement with those of previous studies using standard clinical methods. The new method is simple, rapid in performance, economical in terms of cost and time, and has been shown to be sensitive and reproducible. The results indicate that there are no significant differences in efficacy between the currently available non-steroidal, anti-inflammatory analgesic drugs, in the treatment of rheumatoid arthritis. | Evaluation of analgesic action and efficacy of antirheumatic drugs. Study of 10 drugs in 684 patients with rheumatoid arthritis. A single-blind non-crossover method for assessing the potential effectiveness of antirheumatic drugs has been described. The method employs entirely subjective indices and incorporates a daily pain chart for measuring the pain response over the duration of the trial. In addition, the mean number of days withdrawn and patients' satisfaction rating are measured. The statistical method can correct for initial imbalances between groups and allows for the valid comparison of drugs from separate trials. Ten antirheumatic medications were evaluated using this technique in 684 patients with rheumatoid arthritis, and the results are in agreement with those of previous studies using standard clinical methods. The new method is simple, rapid in performance, economical in terms of cost and time, and has been shown to be sensitive and reproducible. The results indicate that there are no significant differences in efficacy between the currently available non-steroidal, anti-inflammatory analgesic drugs, in the treatment of rheumatoid arthritis. | [
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PMID:10441 | Sympathomimetic amines having a carbostyril nucleus. | A series of new sympathomimetic amines containing an 8-hydroxycarbostyril moiety was synthesized. These compounds probably exist as resonance hybrids having two acidic hydrogen atoms in locations approximating to those of the hydroxyl groups of catechol-containing adrenergic agents. In an in vitro test, many of these compounds showed potent activity for relaxation of guinea pig tracheal smooth muscle. One of the compounds was 24 000 times more potent than isoproterenol. Their actions on cardiac muscle were also examined in vitro by measuring increase in the beating rate of the right atria of guinea pigs. Several of the compounds appeared to be beta-selective. Some of the compounds seem suitable for use as bronchodilators. The structure-activity relationships of these compounds were discussed in comparison with those of catecholamines. | Sympathomimetic amines having a carbostyril nucleus. A series of new sympathomimetic amines containing an 8-hydroxycarbostyril moiety was synthesized. These compounds probably exist as resonance hybrids having two acidic hydrogen atoms in locations approximating to those of the hydroxyl groups of catechol-containing adrenergic agents. In an in vitro test, many of these compounds showed potent activity for relaxation of guinea pig tracheal smooth muscle. One of the compounds was 24 000 times more potent than isoproterenol. Their actions on cardiac muscle were also examined in vitro by measuring increase in the beating rate of the right atria of guinea pigs. Several of the compounds appeared to be beta-selective. Some of the compounds seem suitable for use as bronchodilators. The structure-activity relationships of these compounds were discussed in comparison with those of catecholamines. | [
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PMID:10444 | Fusion in phospholipid spherical membranes. II. Effect of cholesterol, divalent ions and pH. | Effect of cholesterol, divalent ions and pH on spherical bilayer membrane fusion was studied as a function of increasing temperature. Spherical bilayer membranes were composed of natural [phosphatidylcholine (PC) and phosphatidylserine (PS)] as well as synthetic (dipalmitoyl-PC, dimyristoyl-PC and dioleoyl-PC) phospholipids. Incorporation of cholesterol into the membrane (33% by weight) suppressed the fusion temperature and also greatly reduced the percentage of membrane fusion. The presence of 1 mM divalent ions (Ca++, Mg++ or Mn++) on both sides or one side of the PC membrane did not affect appreciably its fusion characteristic with temperature, but the PS membrane fusion with temperature was greatly enhanced by the presence of divalent ions. The variation of pH of the environmental solution in the range of 5.5 approximately 7.0 did not affect the membrane fusion characteristic. However, at pH 8.5, the fusion with respect to temperature was shifted toward the lower temperature by approximately 3degreesC for PC and PS membranes, and at pH 3.0 the opposite situation was observed as the fusion temperature was increased by 6degreesC for PS membranes and by 4degreesC for PC membranes The results seem to indicate that membrane fluidity and structural instability in the bilayer are important for membrane fusion to occur. | Fusion in phospholipid spherical membranes. II. Effect of cholesterol, divalent ions and pH. Effect of cholesterol, divalent ions and pH on spherical bilayer membrane fusion was studied as a function of increasing temperature. Spherical bilayer membranes were composed of natural [phosphatidylcholine (PC) and phosphatidylserine (PS)] as well as synthetic (dipalmitoyl-PC, dimyristoyl-PC and dioleoyl-PC) phospholipids. Incorporation of cholesterol into the membrane (33% by weight) suppressed the fusion temperature and also greatly reduced the percentage of membrane fusion. The presence of 1 mM divalent ions (Ca++, Mg++ or Mn++) on both sides or one side of the PC membrane did not affect appreciably its fusion characteristic with temperature, but the PS membrane fusion with temperature was greatly enhanced by the presence of divalent ions. The variation of pH of the environmental solution in the range of 5.5 approximately 7.0 did not affect the membrane fusion characteristic. However, at pH 8.5, the fusion with respect to temperature was shifted toward the lower temperature by approximately 3degreesC for PC and PS membranes, and at pH 3.0 the opposite situation was observed as the fusion temperature was increased by 6degreesC for PS membranes and by 4degreesC for PC membranes The results seem to indicate that membrane fluidity and structural instability in the bilayer are important for membrane fusion to occur. | [
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PMID:10447 | Properties of two isolated antigens associated with bovine leukemia virus infection. | We isolated an ether-resistant internal antigen and an ether-sensitive antigen previously described in relation to bovine leukemia virus infection. These two antigens have now been isolated by isoelectric focusing and concanavalin A affinity chromatography, respectively. The ether-resistant antigen exhibited isoelectric heterogeneity with a major peak at pH 7.2 and a minor peak at pH 6.2. Its molecular weight, estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), was 23,000 (p23), and it gave a sedimentation value of 2.3s. For material containing ether-sensitive antigen, analyzed by SDS-PAGE, protein staining revealed four components with molecular weights of 18,000, 25,000, 45,000, and 55,000. Two of these [45,000 (gp45) and 55,000 (gp55)] were stained by periodic acid-Schiff reagent. Isoelectric point and sedimentation value of the major glycoprotein (gp45) were pH 5.0 and 3.4s, respectively; no immunologic cross-reactivity was found between p23 and glycoprotein antigen. | Properties of two isolated antigens associated with bovine leukemia virus infection. We isolated an ether-resistant internal antigen and an ether-sensitive antigen previously described in relation to bovine leukemia virus infection. These two antigens have now been isolated by isoelectric focusing and concanavalin A affinity chromatography, respectively. The ether-resistant antigen exhibited isoelectric heterogeneity with a major peak at pH 7.2 and a minor peak at pH 6.2. Its molecular weight, estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), was 23,000 (p23), and it gave a sedimentation value of 2.3s. For material containing ether-sensitive antigen, analyzed by SDS-PAGE, protein staining revealed four components with molecular weights of 18,000, 25,000, 45,000, and 55,000. Two of these [45,000 (gp45) and 55,000 (gp55)] were stained by periodic acid-Schiff reagent. Isoelectric point and sedimentation value of the major glycoprotein (gp45) were pH 5.0 and 3.4s, respectively; no immunologic cross-reactivity was found between p23 and glycoprotein antigen. | [
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PMID:10448 | Glutathione and gamma glutamyl transpeptidase in rat liver during chemical carcinogenesis. | Continued administration of several hepatocarcinogens led to an increase in the concentration of glutathione (GSH) in the livers of intact, but not of hypophysectomized or adrenalectomized rats. The concentration of GSH remained high untill the development of hyperplastic nodules. Subsequently, the concentration of GSH dropped to the normal level or below. A single dose of 3'-methyl-4-dimethylaminoazobenzene (3'-Me-DAB) produced an increase of GSH which, within a certain range, depended upon the amount of the carcinogen. In well differentiated, slowly growing hepatomas, the concentration of GSH approached the level in normal adult rat liver. On the other hand, in nondifferentiated and rapidly growing hepatomas, GSH was only 30-40% of that in normal liver. The activity of gamma-glutamyl transpeptidase (GTase) increased within 24-48 hours after a single large dose of 3'-Me-DAB. Continued feeding of 3'-Me-DAB led to an exponential increase of GTase. During hepatocarcinogenesis, the level of GTase activity corresponded to the degree and size of pathologic changes produced in rat liver. Chloramphenicol partially inhibited the increase of GTase induced by 2-acetylaminofluorene. Pretreatment with 3-methylcholanthrene partially inhibited the increase of GTase that had been induced by a single dose of 3'-Me-DAB. Puromycin partially inhibited the increase of GTase induced by several doses of dimethylnitrosamine. These observations indicated a close connection between the activation of GTase and chemical carcinogenesis in rat liver. Measurements of GTase activity in 12 Morris hepatomas supported this conclusion; their GTase levels were greatly elevated compared with that in normal adult rat liver. | Glutathione and gamma glutamyl transpeptidase in rat liver during chemical carcinogenesis. Continued administration of several hepatocarcinogens led to an increase in the concentration of glutathione (GSH) in the livers of intact, but not of hypophysectomized or adrenalectomized rats. The concentration of GSH remained high untill the development of hyperplastic nodules. Subsequently, the concentration of GSH dropped to the normal level or below. A single dose of 3'-methyl-4-dimethylaminoazobenzene (3'-Me-DAB) produced an increase of GSH which, within a certain range, depended upon the amount of the carcinogen. In well differentiated, slowly growing hepatomas, the concentration of GSH approached the level in normal adult rat liver. On the other hand, in nondifferentiated and rapidly growing hepatomas, GSH was only 30-40% of that in normal liver. The activity of gamma-glutamyl transpeptidase (GTase) increased within 24-48 hours after a single large dose of 3'-Me-DAB. Continued feeding of 3'-Me-DAB led to an exponential increase of GTase. During hepatocarcinogenesis, the level of GTase activity corresponded to the degree and size of pathologic changes produced in rat liver. Chloramphenicol partially inhibited the increase of GTase induced by 2-acetylaminofluorene. Pretreatment with 3-methylcholanthrene partially inhibited the increase of GTase that had been induced by a single dose of 3'-Me-DAB. Puromycin partially inhibited the increase of GTase induced by several doses of dimethylnitrosamine. These observations indicated a close connection between the activation of GTase and chemical carcinogenesis in rat liver. Measurements of GTase activity in 12 Morris hepatomas supported this conclusion; their GTase levels were greatly elevated compared with that in normal adult rat liver. | [
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PMID:10449 | Cyclic nucleotides in the regulation of expression of differentiated functions in neuroblastoma cells. | Adenosine 3',5'-cyclic monophosphate (cAMP) may be one of the important factors in regulating the expression of many differentiated functions in neuroblastoma cells, but some of these functions can be induced by agents that do not increase the intracellular level of cAMP. An elevation of the intracellular level of guanosine 3',5'-cyclic monophosphate (cGMP) neither induced differentiation nor antagonized the effects of cAMP. Neuroblastoma cells increased the level of cAMP-binding proteins during differentiation, whereas glial cells and L-cells did not. This might have accounted in part for an increase in the intracellular level of cAMP even in the presence of high phosphodiesterase activity in neuroblastoma cells, since the protein-bound with the same proteins, but cAMP had about 10 times higher affinity than did cGMP. cAMP promoted the organization of microtubules and microfilaments necessary for the expression of differentiated phenotypes. The extension of neurites required the synthesis of new protein, but it did not need the synthesis of new RNA. cAMP induced differentiation in neuroblastoma cells by increasing the expression of some genetic information while suppressing the expression of others; e.g., the activities of neural enzymes increased, whereas the synthesis of histone and the phosphorylation of H1-histone markedly decreased in differentiated cells. A hypothesis was offered: An increase in cAMP phosphodiesterase activity as a result of mutation in the regulatory gene for phosphodiesterase in a single, or group of, dividing nerve cell(s) is the primary lesion that leads to malignancy. Based on the concept that selective cytocytoxic drugs should be used with agents that cause differentiation, a new therapeutic approach was suggested for the treatment of neuroblastoma. This involved administration of sodium butyrate followed by L-DOPA or prostaglandin E1 in the presence of cAMP phosphodiesterase inhibitor followed by the less immunosuppressive vincristine and 5-(3,3-dimethyl-1-triazeno)imidazole-4-carboxamide. | Cyclic nucleotides in the regulation of expression of differentiated functions in neuroblastoma cells. Adenosine 3',5'-cyclic monophosphate (cAMP) may be one of the important factors in regulating the expression of many differentiated functions in neuroblastoma cells, but some of these functions can be induced by agents that do not increase the intracellular level of cAMP. An elevation of the intracellular level of guanosine 3',5'-cyclic monophosphate (cGMP) neither induced differentiation nor antagonized the effects of cAMP. Neuroblastoma cells increased the level of cAMP-binding proteins during differentiation, whereas glial cells and L-cells did not. This might have accounted in part for an increase in the intracellular level of cAMP even in the presence of high phosphodiesterase activity in neuroblastoma cells, since the protein-bound with the same proteins, but cAMP had about 10 times higher affinity than did cGMP. cAMP promoted the organization of microtubules and microfilaments necessary for the expression of differentiated phenotypes. The extension of neurites required the synthesis of new protein, but it did not need the synthesis of new RNA. cAMP induced differentiation in neuroblastoma cells by increasing the expression of some genetic information while suppressing the expression of others; e.g., the activities of neural enzymes increased, whereas the synthesis of histone and the phosphorylation of H1-histone markedly decreased in differentiated cells. A hypothesis was offered: An increase in cAMP phosphodiesterase activity as a result of mutation in the regulatory gene for phosphodiesterase in a single, or group of, dividing nerve cell(s) is the primary lesion that leads to malignancy. Based on the concept that selective cytocytoxic drugs should be used with agents that cause differentiation, a new therapeutic approach was suggested for the treatment of neuroblastoma. This involved administration of sodium butyrate followed by L-DOPA or prostaglandin E1 in the presence of cAMP phosphodiesterase inhibitor followed by the less immunosuppressive vincristine and 5-(3,3-dimethyl-1-triazeno)imidazole-4-carboxamide. | [
