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pubmed23n0001_300

[Effect of peripheral counterpulsation on the body of an animal with intact heart].

The effect of lasting peripheral counterpulsation upon the haemodynamics and the main biochemical factors of the blood was studied in 14 dogs with intact hearts. In cases of significant tachycardia counterpulsation in a 1:2 regimen results in only a partial reduction of the resistance to the cardiac output. This does not always permit to prevent the formation of the phenomenon of an elevated myocardial contractility. The haemodynamic conditions are most optimal in a 1:1 regimen of counterpulsation. Slowing down of the cardiac rhythm was achieved by means of hypothermia.

[Effect of peripheral counterpulsation on the body of an animal with intact heart]. The effect of lasting peripheral counterpulsation upon the haemodynamics and the main biochemical factors of the blood was studied in 14 dogs with intact hearts. In cases of significant tachycardia counterpulsation in a 1:2 regimen results in only a partial reduction of the resistance to the cardiac output. This does not always permit to prevent the formation of the phenomenon of an elevated myocardial contractility. The haemodynamic conditions are most optimal in a 1:1 regimen of counterpulsation. Slowing down of the cardiac rhythm was achieved by means of hypothermia.

pubmed23n0001_301

[Regulation of interrelation between pulmonary ventilation and circulation].

In acute experiments on 92 cats anesthetized with Urethane and kept under controlled respiration the mechanism of tonic activity of the pulmonary vessels was studied in the presence of a decreased partial pressure of oxygen in the alveoli. The tonicity of the pulmonary vessels was recorded during autoperfusion of the vessels of the posterior lobe of the left lung by means of a perfusion pump. Simultaneously, the pressure in the common carotid artery was recorded and the oxygen saturation of the blood was measured. Pharmacological analysis was used for the study of the mechanism of the pressor reaction of the pulmonary vessels under hypoxic hypoxy, and it demonstrated that the pression of the pulmonary vessels that develops under alveolar hypoxy is less distinct under the effect of the ganglioblocking agent Benzo-hexonium, as well as the myotropic agents Papaverine and Chloracizine. The above reaction was significantly inhibited by the blocking agents of the D- and M-serotonin-reactive structures -- Dihydroergothamine, Morphine and Novocain, and it was completely lacking against the background of the action of Izadrine and Dimedrol -- blocking agents of serotonin- and histamine-reactive structures. It can be supposed that the D- and M-serotonin-reactive, and probably also the histamine-reactive structures participate in the regulation of the interrelationship of ventilation and pulmonary circulation.

[Regulation of interrelation between pulmonary ventilation and circulation]. In acute experiments on 92 cats anesthetized with Urethane and kept under controlled respiration the mechanism of tonic activity of the pulmonary vessels was studied in the presence of a decreased partial pressure of oxygen in the alveoli. The tonicity of the pulmonary vessels was recorded during autoperfusion of the vessels of the posterior lobe of the left lung by means of a perfusion pump. Simultaneously, the pressure in the common carotid artery was recorded and the oxygen saturation of the blood was measured. Pharmacological analysis was used for the study of the mechanism of the pressor reaction of the pulmonary vessels under hypoxic hypoxy, and it demonstrated that the pression of the pulmonary vessels that develops under alveolar hypoxy is less distinct under the effect of the ganglioblocking agent Benzo-hexonium, as well as the myotropic agents Papaverine and Chloracizine. The above reaction was significantly inhibited by the blocking agents of the D- and M-serotonin-reactive structures -- Dihydroergothamine, Morphine and Novocain, and it was completely lacking against the background of the action of Izadrine and Dimedrol -- blocking agents of serotonin- and histamine-reactive structures. It can be supposed that the D- and M-serotonin-reactive, and probably also the histamine-reactive structures participate in the regulation of the interrelationship of ventilation and pulmonary circulation.

pubmed23n0001_302

Multiple cyclic nucleotide phosphodiesterases in rat kidney.

Using DEAE-cellulose chromatography and Agarose gel filtration we have partially purified a low Km cyclic adenosine monophosphate (AMP) phosphodiesterase from the 100,000 X g supernatant of rat kidneys. The characteristics of this enzyme included a Km of approximately 4 muM a pH optimum of around 8.0 and a requirement for magnesium. This preparation should be suitable for investigation of possible effects of hormones, drugs and cellular constituents on the cyclic AMP pathway through any direct effects on the low Km enzyme. We have also demonstrated a nonspecific, high Km cyclic nucleotide phosphodiesterase and possibly a specific cyclic guanosine monophosphate (GMP) phosphodiesterase in the soluble fraction from rat kidneys.

Multiple cyclic nucleotide phosphodiesterases in rat kidney. Using DEAE-cellulose chromatography and Agarose gel filtration we have partially purified a low Km cyclic adenosine monophosphate (AMP) phosphodiesterase from the 100,000 X g supernatant of rat kidneys. The characteristics of this enzyme included a Km of approximately 4 muM a pH optimum of around 8.0 and a requirement for magnesium. This preparation should be suitable for investigation of possible effects of hormones, drugs and cellular constituents on the cyclic AMP pathway through any direct effects on the low Km enzyme. We have also demonstrated a nonspecific, high Km cyclic nucleotide phosphodiesterase and possibly a specific cyclic guanosine monophosphate (GMP) phosphodiesterase in the soluble fraction from rat kidneys.

pubmed23n0001_303

Effect on aging on plasma renin and aldosterone in normal man.

The influence of aging on the renin-angiotensin-aldosterone system was evaluated by comparing young (20 to 30 yr) with elderly (62 to 70 yr) healthy subjects. Despite comparable body sodium-fluid balance in the two age groups, serum renin concentration, plasma renin activity and aldosterone concentrations were lower in the elderly. The age-related decreases in circulating renin and aldosterone concentrations were slight while subjects were supine and receiving normal sodium intake; when upright and during sodium depletion, they were more pronounced. Inverse renin-blood pressure interrelations were noted during two of four study conditions involving normal sodium intake or mild sodium depletion (r = --0.44 and --0.47, respectively), but not during progressive sodium depletion. Plasma renin levels were decreased in the elderly regardless of the presence or absence of an inverse relationship with blood pressure. Aldosterone and cortisol responses to corticotropin infusion were unaltered in the elderly. It is concluded that aging may cause a decrease in circulating renin, with parallel lowering of plasma aldosterone concentrations.

Effect on aging on plasma renin and aldosterone in normal man. The influence of aging on the renin-angiotensin-aldosterone system was evaluated by comparing young (20 to 30 yr) with elderly (62 to 70 yr) healthy subjects. Despite comparable body sodium-fluid balance in the two age groups, serum renin concentration, plasma renin activity and aldosterone concentrations were lower in the elderly. The age-related decreases in circulating renin and aldosterone concentrations were slight while subjects were supine and receiving normal sodium intake; when upright and during sodium depletion, they were more pronounced. Inverse renin-blood pressure interrelations were noted during two of four study conditions involving normal sodium intake or mild sodium depletion (r = --0.44 and --0.47, respectively), but not during progressive sodium depletion. Plasma renin levels were decreased in the elderly regardless of the presence or absence of an inverse relationship with blood pressure. Aldosterone and cortisol responses to corticotropin infusion were unaltered in the elderly. It is concluded that aging may cause a decrease in circulating renin, with parallel lowering of plasma aldosterone concentrations.

pubmed23n0001_304

[Immune mechanisms in uremia].

There is both clinical and experimental evidence that cellular and humoral immunity are suppressed in patients with renal insufficiency: observations in organ transplantation and in vitro stimulation of lymphocytes from uraemic patients, investigations of acute and late hypersensitivity reactions, the immune response after active immunization as well as changes of immunoglobulins and lymphatic organs in uraemia are discussed in the paper. The underlying mechanisms are complex and not yet fully understood. Lymphopenia, atrophy of the thymus gland, toxic serum factors, induction of enhancing mechanisms by certain serum fractions and metabolic defects of lymphocytes--all were shown to be involved or at least considered to be. At present, however, it is impossible to define their rank of importance and the exact place they may occupy in the genesis of this type of "natural immunosuppression".

[Immune mechanisms in uremia]. There is both clinical and experimental evidence that cellular and humoral immunity are suppressed in patients with renal insufficiency: observations in organ transplantation and in vitro stimulation of lymphocytes from uraemic patients, investigations of acute and late hypersensitivity reactions, the immune response after active immunization as well as changes of immunoglobulins and lymphatic organs in uraemia are discussed in the paper. The underlying mechanisms are complex and not yet fully understood. Lymphopenia, atrophy of the thymus gland, toxic serum factors, induction of enhancing mechanisms by certain serum fractions and metabolic defects of lymphocytes--all were shown to be involved or at least considered to be. At present, however, it is impossible to define their rank of importance and the exact place they may occupy in the genesis of this type of "natural immunosuppression".

pubmed23n0001_305

Improvement of renin determination in human plasma using a commonly available renin standard in a radioimmunological method.

A new method for the measurement of renin in human plasma is described. The method is based on the introduction of the internationally available renin standard of the Medical Research Council (MRC) London, as a calibration system. Thus, some principal disadvantages of methods expressing results in renin reaction velocity (angiotensin generation rate) only are avoided. Both renins, unknown and standard, react with a sheep substrate preparation and are handled identically throughout the whole procedure including the angiotensin I radioimmunoassay (RIA). The plasma renin concentration (PRC) is given in 10(-6) MRC-renin units (muM/ml). the renin standard is free of angiotensin, angiotensinases, and angiotensinogen; it is stable on storage. Identical enzyme kinetics are shown for both renins. An interference between endogenous and exogenous substrate could be avoided. The potentially harmful influences of proteins from the enzyme incubation mixture of the RIA dose response curve are shown. The use of an angiotensin I calibration system could be omitted. Using a standard renin dilution from 250-0.9 muU/ml also the full biological range is covered. When giving an unrestricted diet the preliminary normal values of PRC are 21.9 +/- 12.6 muU/ml in recumbent and 40.1 +/- 19.8 muU/ml in upright position (n = 16,x +/- s, age 20-35 years). Earlier findings of age-dependency of PRC were confirmed.