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PMID:10450 | Catecholamine metabolism in neuroblastoma. | Previous studies indicating the importance of catecholamine metabolism in neuroblastoma were briefly reviewed. Metabolic pathways were presented showing how the major urinary metabolites 3-methoxy-4-hydroxymandelic acid (VMA) and 3-methoxy-4-hydroxy-phenylacetic acid (HVA) are formed from norepinephrine and from dopamine plus 3,4-dihydroxyphenylalanine (DOPA), respectively. For 289 neuroblastoma patients at the time of diagnosis, the urinary excretion of VMA was significantly elevated in 75%, and HVA was elevated in 80%. Periodic assay of these metabolites during the course of the disease revealed that the excretion trends were of prognostic value with 80-90% reliability. By contrast, when the excretion in only the initial urine specimens was considered, the survival rate was the same for patients with normal, and with significantly elevated, excretion. Review of the results of tracer studies aimed at elucidating the in vivo metabolic origins of the urinary metabolites suggested that a) in neuroblastoma, the catecholamines were largely inactivated by intracellular metabolism in the tumor cells; b) there was excess production and excretion of the norepinephrine precursors, DOPA and dopamine; and c) in the tumors of most neuroblastoma patients, the initial enzyme in catecholamine synthesis, tyrosine hydroxylase, had an activity comparable with that in normal adrenal glands. The importance of the metabolism of catecholamines in patients with neuroblastoma was stressed: a) The excretion of elevated levels of urinary catecholamine metabolites were useful in diagnosis and in following the course of the disease, and b) study of the catecholamine metabolism in these patients permitted examination of possible relationships between the activity of the enzymes involved in catecholamine synthesis and the malignancy of this tumor. | Catecholamine metabolism in neuroblastoma. Previous studies indicating the importance of catecholamine metabolism in neuroblastoma were briefly reviewed. Metabolic pathways were presented showing how the major urinary metabolites 3-methoxy-4-hydroxymandelic acid (VMA) and 3-methoxy-4-hydroxy-phenylacetic acid (HVA) are formed from norepinephrine and from dopamine plus 3,4-dihydroxyphenylalanine (DOPA), respectively. For 289 neuroblastoma patients at the time of diagnosis, the urinary excretion of VMA was significantly elevated in 75%, and HVA was elevated in 80%. Periodic assay of these metabolites during the course of the disease revealed that the excretion trends were of prognostic value with 80-90% reliability. By contrast, when the excretion in only the initial urine specimens was considered, the survival rate was the same for patients with normal, and with significantly elevated, excretion. Review of the results of tracer studies aimed at elucidating the in vivo metabolic origins of the urinary metabolites suggested that a) in neuroblastoma, the catecholamines were largely inactivated by intracellular metabolism in the tumor cells; b) there was excess production and excretion of the norepinephrine precursors, DOPA and dopamine; and c) in the tumors of most neuroblastoma patients, the initial enzyme in catecholamine synthesis, tyrosine hydroxylase, had an activity comparable with that in normal adrenal glands. The importance of the metabolism of catecholamines in patients with neuroblastoma was stressed: a) The excretion of elevated levels of urinary catecholamine metabolites were useful in diagnosis and in following the course of the disease, and b) study of the catecholamine metabolism in these patients permitted examination of possible relationships between the activity of the enzymes involved in catecholamine synthesis and the malignancy of this tumor. | [
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PMID:10451 | Exonuclease associated with bacteriophage T5-Induced DNA polymerase. | T-5-induced DNA polymerase has been shown to possess a 3' leads to 5'-exonucleolytic activity. The exonuclease acts on both native and denatured DNA, but the apparent rate of degradation of denatured DNA is about five times faster than that for native DNA. The enzyme appears to act only on 3'-OH ends and produces mainly 5'-dNMP's. Like polymerase activity, exonuclease activity shows a pH optimum around 8.6. Mg2+, dithiothreitol, and N-ethylmaleimide had identical effects on both the activities. Nicked DNA was almost totally protected from exonuclease action under synthetic conditions, i.e., in the presence of 4dNTP's. Denatured DNA was partly degraded in the early phase of incubation with 4dNTP's, presumably due to unhybridized tails at the 3'-OH primer ends. However, the exonuclease activity was operative in both cases under synthetic conditions, as evidenced by template-dependent conversion of [3H]dTTP to [3H]dTMP. | Exonuclease associated with bacteriophage T5-Induced DNA polymerase. T-5-induced DNA polymerase has been shown to possess a 3' leads to 5'-exonucleolytic activity. The exonuclease acts on both native and denatured DNA, but the apparent rate of degradation of denatured DNA is about five times faster than that for native DNA. The enzyme appears to act only on 3'-OH ends and produces mainly 5'-dNMP's. Like polymerase activity, exonuclease activity shows a pH optimum around 8.6. Mg2+, dithiothreitol, and N-ethylmaleimide had identical effects on both the activities. Nicked DNA was almost totally protected from exonuclease action under synthetic conditions, i.e., in the presence of 4dNTP's. Denatured DNA was partly degraded in the early phase of incubation with 4dNTP's, presumably due to unhybridized tails at the 3'-OH primer ends. However, the exonuclease activity was operative in both cases under synthetic conditions, as evidenced by template-dependent conversion of [3H]dTTP to [3H]dTMP. | [
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PMID:10452 | Structural proteins of polyoma virus: proteolytic degradation of virion proteins by exogenous and by virion-associated proteases. | A model has previously been proposed for the genetic relatedness of the structural proteins of polyoma virus, based upon similarities in the peptide maps of the major capsid protein VP1 with the virion proteins VP2 and VP3. Newer evidence suggests that this model is incorrect, and that protein VP1 is a product of one viral gene and that the multiple components of VP2 and VP3 are products of a second viral gene. Two-dimensional peptide maps of several preparations of polyoma purified separately from four separate infected-cell lysates has shown a variable content of VP1 peptides in proteins VP2 and VP3, with some preparations being free of detectable VP1 material in VP2 and VP3. An alternative explanation for the presence of VP1 peptides in the regions of VP2 and VP3 of some polyoma preparations involves the cleavage of proteins of polyoma virions during exposure to proteolytic enzymes in lysates of infected cells or to endogenous proteolytic activity of virions. Prolonged incubation of infected-cell lysates at 37 degrees C leads to an increase in the amount of 86,000-dalton dimer of VP1, a decrease in the relative amount of VP1, a decrease in or a loss of the lower band of VP2, and the appearance of a new major protein band of approximately 29,000 daltons. Two-dimensional peptide maps of the new 29,000-dalton protein show that it contains some VP1 peptides, indicating that this protein is derived from proteolytic cleavage of VP1. In addition, extensively purified polyoma virus contains a proteolytic activity that can be activated during disruption of the virus with 0.2 M Na2CO3-NaHCO3 (pH 10.6) in the presence of 5 X 10(-3) M dithiothreitol. | Structural proteins of polyoma virus: proteolytic degradation of virion proteins by exogenous and by virion-associated proteases. A model has previously been proposed for the genetic relatedness of the structural proteins of polyoma virus, based upon similarities in the peptide maps of the major capsid protein VP1 with the virion proteins VP2 and VP3. Newer evidence suggests that this model is incorrect, and that protein VP1 is a product of one viral gene and that the multiple components of VP2 and VP3 are products of a second viral gene. Two-dimensional peptide maps of several preparations of polyoma purified separately from four separate infected-cell lysates has shown a variable content of VP1 peptides in proteins VP2 and VP3, with some preparations being free of detectable VP1 material in VP2 and VP3. An alternative explanation for the presence of VP1 peptides in the regions of VP2 and VP3 of some polyoma preparations involves the cleavage of proteins of polyoma virions during exposure to proteolytic enzymes in lysates of infected cells or to endogenous proteolytic activity of virions. Prolonged incubation of infected-cell lysates at 37 degrees C leads to an increase in the amount of 86,000-dalton dimer of VP1, a decrease in the relative amount of VP1, a decrease in or a loss of the lower band of VP2, and the appearance of a new major protein band of approximately 29,000 daltons. Two-dimensional peptide maps of the new 29,000-dalton protein show that it contains some VP1 peptides, indicating that this protein is derived from proteolytic cleavage of VP1. In addition, extensively purified polyoma virus contains a proteolytic activity that can be activated during disruption of the virus with 0.2 M Na2CO3-NaHCO3 (pH 10.6) in the presence of 5 X 10(-3) M dithiothreitol. | [
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PMID:10455 | Purification and properties of glucose-6-phosphate dehydrogenase from Bacillus subtilis spores. | Glucose-6-phosphate dehydrogenase [D-glucose-6-phosphate: NADP oxidoreductase, EC. 1. 1. 1. 49] obtained from spores of Bacillus subtilis PCI 219 strain was partially purified by filtration on Sephadex G-200, ammonium sulfate fractionation and chromatography on DEAE-Sephadex A-25 (about 54-fold). The optimum pH for stability of this enzyme was about 6.3 and the optimum pH for the reaction about 8.3. The apparent Km values of the enzyme were 5.7 X 10(-4) M for glucose-6-phosphate and 2.4 X 10(-4) M for nicotinamide adenine dinucleotide phosphate (NADP). The isoelectric point was about pH 3.9. The enzyme activity was unaffected by the addition of Mg++ or Ca++. The inactive glucose-6-phosphate dehydrogenase obtained from the spores heated at 85 C for 30 min was not reactivated by the addition of ethylenediaminetetraacetic acid, dipicolinic acid or some salts unlike inactive glucose dehydrogenase. | Purification and properties of glucose-6-phosphate dehydrogenase from Bacillus subtilis spores. Glucose-6-phosphate dehydrogenase [D-glucose-6-phosphate: NADP oxidoreductase, EC. 1. 1. 1. 49] obtained from spores of Bacillus subtilis PCI 219 strain was partially purified by filtration on Sephadex G-200, ammonium sulfate fractionation and chromatography on DEAE-Sephadex A-25 (about 54-fold). The optimum pH for stability of this enzyme was about 6.3 and the optimum pH for the reaction about 8.3. The apparent Km values of the enzyme were 5.7 X 10(-4) M for glucose-6-phosphate and 2.4 X 10(-4) M for nicotinamide adenine dinucleotide phosphate (NADP). The isoelectric point was about pH 3.9. The enzyme activity was unaffected by the addition of Mg++ or Ca++. The inactive glucose-6-phosphate dehydrogenase obtained from the spores heated at 85 C for 30 min was not reactivated by the addition of ethylenediaminetetraacetic acid, dipicolinic acid or some salts unlike inactive glucose dehydrogenase. | [
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PMID:10456 | A new analgesic testing method using ultrasonic stimulation. I. Effects of narcotic and nonnarcotic analgesics. | A quantitative method for measuring pain threshold by the use of ultrasonic stimulation in mice has been designed. The method had the advantage of precision, simplicity of technique, rapidity of measurement, and the fact that the stimuli is innocuous upon repeated application. The nature of the senstaions induced by ultrasonic stimulus is somewhat like that felt with a prick type of pain. Pentazocine (30, 100, 150 mg/kg i.p.) aminopyrine (15,50, 100, 150 mg/kg i.p.), phenacetin (100,150, 200, 250 mg/kg i.p.) sodium salicylate (150, 200, 250 mg/kg i.p.) and other antipyretic analgesics were active in a wide range of doses indicating that this technique is sensitive to the narcotic antagonist and to the weak analgesics as well as to the narcotic analgesics as well as to the narcotic analgesics such as morphine (2.5, 5, 10, 15 mg/kg i.p.), codeine (10, 20, 25, 30, 50 mg/kg i.p.) and pethidine (5,10, 15, 20, 25 mg/kg i.p.). The ultrasonic method is, therefore, applicable in screening procedures when attempting to evaluate the analgesic potency of a wide variety of chemical agents. | A new analgesic testing method using ultrasonic stimulation. I. Effects of narcotic and nonnarcotic analgesics. A quantitative method for measuring pain threshold by the use of ultrasonic stimulation in mice has been designed. The method had the advantage of precision, simplicity of technique, rapidity of measurement, and the fact that the stimuli is innocuous upon repeated application. The nature of the senstaions induced by ultrasonic stimulus is somewhat like that felt with a prick type of pain. Pentazocine (30, 100, 150 mg/kg i.p.) aminopyrine (15,50, 100, 150 mg/kg i.p.), phenacetin (100,150, 200, 250 mg/kg i.p.) sodium salicylate (150, 200, 250 mg/kg i.p.) and other antipyretic analgesics were active in a wide range of doses indicating that this technique is sensitive to the narcotic antagonist and to the weak analgesics as well as to the narcotic analgesics as well as to the narcotic analgesics such as morphine (2.5, 5, 10, 15 mg/kg i.p.), codeine (10, 20, 25, 30, 50 mg/kg i.p.) and pethidine (5,10, 15, 20, 25 mg/kg i.p.). The ultrasonic method is, therefore, applicable in screening procedures when attempting to evaluate the analgesic potency of a wide variety of chemical agents. | [
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PMID:10457 | Studies on the mode of antagonism between adrenergic beta-mimetics and beta-blocking agents (III). Functional antagonism between beta-mimetics and spasmogens. | New equations which can explain the following characteristic phenomena in the functional antagonism between isoproterenol (ISO) and spasmogens in guinea-pig trachea are proposed: (1) The amplitude of relaxation of the muscle induced by ISO varied depending on the concentration of a spasmogen used (histamine or carbachol). (2) The dose-response curves for the relaxation by ISO shifted to the right in a parallel manner as the concentration of the spasmogen increased, and became stationary at high concentrations of the spasmogen. (3) The slope of the dose-response curve became steeper with increasing concentrations of the spasmogen. When the saturable uptake process of ISO was taken into consideration, a satisfactory parallel was seen between the theoretical dose-response curves and the present experimental results. | Studies on the mode of antagonism between adrenergic beta-mimetics and beta-blocking agents (III). Functional antagonism between beta-mimetics and spasmogens. New equations which can explain the following characteristic phenomena in the functional antagonism between isoproterenol (ISO) and spasmogens in guinea-pig trachea are proposed: (1) The amplitude of relaxation of the muscle induced by ISO varied depending on the concentration of a spasmogen used (histamine or carbachol). (2) The dose-response curves for the relaxation by ISO shifted to the right in a parallel manner as the concentration of the spasmogen increased, and became stationary at high concentrations of the spasmogen. (3) The slope of the dose-response curve became steeper with increasing concentrations of the spasmogen. When the saturable uptake process of ISO was taken into consideration, a satisfactory parallel was seen between the theoretical dose-response curves and the present experimental results. | [
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PMID:10458 | Studies on the mode of antagonism between adrenergic beta-mimetics and beta-blocking agents (IV). Influence of functional antagonism by spasmogens. | A new theory is presented to describe the effect of functional antagonism on the competitive antagonism between adrenergic beta-mimetics and beta-blockers. According to this theory the shape of the log (dose ratio-1) vs.--log [B] curve and the apparent pA2 value in competitive antagonism should be affected by functional antagonism when the agonist is taken up by the saturable uptake process, and this was experimentally confirmed. The competitive antagonism between isoproterenol (ISO) and propranolol (Prop) was influenced by the functional antagonism between ISO and spasmogens (histamine and carbachol). The log (dose ratio-1) vs. --log [B] curve is ISO-Prop competitive antagonism was shifted variously depending on the concentration of a spasmogen used. Theoretical predictions and experimental results were in good parallel. | Studies on the mode of antagonism between adrenergic beta-mimetics and beta-blocking agents (IV). Influence of functional antagonism by spasmogens. A new theory is presented to describe the effect of functional antagonism on the competitive antagonism between adrenergic beta-mimetics and beta-blockers. According to this theory the shape of the log (dose ratio-1) vs.--log [B] curve and the apparent pA2 value in competitive antagonism should be affected by functional antagonism when the agonist is taken up by the saturable uptake process, and this was experimentally confirmed. The competitive antagonism between isoproterenol (ISO) and propranolol (Prop) was influenced by the functional antagonism between ISO and spasmogens (histamine and carbachol). The log (dose ratio-1) vs. --log [B] curve is ISO-Prop competitive antagonism was shifted variously depending on the concentration of a spasmogen used. Theoretical predictions and experimental results were in good parallel. | [
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PMID:10460 | HLA antigens in arterial occlusive diseases in Japan. | Using a NIH standard lymphocytotoxicity test, a possible Japanese specific HLA antigen, HLA-BJW 22.2 was identified in 17 out of 48 patients with thromboangiitis obliterans (35.4 per cent), in 5 out of 15 patients with Takayasu's arteritis (33.3 per cent) and in 11 out of 113 normal controls (9.7 per cent). On the other hand, HLA-CWl was found in 4 out of 47 patients with arteriosclerosis obliterans (8.5 per cent) and in 41 out of 113 normal controls (36.3 per cent). | HLA antigens in arterial occlusive diseases in Japan. Using a NIH standard lymphocytotoxicity test, a possible Japanese specific HLA antigen, HLA-BJW 22.2 was identified in 17 out of 48 patients with thromboangiitis obliterans (35.4 per cent), in 5 out of 15 patients with Takayasu's arteritis (33.3 per cent) and in 11 out of 113 normal controls (9.7 per cent). On the other hand, HLA-CWl was found in 4 out of 47 patients with arteriosclerosis obliterans (8.5 per cent) and in 41 out of 113 normal controls (36.3 per cent). | [
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PMID:10463 | [Comparative evaluation of the clinical action of a series of beta-adrenergic blockaders]. | A comparative clinical study of the efficacy of Benzoral, Trasicor, Viskene, Aptene, Eraldine and Inderal was conducted in the ischaemic heart disease patients. Their antiarrhythmic and antianginal effect was determined, as well as their optimum therapeutic dosages, the activity of their specific beta-adrenolytic properties, the effect of the drugs on the bronchi and the peripheral venous tone. Apart from the clinical study, electro- and polycardiography, functional pulmonary tests and the Schellong orthostatic test were used. All the drugs in question were found to produce a distinct specific beta-blocking effect. They are effective in cases of atrial and ventricular extrasystole, paroxysmal tachycardia, sinus tachycardia and tachyarrhythmic fibrillation, as well as for the prevention of anginal attacks and arrhythmic fibrillation. All the drugs produce a negative inotropic effect, Inderal--the strongest, Viskene and Benzoral--the weakest. All beta-blockers can impair bronchial patency in patients with bronchial obstruction. This effect is least pronounced with Eraldine that may be used as the drug of choice in such cases. In most cases the beta-blockers do not affect the peripheral venous tone, but in some cases they may reduce it. | [Comparative evaluation of the clinical action of a series of beta-adrenergic blockaders]. A comparative clinical study of the efficacy of Benzoral, Trasicor, Viskene, Aptene, Eraldine and Inderal was conducted in the ischaemic heart disease patients. Their antiarrhythmic and antianginal effect was determined, as well as their optimum therapeutic dosages, the activity of their specific beta-adrenolytic properties, the effect of the drugs on the bronchi and the peripheral venous tone. Apart from the clinical study, electro- and polycardiography, functional pulmonary tests and the Schellong orthostatic test were used. All the drugs in question were found to produce a distinct specific beta-blocking effect. They are effective in cases of atrial and ventricular extrasystole, paroxysmal tachycardia, sinus tachycardia and tachyarrhythmic fibrillation, as well as for the prevention of anginal attacks and arrhythmic fibrillation. All the drugs produce a negative inotropic effect, Inderal--the strongest, Viskene and Benzoral--the weakest. All beta-blockers can impair bronchial patency in patients with bronchial obstruction. This effect is least pronounced with Eraldine that may be used as the drug of choice in such cases. In most cases the beta-blockers do not affect the peripheral venous tone, but in some cases they may reduce it. | [
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PMID:10464 | [Change in the central and regional (brain) hemodynamics in the treatment of hypertension using beta-adrenergic blockaders]. | The haemodynamic parameters were studied dynamically by way of noninvasive techniques--tetrapolar thoracic rheography in the course of treatment of 48 patients with essential hypertension Stage IB and IIA urth beta-blocking agents Obsidan and Visken. It was found that as early as by the 5th day of therapy the stroke volume and heart rate decreased, the maximum fall being noted after 2-3 weeks of treatment. The reflex elevation of the total peripheral resistance was moderate, as a result of which a gradual reduction of the arterial pressure developed. The state of the cerebral circulation significantly improved in the course of the treatment (as shown by rheoencephalography) and the tone of the cerebral arteries decreased. After 2-3 weeks of beta-blockers therapy favourable shifts were observed in the response of the general and cerebral haemodynamics to the orthostatic test. | [Change in the central and regional (brain) hemodynamics in the treatment of hypertension using beta-adrenergic blockaders]. The haemodynamic parameters were studied dynamically by way of noninvasive techniques--tetrapolar thoracic rheography in the course of treatment of 48 patients with essential hypertension Stage IB and IIA urth beta-blocking agents Obsidan and Visken. It was found that as early as by the 5th day of therapy the stroke volume and heart rate decreased, the maximum fall being noted after 2-3 weeks of treatment. The reflex elevation of the total peripheral resistance was moderate, as a result of which a gradual reduction of the arterial pressure developed. The state of the cerebral circulation significantly improved in the course of the treatment (as shown by rheoencephalography) and the tone of the cerebral arteries decreased. After 2-3 weeks of beta-blockers therapy favourable shifts were observed in the response of the general and cerebral haemodynamics to the orthostatic test. | [
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PMID:10466 | [Assay of carbromal and its main metabolite (2-ethyl-butyryl-urea) in biological fluids (author's transl)]. | The materials mentioned above are reextracted from an ether solution by 2 N sodium hydroxide. The UV extinction of the aqueous layer at 232 nm and 30 degrees C is followed by means of a slave recorder. The original extinction is found by back-extrapolating to the time of re-extraction. It has to be corrected for the "biological matrix" by subtracting the extinction value after complete hydrolysis. Most interfering substances in the ether solution (carboxylic acids, barbiturates) may be removed previously by washing with aqueous buffers (pH 7.0 and 10.5). | [Assay of carbromal and its main metabolite (2-ethyl-butyryl-urea) in biological fluids (author's transl)]. The materials mentioned above are reextracted from an ether solution by 2 N sodium hydroxide. The UV extinction of the aqueous layer at 232 nm and 30 degrees C is followed by means of a slave recorder. The original extinction is found by back-extrapolating to the time of re-extraction. It has to be corrected for the "biological matrix" by subtracting the extinction value after complete hydrolysis. Most interfering substances in the ether solution (carboxylic acids, barbiturates) may be removed previously by washing with aqueous buffers (pH 7.0 and 10.5). | [
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PMID:10467 | Histamine H2-receptor antagonists and gastric acid secretion -- a progress report. | Histamine H2-receptor antagonists, including burimamide, metiamide and cimetidine, are effective antagonists of histamine-stimulated acid secretion from mammalian, avian or reptilian gastric mucosa. Acid secretion stimulated by gastrin or pentagastrin is also inhibited by these drugs, but there is disagreement about the effects of these drugs on acid secretion resulting from activation of acetylcholine receptors. Based on the pharmacological evidence possibilities of treatment by these drugs were discussed in cases with excessive stimulation of acid secretion due to high blood levels of histamine or gastrin. The positive results in several trials on Zollinger-Ellison syndrome and peptic ulcer were very impressive. Some practical problems have still to be solved, for example the appropriate phase for applying the drugs. The demonstrated clinical effectiveness, however, against peptic ulceration offers a clear alternative to surgery for many patients. | Histamine H2-receptor antagonists and gastric acid secretion -- a progress report. Histamine H2-receptor antagonists, including burimamide, metiamide and cimetidine, are effective antagonists of histamine-stimulated acid secretion from mammalian, avian or reptilian gastric mucosa. Acid secretion stimulated by gastrin or pentagastrin is also inhibited by these drugs, but there is disagreement about the effects of these drugs on acid secretion resulting from activation of acetylcholine receptors. Based on the pharmacological evidence possibilities of treatment by these drugs were discussed in cases with excessive stimulation of acid secretion due to high blood levels of histamine or gastrin. The positive results in several trials on Zollinger-Ellison syndrome and peptic ulcer were very impressive. Some practical problems have still to be solved, for example the appropriate phase for applying the drugs. The demonstrated clinical effectiveness, however, against peptic ulceration offers a clear alternative to surgery for many patients. | [
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PMID:10470 | [Selection of a urine preservative agent in relation to oxidative-catalytic methods of water regeneration]. | Various chemicals were assayed as possible urine conservers to the used in space water reclamation systems. With respect to the antimicrobial activity and the quality of reclaimed water formalin at a concentration of 0.1% was selected as the best conserver. The reagent ensures urine preservation for 3 months. | [Selection of a urine preservative agent in relation to oxidative-catalytic methods of water regeneration]. Various chemicals were assayed as possible urine conservers to the used in space water reclamation systems. With respect to the antimicrobial activity and the quality of reclaimed water formalin at a concentration of 0.1% was selected as the best conserver. The reagent ensures urine preservation for 3 months. | [
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PMID:10478 | [Hypoxia and polytrauma]. | The importance of polytrauma and hypoxia, resp., is discussed by the authors with regard to the mortality and the common effect of these is analysed. It was found that in the mortality of the patients suffered polytraumatism the first place (59,4%) is taken by such lesions, in which simultaneous lesion of the skull and the thorax occurs. The values of the blood gas and the acid-base balance of the patients who had suffered polytraumatism, were examined in the 12th--24th hours proceding the death. It was found that whereas--using the possibilities of the intensive therapy--the values of pH, paCO2 and BE could be kept in a part of the cases between the physiological limits--the paO2 value was under the normal value in all cases--moreover 2/3 of the values fell into the domain between 40--60 mmHg--signifying severe hypoxia. On the basis of the analysis it may be presumed that in the process leading to death of the patients who had suffered polytraumatism the anoxic hypoxy plays considerable role. This is supported also by the earlier data published by the institute, according to which the mortality in itself high--33.8%--of polytraumatism increases to 71.4%, if it is associated hypoxy. Consequently the aim of the therapy must be to exert the greatest activity in order to prevent or to combat, resp., the anoxic hypoxy. | [Hypoxia and polytrauma]. The importance of polytrauma and hypoxia, resp., is discussed by the authors with regard to the mortality and the common effect of these is analysed. It was found that in the mortality of the patients suffered polytraumatism the first place (59,4%) is taken by such lesions, in which simultaneous lesion of the skull and the thorax occurs. The values of the blood gas and the acid-base balance of the patients who had suffered polytraumatism, were examined in the 12th--24th hours proceding the death. It was found that whereas--using the possibilities of the intensive therapy--the values of pH, paCO2 and BE could be kept in a part of the cases between the physiological limits--the paO2 value was under the normal value in all cases--moreover 2/3 of the values fell into the domain between 40--60 mmHg--signifying severe hypoxia. On the basis of the analysis it may be presumed that in the process leading to death of the patients who had suffered polytraumatism the anoxic hypoxy plays considerable role. This is supported also by the earlier data published by the institute, according to which the mortality in itself high--33.8%--of polytraumatism increases to 71.4%, if it is associated hypoxy. Consequently the aim of the therapy must be to exert the greatest activity in order to prevent or to combat, resp., the anoxic hypoxy. | [
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PMID:10479 | [Classification and diagnosis of ankle injuries]. | A new method for the classification of the injuries of the ankle is recommended by the author. The main types according to his classifixation are the following: pronation-flexion, pronation-extension, supination-extension, supination-flexion and supination-extension types. His classification is compared with Lauge-Hansen's and Weber's classification. Critical analysis of these two last classifications is given. The aim of the author's classification is to render help to the doctors for their every-days' curative work. The characteristic symptoms of the pronation and supination, resp., injuries are described. Attention is drawn to "Weber's lace"--this denomination is proposed by the author, since the first description is due to Weber. On the basis of the author's examinations described in his candidate's dissertation "syndesmolysis trigonum"--pathognostic for syndesmolysis--is dealt with. The "reclined" roentgenograms are dealt with. The so-called pronation reclined roentgenogram visualize the rupture of the deltoid ligament and the syndesmolysis in the same time. The sagittal reclined roentgenogram is dealth with separately, by means of which the "table-drawer" symptom may be produced. | [Classification and diagnosis of ankle injuries]. A new method for the classification of the injuries of the ankle is recommended by the author. The main types according to his classifixation are the following: pronation-flexion, pronation-extension, supination-extension, supination-flexion and supination-extension types. His classification is compared with Lauge-Hansen's and Weber's classification. Critical analysis of these two last classifications is given. The aim of the author's classification is to render help to the doctors for their every-days' curative work. The characteristic symptoms of the pronation and supination, resp., injuries are described. Attention is drawn to "Weber's lace"--this denomination is proposed by the author, since the first description is due to Weber. On the basis of the author's examinations described in his candidate's dissertation "syndesmolysis trigonum"--pathognostic for syndesmolysis--is dealt with. The "reclined" roentgenograms are dealt with. The so-called pronation reclined roentgenogram visualize the rupture of the deltoid ligament and the syndesmolysis in the same time. The sagittal reclined roentgenogram is dealth with separately, by means of which the "table-drawer" symptom may be produced. | [
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PMID:10480 | [The effect of ligament calcification following syndesmolysis on pseudoarthrosis of the median malleolus]. | The development of the false joint of the medial malleolus of 16 patients has been followed by the authors at the least for 13 years. It was found that in the cases, in which the fracture of the medial malleolus was concomitant with syndesmolysis, recovery of the false joint occured in the cases, in which in the place of the syndesmolysis calcification appeared, which brought the lateral cutting effect in the talo-crural joint to an end. | [The effect of ligament calcification following syndesmolysis on pseudoarthrosis of the median malleolus]. The development of the false joint of the medial malleolus of 16 patients has been followed by the authors at the least for 13 years. It was found that in the cases, in which the fracture of the medial malleolus was concomitant with syndesmolysis, recovery of the false joint occured in the cases, in which in the place of the syndesmolysis calcification appeared, which brought the lateral cutting effect in the talo-crural joint to an end. | [
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PMID:10481 | [Conservative and surgical treatment of open fractures of the forearm]. | On the basis of the 10 years' material of the Hugarian National Institute of Traumatology the possibilities of treatment of the open fractures of the forearm are discussed. It is stated by the authors that operative treatment of the open fractures of the forearm is more and more frequent, and--chiefly in the last time--the surgeon strives to obtain rigid internal fixation. If the individual and objective conditions of the internal fixation are not given or the operation seems risky for other reasons--intramedullary splinting--completing the conservative treatment--or wire fixation are the method of choice. In connection with fractures of different type the authors' method is discussed and a few cases are reported. | [Conservative and surgical treatment of open fractures of the forearm]. On the basis of the 10 years' material of the Hugarian National Institute of Traumatology the possibilities of treatment of the open fractures of the forearm are discussed. It is stated by the authors that operative treatment of the open fractures of the forearm is more and more frequent, and--chiefly in the last time--the surgeon strives to obtain rigid internal fixation. If the individual and objective conditions of the internal fixation are not given or the operation seems risky for other reasons--intramedullary splinting--completing the conservative treatment--or wire fixation are the method of choice. In connection with fractures of different type the authors' method is discussed and a few cases are reported. | [