Improvement of renin determination in human plasma using a commonly available renin standard in a radioimmunological method. A new method for the measurement of renin in human plasma is described. The method is based on the introduction of the internationally available renin standard of the Medical Research Council (MRC) London, as a calibration system. Thus, some principal disadvantages of methods expressing results in renin reaction velocity (angiotensin generation rate) only are avoided. Both renins, unknown and standard, react with a sheep substrate preparation and are handled identically throughout the whole procedure including the angiotensin I radioimmunoassay (RIA). The plasma renin concentration (PRC) is given in 10(-6) MRC-renin units (muM/ml). the renin standard is free of angiotensin, angiotensinases, and angiotensinogen; it is stable on storage. Identical enzyme kinetics are shown for both renins. An interference between endogenous and exogenous substrate could be avoided. The potentially harmful influences of proteins from the enzyme incubation mixture of the RIA dose response curve are shown. The use of an angiotensin I calibration system could be omitted. Using a standard renin dilution from 250-0.9 muU/ml also the full biological range is covered. When giving an unrestricted diet the preliminary normal values of PRC are 21.9 +/- 12.6 muU/ml in recumbent and 40.1 +/- 19.8 muU/ml in upright position (n = 16,x +/- s, age 20-35 years). Earlier findings of age-dependency of PRC were confirmed.

pubmed23n0001_306

[Validity of pH measurements by means of micro pH combination electrodes in blood and other biological fluids (author's transl)].

pH measurements in blood or in media containing either proteins or polypeptides and performed by means of micro pH combination electrodes type N 58 (Schott & Gen., Mainz) yield in a systematic error according to the regression line y = 1.135 chi - 0.842, in the range between pH 5.3 and 8.3. This deviation from the real pH value is independent of the protein concentration and amounts to 0.1-0.2 pH units in the physiological range. The error does not occur if the pH measurements are performed in media which are free of proteins and polypeptides, respectively. If the electrolyte solution within the reference electrode is replaced (NaCl solution instead of KCl solution) the error is distinctly reduced. For this reason, this deviation should be caused by the variation of the diffusion potential across the platinum junctions.

[Validity of pH measurements by means of micro pH combination electrodes in blood and other biological fluids (author's transl)]. pH measurements in blood or in media containing either proteins or polypeptides and performed by means of micro pH combination electrodes type N 58 (Schott & Gen., Mainz) yield in a systematic error according to the regression line y = 1.135 chi - 0.842, in the range between pH 5.3 and 8.3. This deviation from the real pH value is independent of the protein concentration and amounts to 0.1-0.2 pH units in the physiological range. The error does not occur if the pH measurements are performed in media which are free of proteins and polypeptides, respectively. If the electrolyte solution within the reference electrode is replaced (NaCl solution instead of KCl solution) the error is distinctly reduced. For this reason, this deviation should be caused by the variation of the diffusion potential across the platinum junctions.

pubmed23n0001_307

The effects of drinking on offspring: an historical survey of the American and British literature.

Current research on the effects on offspring of drinking during pregnancy has revived interest in an extremely old topic. Observations made during England's Gin Epidemic (1720-1750) were followed by warnings of 19th-century medical writers that parental drinking could damage the fetus. Many concurring studies were reported in the medical literature from 1865 to 1920. Research interest declined during Prohibition, and authorities later discounted the previous work. Recently a relationship between maternal drinking and abnormal morphogenesis has been again described.

The effects of drinking on offspring: an historical survey of the American and British literature. Current research on the effects on offspring of drinking during pregnancy has revived interest in an extremely old topic. Observations made during England's Gin Epidemic (1720-1750) were followed by warnings of 19th-century medical writers that parental drinking could damage the fetus. Many concurring studies were reported in the medical literature from 1865 to 1920. Research interest declined during Prohibition, and authorities later discounted the previous work. Recently a relationship between maternal drinking and abnormal morphogenesis has been again described.

pubmed23n0001_308

[Glutathione (author's transl)].

Glutathione plays an important role in biology and medicine. Most cells of plants and animals contain high concentrations of reduced glutathione and a much smaller amount of oxidised glutathione. GSH is important for several metabolic functions of live cells, e.g. the protection of oxidative stress by peroxides, mediation of enzyme reactions, regulation of metabolic events, transport of amino acids across cell membranes via the gamma-glutamyl cycle, elimination of foreign compounds by GSH-conjugation, release of neurotransmitter substances. Irreversible perturbations of the glutathione metabolism may be the reason for severe clinical symptoms of hemolytic anemia or, perhaps, of central nervous disease.

[Glutathione (author's transl)]. Glutathione plays an important role in biology and medicine. Most cells of plants and animals contain high concentrations of reduced glutathione and a much smaller amount of oxidised glutathione. GSH is important for several metabolic functions of live cells, e.g. the protection of oxidative stress by peroxides, mediation of enzyme reactions, regulation of metabolic events, transport of amino acids across cell membranes via the gamma-glutamyl cycle, elimination of foreign compounds by GSH-conjugation, release of neurotransmitter substances. Irreversible perturbations of the glutathione metabolism may be the reason for severe clinical symptoms of hemolytic anemia or, perhaps, of central nervous disease.

pubmed23n0001_309

Hyperexcitability in the neural substrate of emotional behavior in cats after alcohol withdrawal. Evidence of a rapid development of alcohol dependence.

Substantial and prolonged withdrawal hyperexcitability in the neural substrate for affective defense was revealed by behavioral and electrophysiological measures in cats exposed to moderate to heavy doses of alcohol for periods ranging from 6 to 72 hours. The data are interpreted as indicating a rapid development of physical dependence on alcohol in this portion of the central nervous system.

Hyperexcitability in the neural substrate of emotional behavior in cats after alcohol withdrawal. Evidence of a rapid development of alcohol dependence. Substantial and prolonged withdrawal hyperexcitability in the neural substrate for affective defense was revealed by behavioral and electrophysiological measures in cats exposed to moderate to heavy doses of alcohol for periods ranging from 6 to 72 hours. The data are interpreted as indicating a rapid development of physical dependence on alcohol in this portion of the central nervous system.

pubmed23n0001_310

Interactions of age, sex and long-term alcohol intake in selectively bred strains of rats.

The alcohol consumption by five genotypes of rats was studied in two experiments. Alcohol intake was age-dependent in rats bred for high emotional reactivity and avoidance conditionability. Differences in consumption by sex appeared to be primarily due to differences in body weight.

Interactions of age, sex and long-term alcohol intake in selectively bred strains of rats. The alcohol consumption by five genotypes of rats was studied in two experiments. Alcohol intake was age-dependent in rats bred for high emotional reactivity and avoidance conditionability. Differences in consumption by sex appeared to be primarily due to differences in body weight.

pubmed23n0001_311

Regional study of acid hydrolases and lysosomal membrane properties in the normal human brain at various ages.

Acid hydrolases and lysosomal membrane properties were studied at various ages in the normal human brain. In CSF and four brain regions, the inferior olive, the cerebellar cortex, the caudate nucleus and the frontal cortex were thus beta-galactosidase, beta-glucosidase, alpha-mannosidase, hexosaminidase and acid phosphatase biochemically quantitated at ages varying between 2 and 89 years of age. Also the membrane latency for acid phosphatase was studied in these regions. No major regional quantitative differences were found with regard to the enzymes studied. Their kinetic properties were also defined. There appeared to exist a regional and intra-areal variation in lysosomal membrane permeability. There was, however, no age related increase in total enzyme contents. The possibility significance of these findings are discussed with reference to the aging process.

Regional study of acid hydrolases and lysosomal membrane properties in the normal human brain at various ages. Acid hydrolases and lysosomal membrane properties were studied at various ages in the normal human brain. In CSF and four brain regions, the inferior olive, the cerebellar cortex, the caudate nucleus and the frontal cortex were thus beta-galactosidase, beta-glucosidase, alpha-mannosidase, hexosaminidase and acid phosphatase biochemically quantitated at ages varying between 2 and 89 years of age. Also the membrane latency for acid phosphatase was studied in these regions. No major regional quantitative differences were found with regard to the enzymes studied. Their kinetic properties were also defined. There appeared to exist a regional and intra-areal variation in lysosomal membrane permeability. There was, however, no age related increase in total enzyme contents. The possibility significance of these findings are discussed with reference to the aging process.

pubmed23n0001_312

The late effects of selected immunosuppressants on immunocompetence, disease incidence, and mean life-span. II. Cell-mediated immune activity.

The late effects of various immunosuppressive insults on cell-mediated immunity in mice were studied in an attempt to assess the role of immune surveillance in the aging process. Results were obtained using susceptibility to allogeneic tumor cell challenge, graft-versus-host reaction (GVHR), blastogenic response to PHA, a thymus derived T cell-specific plant mitogen, and cytolytic activity against allogeneic tumor cells as measures of immunologic activity. In vivo studies late in life show that resistance to allogeneic tumor cells is significantly decreased in thymectomized mice, whereas those treated with cortisone, cyclophosphamide and sublethal X-ray remain unchanged. Spleen cells from only the thymectomized and the sublethally irradiated mice show reduced activity in the GVHR. No difference is seen in the activity of bone marrow cells. Results consistent with these findings were obtained in in vitro studies. Thus spleen cells from thymectomized or sublethally irradiated mice show decreased activity is response to PHA, whereas no change is seen in spleen cells from other treated groups. Hence, surgical and physical insults are more likely to induce long-lasting immunosuppression in those immunocompetent tissues whose activity normally diminishes with advancing age. Furthermore, the degree of immunosuppression seen in this study is not of the order of magnitude that one could reasonably predict a significant decrease in mean life-span.