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PMID:10477 | Helix-coil stability constants for the naturally occurring amino acids in water. 11. Lysine parameters from random poly(hydroxybutylglutamine-co-L-lysine). | The synthesis and characterization of water-soluble random copolymers containing L-lysine with N5-(4-hydroxybutyl)-L-glutamine, and the thermally induced helix-coil transitions of these copolymers in water, are described. The incorporation of L-lysine was found to decrease the helix content of the polymers at neutral pH. The Zimm-Bragg parameters sigma and s for the helix-coil transition in poly(L-lysine) in water were deduced from an analysis of the melting curves in the manner described in earlier papers. The computed values of s indicate that, in the temperature range of 0-60 degrees C, lysine has a tendency to destabilize helical sequences, this tendency being minimal at approximately 25 degrees C and increasing at lower and higher temperatures. | Helix-coil stability constants for the naturally occurring amino acids in water. 11. Lysine parameters from random poly(hydroxybutylglutamine-co-L-lysine). The synthesis and characterization of water-soluble random copolymers containing L-lysine with N5-(4-hydroxybutyl)-L-glutamine, and the thermally induced helix-coil transitions of these copolymers in water, are described. The incorporation of L-lysine was found to decrease the helix content of the polymers at neutral pH. The Zimm-Bragg parameters sigma and s for the helix-coil transition in poly(L-lysine) in water were deduced from an analysis of the melting curves in the manner described in earlier papers. The computed values of s indicate that, in the temperature range of 0-60 degrees C, lysine has a tendency to destabilize helical sequences, this tendency being minimal at approximately 25 degrees C and increasing at lower and higher temperatures. | [
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PMID:10483 | [Substitution of the frontal bone with silicon]. | The good results obtained with silicon implants used in the frontal region are reported by the authors. After the correct primary treatment the reconstructive operations have been performed at the least 6 months after the lesion, if the environment of the defect was free from any reaction during this time. 14 implantations have been performed in 2 years. Satisfactory functional and cosmetic results were obtained and the implant was to be removed in no case. | [Substitution of the frontal bone with silicon]. The good results obtained with silicon implants used in the frontal region are reported by the authors. After the correct primary treatment the reconstructive operations have been performed at the least 6 months after the lesion, if the environment of the defect was free from any reaction during this time. 14 implantations have been performed in 2 years. Satisfactory functional and cosmetic results were obtained and the implant was to be removed in no case. | [
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PMID:10484 | [Treatment of injuries of the penis and scrotum complicated by total skin loss]. | In connection with two healed cases the free transplantation of semi-thick skin is recommended by the authors for the treatment of the above-mentioned injuries, on the basis of the following reasons: 1. The most ideal results may be expected from this method. 2. The objective conditions (dermatome, Humbey's knife, sponge etc.) age generally given. 3. The semi-thick skin plasty is one of the mostly used and well-known methods. 4. In the case of failure the method may be repeated or other method may be chosen. 5. Semi-thick skin plasty is the easiest way to observe the principle: "Nil nocere". | [Treatment of injuries of the penis and scrotum complicated by total skin loss]. In connection with two healed cases the free transplantation of semi-thick skin is recommended by the authors for the treatment of the above-mentioned injuries, on the basis of the following reasons: 1. The most ideal results may be expected from this method. 2. The objective conditions (dermatome, Humbey's knife, sponge etc.) age generally given. 3. The semi-thick skin plasty is one of the mostly used and well-known methods. 4. In the case of failure the method may be repeated or other method may be chosen. 5. Semi-thick skin plasty is the easiest way to observe the principle: "Nil nocere". | [
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PMID:10485 | [Acute arteria mesenterica superior syndrome caused by severe electric injury]. | The case of acute arteria mesenterica superior syndrome--occurred in a boy aged 10, as rare complication of high voltage electro-trauma--is reported. The aetiology of the syndrome, as well as a possible new pathogenic factor are discussed. The clinical picture and the therapeutic principles are dealt with. | [Acute arteria mesenterica superior syndrome caused by severe electric injury]. The case of acute arteria mesenterica superior syndrome--occurred in a boy aged 10, as rare complication of high voltage electro-trauma--is reported. The aetiology of the syndrome, as well as a possible new pathogenic factor are discussed. The clinical picture and the therapeutic principles are dealt with. | [
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PMID:10486 | [Substitution of an injured finger joint by silicone rubber prosthesis]. | Between 1973 and the first semester of 1975, on the injured and destroyed digital articulation 27 patients have been operated on by means silicon rubber prosthesis of Swanson type. The prosthesis plasty and the field of its use are shortly resumed, pointing out the importance of its adaptation on the injured articulation. The operative technique is described, as well as the results obtained and the aspects of the evaluation are discussed. The results obtained with the silicon rubber prosthesis are very promising. In the case of failure arthrodesis may be performed at any time. | [Substitution of an injured finger joint by silicone rubber prosthesis]. Between 1973 and the first semester of 1975, on the injured and destroyed digital articulation 27 patients have been operated on by means silicon rubber prosthesis of Swanson type. The prosthesis plasty and the field of its use are shortly resumed, pointing out the importance of its adaptation on the injured articulation. The operative technique is described, as well as the results obtained and the aspects of the evaluation are discussed. The results obtained with the silicon rubber prosthesis are very promising. In the case of failure arthrodesis may be performed at any time. | [
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] |
PMID:10487 | [Phlebography of the talus following fracture and dislocation (preliminary report)]. | After dislocation or dislocated fracture of the talus late arthrosis is a frequent complication, in which primary bone and cartilage damage play a role. For the demonstration and preclusion, resp., of the vascular damage phlebography performed 3--6 months after the injury is recommended by the authors--by means of which favourable initial experiences could be obtained. | [Phlebography of the talus following fracture and dislocation (preliminary report)]. After dislocation or dislocated fracture of the talus late arthrosis is a frequent complication, in which primary bone and cartilage damage play a role. For the demonstration and preclusion, resp., of the vascular damage phlebography performed 3--6 months after the injury is recommended by the authors--by means of which favourable initial experiences could be obtained. | [
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PMID:10490 | Pharmacotherapy of myocardial ischemia. | The cornerstones of pharmacotherapy for myocardial ischemia are the nitrites and the beta-adrenergic blocking agents. These drugs not only inhibit cardiac mechanical activity (and therefore energy requirements) in a variety of ways but also redistribute available blood flow to the potentially ischemic segments of cardiac muscle. The least effective dose of nitroglycerin and the most tolerated (or blocking) dose of propranolol provide the optimum in management. There is increasing evidence that certain orally administered nitrates at larger than usual dosage can further increase the tolerance to effort. Amelioration of hypertension or congestive failure may play a significant role in selected patients. | Pharmacotherapy of myocardial ischemia. The cornerstones of pharmacotherapy for myocardial ischemia are the nitrites and the beta-adrenergic blocking agents. These drugs not only inhibit cardiac mechanical activity (and therefore energy requirements) in a variety of ways but also redistribute available blood flow to the potentially ischemic segments of cardiac muscle. The least effective dose of nitroglycerin and the most tolerated (or blocking) dose of propranolol provide the optimum in management. There is increasing evidence that certain orally administered nitrates at larger than usual dosage can further increase the tolerance to effort. Amelioration of hypertension or congestive failure may play a significant role in selected patients. | [
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PMID:10491 | Principles in selection of therapy. | The physician today is presented with a plethora of possibilities in the therapy of each of the aspects of ischemic heart disease (Fig. 15-5). There is the temptation to recommend complex and impossible dietary prescriptions coupled with several pharmaceutical agents for control of anginal pain, hypertension, arrhythmias, hypercholesterolemia, and clinical congestive heart failure. While each of the objectives may be in part valid, the burden on the patient of following such a constraining and difficult life may make it virtually impossible either to enjoy life or to follow the physician's recommendations explicitly. Often a compromise must be reached between theoretically optimal therapy and that which is reasonable and acceptable to the patient. Again, a review of each aspect of the program with the patient may aid in establishing that which is possible rather than that which is ideal. | Principles in selection of therapy. The physician today is presented with a plethora of possibilities in the therapy of each of the aspects of ischemic heart disease (Fig. 15-5). There is the temptation to recommend complex and impossible dietary prescriptions coupled with several pharmaceutical agents for control of anginal pain, hypertension, arrhythmias, hypercholesterolemia, and clinical congestive heart failure. While each of the objectives may be in part valid, the burden on the patient of following such a constraining and difficult life may make it virtually impossible either to enjoy life or to follow the physician's recommendations explicitly. Often a compromise must be reached between theoretically optimal therapy and that which is reasonable and acceptable to the patient. Again, a review of each aspect of the program with the patient may aid in establishing that which is possible rather than that which is ideal. | [
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PMID:10504 | Triacetylated insulin: biologic activity and resistance to degradation. | Tritiated N-hydroxysuccinimide acetate was prepared with specific activities up to 5 Ci/mmole and utilized to prepare tritiated triacetyl insulin. Binding of triacetyl insulin to liver plasma membranes was measured by its capacity to displace 125I-monoiodoinsulin. At low concentrations, less than 10 ng/ml triacetyl insulin appears to be as effective as native insulin in reducing the binding of 125I-monoiodoinsulin to plasma membranes. At concentrations of 20 ng/ml and higher, triacetyl insulin is significantly less effective than native insulin in displacing binding of 125I-monoiodoinsulin to plasma membranes. The properties of triacetyl insulin in this system are not ascribable to deacetylation and conversion of the substituted product to native insulin. Biologic activity of triacetylated insulin was studied in two other in vitro systmes. A comparison was made of the capacity of native beef insulin and its triacetyl derivative to stimulate glucose oxidation by epididymal fat pads. At all three concentrations tested (2, 6, and 18 ng/ml), triacetyl insulin exerted considerable activity, although its potency was significantly less than that of native insulin. Similar effects were observed when biologic activity was measured by induction of tyrosine-alpha-ketoglutarate transaminase in a cultured liver cell system where significant activity of triacetyl insulin was found at concentrations of 10(-9)-10(-7) M. In all systems tested, the activity of triacetylated insulin could not be accounted for by deacetylation and conversion to native insulin. In all systems studied, triacetyl insulin was more resistant to degradation than was monoiodoinsulin. | Triacetylated insulin: biologic activity and resistance to degradation. Tritiated N-hydroxysuccinimide acetate was prepared with specific activities up to 5 Ci/mmole and utilized to prepare tritiated triacetyl insulin. Binding of triacetyl insulin to liver plasma membranes was measured by its capacity to displace 125I-monoiodoinsulin. At low concentrations, less than 10 ng/ml triacetyl insulin appears to be as effective as native insulin in reducing the binding of 125I-monoiodoinsulin to plasma membranes. At concentrations of 20 ng/ml and higher, triacetyl insulin is significantly less effective than native insulin in displacing binding of 125I-monoiodoinsulin to plasma membranes. The properties of triacetyl insulin in this system are not ascribable to deacetylation and conversion of the substituted product to native insulin. Biologic activity of triacetylated insulin was studied in two other in vitro systmes. A comparison was made of the capacity of native beef insulin and its triacetyl derivative to stimulate glucose oxidation by epididymal fat pads. At all three concentrations tested (2, 6, and 18 ng/ml), triacetyl insulin exerted considerable activity, although its potency was significantly less than that of native insulin. Similar effects were observed when biologic activity was measured by induction of tyrosine-alpha-ketoglutarate transaminase in a cultured liver cell system where significant activity of triacetyl insulin was found at concentrations of 10(-9)-10(-7) M. In all systems tested, the activity of triacetylated insulin could not be accounted for by deacetylation and conversion to native insulin. In all systems studied, triacetyl insulin was more resistant to degradation than was monoiodoinsulin. | [
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PMID:10505 | Effects of arginine infusion in infants: increased urea synthesis associated with unchanged ammonia blood levels. | Infusion of L-arginine hydrochloride in infants and children (ages ranging from 1 day to 12 yr) at a dosage of 0.5 g/kg body weight resulted in a dramatic increase in the arginine plasma concentration, with highest values of approximately 7 mmole/liter immediately after the end of the infusion; 120 min later the mean plasma level of the amino acid had decreased to mean values of 1 mmole/liter. These fluctuations were paralleled by increased ornithine concentrations, although the mean plasma levels of this amino acid remained far below those of arginine, i.e., 0.73 and 0.22 mmole/liter after 30 and 90 min, respectively. When expressed on a molar basis, arginine administration resulted in an almost stoichiometric rise in urinary urea excretion. These findings indicate that arginine is rapidly metabolized via urea and ornithine, the latter being transformed to glucose, as evidenced by a significant rise in the blood glucose concentration. Blood gas analyses and serum urea and blood ammonia concentrations determined after the load showed no significant deviations from preinfusion levels. Thus, in contrast to the effects to be expected form studies with tissue culture homogenates, even when administered to newborn infants, arginine does not impair the turnover of the urea cycle. | Effects of arginine infusion in infants: increased urea synthesis associated with unchanged ammonia blood levels. Infusion of L-arginine hydrochloride in infants and children (ages ranging from 1 day to 12 yr) at a dosage of 0.5 g/kg body weight resulted in a dramatic increase in the arginine plasma concentration, with highest values of approximately 7 mmole/liter immediately after the end of the infusion; 120 min later the mean plasma level of the amino acid had decreased to mean values of 1 mmole/liter. These fluctuations were paralleled by increased ornithine concentrations, although the mean plasma levels of this amino acid remained far below those of arginine, i.e., 0.73 and 0.22 mmole/liter after 30 and 90 min, respectively. When expressed on a molar basis, arginine administration resulted in an almost stoichiometric rise in urinary urea excretion. These findings indicate that arginine is rapidly metabolized via urea and ornithine, the latter being transformed to glucose, as evidenced by a significant rise in the blood glucose concentration. Blood gas analyses and serum urea and blood ammonia concentrations determined after the load showed no significant deviations from preinfusion levels. Thus, in contrast to the effects to be expected form studies with tissue culture homogenates, even when administered to newborn infants, arginine does not impair the turnover of the urea cycle. | [