The late effects of selected immunosuppressants on immunocompetence, disease incidence, and mean life-span. II. Cell-mediated immune activity. The late effects of various immunosuppressive insults on cell-mediated immunity in mice were studied in an attempt to assess the role of immune surveillance in the aging process. Results were obtained using susceptibility to allogeneic tumor cell challenge, graft-versus-host reaction (GVHR), blastogenic response to PHA, a thymus derived T cell-specific plant mitogen, and cytolytic activity against allogeneic tumor cells as measures of immunologic activity. In vivo studies late in life show that resistance to allogeneic tumor cells is significantly decreased in thymectomized mice, whereas those treated with cortisone, cyclophosphamide and sublethal X-ray remain unchanged. Spleen cells from only the thymectomized and the sublethally irradiated mice show reduced activity in the GVHR. No difference is seen in the activity of bone marrow cells. Results consistent with these findings were obtained in in vitro studies. Thus spleen cells from thymectomized or sublethally irradiated mice show decreased activity is response to PHA, whereas no change is seen in spleen cells from other treated groups. Hence, surgical and physical insults are more likely to induce long-lasting immunosuppression in those immunocompetent tissues whose activity normally diminishes with advancing age. Furthermore, the degree of immunosuppression seen in this study is not of the order of magnitude that one could reasonably predict a significant decrease in mean life-span.

pubmed23n0001_313

Lipoxygenase isozymes of peanut.

Lipoxygenase was isolated and partially purified from peanut seed by ammonium sulfate precipitation, gel filtration, and ion exchange column chromatography. Three isozymes of lipoxygenase were identified. Two had pH optima of 6.2, and the other an optimum of 8.3. Molecular weight of each isozyme was 7.3 x 10(4), as determined by gel filtration. The alkaline optimum isozyme was not inhibited by NaCN and was inhibited by CaCl2 except at very low concentrations. The acid optimum isozymes were inhibited by NaCN and were stimulated by CaCl2 concentrations up to ca. 0.7 mM.

Lipoxygenase isozymes of peanut. Lipoxygenase was isolated and partially purified from peanut seed by ammonium sulfate precipitation, gel filtration, and ion exchange column chromatography. Three isozymes of lipoxygenase were identified. Two had pH optima of 6.2, and the other an optimum of 8.3. Molecular weight of each isozyme was 7.3 x 10(4), as determined by gel filtration. The alkaline optimum isozyme was not inhibited by NaCN and was inhibited by CaCl2 except at very low concentrations. The acid optimum isozymes were inhibited by NaCN and were stimulated by CaCl2 concentrations up to ca. 0.7 mM.

pubmed23n0001_314

Crassin acetate, the principal antineoplastic agent in four gorgonians of the Pseudoplexaura genus.

Crassin acetate, a lactonic cembrane diterpene, has been shown to be the principal antineoplastic agent present in the marine invertebrates (gorgonians) Pseudoplexaura porosa, P. flagellosa, P. wagenaari and P. crucis.

Crassin acetate, the principal antineoplastic agent in four gorgonians of the Pseudoplexaura genus. Crassin acetate, a lactonic cembrane diterpene, has been shown to be the principal antineoplastic agent present in the marine invertebrates (gorgonians) Pseudoplexaura porosa, P. flagellosa, P. wagenaari and P. crucis.

pubmed23n0001_315

[Computer experience and further developments in the respiratory function laboratory (author's transl)].

Reported is on satisfactory results obtained with a small-size computer consisting of punching and scanning device, as well as plain writing machine in the respiratory function laboratory. Developed in on- as well as off-line processing by an own technical staff, a diagnostic and teaching program was established for all respiratory function routine methods with the advantages of a large number of cases examined, elimination of sources of error, considerable supply of data and information, automatic documentation and filing, plain writing, interpretation and evaluation of findings. In continuation of such works also the blood gas analysis has been included. These values as the total of disturbances of the pathophysiological acid-base status are considered and interpreted. Clinical correction is forced in this man-machine dialogue by automatic stops of the whole machinery before going on. Subsequently and in addition are computer alveolar-arterial oxygen pressure gradient, venous shunt and oxygen saturation and expressed utilizing the capacity of the small-size computer. Further developments in the respiratory function diagnostic- and teaching program for small-size computers--not too expensive in the building block principle - are intended.

[Computer experience and further developments in the respiratory function laboratory (author's transl)]. Reported is on satisfactory results obtained with a small-size computer consisting of punching and scanning device, as well as plain writing machine in the respiratory function laboratory. Developed in on- as well as off-line processing by an own technical staff, a diagnostic and teaching program was established for all respiratory function routine methods with the advantages of a large number of cases examined, elimination of sources of error, considerable supply of data and information, automatic documentation and filing, plain writing, interpretation and evaluation of findings. In continuation of such works also the blood gas analysis has been included. These values as the total of disturbances of the pathophysiological acid-base status are considered and interpreted. Clinical correction is forced in this man-machine dialogue by automatic stops of the whole machinery before going on. Subsequently and in addition are computer alveolar-arterial oxygen pressure gradient, venous shunt and oxygen saturation and expressed utilizing the capacity of the small-size computer. Further developments in the respiratory function diagnostic- and teaching program for small-size computers--not too expensive in the building block principle - are intended.

pubmed23n0001_316

Phosphate transport in rat liver mitochondria. Kinetics, inhibitor sensitivity, energy requirements, and labeled components.

Experiments were carried out to define the kinetic parameters of the major phosphate transport processes of rat liver mitochondria, and to obtain information about the molecular properties of these systems.

Phosphate transport in rat liver mitochondria. Kinetics, inhibitor sensitivity, energy requirements, and labeled components. Experiments were carried out to define the kinetic parameters of the major phosphate transport processes of rat liver mitochondria, and to obtain information about the molecular properties of these systems.

pubmed23n0001_317

L-tyrosine: 2-oxoglutarate aminotransferase induction by hydrocortisone in the thymus of the white rat.

Hydrocortisone hemisuccinate within 4 hours after in vivo administration produced an increase in precursor incoporation into rat thymus RNA and proteins in the whole animal. From these results, together with information obtained from measurements of the tyrosine aminotransferase activity and the action of mitomycin C administered one hour before the injection of hydrocortisone, it can be concluded that the increase in tissue level of the enzyme, consequent to hydrocortisone treatment, results from an increased rate of biosynthesis of the enzyme, which participates in the catabolic processes of proteins in glucocorticoid sensitive thymus cells.

L-tyrosine: 2-oxoglutarate aminotransferase induction by hydrocortisone in the thymus of the white rat. Hydrocortisone hemisuccinate within 4 hours after in vivo administration produced an increase in precursor incoporation into rat thymus RNA and proteins in the whole animal. From these results, together with information obtained from measurements of the tyrosine aminotransferase activity and the action of mitomycin C administered one hour before the injection of hydrocortisone, it can be concluded that the increase in tissue level of the enzyme, consequent to hydrocortisone treatment, results from an increased rate of biosynthesis of the enzyme, which participates in the catabolic processes of proteins in glucocorticoid sensitive thymus cells.

pubmed23n0001_318

Identification of the 30 S protein adjacent to peptidyl transferase catalytic center of Escherichia coli ribosomes.

Iodoacetylphenylalanyl-tRNAPhe was used as an affinity label to localize the ribosomal components involved in the peptidyl transferase catalytic center of Escherichia coli ribosomes. When labeling was carried out at pH 5.0, the affinity label could specifically label the ribosomal components which comprise the catalytic center. Analysis of ribosomal proteins which had reacted with the affinity label revealed that a 30 S subunit protein, S 20, was located at or near to the ribosomal binding site of the 3'-terminus of aminoacyl- or peptidyl-tRNA.

Identification of the 30 S protein adjacent to peptidyl transferase catalytic center of Escherichia coli ribosomes. Iodoacetylphenylalanyl-tRNAPhe was used as an affinity label to localize the ribosomal components involved in the peptidyl transferase catalytic center of Escherichia coli ribosomes. When labeling was carried out at pH 5.0, the affinity label could specifically label the ribosomal components which comprise the catalytic center. Analysis of ribosomal proteins which had reacted with the affinity label revealed that a 30 S subunit protein, S 20, was located at or near to the ribosomal binding site of the 3'-terminus of aminoacyl- or peptidyl-tRNA.

pubmed23n0001_319

The value of a histamine H2-receptor antagonist in the management of patients with the Zollinger-Ellison syndrome.

Inhibition of acid secretion by an H2-receptor antagonist (metiamide) was assessed in three patients with the Zollinger-Ellison syndrome. Metiamide (200 or 300 mg) inhibited acid secretion transiently (2 1/2 hours) by 85 to 100 per cent in all patients. Although anticholinergic drugs alone inhibited acid secretion by only 0 to 35 per cent in these patients, the combination of metiamide and anticholinergic markedly prolonged the inhibitory effect of metiamide. Total gastrectomy was refused by one patient, and was impossible in another; both were treated with metiamide and anticholinergic for five and 10 months. A third patient was treated with metiamide and anticholinergic for three weeks in preparation for total gastrectomy. Ulcer pain and diarrhea disappeared, and each gained weight. H2-receptor antagonists may be useful in the treatment of some patients with the Zollinger-Ellison syndrome.

The value of a histamine H2-receptor antagonist in the management of patients with the Zollinger-Ellison syndrome. Inhibition of acid secretion by an H2-receptor antagonist (metiamide) was assessed in three patients with the Zollinger-Ellison syndrome. Metiamide (200 or 300 mg) inhibited acid secretion transiently (2 1/2 hours) by 85 to 100 per cent in all patients. Although anticholinergic drugs alone inhibited acid secretion by only 0 to 35 per cent in these patients, the combination of metiamide and anticholinergic markedly prolonged the inhibitory effect of metiamide. Total gastrectomy was refused by one patient, and was impossible in another; both were treated with metiamide and anticholinergic for five and 10 months. A third patient was treated with metiamide and anticholinergic for three weeks in preparation for total gastrectomy. Ulcer pain and diarrhea disappeared, and each gained weight. H2-receptor antagonists may be useful in the treatment of some patients with the Zollinger-Ellison syndrome.

pubmed23n0001_320

Histamine receptors in the vasculature of the rabbit ear.

Histamine has a dual action on the isolated perfused ear preparation of the rabbit. The amine induced a dose-dependent rise in perfusion pressure when the preparation was perfused with Krebs' solution. This pressor response was reversed to a depressor effect when meypramine was added to the perfusion fluid. This depressor effect of the amine was also dose-related. Metiamide competitively inhibited the depressor effect of histamine. Prior treatment of the ear vessels with metiamide alone caused an increase in histamine-induced perfusion pressure. From these results it was concluded that the predominant pressor effect of histamine on the vascular bed of the rabbit ear is mediated through the H1-receptors and the depressor effect of the amine through histamine H2-receptors.

Histamine receptors in the vasculature of the rabbit ear. Histamine has a dual action on the isolated perfused ear preparation of the rabbit. The amine induced a dose-dependent rise in perfusion pressure when the preparation was perfused with Krebs' solution. This pressor response was reversed to a depressor effect when meypramine was added to the perfusion fluid. This depressor effect of the amine was also dose-related. Metiamide competitively inhibited the depressor effect of histamine. Prior treatment of the ear vessels with metiamide alone caused an increase in histamine-induced perfusion pressure. From these results it was concluded that the predominant pressor effect of histamine on the vascular bed of the rabbit ear is mediated through the H1-receptors and the depressor effect of the amine through histamine H2-receptors.

pubmed23n0001_321

Relative pre- and postsynaptic potencies of alpha-adrenoceptor agonists in the rabbit pulmonary artery.

The rabbit pulmonary artery contains postsynaptic alpha-adrenoceptors which meidate smooth muscle contraction; its noradrenergic nerves contain presynaptic alpha-adrenoceptors which mediate inhibition of the release of the transmitter evoked by nerve impulses. Dose-response curves for the pre- and postsynaptic effects of eight alpha-receptor agonists were determined on superfused strips of the artery in the presence of cocaine, corticosterone and propranolo. 1. According to the concentrations which caused 20% of the maximal contraction (EC20 post), the postsynaptic rank order of potency was: adrenaline greater than noradrenaline greater than oxymetazoline greater than naphazoline greater than phenylephrine greater than tramazoline greater than alpha-methylnoradrenaline greater than methoxamine. The pA2 values of phentolamine againstoxymethazoline, phenylephrine, alpha-methylnoradrenaline and methoxamine were 7.43, 7.48, 7.59 and 7.69, respectively. 2. For the investigation of presynaptic effects, the arteries were preincubated with 3H-noradrenaline. All agonists inhibited the overflow of tritium evoked by transmural sympathetic nerve stimulation. According to the concentrations which reduced the stimulation-induced overflow by 20% (EC20 pre), the rank order of potency was: adrenaline greater than oxymetazoline greater than tramazoline greater than alpha-methylnoradrenaline greater than noradrenaline greater than naphazoline greater than phenylephrine greater than methoxamine. 10(-5) M phentolamine shifted the presynaptic dose-response curves for moradrenaline and oxymethazoline to the right. 3. The ratio EC20 pre/EC20 post was calculated for each agonist as an index of its relative post- and presynaptic potency. According to the ratios, the agonists were arbitrarily classified into three groups. Group 1 (ratio about 30: preferentially postsynaptic agonists) comprised methoxamine and phenylephrine; group 2 (ratio near 1; similar pre- and postsynaptic potencies) comprised noradrenaline, adrenaline and naphazoline; group 3 (ratio below 0.2; preferentially presynaptic agonists) comprised oxymetazoline, alpha-methylnoradrenaline and tramazoline (as well as clonidine). 4. Preferentially presynaptic and preferentially postsynaptic agonists had opposite effects on the basoconstrictor response to nerve stimulation. Methoxamine and phenylephrine either did not change or enhanced, but never reduced, the response. In contrast, oxymetazoline, alpha-methylnoradrenaline and tramazoline at low concentrations selectively inhibited the response to stimulation at low frequency (0.25-2Hz). 5. It is concluded that alpha-adrenoceptor agonists vary widely in their relative pre- and postsynaptic potencies, possibly because of structural differences between pre- and postsynaptic alpha-receptors. Pre- and postsynaptic components contribute to their overll postsynaptic effec in actively transmitting synapses. The preferential activation of presynaptic alpha-receptors results in alpha-adrenergic inhibition of synaptic transmission.

Relative pre- and postsynaptic potencies of alpha-adrenoceptor agonists in the rabbit pulmonary artery. The rabbit pulmonary artery contains postsynaptic alpha-adrenoceptors which meidate smooth muscle contraction; its noradrenergic nerves contain presynaptic alpha-adrenoceptors which mediate inhibition of the release of the transmitter evoked by nerve impulses. Dose-response curves for the pre- and postsynaptic effects of eight alpha-receptor agonists were determined on superfused strips of the artery in the presence of cocaine, corticosterone and propranolo. 1. According to the concentrations which caused 20% of the maximal contraction (EC20 post), the postsynaptic rank order of potency was: adrenaline greater than noradrenaline greater than oxymetazoline greater than naphazoline greater than phenylephrine greater than tramazoline greater than alpha-methylnoradrenaline greater than methoxamine. The pA2 values of phentolamine againstoxymethazoline, phenylephrine, alpha-methylnoradrenaline and methoxamine were 7.43, 7.48, 7.59 and 7.69, respectively. 2. For the investigation of presynaptic effects, the arteries were preincubated with 3H-noradrenaline. All agonists inhibited the overflow of tritium evoked by transmural sympathetic nerve stimulation. According to the concentrations which reduced the stimulation-induced overflow by 20% (EC20 pre), the rank order of potency was: adrenaline greater than oxymetazoline greater than tramazoline greater than alpha-methylnoradrenaline greater than noradrenaline greater than naphazoline greater than phenylephrine greater than methoxamine. 10(-5) M phentolamine shifted the presynaptic dose-response curves for moradrenaline and oxymethazoline to the right. 3. The ratio EC20 pre/EC20 post was calculated for each agonist as an index of its relative post- and presynaptic potency. According to the ratios, the agonists were arbitrarily classified into three groups. Group 1 (ratio about 30: preferentially postsynaptic agonists) comprised methoxamine and phenylephrine; group 2 (ratio near 1; similar pre- and postsynaptic potencies) comprised noradrenaline, adrenaline and naphazoline; group 3 (ratio below 0.2; preferentially presynaptic agonists) comprised oxymetazoline, alpha-methylnoradrenaline and tramazoline (as well as clonidine). 4. Preferentially presynaptic and preferentially postsynaptic agonists had opposite effects on the basoconstrictor response to nerve stimulation. Methoxamine and phenylephrine either did not change or enhanced, but never reduced, the response. In contrast, oxymetazoline, alpha-methylnoradrenaline and tramazoline at low concentrations selectively inhibited the response to stimulation at low frequency (0.25-2Hz). 5. It is concluded that alpha-adrenoceptor agonists vary widely in their relative pre- and postsynaptic potencies, possibly because of structural differences between pre- and postsynaptic alpha-receptors. Pre- and postsynaptic components contribute to their overll postsynaptic effec in actively transmitting synapses. The preferential activation of presynaptic alpha-receptors results in alpha-adrenergic inhibition of synaptic transmission.

pubmed23n0001_322

Investigation into some imidazoline compounds, with respect to peripheral alpha-adrenoceptor stimulation and depression of cardiovascular centers.

Peripheral alpha-adrenoceptor stimulation was tested by means of hypertensive effects of the drugs following i.v. injection in spinal rats. Naphazoline (NP), oxymetazoline (OM), St 91-2-(2,6-diethylphenylimino)-2-imidazolidine--and St 1697--2-(2-ethyl, 6-methylphenylimino)-2-imidazolidine--were 3 to 5 times more potent in tthis respect thatn clonidine (CLON) whereas St 363--2-(2,4-dichlorophenylimino)-2-imidazolidine--and xylazine (XY) exerted only approx. 1/20 the effect of that of clonidine. Sympathoinhibitory activity after i.v. injection was tested by the bradycardiac effect in vagotomized rats; St 1697, St 363 and XY were active, approx. 1/10-1/30 of CLON, whereas NP, OM and St 91 were inactive. However, following intracisternal (i.ci.) injection of cardiovascular depression, typical for clonidine: (1) in dogs with blocked beta-adrenoceptors, the drugs facilitated the vagally meditated cardiodepressor reflex in response to baroreceptor stimulation by i.v. injection of angiotensin; (2) in dogs treated with atropine and in (3) vagotomized cats (only NP, OM and St 363) a long lasting decrease in heart rate was observed. Some of the experiments were complicated by increases in blood pressure, due to the "leakage" of small amounts of the highly vasopressor active drugs, from the cisternal spaces into the peripheral circulation. The majority of results indicated, that the central cardiovascular depressor effects of the tested drugs depend on their alpha-adrenoreceptor stimulating potency and on their ability to penetrate from cerebrospinal fluid or from the blood to cardiovascular centers. Relationships between the ability for penetration and the lipoid affinity are discussed.

Investigation into some imidazoline compounds, with respect to peripheral alpha-adrenoceptor stimulation and depression of cardiovascular centers. Peripheral alpha-adrenoceptor stimulation was tested by means of hypertensive effects of the drugs following i.v. injection in spinal rats. Naphazoline (NP), oxymetazoline (OM), St 91-2-(2,6-diethylphenylimino)-2-imidazolidine--and St 1697--2-(2-ethyl, 6-methylphenylimino)-2-imidazolidine--were 3 to 5 times more potent in tthis respect thatn clonidine (CLON) whereas St 363--2-(2,4-dichlorophenylimino)-2-imidazolidine--and xylazine (XY) exerted only approx. 1/20 the effect of that of clonidine. Sympathoinhibitory activity after i.v. injection was tested by the bradycardiac effect in vagotomized rats; St 1697, St 363 and XY were active, approx. 1/10-1/30 of CLON, whereas NP, OM and St 91 were inactive. However, following intracisternal (i.ci.) injection of cardiovascular depression, typical for clonidine: (1) in dogs with blocked beta-adrenoceptors, the drugs facilitated the vagally meditated cardiodepressor reflex in response to baroreceptor stimulation by i.v. injection of angiotensin; (2) in dogs treated with atropine and in (3) vagotomized cats (only NP, OM and St 363) a long lasting decrease in heart rate was observed. Some of the experiments were complicated by increases in blood pressure, due to the "leakage" of small amounts of the highly vasopressor active drugs, from the cisternal spaces into the peripheral circulation. The majority of results indicated, that the central cardiovascular depressor effects of the tested drugs depend on their alpha-adrenoreceptor stimulating potency and on their ability to penetrate from cerebrospinal fluid or from the blood to cardiovascular centers. Relationships between the ability for penetration and the lipoid affinity are discussed.

pubmed23n0001_323

The regulation of striatal tyrosine hydroxylase. Effects of gamma hydroxybutric acid and healperidol.