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PMID:10506 | Effect of hydrogen ion buffers on photosynthetic oxygen evolution in the blue-green alga, Agmenellum quadruplicatum. | The photosynthetic oxygen evolution capacity of Agmenelium quadruplication suspended in four hydrogen ion buffers (pH 7.4, 0.05 M) and its synthetic marine growth medium was measured with an oxygen electrode. High rates of oxygen evolution were obtained in the growth medium and N-tris(hydroxymethyl)-methylglycine (Tricine) buffer. Compared to oxygen evolution in the growth medium, rates in phosphate buffer and N-tris(hydroxymethyl)-2-aminoethanesulphonic acid (TES) buffer were sometimes reduced by up to 30% and rates in tris (hydroxymethyl) amino-methane (Tris) were consistently reduced by 50%. An incubation-rinsing procedure caused inhibition of oxygen evolution in TES, phosphate, and Tris by 50 to 100%. Oxygen evolution could be restored to cells rinsed in TES or phosphate by resuspension in growth medium or in buffer plus magnesium and calcium ions. Bezoquinone-supported oxygen evolution was not affected by rinsing with any buffer tested except Tris. Ferricyanide was photoreduced at a low rate by cells rinsed in Tes but at a high rate in TES plus magnesium and calcium ions. We interpreted our results to mean that, in Agmenellum quadruplicatum, inhibition of photosynthetic oxygen evolution by Tris occurs at the level of photosystem 2 while the effects of TES and phosphate are on electron-transport occurring after the rate-limiting reaction. | Effect of hydrogen ion buffers on photosynthetic oxygen evolution in the blue-green alga, Agmenellum quadruplicatum. The photosynthetic oxygen evolution capacity of Agmenelium quadruplication suspended in four hydrogen ion buffers (pH 7.4, 0.05 M) and its synthetic marine growth medium was measured with an oxygen electrode. High rates of oxygen evolution were obtained in the growth medium and N-tris(hydroxymethyl)-methylglycine (Tricine) buffer. Compared to oxygen evolution in the growth medium, rates in phosphate buffer and N-tris(hydroxymethyl)-2-aminoethanesulphonic acid (TES) buffer were sometimes reduced by up to 30% and rates in tris (hydroxymethyl) amino-methane (Tris) were consistently reduced by 50%. An incubation-rinsing procedure caused inhibition of oxygen evolution in TES, phosphate, and Tris by 50 to 100%. Oxygen evolution could be restored to cells rinsed in TES or phosphate by resuspension in growth medium or in buffer plus magnesium and calcium ions. Bezoquinone-supported oxygen evolution was not affected by rinsing with any buffer tested except Tris. Ferricyanide was photoreduced at a low rate by cells rinsed in Tes but at a high rate in TES plus magnesium and calcium ions. We interpreted our results to mean that, in Agmenellum quadruplicatum, inhibition of photosynthetic oxygen evolution by Tris occurs at the level of photosystem 2 while the effects of TES and phosphate are on electron-transport occurring after the rate-limiting reaction. | [
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PMID:10508 | A new form of antihistamine--the H2-receptor antagonist. | A new group of drugs, the histamine2 (H2)-receptor antagonists, act on receptors in the stomach to reduce acid secretion when this is stimulated by histamine, pentagastrin, the vagus nerve or food. The reduction in acid secretion is profound and may approach the degree of reduction brought about by gastric surgery. The H2-receptor antagonist metiamide, administered orally, has been used successfully in the treatment of duodenal ulcer and the Zollinger-Ellison syndrome, but it has been shown to cause agranulocytosis. Trials are in process with an analogue, cimetidine (Tagamet, SKF), which has a different chemical structure from metiamide and has not caused haematological changes in animals or man. These drugs offer the prospect of successful medical management of duodenal ulcer, while a study of their effects on H2-receptors elsewhere in the body may reveal other therapeutic benefits. | A new form of antihistamine--the H2-receptor antagonist. A new group of drugs, the histamine2 (H2)-receptor antagonists, act on receptors in the stomach to reduce acid secretion when this is stimulated by histamine, pentagastrin, the vagus nerve or food. The reduction in acid secretion is profound and may approach the degree of reduction brought about by gastric surgery. The H2-receptor antagonist metiamide, administered orally, has been used successfully in the treatment of duodenal ulcer and the Zollinger-Ellison syndrome, but it has been shown to cause agranulocytosis. Trials are in process with an analogue, cimetidine (Tagamet, SKF), which has a different chemical structure from metiamide and has not caused haematological changes in animals or man. These drugs offer the prospect of successful medical management of duodenal ulcer, while a study of their effects on H2-receptors elsewhere in the body may reveal other therapeutic benefits. | [
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PMID:10509 | Streptococcal sore throat in general practice--a controlled study. | A controlled study was undertaken to determine the incidence of Group A haemolytic streptococci and other bacterial pathogens in throat swabs from patients presenting with sore throats. The isolation rate (38-8%) of Group A haemolytic streptococci from patients with acute tonsillitis was significantly higher than the isolation rate (8-9%) in a control group, but there was no significant difference in the isolation rates in patients diagnosed as having acute pharyngitis compared with the control group. There was no difference in the isolation rates of the other bacterial pathogens in the patient and control groups. In-vitro antibiotic studies were performed, and a comparison was made of the results of these tests and the treatment prescribed. | Streptococcal sore throat in general practice--a controlled study. A controlled study was undertaken to determine the incidence of Group A haemolytic streptococci and other bacterial pathogens in throat swabs from patients presenting with sore throats. The isolation rate (38-8%) of Group A haemolytic streptococci from patients with acute tonsillitis was significantly higher than the isolation rate (8-9%) in a control group, but there was no significant difference in the isolation rates in patients diagnosed as having acute pharyngitis compared with the control group. There was no difference in the isolation rates of the other bacterial pathogens in the patient and control groups. In-vitro antibiotic studies were performed, and a comparison was made of the results of these tests and the treatment prescribed. | [
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PMID:10527 | Reflex increase in coronary vascular resistance in patients with ischemic heart disease. | To assess possible coronary vasoconstriction in patients with ischemic heart disease, we measured coronary vascular resistance in 12 patients with normal hearts and 12 with coronary disease before and during the initial 50 seconds of cold pressor test, a stimulus known to produce systemic vasoconstriction. Control coronary vascular resistance was similar in the two groups, and although it did not change in patients with normal vessels, it rose by 27 per cent (P less than 0.005) in the group with coronary disease during the cold pressor test. In three of 12 patients with coronary disease coronary flow actually declined despite an increase in arterial pressure; in four, angina was precipitated. Phentolamine abolished increases in arterial pressure and coronary vascular resistance during the test in three patients with coronary disease. Adrenergically mediated coronary vascular tone may be an important determinant of coronary blood flow and may contribute to ischemia in patients with coronary disease. | Reflex increase in coronary vascular resistance in patients with ischemic heart disease. To assess possible coronary vasoconstriction in patients with ischemic heart disease, we measured coronary vascular resistance in 12 patients with normal hearts and 12 with coronary disease before and during the initial 50 seconds of cold pressor test, a stimulus known to produce systemic vasoconstriction. Control coronary vascular resistance was similar in the two groups, and although it did not change in patients with normal vessels, it rose by 27 per cent (P less than 0.005) in the group with coronary disease during the cold pressor test. In three of 12 patients with coronary disease coronary flow actually declined despite an increase in arterial pressure; in four, angina was precipitated. Phentolamine abolished increases in arterial pressure and coronary vascular resistance during the test in three patients with coronary disease. Adrenergically mediated coronary vascular tone may be an important determinant of coronary blood flow and may contribute to ischemia in patients with coronary disease. | [
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PMID:10531 | Proteolytic activity in liver cells from mouse, rat, Ehrlich ascites carcinoma bearing mouse and in Ehrlich ascites carcinoma cells. | The activity of intracellular proteinases from Ehrlich ascites carcinoma bearing mice were compared with that from liver cells of normal mice and rats. The activity of intracellular proteinases was measured in the supernatant of Ehrlich ascites carcinoma cells homogenate. The activity of intracellular proteinases in normal mouse and normal rat liver were different at pH 3.5, pH 6.0 and pH 7.5. The activity in liver cells from Ehlrich ascites carcinoma bearing mouse at pH 3.5 was not significantly changed from normal mouse or rat liver cells, however at pH 6.0 and pH 7.5 the activity in the affected liver significantly decreased. The proteolytic activity in the supernatant of Ehrlich ascites tumor cell homogenate was 0.110 E750 mmu/mgN at pH 7.5, 0.154 E750 mmu/mgN at pH 3.5. The proteolytic activity at pH 6.0 was not detected in any experiment. | Proteolytic activity in liver cells from mouse, rat, Ehrlich ascites carcinoma bearing mouse and in Ehrlich ascites carcinoma cells. The activity of intracellular proteinases from Ehrlich ascites carcinoma bearing mice were compared with that from liver cells of normal mice and rats. The activity of intracellular proteinases was measured in the supernatant of Ehrlich ascites carcinoma cells homogenate. The activity of intracellular proteinases in normal mouse and normal rat liver were different at pH 3.5, pH 6.0 and pH 7.5. The activity in liver cells from Ehlrich ascites carcinoma bearing mouse at pH 3.5 was not significantly changed from normal mouse or rat liver cells, however at pH 6.0 and pH 7.5 the activity in the affected liver significantly decreased. The proteolytic activity in the supernatant of Ehrlich ascites tumor cell homogenate was 0.110 E750 mmu/mgN at pH 7.5, 0.154 E750 mmu/mgN at pH 3.5. The proteolytic activity at pH 6.0 was not detected in any experiment. | [
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PMID:10538 | Correlation of the oxytocin challenge test with perinatal outcome. | A total of 234 oxytocin challenge tests (OCT) were performed on 100 high-risk patients. The results were negative (N) in 68 of these 100 patients, suspicious (S) in 22, and positive (P) in 10. The incidence of late decelerations during labor was N, 5%; S, 40%; P, 86%; and meconium staining of the amniotic fluid was N, 4%; S, 5%; and P, 30%. The cesarean section rate was N, 16%; S, 36%; and P, 60%; and of these the cesarean section rate for fetal indications was N, 9%; S, 25%; and P, 67%. The overall perinatal mortality in the study group was 2% (N, 1.5%; S, 0%; P, 10%). The results confirm the negative OCT as innocuous and positive OCT as the most ominous. They also indicate that the majority of patients with positive OCT can be delivered vaginally without endangering the fetus if fetal scalp blood pH determinations can be performed. | Correlation of the oxytocin challenge test with perinatal outcome. A total of 234 oxytocin challenge tests (OCT) were performed on 100 high-risk patients. The results were negative (N) in 68 of these 100 patients, suspicious (S) in 22, and positive (P) in 10. The incidence of late decelerations during labor was N, 5%; S, 40%; P, 86%; and meconium staining of the amniotic fluid was N, 4%; S, 5%; and P, 30%. The cesarean section rate was N, 16%; S, 36%; and P, 60%; and of these the cesarean section rate for fetal indications was N, 9%; S, 25%; and P, 67%. The overall perinatal mortality in the study group was 2% (N, 1.5%; S, 0%; P, 10%). The results confirm the negative OCT as innocuous and positive OCT as the most ominous. They also indicate that the majority of patients with positive OCT can be delivered vaginally without endangering the fetus if fetal scalp blood pH determinations can be performed. | [
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PMID:10539 | Microstructure of the solder-casting zone in bridges of dental gold alloys. | The composition and structure were studied in the cast and in the solder of bridges which had failed clinically. Both materials were gold alloys but with different contents of Pt, Ag and Zn. The metallographic investigation revealed defects mostly in the solder, situated in the subsurface layer. SEM studied of the fracture surface revealed large porosites and a structure of dense parallel lines--striations--, indicating that the material had failed from fatigue. The materials in the fractured bridges were identified by micro-probe measurements. It was stated that cast material and solder material used together, were of different composition. | Microstructure of the solder-casting zone in bridges of dental gold alloys. The composition and structure were studied in the cast and in the solder of bridges which had failed clinically. Both materials were gold alloys but with different contents of Pt, Ag and Zn. The metallographic investigation revealed defects mostly in the solder, situated in the subsurface layer. SEM studied of the fracture surface revealed large porosites and a structure of dense parallel lines--striations--, indicating that the material had failed from fatigue. The materials in the fractured bridges were identified by micro-probe measurements. It was stated that cast material and solder material used together, were of different composition. | [
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PMID:10541 | Gingival crevicular fluid: a new diagnostic aid in managing the periodontal patient. | On the whole, the studies on GCF have demonstrated that the flow of this fluid is sufficiently indicative of the inflammatory state that it can be used under a variety of clinical conditions to monitor and control gingival inflammation. Since gingivitis is extremely common, and since some cases of gingivitis presumably do not progress to periodontitis, the question could be posed whether or not a concerted effort to control inflammation (i.e. trying to achieve a GCF flow as near to zero as possible) would be clinically significant. Until there is evidence to the contrary, the answer must be "yes", since few cases are known where periodontitis occurs without being preceded by gingivitis. In other words, the control of all gingivitis, if feasible, should prevent most cases of periodontitis. Although control of all gingivitis would mean the treatment of many cases that would not progress to periodontal breakdown, such efforts would be worth-while if most periodontal destruction were prevented. Even the early destructive lesion exhibiting little or no inflammation may soon be identified, mainly because the minute volume of fluid collected from the gingival crevice can now be measured accurately. Accordingly, the concentration of various constituents in the GCF can be determined, which should lead to the development of tests to differentiate between pockets undergoing active destruction with minimal inflammation from the majority of active lesions that are intimately involved with frank inflammation. Thus, a clinician would measure sub-clinical gingival inflammation by measuring GCF flow, then differentiate destructive from quiescent lesions by analyzing the GCF sample for some constituent(s), chemical or microbial (Listgarten et al. 1975) indicative of the periodontal destructive process. Monitoring the flow of GCF might be of value in other clinical situations. For example, one could monitor the response of gingival tissues to various restorative and prosthetic procedures (Strauss et al. 1975) to ensure that these procedures do not aggravate the periodontal tissues and induce gingivitis or periodontitis. The education of the patient should be easier since patients can read their own numbers on the GCF meter at each examination and self-evaluate their personal periodontal condition and the effectiveness of their home care. Even the education of the dental student should be easier since he or she would have the means of self-evaluating the effectiveness of treatment, and not be as dependent upon the subjective assessment of his efforts by an instructor. Finally, monitoring GCF for various components could provide the dentist with a valuable means of easily screening patients for systemic disease. Obviously, this area of investigation is in its infancy, but does promise an exciting future for the oral diagnostician. | Gingival crevicular fluid: a new diagnostic aid in managing