Gamma-hydroxybutyric acid (GHBA) in doses that increased the striatal dopamine (DA) content of rat brain failed to increase the affinity of striatal tyrosine hydroxylase (TH) for its pterdine cofactor or to change the sensitivity of the enzyme to the inhibition by DA. Haloperidol (1 mg/kg) decreased the apparent Km of striatal TH for the pteridine cofactor. However, when GHBA was injected before haloperidol it prevented the decrease in the apparent Kn of TH, in a dose related manner. In vitro GHBA (10(-4) M) neither changed the stimulation of the striatal adenylyl cyclase by DA nor its inhibition by haloperidol. These results suggest that in striatal dopaminergic terminals the Kn of TH for the pteridine cofactor is regulated by an molecuular mechanism which requires that the impulse flow in the DA neurons is unimpaired.

The regulation of striatal tyrosine hydroxylase. Effects of gamma hydroxybutric acid and healperidol. Gamma-hydroxybutyric acid (GHBA) in doses that increased the striatal dopamine (DA) content of rat brain failed to increase the affinity of striatal tyrosine hydroxylase (TH) for its pterdine cofactor or to change the sensitivity of the enzyme to the inhibition by DA. Haloperidol (1 mg/kg) decreased the apparent Km of striatal TH for the pteridine cofactor. However, when GHBA was injected before haloperidol it prevented the decrease in the apparent Kn of TH, in a dose related manner. In vitro GHBA (10(-4) M) neither changed the stimulation of the striatal adenylyl cyclase by DA nor its inhibition by haloperidol. These results suggest that in striatal dopaminergic terminals the Kn of TH for the pteridine cofactor is regulated by an molecuular mechanism which requires that the impulse flow in the DA neurons is unimpaired.

pubmed23n0001_324

Host-tumor relationship XXXIII. Inhibitor of hyaluronidase in blood serum of cancer patients.

Inhibiting activity of blood serum was determined from the decrease of N-acetyl-hexosamine end groups of hyaluronic acid products released by testicular hyaluronidase. Maximum inhibition was observed within the region of pH from 6.5 to 6.8. Correlation between the serum concentration and its inhibiting effect on hyaluronidase was found within the range of the final dilution of serum (in the reaction mixture) from 9 to 45 X. Blood sera of cancer patients showed statistically significant increase of hyaluronidase inhibitor as compared with that of health people.

Host-tumor relationship XXXIII. Inhibitor of hyaluronidase in blood serum of cancer patients. Inhibiting activity of blood serum was determined from the decrease of N-acetyl-hexosamine end groups of hyaluronic acid products released by testicular hyaluronidase. Maximum inhibition was observed within the region of pH from 6.5 to 6.8. Correlation between the serum concentration and its inhibiting effect on hyaluronidase was found within the range of the final dilution of serum (in the reaction mixture) from 9 to 45 X. Blood sera of cancer patients showed statistically significant increase of hyaluronidase inhibitor as compared with that of health people.

pubmed23n0001_325

Influence of peritoneal dialysis on factors affecting oxygen transport.

To determine the effect of changing concentrations of uremic metabolites on factors affecting oxygen transport, without the effects of extracorporeal blood pumping, we studied five patients before, during and after peritoneal dialysis. Significant decreases in serum urea, creatinine and phosphate and increase in serum bicarbonate were not associated with changes in P50, a reflection of hemoglobin-oxygen affinity. High erythrocyte 2,3-DPG concentrations decreased only slightly. Arterial pO2 increased slightly as negative fluid balance was achieved. The slight changes in oxygen transport parameters with dialysis suggest an interplay of compensatory factors and do not warrant modifying dialysis to limit the correction on acidosis or hyperphosphatemia. Effects on hemoglobin and pO2 resulting from fluid loss can be the dominant influence of peritoneal dialysis on tissue oxygenation.

Influence of peritoneal dialysis on factors affecting oxygen transport. To determine the effect of changing concentrations of uremic metabolites on factors affecting oxygen transport, without the effects of extracorporeal blood pumping, we studied five patients before, during and after peritoneal dialysis. Significant decreases in serum urea, creatinine and phosphate and increase in serum bicarbonate were not associated with changes in P50, a reflection of hemoglobin-oxygen affinity. High erythrocyte 2,3-DPG concentrations decreased only slightly. Arterial pO2 increased slightly as negative fluid balance was achieved. The slight changes in oxygen transport parameters with dialysis suggest an interplay of compensatory factors and do not warrant modifying dialysis to limit the correction on acidosis or hyperphosphatemia. Effects on hemoglobin and pO2 resulting from fluid loss can be the dominant influence of peritoneal dialysis on tissue oxygenation.

pubmed23n0001_326

Some effects of ammonium salts on renal histology and function in the dog.

NH4Cl was infused into the left renal artery of anesthetized dogs at 50-125 mum/kg/min for up to 110 min. Renal blood flow declined early then increased to supra-control levels during infusion. Kidneys perfused at 125 mum/kg/min for 90 min showed patchy to confluent mixtures of cortical necrosis and tubular necrosis. Experimental kidneys invariably showed lower urine osmolality than contralateral controls 48 h after perfusion. Kidneys with necrosis showed depressed creatinine clearance as well. Renal artery infusion of NH4 acetate or intravenous infusion of NaHCO3 during arterial infusion of NH4Cl prevented significant acidosis and caused minimal histological changes, but depression of urine osmolality was not prevented. It is concluded that renal ammonium concentrations up to 40 mum/liter for 90 min does not cause tubular necrosis but does impair urine concentration. Severe tissue damage followed renal exposure to high ammonium concentrations in the presence of metabolic or renal acidosis.

Some effects of ammonium salts on renal histology and function in the dog. NH4Cl was infused into the left renal artery of anesthetized dogs at 50-125 mum/kg/min for up to 110 min. Renal blood flow declined early then increased to supra-control levels during infusion. Kidneys perfused at 125 mum/kg/min for 90 min showed patchy to confluent mixtures of cortical necrosis and tubular necrosis. Experimental kidneys invariably showed lower urine osmolality than contralateral controls 48 h after perfusion. Kidneys with necrosis showed depressed creatinine clearance as well. Renal artery infusion of NH4 acetate or intravenous infusion of NaHCO3 during arterial infusion of NH4Cl prevented significant acidosis and caused minimal histological changes, but depression of urine osmolality was not prevented. It is concluded that renal ammonium concentrations up to 40 mum/liter for 90 min does not cause tubular necrosis but does impair urine concentration. Severe tissue damage followed renal exposure to high ammonium concentrations in the presence of metabolic or renal acidosis.

pubmed23n0001_327

Papillary necrosis associated with calyceal arteritis.

The renal papilla has a double blood supply - from both the vasa recta and the calyceal arteries. The importance of the latter supply is not established. A case of polyarteritis associated with papillary necrosis is reported, in which the calyceal vessels, supplying the area, show acute necrotizing arteritis and occlusion. The pathophysiological and clinical implications are discussed.

Papillary necrosis associated with calyceal arteritis. The renal papilla has a double blood supply - from both the vasa recta and the calyceal arteries. The importance of the latter supply is not established. A case of polyarteritis associated with papillary necrosis is reported, in which the calyceal vessels, supplying the area, show acute necrotizing arteritis and occlusion. The pathophysiological and clinical implications are discussed.

pubmed23n0001_328

Fasting uric acid and phosphate in urine and plasma of renal calcium-stone formers.

Clearance experiments in calcium-stone patients (n = 60) and controls (n = 60) demonstrated significantly higher urinary uric acid (UA) in younger (less 40 years) stone patients than controls (median: 480 vs. 351 mug/min) but not in older (greater than 40 years) patients. Serum UA and urinary oxypurines were comparable in health and stone disease. Conversely urinary phosphate was significantly lower in younger patients than matched controls (males: 224 vs. 304 mug) and presumably is responsible for the more alkaline pH. It is suggested that calcium-stone formation in humans is represented by two different populations.

Fasting uric acid and phosphate in urine and plasma of renal calcium-stone formers. Clearance experiments in calcium-stone patients (n = 60) and controls (n = 60) demonstrated significantly higher urinary uric acid (UA) in younger (less 40 years) stone patients than controls (median: 480 vs. 351 mug/min) but not in older (greater than 40 years) patients. Serum UA and urinary oxypurines were comparable in health and stone disease. Conversely urinary phosphate was significantly lower in younger patients than matched controls (males: 224 vs. 304 mug) and presumably is responsible for the more alkaline pH. It is suggested that calcium-stone formation in humans is represented by two different populations.

pubmed23n0001_329

Effects of varying dialysate calcium concentrations on the plasma calcium fractions in patients on dialysis.

The plasma-ionized calcium levels decreased during haemodialysis when a dialysate calcium concentration of 5 mg/100 ml was used. When dialysis was performed with a bath calcium concentration of 7.5 mg/100 ml, there was a significant increase in the plasm-ionized calcium levels in the post-dialysis period. These results are discussed in relation of the optimal dialysate calcium concentrations and development of dialytic bone disease.

Effects of varying dialysate calcium concentrations on the plasma calcium fractions in patients on dialysis. The plasma-ionized calcium levels decreased during haemodialysis when a dialysate calcium concentration of 5 mg/100 ml was used. When dialysis was performed with a bath calcium concentration of 7.5 mg/100 ml, there was a significant increase in the plasm-ionized calcium levels in the post-dialysis period. These results are discussed in relation of the optimal dialysate calcium concentrations and development of dialytic bone disease.

pubmed23n0001_330

[Borderline between antiepileptic and psychopharmacological drugs (author's transl)].