the periodontal patient. On the whole, the studies on GCF have demonstrated that the flow of this fluid is sufficiently indicative of the inflammatory state that it can be used under a variety of clinical conditions to monitor and control gingival inflammation. Since gingivitis is extremely common, and since some cases of gingivitis presumably do not progress to periodontitis, the question could be posed whether or not a concerted effort to control inflammation (i.e. trying to achieve a GCF flow as near to zero as possible) would be clinically significant. Until there is evidence to the contrary, the answer must be "yes", since few cases are known where periodontitis occurs without being preceded by gingivitis. In other words, the control of all gingivitis, if feasible, should prevent most cases of periodontitis. Although control of all gingivitis would mean the treatment of many cases that would not progress to periodontal breakdown, such efforts would be worth-while if most periodontal destruction were prevented. Even the early destructive lesion exhibiting little or no inflammation may soon be identified, mainly because the minute volume of fluid collected from the gingival crevice can now be measured accurately. Accordingly, the concentration of various constituents in the GCF can be determined, which should lead to the development of tests to differentiate between pockets undergoing active destruction with minimal inflammation from the majority of active lesions that are intimately involved with frank inflammation. Thus, a clinician would measure sub-clinical gingival inflammation by measuring GCF flow, then differentiate destructive from quiescent lesions by analyzing the GCF sample for some constituent(s), chemical or microbial (Listgarten et al. 1975) indicative of the periodontal destructive process. Monitoring the flow of GCF might be of value in other clinical situations. For example, one could monitor the response of gingival tissues to various restorative and prosthetic procedures (Strauss et al. 1975) to ensure that these procedures do not aggravate the periodontal tissues and induce gingivitis or periodontitis. The education of the patient should be easier since patients can read their own numbers on the GCF meter at each examination and self-evaluate their personal periodontal condition and the effectiveness of their home care. Even the education of the dental student should be easier since he or she would have the means of self-evaluating the effectiveness of treatment, and not be as dependent upon the subjective assessment of his efforts by an instructor. Finally, monitoring GCF for various components could provide the dentist with a valuable means of easily screening patients for systemic disease. Obviously, this area of investigation is in its infancy, but does promise an exciting future for the oral diagnostician. | [
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PMID:10544 | [Neuroaminergic control of anterior pituitary secretions (author's transl)]. | The demonstration and identification of monoamines and of aminergic tracts in the central nervous system has permitted a study of their role in the control of the liberation of hypothalamic releasing hormones. Knowledge of the role of these hypothalamic neurohormones in the release of pituitary hormones is at present under study. The role of monoamines in the control of pituitary hormone functions depends narrowly on pharmacological methods intervening either in the synthesis of neuroamines or in their action on a specific receptor. The authors consider successively the implication of monoamines in the control of liberation of ACTH, GH, TSH, prolactin and gonadotropic hormones. The role of aminergic mechanisms in the physiology of pituitary releasing hormones forms an integral part of homeostasis. Knowledge of these mechanisms leads to a clinical study of their role in disorders of hypothalamo-pituitary function. | [Neuroaminergic control of anterior pituitary secretions (author's transl)]. The demonstration and identification of monoamines and of aminergic tracts in the central nervous system has permitted a study of their role in the control of the liberation of hypothalamic releasing hormones. Knowledge of the role of these hypothalamic neurohormones in the release of pituitary hormones is at present under study. The role of monoamines in the control of pituitary hormone functions depends narrowly on pharmacological methods intervening either in the synthesis of neuroamines or in their action on a specific receptor. The authors consider successively the implication of monoamines in the control of liberation of ACTH, GH, TSH, prolactin and gonadotropic hormones. The role of aminergic mechanisms in the physiology of pituitary releasing hormones forms an integral part of homeostasis. Knowledge of these mechanisms leads to a clinical study of their role in disorders of hypothalamo-pituitary function. | [
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PMID:10545 | Diagnosis and treatment of duodenal ulcer in infancy and childhood. | An awareness of the modes of presentation in childhood duodenal ulcer disease is necessary for prompt diagnosis and treatment. Therapy should be based on the rationale of decreasing acid secretion and providing adequate buffering of gastric acid throughout the day. Because of the high incidence of recurrence in children, aggressive diagnosis and treatment of duodenal ulcer are indicated in any child suspected of having the disease. | Diagnosis and treatment of duodenal ulcer in infancy and childhood. An awareness of the modes of presentation in childhood duodenal ulcer disease is necessary for prompt diagnosis and treatment. Therapy should be based on the rationale of decreasing acid secretion and providing adequate buffering of gastric acid throughout the day. Because of the high incidence of recurrence in children, aggressive diagnosis and treatment of duodenal ulcer are indicated in any child suspected of having the disease. | [
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PMID:10546 | Genetic variation of lysosomal acid lipase. | Lysosomal acid lipase (LAL) activity was measured using a new fluorometric assay in cultured skin fibroblasts from eight control subjects, two obligate heterozygotes for Wolman's disease (WD), one patient with WD, and one patient with cholesteryl ester storage disease (CESD). The LAL activities (mean+/-SD) were 25.8+/-8.2, 13.2+/-0.1,1.1, and 1.4 nmol 4-methylumbelliferyl oleate (4-MUO) hydrolyzed/min/mg protein, respectively. These results compare favorably with those obtained using standard radioassays. The LAL activities of two cultures of amniotic fluid cells were 12.1 and 10.5. The LAL activity (mean+/-SD) of peripheral leukocytes obtained from 34 laboratory volunteers (19 females, 15 males) was 4.0+/-1.8. Partially purified lymphocytes contained about 25 times as much LAL activity as did granulocytes. Cellogel electrophoresis, followed by staining with 4-MUO, showed at least two bands of LAL (A and B) from normal fibroblasts, amniotic fluid cells, and lymphocytes. Band A was absent from WD and CESD fibroblasts and was reduced in fibroblasts of the WD heterozygotes. | Genetic variation of lysosomal acid lipase. Lysosomal acid lipase (LAL) activity was measured using a new fluorometric assay in cultured skin fibroblasts from eight control subjects, two obligate heterozygotes for Wolman's disease (WD), one patient with WD, and one patient with cholesteryl ester storage disease (CESD). The LAL activities (mean+/-SD) were 25.8+/-8.2, 13.2+/-0.1,1.1, and 1.4 nmol 4-methylumbelliferyl oleate (4-MUO) hydrolyzed/min/mg protein, respectively. These results compare favorably with those obtained using standard radioassays. The LAL activities of two cultures of amniotic fluid cells were 12.1 and 10.5. The LAL activity (mean+/-SD) of peripheral leukocytes obtained from 34 laboratory volunteers (19 females, 15 males) was 4.0+/-1.8. Partially purified lymphocytes contained about 25 times as much LAL activity as did granulocytes. Cellogel electrophoresis, followed by staining with 4-MUO, showed at least two bands of LAL (A and B) from normal fibroblasts, amniotic fluid cells, and lymphocytes. Band A was absent from WD and CESD fibroblasts and was reduced in fibroblasts of the WD heterozygotes. | [
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PMID:10549 | Influence of low extracellular pH upon the Ca inward current and isometric contractile force in mammalian ventricular myocardium. | In isolated papillary muscles of cats the changes in Ca inward current and isometric contractile force following a decrease of extracellular pH from 7.4 to 5.5 were studied. The Ca current was analyzed (a) by measuring the upstroke velocity of Ca-mediated action potentials and (b) in voltage clamp experiments using the double sucrose gap technique. 1. At a pH of 5.5 the upstroke velocity of the Ca-mediated action potential decreased to 65% of the control, while overshoot and action potential duration remained almost unchanged. Furthermore, the relative refractory period was prolonged and in some cases, a "Wenckebach-like" phenomenon occurred. In voltage clamp experiments, the slow inward current was found to be diminished to 50-60% of the initial control value and over a broad voltage range the current voltage relationship curve was shifted to weaker currents. Acidosis did not influence the steady state inactivation but altered the kinetics of inactivation of the slow inward current and induced an increase of tauinactivation and taurecovery. This indicates that acidosis exerts a complex effect on the slow membrane channel. 2. The normal response of the Ca current towards variations of the extracellular Ca concentration (0.5-4 mM) or towards the addition of the beta-stimulating compound isoproterenol (2 mg/l) was not altered by the lowered extracellular pH. 3. In the acid medium, isometric contractile force declined to 40% of the control value within 25 min and, thus, reacted stronger than the Ca current. This indicates that those forms of acidosis used in the present experiments caused their negative inotropic effect not exclusively via a depression of the Ca current. Rather an additional intracellular effect has to be assumed which finally leads to a reduced activity of the contractile system. 4. At pH 5.5 excess Ca (4 mM) induced the same quantitative response of the contractile system as obtained at normal pH. In contrast, the positive-inotropic effect of 2 mg/l isoproterenol was more pronounced, whilst the sensitivity of the Ca inward current towards this beta-stimulating compound remained unchanged. | Influence of low extracellular pH upon the Ca inward current and isometric contractile force in mammalian ventricular myocardium. In isolated papillary muscles of cats the changes in Ca inward current and isometric contractile force following a decrease of extracellular pH from 7.4 to 5.5 were studied. The Ca current was analyzed (a) by measuring the upstroke velocity of Ca-mediated action potentials and (b) in voltage clamp experiments using the double sucrose gap technique. 1. At a pH of 5.5 the upstroke velocity of the Ca-mediated action potential decreased to 65% of the control, while overshoot and action potential duration remained almost unchanged. Furthermore, the relative refractory period was prolonged and in some cases, a "Wenckebach-like" phenomenon occurred. In voltage clamp experiments, the slow inward current was found to be diminished to 50-60% of the initial control value and over a broad voltage range the current voltage relationship curve was shifted to weaker currents. Acidosis did not influence the steady state inactivation but altered the kinetics of inactivation of the slow inward current and induced an increase of tauinactivation and taurecovery. This indicates that acidosis exerts a complex effect on the slow membrane channel. 2. The normal response of the Ca current towards variations of the extracellular Ca concentration (0.5-4 mM) or towards the addition of the beta-stimulating compound isoproterenol (2 mg/l) was not altered by the lowered extracellular pH. 3. In the acid medium, isometric contractile force declined to 40% of the control value within 25 min and, thus, reacted stronger than the Ca current. This indicates that those forms of acidosis used in the present experiments caused their negative inotropic effect not exclusively via a depression of the Ca current. Rather an additional intracellular effect has to be assumed which finally leads to a reduced activity of the contractile system. 4. At pH 5.5 excess Ca (4 mM) induced the same quantitative response of the contractile system as obtained at normal pH. In contrast, the positive-inotropic effect of 2 mg/l isoproterenol was more pronounced, whilst the sensitivity of the Ca inward current towards this beta-stimulating compound remained unchanged. | [
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PMID:10550 | [Acute kidney failure during periarteritis nodosa]. | In 9 patients aged 16 to 71 years with acute renal failure, histological examination disclosed polyarteritis nodosa. Symptoms of rapidly progressive glomerulonephritis (GN) were present in 7 cases. Hypertension was a constant feature. Fever and muscular articular and cutaneous signs occured at times simulating Henoch-Schönlein purpura. Necrotizing angiitis was a constant finding, with perivascular granuloma in 7 cases. Lesions affecting smaller arteries were associated with diffuse or segmental extracapillary GN, and lesions of larger arteries with ischaemic appearance of the glomeruli. Diagnosis was obtained during life by percutaneous renal biopsy in 5 patients and muscular or cutaneous biopsy in 2 other patients, whereas the evidence was only obtained on post mortem examination in the 2 remaining patients. In the 6 renal biopsies examined by immunofluorescence, fibrinogen was always present in glomeruli and/or arteries. In one patient there was a linear deposit of IgG along glomerular basement membrane (GBM) and there were circulating antibodies directed against GBM. Arteries of different sizes may be damaged in various organs, as showed by arteriography and autopsy. Steroid therapy and immunosuppressive drugs had no effect on renal symptoms. 8 out of 9 patients died rapidly, in 6 the death was due to extrarenal localization of the disease, even though uraemia was controlled by haemodialysis. The last patient died 2 years after onset of the disease, while being on maintenance haemodialysis treatment. | [Acute kidney failure during periarteritis nodosa]. In 9 patients aged 16 to 71 years with acute renal failure, histological examination disclosed polyarteritis nodosa. Symptoms of rapidly progressive glomerulonephritis (GN) were present in 7 cases. Hypertension was a constant feature. Fever and muscular articular and cutaneous signs occured at times simulating Henoch-Schönlein purpura. Necrotizing angiitis was a constant finding, with perivascular granuloma in 7 cases. Lesions affecting smaller arteries were associated with diffuse or segmental extracapillary GN, and lesions of larger arteries with ischaemic appearance of the glomeruli. Diagnosis was obtained during life by percutaneous renal biopsy in 5 patients and muscular or cutaneous biopsy in 2 other patients, whereas the evidence was only obtained on post mortem examination in the 2 remaining patients. In the 6 renal biopsies examined by immunofluorescence, fibrinogen was always present in glomeruli and/or arteries. In one patient there was a linear deposit of IgG along glomerular basement membrane (GBM) and there were circulating antibodies directed against GBM. Arteries of different sizes may be damaged in various organs, as showed by arteriography and autopsy. Steroid therapy and immunosuppressive drugs had no effect on renal symptoms. 8 out of 9 patients died rapidly, in 6 the death was due to extrarenal localization of the disease, even though uraemia was controlled by haemodialysis. The last patient died 2 years after onset of the disease, while being on maintenance haemodialysis treatment. | [
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PMID:10562 | Treatment of angina pectoris. Pharmacologic approaches. | The short-acting nitrite sublingual nitroglycerin is the cornerstone of drug treatment of angina pectoris. It is most useful when given two to three minutes before activity that may precipitate an anginal attack. If disabling angina persists despite medical management and there is no contraindication, the beta-adrenergic blocking agent propranolol should be tried before coronary artery surgery is considered. Newer bera-adrenergic blocking agents do not appear to be more effective than propranolol. Digitalis may be beneficial in patients with congestive heart failure or with cardiac arrhythmias responsive to digitalis and in some patients with radiographic evidence of left ventricular enlargement or with nocturnal angina resulting from increased left ventricular end-diastolic volume. If bypass graft surgery is done, medical management must be continued postoperatively. | Treatment of angina pectoris. Pharmacologic approaches. The short-acting nitrite sublingual nitroglycerin is the cornerstone of drug treatment of angina pectoris. It is most useful when given two to three minutes before activity that may precipitate an anginal attack. If disabling angina persists despite medical management and there is no contraindication, the beta-adrenergic blocking agent propranolol should be tried before coronary artery surgery is considered. Newer bera-adrenergic blocking agents do not appear to be more effective than propranolol. Digitalis may be beneficial in patients with congestive heart failure or with cardiac arrhythmias responsive to digitalis and in some patients with radiographic evidence of left ventricular enlargement or with nocturnal angina resulting from increased left ventricular end-diastolic volume. If bypass graft surgery is done, medical management must be continued postoperatively. | [
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PMID:10564 | An eighteen months' study of the clinical response to metoprolol, a selective beta1-receptor blocking agent, in patients with angina pectoris. | Following an initial dose response study, metoprolol, a selective beta1-receptor blocking agent, was compared with equipotent dosages of propanolol in a double blind cross-over study, including exercise tolerance tests, on fourteen patients with angina pectoris. Long term therapy with metoprolol then followed until the seventy-second week. Patients performed 8% more total work on metoprolol with 15% more work recorded up to the onset of S-T depression, in comparison with propranolol. In the long term, ther was no significant difference in work performed when the daily dosage of metoprolol was changed from a q.i.d. to a b.d. regime. Metoprolol was shown to be an effective anti-anginal compound with good tolerance and safety, with gradual improvement in underlying myocardial ischaemia during long term treatment. | An eighteen months' study of the clinical response to metoprolol, a selective beta1-receptor blocking agent, in patients with angina pectoris. Following an initial dose response study, metoprolol, a selective beta1-receptor blocking agent, was compared with equipotent dosages of propanolol in a double blind cross-over study, including exercise tolerance tests, on fourteen patients with angina pectoris. Long term therapy with metoprolol then followed until the seventy-second week. Patients performed 8% more total work on metoprolol with 15% more work recorded up to the onset of S-T depression, in comparison with propranolol. In the long term, ther was no significant difference in work performed when the daily dosage of metoprolol was changed from a q.i.d. to a b.d. regime. Metoprolol was shown to be an effective anti-anginal compound with good tolerance and safety, with gradual improvement in underlying myocardial ischaemia during long term treatment. | [
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PMID:10560 | Piperazine derivatives of dimethylxanthines. IV. Reaction of beta,gamma-epoxypropyldimethylxanthines with piperazines. | In the reaction of 7-resp. 1-beta,gamma-epoxyproplderivatives of theophylline (Th) or theobromine (Tb) with piperazine and N-monoalkyl or monoarylpiperazines the corresponding 7-resp. 1-beta-hydroxy-gamma-piperazinopropyl-Th (1-8) resp. Tb (9-16) were formed. In preliminary screening some of the new compounds showed strong antagonism with histamine H1 receptor and with endogenous histamine. | Piperazine derivatives of dimethylxanthines. IV. Reaction of beta,gamma-epoxypropyldimethylxanthines with piperazines. In the reaction of 7-resp. 1-beta,gamma-epoxyproplderivatives of theophylline (Th) or theobromine (Tb) with piperazine and N-monoalkyl or monoarylpiperazines the corresponding 7-resp. 1-beta-hydroxy-gamma-piperazinopropyl-Th (1-8) resp. Tb (9-16) were formed. In preliminary screening some of the new compounds showed strong antagonism with histamine H1 receptor and with endogenous histamine. | [
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] |
PMID:10565 | Pneumococcal antigen in pneumonia. A post-mortem study with the histological and bacteriological findings. | Pneumococcal capsular antigens can be detected in lung tissue by counter-current immunoelectrophoresis even when, following antibiotics, post-mortem bacterilogy suggests that Escherichia coli has replaced pneumococci. The results suggest that antipneumococcal therapy would benefit at least 55% of patients critically ill with lung infection and that the potentially toxic drugs directed at coliform bacteria may be unnecessary. | Pneumococcal antigen in pneumonia. A post-mortem study with the histological and bacteriological findings. Pneumococcal capsular antigens can be detected in lung tissue by counter-current immunoelectrophoresis even when, following antibiotics, post-mortem bacterilogy suggests that Escherichia coli has replaced pneumococci. The results suggest that antipneumococcal therapy would benefit at least 55% of patients critically ill with lung infection and that the potentially toxic drugs directed at coliform bacteria may be unnecessary. | [
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PMID:10561 | The stability of N-pyrrolidinomethyltetracycline in solutions. | In aqueous solution N-pyrrolidinomethyltetracycline (PTC) undergoes decomposition into tetracycline, epitetracylcine, anhydroepitetracycline and ynhydrotetracycline. The decomposition products were identified and separated by TLC and their activity was estimated microbiologically. | The stability of N-pyrrolidinomethyltetracycline in solutions. In aqueous solution N-pyrrolidinomethyltetracycline (PTC) undergoes decomposition into tetracycline, epitetracylcine, anhydroepitetracycline and ynhydrotetracycline. The decomposition products were identified and separated by TLC and their activity was estimated microbiologically. | [
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PMID:10570 | Catecholamine-stimulated cyclic GMP accumulation in the rat pineal: apparent presynaptic site of action. | Guanosine 3':5'-cyclic monophosphate (cGMP) increased 7-fold in rat pineal glands incubated in the presence of l-norepinephrine. This response consisted of two components-one was stereospecific and inhibited by alpha-adrenergic antagonists while the other was not stereospecific and not readily inhibited by antagonists. Although l-isoproterenol was more potent than l-norepinephrine it had less intrinsic activity and its action was not stereospecifc. The increase in cGMP caused by these catecholamines, unlike that of adenosine 3':5'-cyclic monophosphate (cAMP), was dependent upon extracellular calcium. Ouabain and high levels of potassium produced a marked, calcium-dependent increase in pineal cGMP, without affecting cAMP. There was no effect of cholinergic agonists on cGMP. Surgical denervation markedly reduced the cGMP response to stimulation by l-norepinephrine, potassium, or ouabain. This was in contrast to the enhanced response of cAMP in denervated glands. The nonspecific increase in cGMP caused by l-isoproterenol, however, was not affected by denervation. These data demonstrate the existence of a calcium-dependent presynaptic mechanism for the generation of cGMP which may be mediated by an alpha-adrenergic-like receptor. In addition, the mechanisms regulating pineal cGMP appear to be physiologically distinct from those regulating cAMP. | Catecholamine-stimulated cyclic GMP accumulation in the rat pineal: apparent presynaptic site of action. Guanosine 3':5'-cyclic monophosphate (cGMP) increased 7-fold in rat pineal glands incubated in the presence of l-norepinephrine. This response consisted of two components-one was stereospecific and inhibited by alpha-adrenergic antagonists while the other was not stereospecific and not readily inhibited by antagonists. Although l-isoproterenol was more potent than l-norepinephrine it had less intrinsic activity and its action was not stereospecifc. The increase in cGMP caused by these catecholamines, unlike that of adenosine 3':5'-cyclic monophosphate (cAMP), was dependent upon extracellular calcium. Ouabain and high levels of potassium produced a marked, calcium-dependent increase in pineal cGMP, without affecting cAMP. There was no effect of cholinergic agonists on cGMP. Surgical denervation markedly reduced the cGMP response to stimulation by l-norepinephrine, potassium, or ouabain. This was in contrast to the enhanced response of cAMP in denervated glands. The nonspecific increase in cGMP caused by l-isoproterenol, however, was not affected by denervation. These data demonstrate the existence of a calcium-dependent presynaptic mechanism for the generation of cGMP which may be mediated by an alpha-adrenergic-like receptor. In addition, the mechanisms regulating pineal cGMP appear to be physiologically distinct from those regulating cAMP. | [
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PMID:10571 | Erythrocyte membranes undergo cooperative, pH-sensitive state transitions in the physiological temperature range: evidence from Raman spectroscopy. | We have examined the Raman scattering from erythrocyte ghosts at 2700 to 3000 cm-1 (CH-stretching region). Plots of the intensity (I) of the 2930 cm-1 band relative to the intensity of the thermally stable 2850 cm-1 band, i.e., the [I2930/I2850] ratio, as a function of temperature reveal a sharp discontinuity, which at pH 7.4 has a lower limit of 38 degrees and is irreversible above 42 degrees. [I2930/I2850] is stable between pH 7.0 and pH 7.4, but increases or decreases sharply below pH 7.0 or above pH 7.5, respectively. Reduction of pH to 6.5 lowers the transition temperature by about 16 degrees, and a shift to pH 6.0 drops the transition range to 0 to 7 degrees. The above effects of temperature and pH on Raman scattering closely correspond to those detected by studies on the interaction of membrane protein fluorophores and lipid-soluble fluorescence quenchers [Bieri, V. and Wallach, D.F.H. (1975) Biochim. Biophys. Acta 406, 415-423]. Taken together, these results suggest that the transitions represent concerted process, involving hydrophobic amino acid residues and lipid chains at apolar protein-lipid boundaries. | Erythrocyte membranes undergo cooperative, pH-sensitive state transitions in the physiological temperature range: evidence from Raman spectroscopy. We have examined the Raman scattering from erythrocyte ghosts at 2700 to 3000 cm-1 (CH-stretching region). Plots of the intensity (I) of the 2930 cm-1 band relative to the intensity of the thermally stable 2850 cm-1 band, i.e., the [I2930/I2850] ratio, as a function of temperature reveal a sharp discontinuity, which at pH 7.4 has a lower limit of 38 degrees and is irreversible above 42 degrees. [I2930/I2850] is stable between pH 7.0 and pH 7.4, but increases or decreases sharply below pH 7.0 or above pH 7.5, respectively. Reduction of pH to 6.5 lowers the transition temperature by about 16 degrees, and a shift to pH 6.0 drops the transition range to 0 to 7 degrees. The above effects of temperature and pH on Raman scattering closely correspond to those detected by studies on the interaction of membrane protein fluorophores and lipid-soluble fluorescence quenchers [Bieri, V. and Wallach, D.F.H. (1975) Biochim. Biophys. Acta 406, 415-423]. Taken together, these results suggest that the transitions represent concerted process, involving hydrophobic amino acid residues and lipid chains at apolar protein-lipid boundaries. | [
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PMID:10572 | Central regulation of sympathetic neuron development. | The sixth lumbar (L-6) ganglion has been used to study the central regulation of peripheral sympathetic neuron development. During post-natal ontogeny, tyrosine hydroxylase [tyrosine 3-monooxygenase, L-tyrosine, tetrahydropteridine: oxygen oxidoreductase (3-hydroxylating), EC 1.14.16.2] activity increased 60-fold, while total protein rose 10-fold in the ganglion. Transection of the spinal cord at the fifth thoracic (T-5) segment in neonatal rats prevented the normal developmental increase in tyrosine hydroxylase activity of the L-6 ganglion. However, spinal transection did not alter the ontogeny of tyrosine hydroxylase in the superior cervical ganglion, which derives its innervation from spinal segments rostral to the surgical lesion. Thus, spinal transection interfered with the maturation of sympathetic neurons distal to, but not proximal to, the lesion. The effect of transection on the L-6 ganglion persisted for at least one month, the longest time tested. Our observations suggest that trans-synaptic regulation of adrenergic maturation in the periphery is governed by suprasegmental mechanisms in the central nervous system. | Central regulation of sympathetic neuron development. The sixth lumbar (L-6) ganglion has been used to study the central regulation of peripheral sympathetic neuron development. During post-natal ontogeny, tyrosine hydroxylase [tyrosine 3-monooxygenase, L-tyrosine, tetrahydropteridine: oxygen oxidoreductase (3-hydroxylating), EC 1.14.16.2] activity increased 60-fold, while total protein rose 10-fold in the ganglion. Transection of the spinal cord at the fifth thoracic (T-5) segment in neonatal rats prevented the normal developmental increase in tyrosine hydroxylase activity of the L-6 ganglion. However, spinal transection did not alter the ontogeny of tyrosine hydroxylase in the superior cervical ganglion, which derives its innervation from spinal segments rostral to the surgical lesion. Thus, spinal transection interfered with the maturation of sympathetic neurons distal to, but not proximal to, the lesion. The effect of transection on the L-6 ganglion persisted for at least one month, the longest time tested. Our observations suggest that trans-synaptic regulation of adrenergic maturation in the periphery is governed by suprasegmental mechanisms in the central nervous system. | [
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PMID:10569 | [Purification and properties of L-glutamine and L-asparagine deaminase from Pseudomonas aurantiaca IBPM-14]. | An enzymic preparation of L-glutamine and L-asparagine deamidase was obtained from Pseudomonas auractiaca IBPM B-14. The preparation was purified 100--150-fold by thermal treatment and chromotography on columns with biogel P-150 and DEAE-cellulose. The enzymic activity was measured by the methods of hydroxylaminolysis and direct nesslerization. The deamidase preparation had an activity of 51 i. u. by glutamine and 15 i. e. by asparagine. Evidence on the pH effect on the deamidase activity was accumulated. | [Purification and properties of L-glutamine and L-asparagine deaminase from Pseudomonas aurantiaca IBPM-14]. An enzymic preparation of L-glutamine and L-asparagine deamidase was obtained from Pseudomonas auractiaca IBPM B-14. The preparation was purified 100--150-fold by thermal treatment and chromotography on columns with biogel P-150 and DEAE-cellulose. The enzymic activity was measured by the methods of hydroxylaminolysis and direct nesslerization. The deamidase preparation had an activity of 51 i. u. by glutamine and 15 i. e. by asparagine. Evidence on the pH effect on the deamidase activity was accumulated. | [
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PMID:10573 | Use of thyrotropin and cholera toxin to probe the mechanism by which interferon initiates its antiviral activity. | Thyrotropin (10 muM) inhibited the antiviral activity of interferon. When added after interferon, thyrotropin (TSH) had no effect on antiviral activity. There was also no inhibition of interferon action in cells washed with medium between incubations with TSH and interferon. 125I-Labeled TSH and 125I-labeled cholera toxin could bind to preparations of mouse L-cell plasma membranes. The binding was specific in that it was prevented by unlabeled thyrotropin or cholera toxin, but not by insulin, glucagon, prolactin, growth hormone, human chorionic gonadotropin, or luteinizing hormone. Mouse interferon inhibited 125I-labeled TSH binding to L-cell plasma membranes. The effect of mouse interferon on 125I-labeled cholera toxon binding was more complex, inhibition occurring only after an initial enhancement at low interferon concentrations. A 10-fold higher concentration of interferon was required to inhibit 125I-labeled TSH binding. Mouse interferon was also able to displace bound 125I-labeled TSH, but not bound 125I-labeled cholera toxin. The interferon interaction with cell membranes was temperature-sensitive. Human interferon could induce changes in binding of 125I-labeled TSH and 125I-labeled cholera toxin to mouse L-cell plasma membranes similar to those induced by mouse interferon. Mouse interferon induced similar changes in plasma membranes of human KB-3 cells, which are insensitive to both human and mouse interferons. In view of these results, the species specificity of interferons does not appear to reside solely at the point of the initial interaction with their binding sites. | Use of thyrotropin and cholera toxin to probe the mechanism by which interferon initiates its antiviral activity. Thyrotropin (10 muM) inhibited the antiviral activity of interferon. When added after interferon, thyrotropin (TSH) had no effect on antiviral activity. There was also no inhibition of interferon action in cells washed with medium between incubations with TSH and interferon. 125I-Labeled TSH and 125I-labeled cholera toxin could bind to preparations of mouse L-cell plasma membranes. The binding was specific in that it was prevented by unlabeled thyrotropin or cholera toxin, but not by insulin, glucagon, prolactin, growth hormone, human chorionic gonadotropin, or luteinizing hormone. Mouse interferon inhibited 125I-labeled TSH binding to L-cell plasma membranes. The effect of mouse interferon on 125I-labeled cholera toxon binding was more complex, inhibition occurring only after an initial enhancement at low interferon concentrations. A 10-fold higher concentration of interferon was required to inhibit 125I-labeled TSH binding. Mouse interferon was also able to displace bound 125I-labeled TSH, but not bound 125I-labeled cholera toxin. The interferon interaction with cell membranes was temperature-sensitive. Human interferon could induce changes in binding of 125I-labeled TSH and 125I-labeled cholera toxin to mouse L-cell plasma membranes similar to those induced by mouse interferon. Mouse interferon induced similar changes in plasma membranes of human KB-3 cells, which are insensitive to both human and mouse interferons. In view of these results, the species specificity of interferons does not appear to reside solely at the point of the initial interaction with their binding sites. | [