From the chemical and pharmalogical point of view it is impossible to separate strictly between antiepileptic and psychopharmalogical drugs. Inspite of that and several clinical overlappings it is possible and helpful finding differences between the two groups of drugs. Basing on the indication we try to demonstrate the use of these drug-groups. 1. Antiepileptic drugs in behaviour disorders. A. In spite of various publications we recommend carefullness and restriction. B. Only in case of behaviour disorders due to epileptic origin antiepileptic therapy is indicated. 2. Psychopharmalogical drugs in epilepsies A. The frequency of seizures may be reduced B. Psychical disorders connected with seizures may be influnced.

[Borderline between antiepileptic and psychopharmacological drugs (author's transl)]. From the chemical and pharmalogical point of view it is impossible to separate strictly between antiepileptic and psychopharmalogical drugs. Inspite of that and several clinical overlappings it is possible and helpful finding differences between the two groups of drugs. Basing on the indication we try to demonstrate the use of these drug-groups. 1. Antiepileptic drugs in behaviour disorders. A. In spite of various publications we recommend carefullness and restriction. B. Only in case of behaviour disorders due to epileptic origin antiepileptic therapy is indicated. 2. Psychopharmalogical drugs in epilepsies A. The frequency of seizures may be reduced B. Psychical disorders connected with seizures may be influnced.

pubmed23n0001_331

Identification, purification and properties of clone-specific glycoprotein antigens constituting the surface coat of Trypanosoma brucei.

Soluble glycoproteins have been purified from a series of clones of Trypanosoma brucei 427. Each clone yielded a characteristic predominant glycoprotein which induced clone-specific immunity to trypanosome infection in mice. These glycoproteins were shown by specific labelling and enzyme digestion of cells to be the major components of the trypanosome surface coat. Each glycoprotein consisted of a single polypeptide chain having an apparent molecular weight of 65 000 (as measured by SDS-polyacrylamide gel electrophoresis) and containing around 600 amino acid and 20 monosaccharide residues. Preliminary structural studies indicated large changes in amino acid sequence dispersed over a considerable length of the polypeptide chain. Proteolytic activity was demonstrated in semi-purified trypanosome extracts, providing one reason for the heterogeneity sometimes observed in surface glycoprotein antigen preparations.

Identification, purification and properties of clone-specific glycoprotein antigens constituting the surface coat of Trypanosoma brucei. Soluble glycoproteins have been purified from a series of clones of Trypanosoma brucei 427. Each clone yielded a characteristic predominant glycoprotein which induced clone-specific immunity to trypanosome infection in mice. These glycoproteins were shown by specific labelling and enzyme digestion of cells to be the major components of the trypanosome surface coat. Each glycoprotein consisted of a single polypeptide chain having an apparent molecular weight of 65 000 (as measured by SDS-polyacrylamide gel electrophoresis) and containing around 600 amino acid and 20 monosaccharide residues. Preliminary structural studies indicated large changes in amino acid sequence dispersed over a considerable length of the polypeptide chain. Proteolytic activity was demonstrated in semi-purified trypanosome extracts, providing one reason for the heterogeneity sometimes observed in surface glycoprotein antigen preparations.

pubmed23n0001_332

The determination of gamma-glutamyl transpeptidase by reaction rate assay at 37 degrees C.

The determination of gamma-glutamyl transpeptidase by a reaction rate assay using optimal reaction conditions at 37 degrees is described. Specific conditions and instrument settings are described for the LKB Reaction Rate Analyser but the actual assay conditions are applicable to any similar reaction rate system. The precision of the method has been evaluated and a reference range for normal male (less than 45 U/l) and female (less than 30 U/l) subjects determined.

The determination of gamma-glutamyl transpeptidase by reaction rate assay at 37 degrees C. The determination of gamma-glutamyl transpeptidase by a reaction rate assay using optimal reaction conditions at 37 degrees is described. Specific conditions and instrument settings are described for the LKB Reaction Rate Analyser but the actual assay conditions are applicable to any similar reaction rate system. The precision of the method has been evaluated and a reference range for normal male (less than 45 U/l) and female (less than 30 U/l) subjects determined.

pubmed23n0001_333

The influence of cortisol on the enzymes of fatty acid synthesis in developing mammalian lung and brain.

Administration of cortisol to fetal rabbits resulted in a 42% inhibition of pulmonary de novo fatty acid synthesis from acetyl coenzyme A (CoA) (P = less than 0.025). This was associated with inhibition of acetyl-CoA carboxylase (EC. 6.4.1.2.) activity (P = less than 0.01) and a tendency towards decreased activity of fatty acid synthetase. There was no effect on pulmonary microsomal fatty acid elongation activity. Light and electron microscopic examination of the apex of the right lung of control and cortisol-treated animals revealed changes consistent with accelerated lung maturation in the treated animals. The in vitro activities of acetyl-CoA carboxylase and fatty acid synthetase were similar in rabbit lung and thus acetyl-CoA carboxylase activity does not appear to be rate limiting for de novo fatty acid synthesis in lung. No significant change in the activity of enzymes associated with de novo fatty acid synthesis of microsomal fatty acid elongation was found in fetal brain after cortisol exposure. However, in a parallel study on fatty acid synthesis in fetal liver, cortisol administration resulted in a 30% increase in fatty acid synthetase activity (P less than 0.025). The finding of cortisol-induced inhibition of de novo fatty acid synthesis in fetal rabbit lung may be related to the known inhibitory effect of cortisol on lung growth in the fetus.

The influence of cortisol on the enzymes of fatty acid synthesis in developing mammalian lung and brain. Administration of cortisol to fetal rabbits resulted in a 42% inhibition of pulmonary de novo fatty acid synthesis from acetyl coenzyme A (CoA) (P = less than 0.025). This was associated with inhibition of acetyl-CoA carboxylase (EC. 6.4.1.2.) activity (P = less than 0.01) and a tendency towards decreased activity of fatty acid synthetase. There was no effect on pulmonary microsomal fatty acid elongation activity. Light and electron microscopic examination of the apex of the right lung of control and cortisol-treated animals revealed changes consistent with accelerated lung maturation in the treated animals. The in vitro activities of acetyl-CoA carboxylase and fatty acid synthetase were similar in rabbit lung and thus acetyl-CoA carboxylase activity does not appear to be rate limiting for de novo fatty acid synthesis in lung. No significant change in the activity of enzymes associated with de novo fatty acid synthesis of microsomal fatty acid elongation was found in fetal brain after cortisol exposure. However, in a parallel study on fatty acid synthesis in fetal liver, cortisol administration resulted in a 30% increase in fatty acid synthetase activity (P less than 0.025). The finding of cortisol-induced inhibition of de novo fatty acid synthesis in fetal rabbit lung may be related to the known inhibitory effect of cortisol on lung growth in the fetus.

pubmed23n0001_334

The relation between myosin adenosinetriphosphatase activity and inactivation of myosin under alkaline conditions of heart muscles in mammals of different size.

ATPase activity of myosin in the heart muscle of the mouse, rat, guinea-pig, rabbit and pig was studied at neutral pH and under mild alkaline conditions. At neutral pH the ATPase activity of myosin is inversely related to body size of the animal species. The decrease of ATPase activity of myosin after alkaline preincubation depends on the degree of ATPase activity of intact myosin, i.e. myosin from the heart of the mouse exhibits high ATPase activity ae same relationship was found, when comparing myosin of new-born and adult heart muscle. It is concluded that the rate of alkaline inactivation of heart myosin is directly related to the degree of ATPase activity of intact myosin in all animals.

The relation between myosin adenosinetriphosphatase activity and inactivation of myosin under alkaline conditions of heart muscles in mammals of different size. ATPase activity of myosin in the heart muscle of the mouse, rat, guinea-pig, rabbit and pig was studied at neutral pH and under mild alkaline conditions. At neutral pH the ATPase activity of myosin is inversely related to body size of the animal species. The decrease of ATPase activity of myosin after alkaline preincubation depends on the degree of ATPase activity of intact myosin, i.e. myosin from the heart of the mouse exhibits high ATPase activity ae same relationship was found, when comparing myosin of new-born and adult heart muscle. It is concluded that the rate of alkaline inactivation of heart myosin is directly related to the degree of ATPase activity of intact myosin in all animals.

pubmed23n0001_335

Carbon dioxide response curves during hypothermia.

The responsiveness of the medullary chemoreceptors, measured by the ventilatory response to hypercapnia given in an hyperoxic gas mixture in intact anesthetized dogs has been evaluated during normothermia and at two levels of hypothermia. The response was studied in: 1) 20 dogs during normothermia, 2) 10 of these dogs at a blood temperature of 32-33 degrees C, and 3) in the other 10 dogs during deeper hypothermia (28-29 degrees C). The ventilatory response to CO2 decreased while blood temperature was lowered until the response became absent during deep hypothermia. For normothermia and both levels of hypothermia a similar oxygen drive of ventilation was found which was equivalent to approximately one fourth of the spontaneous ventilation. It is suggested, that in the deeply hypothermic animal the normal respiratory drive is apparently of peripheral (arterial) chemoreceptor origin and when this drive is nullified or significantly decreased, gentle shivering could be responsible for stimulating the respiratory center.

Carbon dioxide response curves during hypothermia. The responsiveness of the medullary chemoreceptors, measured by the ventilatory response to hypercapnia given in an hyperoxic gas mixture in intact anesthetized dogs has been evaluated during normothermia and at two levels of hypothermia. The response was studied in: 1) 20 dogs during normothermia, 2) 10 of these dogs at a blood temperature of 32-33 degrees C, and 3) in the other 10 dogs during deeper hypothermia (28-29 degrees C). The ventilatory response to CO2 decreased while blood temperature was lowered until the response became absent during deep hypothermia. For normothermia and both levels of hypothermia a similar oxygen drive of ventilation was found which was equivalent to approximately one fourth of the spontaneous ventilation. It is suggested, that in the deeply hypothermic animal the normal respiratory drive is apparently of peripheral (arterial) chemoreceptor origin and when this drive is nullified or significantly decreased, gentle shivering could be responsible for stimulating the respiratory center.

pubmed23n0001_336

[Zollinger-Ellison syndrome treated medically by an inhibitor of H2 histamine receptors].