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PMID:10574 | Multiple factor analysis of the action of local anesthetics. | The pH jump data of Bianchi and Strobel [(1968) Trans. N.Y. Acad. Sci. Ser. II, 30, 1082-1092] on desheathed frog sciatic nerve are fitted to rate equations. A general quantitation of synergism, summation, and antagonism of anesthetics and of excitation is given. | Multiple factor analysis of the action of local anesthetics. The pH jump data of Bianchi and Strobel [(1968) Trans. N.Y. Acad. Sci. Ser. II, 30, 1082-1092] on desheathed frog sciatic nerve are fitted to rate equations. A general quantitation of synergism, summation, and antagonism of anesthetics and of excitation is given. | [
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PMID:10575 | Rumen function in red deer, hill sheep and reindeer in the scottish highlands. | Red deer, sheep and reindeer grazing on their normal hill ranges were examined at intervals over a period of four years. Samples from the digestive tract were taken at different seasons and processed in the field. The Red deer and reindeer were killed before samples were taken; rumen samples from the sheep were taken by stomach tube, but a number of animals were also killed at different seasons to correlate stomach tube and whole rumen samples. The animals sampled were representative of the general condition of the herds. Examinations were made for parasites and any pathological conditions. In most instances parasitic infections were slight. Apparent seasonal changes were found in the compositions of the diets. The Red deer and sheep ate principally heather and grass, the proportion of heather increasing in the winter. The reindeer ate mainly grass in the summer, with lichens and grass forming the winter diet, and these animals seemed to have a higher nutritional status in the winter than did the other two species. The weights of the animals and of their rumen contents, the concentrations of rumen ammonia and volatile fatty acid, and the rates at which different dietary components were fermented are recorded. Rumen fermentation was low in winter and the diets were generally inadequate for the animals. A lack of nitrogen seemed to be a major factor. Some data on caecal contents are also given. | Rumen function in red deer, hill sheep and reindeer in the scottish highlands. Red deer, sheep and reindeer grazing on their normal hill ranges were examined at intervals over a period of four years. Samples from the digestive tract were taken at different seasons and processed in the field. The Red deer and reindeer were killed before samples were taken; rumen samples from the sheep were taken by stomach tube, but a number of animals were also killed at different seasons to correlate stomach tube and whole rumen samples. The animals sampled were representative of the general condition of the herds. Examinations were made for parasites and any pathological conditions. In most instances parasitic infections were slight. Apparent seasonal changes were found in the compositions of the diets. The Red deer and sheep ate principally heather and grass, the proportion of heather increasing in the winter. The reindeer ate mainly grass in the summer, with lichens and grass forming the winter diet, and these animals seemed to have a higher nutritional status in the winter than did the other two species. The weights of the animals and of their rumen contents, the concentrations of rumen ammonia and volatile fatty acid, and the rates at which different dietary components were fermented are recorded. Rumen fermentation was low in winter and the diets were generally inadequate for the animals. A lack of nitrogen seemed to be a major factor. Some data on caecal contents are also given. | [
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PMID:10577 | Modulation of cycloheximide-resistant memory by sympathomimetic agents. | Amphetamine overcomes the amnesia caused by cycloheximide (CXM) provided it is administered closely following the learning trial. In day-old chickens with one trial passive avoidance learning, there is a short-term, labile memory existing for 90 min following training under the influence of CXM. Amphetamine has been shown to keep the memory at precisely the level exhibited by the labile, cycloheximide-resistant memory trace at the time of injection. Norepinephrine, methoxamine (an alpha adrenergic stimulant) and isoprenaline (a beta adrenergic stimulant) each mimic the amphetamine effect in CXM-pretreated chickens. That the action of amphetamine could be due to its release of norepinephrine is supported by the finding that it could be blocked by both alpha adrenergic (piperoxane) and beta adrenergic antagonists (propranolol). It has been suggested that this labile memory trace depends on the functioning of a sodium pump. Norepinephrine may be modulating memory formation by an action on the sodium pump since in preliminary biochemical assays norepinephrine stimulated the sodium pump (Na+/K+ ATPase) activity in chicken forebrain total homogenate. | Modulation of cycloheximide-resistant memory by sympathomimetic agents. Amphetamine overcomes the amnesia caused by cycloheximide (CXM) provided it is administered closely following the learning trial. In day-old chickens with one trial passive avoidance learning, there is a short-term, labile memory existing for 90 min following training under the influence of CXM. Amphetamine has been shown to keep the memory at precisely the level exhibited by the labile, cycloheximide-resistant memory trace at the time of injection. Norepinephrine, methoxamine (an alpha adrenergic stimulant) and isoprenaline (a beta adrenergic stimulant) each mimic the amphetamine effect in CXM-pretreated chickens. That the action of amphetamine could be due to its release of norepinephrine is supported by the finding that it could be blocked by both alpha adrenergic (piperoxane) and beta adrenergic antagonists (propranolol). It has been suggested that this labile memory trace depends on the functioning of a sodium pump. Norepinephrine may be modulating memory formation by an action on the sodium pump since in preliminary biochemical assays norepinephrine stimulated the sodium pump (Na+/K+ ATPase) activity in chicken forebrain total homogenate. | [
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PMID:10578 | Studies on the polymorphism of carbromal. | Specimens of carbromal obtained by crystallization from various solvents were analyzed. After a 9-month storage period, the occurrence of three polymorphic forms (I, II and III) was ascertained. The metastable form III obtained by crystallization from dimethylformamide showed a reasonably high stability. Preliminary pharmacological tests revealed clear-cut differences in the therapeutic effect between a commercial specimen and the polymorphic modification III. The results presented here indicate the importance of polymorphism of carbromal for medicinal use. | Studies on the polymorphism of carbromal. Specimens of carbromal obtained by crystallization from various solvents were analyzed. After a 9-month storage period, the occurrence of three polymorphic forms (I, II and III) was ascertained. The metastable form III obtained by crystallization from dimethylformamide showed a reasonably high stability. Preliminary pharmacological tests revealed clear-cut differences in the therapeutic effect between a commercial specimen and the polymorphic modification III. The results presented here indicate the importance of polymorphism of carbromal for medicinal use. | [
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PMID:10583 | Treatment of depression with tricyclic drugs--pharmacokinetic and pharmacodynamic aspects. | A series of studies on the pharmacokinetic and pharmacodynamic properties of some tricyclic antidepressants is reviewed. During treatment with the same oral dose of these drugs, patients develop widely differing plasma levels. The importance of this variability for the clinical effects has been studied in detail for the monomethylated compound, nortriptyline. There is an association between side-effects and high plasma levels of this drug. In endogenously depressed patients, the relationship between plasma level and effect appears to be curvilinear. The tricyclic antidepressants differ in their capacity to inhibit transmitter uptake into noradrenaline- and serotonin neurons respectively. Nortriptyline is a preferential noradrenaline uptake inhibitor, while the dimethylated compound, chlorimipramine also has a profound influence on serotonin neurons. These differential effects are also reflected in changes in the levels of the transmitter metabolites in cerebrospinal fluid (CSF). The CSF studies have also supported the hypothesis of a biochemical heterogeneity of the depressive syndrome. The levels of the serotonin metabolite, 5-HIAA were bimodally distributed in CSF. In patients with a low level of 5-HIAA there was a significant correlation between the CSF metabolite level and the severity of the depression, and these patients also appeared to be more suicide-prone than those with higher 5-HIAA levels. These patients seemed to be less amenable to treatment with nortriptyline. The effect of chlorimipramine treatment in this subgroup of depressives is presently being explored. | Treatment of depression with tricyclic drugs--pharmacokinetic and pharmacodynamic aspects. A series of studies on the pharmacokinetic and pharmacodynamic properties of some tricyclic antidepressants is reviewed. During treatment with the same oral dose of these drugs, patients develop widely differing plasma levels. The importance of this variability for the clinical effects has been studied in detail for the monomethylated compound, nortriptyline. There is an association between side-effects and high plasma levels of this drug. In endogenously depressed patients, the relationship between plasma level and effect appears to be curvilinear. The tricyclic antidepressants differ in their capacity to inhibit transmitter uptake into noradrenaline- and serotonin neurons respectively. Nortriptyline is a preferential noradrenaline uptake inhibitor, while the dimethylated compound, chlorimipramine also has a profound influence on serotonin neurons. These differential effects are also reflected in changes in the levels of the transmitter metabolites in cerebrospinal fluid (CSF). The CSF studies have also supported the hypothesis of a biochemical heterogeneity of the depressive syndrome. The levels of the serotonin metabolite, 5-HIAA were bimodally distributed in CSF. In patients with a low level of 5-HIAA there was a significant correlation between the CSF metabolite level and the severity of the depression, and these patients also appeared to be more suicide-prone than those with higher 5-HIAA levels. These patients seemed to be less amenable to treatment with nortriptyline. The effect of chlorimipramine treatment in this subgroup of depressives is presently being explored. | [
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PMID:10584 | The contribution of drug research to investigating the nature of endogenous depression. | A relationship between brain monamines and endogenous depression is suggested by observations on the mode of action of drugs producing or alleviating depressive symptoms. For example, reserpine is capable of faithfully mimicking the clinical picture of endogenous depression, which may be related to monoamine depletion. On the other hand, antidepressant drugs, e.g. the monoamine oxidase inhibitors, the tricyclic antidepressants and the monoamine precursors appear to increase the availability of monoamines at postsynaptic receptor sites. The different classes of antidepressant agents in general appear to potentiate each other's actions, according to animal data and clinical observations. Studies on the mode of action of tricyclic antidepressants with different profiles and on monoamine precursors suggest that 5-hydroxytryptamine is primarily involved in the control of mood, and noradrenaline in psychomotor activity. Clincial investigations initiated by the drug studies have demonstrated changes in monoamine metabolism in endogenous depression. The available evidence thus suggests a causal relationship between disturbances in monoamine metabolism and depression. | The contribution of drug research to investigating the nature of endogenous depression. A relationship between brain monamines and endogenous depression is suggested by observations on the mode of action of drugs producing or alleviating depressive symptoms. For example, reserpine is capable of faithfully mimicking the clinical picture of endogenous depression, which may be related to monoamine depletion. On the other hand, antidepressant drugs, e.g. the monoamine oxidase inhibitors, the tricyclic antidepressants and the monoamine precursors appear to increase the availability of monoamines at postsynaptic receptor sites. The different classes of antidepressant agents in general appear to potentiate each other's actions, according to animal data and clinical observations. Studies on the mode of action of tricyclic antidepressants with different profiles and on monoamine precursors suggest that 5-hydroxytryptamine is primarily involved in the control of mood, and noradrenaline in psychomotor activity. Clincial investigations initiated by the drug studies have demonstrated changes in monoamine metabolism in endogenous depression. The available evidence thus suggests a causal relationship between disturbances in monoamine metabolism and depression. | [
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PMID:10585 | Indications for long-term pharmacological treatment of schizophrenic syndromes. | The practice of long term pharmacological treatment of schizophrenic syndromes has become widespread and seems to be currently recommended. The concept appears, however, to be poorly defined, and the assumptions underlying this therapeutic approach are scarcely discussed. In the present paper possible differences as concerns the hypotheses which might motivate a long term pharmacological treatment are schetched and their possible implications discussed. A short review of factors which might influence the decision for long term pharmacological treatment is also presented. Finally some recommendations for an appropriate management of long term psychopharmacological treatment are suggested. | Indications for long-term pharmacological treatment of schizophrenic syndromes. The practice of long term pharmacological treatment of schizophrenic syndromes has become widespread and seems to be currently recommended. The concept appears, however, to be poorly defined, and the assumptions underlying this therapeutic approach are scarcely discussed. In the present paper possible differences as concerns the hypotheses which might motivate a long term pharmacological treatment are schetched and their possible implications discussed. A short review of factors which might influence the decision for long term pharmacological treatment is also presented. Finally some recommendations for an appropriate management of long term psychopharmacological treatment are suggested. | [
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PMID:10586 | Critical study of the use of long acting neuroleptics (depot neuroleptics) in France. | Long acting neuroleptics (L.A.N.) represent 11% of the medical prescription of neuroleptics in France (about 40,000 patients in 1974). Some psychiatrists remain reluctant to prescribe long acting-neuroleptics and many problems related to their use are still solved empirically:1) relation between the posology of the standard Neuroleptic and the corresponding L.A.N. 2)the problem of dose-interval. 3) the necessary prescription of an parkinsonian. The superiority of N.A.P. over the Neuroleptic Standard has received no experimental confirmation in France, but the L.A.N. appear to us as an original therapy, the "pivot chemotherapy" around which psychotherapy and sociotherapy can be arranged without anarchical and ceaseless changes of neuroleptic compounds and of posology. | Critical study of the use of long acting neuroleptics (depot neuroleptics) in France. Long acting neuroleptics (L.A.N.) represent 11% of the medical prescription of neuroleptics in France (about 40,000 patients in 1974). Some psychiatrists remain reluctant to prescribe long acting-neuroleptics and many problems related to their use are still solved empirically:1) relation between the posology of the standard Neuroleptic and the corresponding L.A.N. 2)the problem of dose-interval. 3) the necessary prescription of an parkinsonian. The superiority of N.A.P. over the Neuroleptic Standard has received no experimental confirmation in France, but the L.A.N. appear to us as an original therapy, the "pivot chemotherapy" around which psychotherapy and sociotherapy can be arranged without anarchical and ceaseless changes of neuroleptic compounds and of posology. | [
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