Metiamide an histamine H2-receptors antagonist has been used to treat a case of Zollinger-Ellison syndrome characterized by a long standing diarrhea, an important gastric hypersecretion and a moderatly elevated plasma gastrin but without digestive ulceration. At the dose of 600 mg per day, Metiamide induced a complete suppression of acid secretion, an effect which lasted for 15 days after stopping the drug. Accordingly and since the only finding at time of laparotomy was a small lymph node enlarged with endocrine metastatic tissue, the stomach was left intact and Metiamide pursued. During the first 4 months of chronic administration of Metiamide, acid secretion was maintained at levels below 25 p.cent of initial values. Ulteriorly however, although dosages of Metiamide were increased, acid hypersecretion resumed and a duodenal ulcer developed. Total gastrectomy was then performed 11 months after the beginning of Metiamide. In spite of the failure of Metiamide treatment, the long term follow up of this case of Zollinger-Ellison Syndrome, allowed us to get theoretical and practical informations.

[Zollinger-Ellison syndrome treated medically by an inhibitor of H2 histamine receptors]. Metiamide an histamine H2-receptors antagonist has been used to treat a case of Zollinger-Ellison syndrome characterized by a long standing diarrhea, an important gastric hypersecretion and a moderatly elevated plasma gastrin but without digestive ulceration. At the dose of 600 mg per day, Metiamide induced a complete suppression of acid secretion, an effect which lasted for 15 days after stopping the drug. Accordingly and since the only finding at time of laparotomy was a small lymph node enlarged with endocrine metastatic tissue, the stomach was left intact and Metiamide pursued. During the first 4 months of chronic administration of Metiamide, acid secretion was maintained at levels below 25 p.cent of initial values. Ulteriorly however, although dosages of Metiamide were increased, acid hypersecretion resumed and a duodenal ulcer developed. Total gastrectomy was then performed 11 months after the beginning of Metiamide. In spite of the failure of Metiamide treatment, the long term follow up of this case of Zollinger-Ellison Syndrome, allowed us to get theoretical and practical informations.

pubmed23n0001_337

[Exercise test in the asthmatic patient. Study of 75 patients].

Frequency of exercise-induced asthma has been studied in 75 unselected asthmatic patients (adults and children) by measuring the forced expiratory volume at one second (FEV1) and the vital capacity (VC) before and after a treadmill exercise, continued until heart rate was at least equal to 80 p.cent-85 p.cent of maximum heart rate. In 19 subjects (25 p.cent), a more than 20 p.cent decrease from control value of FEV1 was recorded ten minutes after exercise ended. Exercise-induced bronchial obstruction was relieved by a beta adrenergic bronchodilator aerosol inhalation. In the group of 19 subjects having exercise-induced asthma, a significant positive correlation was found between pre-exercise FEV1 values and post-exercise FEV1 decreases. In the whole group of 75 subjects exercise-induced asthma was related to the severity of asthma and not to other clinical or physical characteristics of the subjects. From the data of other authors it appears that frequency of exercise-induced asthma is variable. The reasons of those differences are discussed.

[Exercise test in the asthmatic patient. Study of 75 patients]. Frequency of exercise-induced asthma has been studied in 75 unselected asthmatic patients (adults and children) by measuring the forced expiratory volume at one second (FEV1) and the vital capacity (VC) before and after a treadmill exercise, continued until heart rate was at least equal to 80 p.cent-85 p.cent of maximum heart rate. In 19 subjects (25 p.cent), a more than 20 p.cent decrease from control value of FEV1 was recorded ten minutes after exercise ended. Exercise-induced bronchial obstruction was relieved by a beta adrenergic bronchodilator aerosol inhalation. In the group of 19 subjects having exercise-induced asthma, a significant positive correlation was found between pre-exercise FEV1 values and post-exercise FEV1 decreases. In the whole group of 75 subjects exercise-induced asthma was related to the severity of asthma and not to other clinical or physical characteristics of the subjects. From the data of other authors it appears that frequency of exercise-induced asthma is variable. The reasons of those differences are discussed.

pubmed23n0001_338

[Anaphylactic accidents due to glaphenine. 5 cases].

The authors report 5 cases of shock type anaphylactic reactions after the ingestion of glafenine. Up to the present, acute tubulo-interstitial nephritis had chiefly been reported. The existence of allergy to this medication remains to be proved by the wider application of allergological studies.

[Anaphylactic accidents due to glaphenine. 5 cases]. The authors report 5 cases of shock type anaphylactic reactions after the ingestion of glafenine. Up to the present, acute tubulo-interstitial nephritis had chiefly been reported. The existence of allergy to this medication remains to be proved by the wider application of allergological studies.

pubmed23n0001_339

Intranasal betamethasone valerate in seasonal rhinitis.

A double-blind study comparing betamethasone valerate and placebo aerosols was carried out in 40 patients with a history of seasonal allergic rhinitis and positive skin tests to grass pollens. Analysis of the symptoms recorded on a daily record card for a period of one month indicated that the mean monthly symptom-score was lower for all symptoms in the group on active therapy and that this reached statistical significance for the symptom of sneezing. Significantly more antihistamine tablets were used by the placebo group as compared with the active group (p less than 0-05). The patients' assessment of their treatment was in favour of betamethasone valerate (p less than 0-05). No clinically significant side-effects were associated with the treatment, which was well tolerated.

Intranasal betamethasone valerate in seasonal rhinitis. A double-blind study comparing betamethasone valerate and placebo aerosols was carried out in 40 patients with a history of seasonal allergic rhinitis and positive skin tests to grass pollens. Analysis of the symptoms recorded on a daily record card for a period of one month indicated that the mean monthly symptom-score was lower for all symptoms in the group on active therapy and that this reached statistical significance for the symptom of sneezing. Significantly more antihistamine tablets were used by the placebo group as compared with the active group (p less than 0-05). The patients' assessment of their treatment was in favour of betamethasone valerate (p less than 0-05). No clinically significant side-effects were associated with the treatment, which was well tolerated.

pubmed23n0001_340

Urinary acidification in renal allografts.

Urinary acid excretion was measured in 35 human renal allograft recipients during the first few days after the onset of diuresis and two months after transplantation. In 16 out of 17 rejection episodes an early significant decrease of renal total net H+ excretion was observed. This urinary acidification impairment preceded the serum creatinine increase. Only few patients showed variations in blood pH and C1-. The acidification defect may be due to the ischaemic changes of rejection. Early impairment of urinary acidification supports a clinical diagnosis of rejection.

Urinary acidification in renal allografts. Urinary acid excretion was measured in 35 human renal allograft recipients during the first few days after the onset of diuresis and two months after transplantation. In 16 out of 17 rejection episodes an early significant decrease of renal total net H+ excretion was observed. This urinary acidification impairment preceded the serum creatinine increase. Only few patients showed variations in blood pH and C1-. The acidification defect may be due to the ischaemic changes of rejection. Early impairment of urinary acidification supports a clinical diagnosis of rejection.

pubmed23n0001_341

A simple test for early detection of severe renal homograft rejection.

Adding urine to a standard buffered fibrinogen solution and then coagulating it with thrombin gives reproducible coagulation times with normal urines. Coagulation of fibrinogen by thrombin is prolonged in acid solutions with a pH below six. Urines of high acidity lower the buffer pH of the fibrinogen solution below a value of six thus rendering the system uncoagulable or significantly prolonging the coagulation time. With this test system we found that out of 16 severe homograft rejections 15 were accompanied by a high acid excretion in six-hour urine specimens. Ten of these acid episodes became apparent 12 to 48 hr before clinical symptoms and before elevation of the serum creatinine could be detected.

A simple test for early detection of severe renal homograft rejection. Adding urine to a standard buffered fibrinogen solution and then coagulating it with thrombin gives reproducible coagulation times with normal urines. Coagulation of fibrinogen by thrombin is prolonged in acid solutions with a pH below six. Urines of high acidity lower the buffer pH of the fibrinogen solution below a value of six thus rendering the system uncoagulable or significantly prolonging the coagulation time. With this test system we found that out of 16 severe homograft rejections 15 were accompanied by a high acid excretion in six-hour urine specimens. Ten of these acid episodes became apparent 12 to 48 hr before clinical symptoms and before elevation of the serum creatinine could be detected.

pubmed23n0001_342

Catalysis by acetylcholinesterase: evidence that the rate-limiting step for acylation with certain substrates precedes general acid-base catalysis.

Inferences about the catalytic mechanism of acetylcholinesterase (acetylcholine hydrolase, EC 3.1.1.7) are frequently made on the basis of a presumed analogy with chymotrypsin, EC 3.4.21.1. Although both enzymes are serine hydrolases, several differences in the steady-state kinetic properties of the two have been observed. In this report particular attention is focused on the second-order reaction constant, kcat/Kapp. While the reported pH dependence and deuterium oxide isotope effect associated with this parameter for chymotrypsin are generally consistent with simple models involving rate-limiting general acid-base catalysis, this study finds a more complicated situation with acetylcholinesterase. The apparent pKa of kcat/Kapp for acetylcholinesterase varies between 5.5 and 6.3 for neutral substrates and involves nonlinear inhibition by [H+]. Deuterium oxide isotope effects for kcat/Kapp range from 1.1 for acetylcholine to 1.9 for p-nitrophenyl acetate. The bimolecular reaction rate appears rate-limiting for acetylcholine at low concentrations, while a rate-limiting induced-fit step is proposed to account for apparent pKa values and low deuterium oxide isotope effects associated with low concentrations of phenyl acetate and isoamyl acetate.

Catalysis by acetylcholinesterase: evidence that the rate-limiting step for acylation with certain substrates precedes general acid-base catalysis. Inferences about the catalytic mechanism of acetylcholinesterase (acetylcholine hydrolase, EC 3.1.1.7) are frequently made on the basis of a presumed analogy with chymotrypsin, EC 3.4.21.1. Although both enzymes are serine hydrolases, several differences in the steady-state kinetic properties of the two have been observed. In this report particular attention is focused on the second-order reaction constant, kcat/Kapp. While the reported pH dependence and deuterium oxide isotope effect associated with this parameter for chymotrypsin are generally consistent with simple models involving rate-limiting general acid-base catalysis, this study finds a more complicated situation with acetylcholinesterase. The apparent pKa of kcat/Kapp for acetylcholinesterase varies between 5.5 and 6.3 for neutral substrates and involves nonlinear inhibition by [H+]. Deuterium oxide isotope effects for kcat/Kapp range from 1.1 for acetylcholine to 1.9 for p-nitrophenyl acetate. The bimolecular reaction rate appears rate-limiting for acetylcholine at low concentrations, while a rate-limiting induced-fit step is proposed to account for apparent pKa values and low deuterium oxide isotope effects associated with low concentrations of phenyl acetate and isoamyl acetate.

pubmed23n0001_343

Light-dependent absorption and selective scattering changes at 518 nm in chloroplast thylakoid membranes.

The light-induced absorbance change at 518 nm of isolated chloroplasts consists of a rapid phase, and a slow phase which is complete in about 20 sec. The slow component of the 518 nm absorbance change correlates with the light-induced change in 90 degrees light scattering at 518 nm. Both show a similar time course, similar pH dependence with a maximum at pH 6.0, and similar sensitivity to inhibitors and to treatment of the chloroplasts with a low concentration of glutaraldehyde. Their light minus dark difference spectra are similar with maxima at about 520 nm. It is concluded that they are manifestations of the same phenomenon, and the slow absorbance increase at 518 nm is due to enhanced scattering. It is proposed that the light-induced changes in scattering at 518 nm reflect alterations in selective dispersion, due to proton uptake and conformational changes in the chloroplast thylakoid membrane.

Light-dependent absorption and selective scattering changes at 518 nm in chloroplast thylakoid membranes. The light-induced absorbance change at 518 nm of isolated chloroplasts consists of a rapid phase, and a slow phase which is complete in about 20 sec. The slow component of the 518 nm absorbance change correlates with the light-induced change in 90 degrees light scattering at 518 nm. Both show a similar time course, similar pH dependence with a maximum at pH 6.0, and similar sensitivity to inhibitors and to treatment of the chloroplasts with a low concentration of glutaraldehyde. Their light minus dark difference spectra are similar with maxima at about 520 nm. It is concluded that they are manifestations of the same phenomenon, and the slow absorbance increase at 518 nm is due to enhanced scattering. It is proposed that the light-induced changes in scattering at 518 nm reflect alterations in selective dispersion, due to proton uptake and conformational changes in the chloroplast thylakoid membrane.

pubmed23n0001_344

phiX174 DNA-dependent DNA synthesis in vitro: requirement for P1 ban protein in dnaB mutant extracts of Escherichia coli.

Ammonium sulfate fractionation of crude extracts of E. coli yields a soluble enzyme fraction (about 25-fold purification) that catalyzes the conversion of phiX174 single-stranded DNA to duplex DNA. The reaction is rifampicin-resistant, requires single-stranded DNA, Mg++, deoxynucleoside triphosphates, and ATP, and is stimulated by KCl. Such soluble enzyme fractions were prepared from E. coli strains carrying the prophage mutant P1bac, in which the viral dnaB analog (ban) protein is expressed constitutively, or P1bacban, in which the expression of ban protein is prevented. DNA-synthesizing activity of ban protein containing fractions from wild-type or dnaB(P1bac) lysogens was more temperature-resistant than that from E. coli containing only wild-type dnaB protein, whereas that from dnaB(P1bacban) lysogens of dnaB cells was extremely thermolabile. It is suggested that the temperature-resistant DNA synthesis with fractions from P1bac lysogens is mediated by the P1 ban protein.

phiX174 DNA-dependent DNA synthesis in vitro: requirement for P1 ban protein in dnaB mutant extracts of Escherichia coli. Ammonium sulfate fractionation of crude extracts of E. coli yields a soluble enzyme fraction (about 25-fold purification) that catalyzes the conversion of phiX174 single-stranded DNA to duplex DNA. The reaction is rifampicin-resistant, requires single-stranded DNA, Mg++, deoxynucleoside triphosphates, and ATP, and is stimulated by KCl. Such soluble enzyme fractions were prepared from E. coli strains carrying the prophage mutant P1bac, in which the viral dnaB analog (ban) protein is expressed constitutively, or P1bacban, in which the expression of ban protein is prevented. DNA-synthesizing activity of ban protein containing fractions from wild-type or dnaB(P1bac) lysogens was more temperature-resistant than that from E. coli containing only wild-type dnaB protein, whereas that from dnaB(P1bacban) lysogens of dnaB cells was extremely thermolabile. It is suggested that the temperature-resistant DNA synthesis with fractions from P1bac lysogens is mediated by the P1 ban protein.

pubmed23n0001_345

Intramolecular arsanilazotyrosine-248-Zn complex of carboxypeptidase A: a monitor of catalytic events.

The intensely chromophoric intramolecular coordination complex formed between arsanilazotyrosine-248 and the active site zinc atom of azocarboxypeptidase A (Johansen, J. T. & Vallee, B. L. (1971) Proc. Nat. Acad. Sci. USA 68, 2532-2535) is a spectrokinetic probe of catalytic events. The interconversion of the azoTyr-248-Zn complex and its constituents is measured by stopped-flow pH and temperature-jump methods. The rate of interconversion, 64,000 sec-1, is orders of magnitude faster than that of the catalytic step itself (about 0.01-100 sec-1). Rapidly turned over peptide and ester substrates disrupt the azoTyr-248-Zn complex before hydrolysis occurs. As a consequence, formation of azoTyr-248, substrate binding, and catalysis can all be monitored while catalysis is actually in progress. The results of these dynamic studies specify a course of catalytic events, different from those postulated based on x-ray structure analysis. If azoTyr-248 is displaced, the direction is opposite to the inward movement postulated on the basis of x-ray studies and is not unique to induction by substrates, since rapid changes in pH also result in analogous spectral changes. AzoTyr-248 carboxypeptidase has all the features which are essential for mechanistic studies: (1) It is enzymatically active; (2) the spectra of the metal complex differ characteristically from those of its constituents; (3) it responds dynamically to environmental factors; and (4) the response time of the probe itself is much more rapid than is required for the measurement of the catalytic step. These combined kinetic and spectral properties of the metal complex render it a powerful spectrokinetic probe to visualize and discern microscopic details of the catalytic process.

Intramolecular arsanilazotyrosine-248-Zn complex of carboxypeptidase A: a monitor of catalytic events. The intensely chromophoric intramolecular coordination complex formed between arsanilazotyrosine-248 and the active site zinc atom of azocarboxypeptidase A (Johansen, J. T. & Vallee, B. L. (1971) Proc. Nat. Acad. Sci. USA 68, 2532-2535) is a spectrokinetic probe of catalytic events. The interconversion of the azoTyr-248-Zn complex and its constituents is measured by stopped-flow pH and temperature-jump methods. The rate of interconversion, 64,000 sec-1, is orders of magnitude faster than that of the catalytic step itself (about 0.01-100 sec-1). Rapidly turned over peptide and ester substrates disrupt the azoTyr-248-Zn complex before hydrolysis occurs. As a consequence, formation of azoTyr-248, substrate binding, and catalysis can all be monitored while catalysis is actually in progress. The results of these dynamic studies specify a course of catalytic events, different from those postulated based on x-ray structure analysis. If azoTyr-248 is displaced, the direction is opposite to the inward movement postulated on the basis of x-ray studies and is not unique to induction by substrates, since rapid changes in pH also result in analogous spectral changes. AzoTyr-248 carboxypeptidase has all the features which are essential for mechanistic studies: (1) It is enzymatically active; (2) the spectra of the metal complex differ characteristically from those of its constituents; (3) it responds dynamically to environmental factors; and (4) the response time of the probe itself is much more rapid than is required for the measurement of the catalytic step. These combined kinetic and spectral properties of the metal complex render it a powerful spectrokinetic probe to visualize and discern microscopic details of the catalytic process.

pubmed23n0001_346

Ubiquinone-mediated coupling of NADH dehydrogenase to active transport in membrane vesicles from Escherichia coli.

Addition of ubiquinone-1 to E. coli ML 308-225 membrane vesicles dramatically increases coupling between NADH oxidation and active transport such that initial rates and steady-state levels of lactose and amino-acid accumulation are comparable to those observed during D-lactate oxidation. Similar but less dramatic effects are observed with the quinone and succinate or L-lactate. In the presence of NADH and ubiquinone-1, the vesicles also generate a membrane potential (interior negative) that is similar in magnitude to that observed in the presence of D-lactate. Stimulation of NADH-dependent transport by ubiquinone-1 cannot be accounted for by increased rates of oxidation of NADH, and the effect of the quinone on NADH-dependent lactose transport is not observed in vesicles depleted of NADH dehydrogenase activity. Thus, it is apparent that ubiquinone-1 shunts electrons from NADH dehydrogenase [NADH:(acceptor)oxidoreductase; EC 1.6.99.3] to the portion of the respiratory chain containing the energy-coupling site. The findings demonstrate unequivocally that inefficient coupling of NADH oxidation to active transport cannot be due to the presence of inverted vesicles. In addition, they provide further support for specific localization of the energy-coupling site.

Ubiquinone-mediated coupling of NADH dehydrogenase to active transport in membrane vesicles from Escherichia coli. Addition of ubiquinone-1 to E. coli ML 308-225 membrane vesicles dramatically increases coupling between NADH oxidation and active transport such that initial rates and steady-state levels of lactose and amino-acid accumulation are comparable to those observed during D-lactate oxidation. Similar but less dramatic effects are observed with the quinone and succinate or L-lactate. In the presence of NADH and ubiquinone-1, the vesicles also generate a membrane potential (interior negative) that is similar in magnitude to that observed in the presence of D-lactate. Stimulation of NADH-dependent transport by ubiquinone-1 cannot be accounted for by increased rates of oxidation of NADH, and the effect of the quinone on NADH-dependent lactose transport is not observed in vesicles depleted of NADH dehydrogenase activity. Thus, it is apparent that ubiquinone-1 shunts electrons from NADH dehydrogenase [NADH:(acceptor)oxidoreductase; EC 1.6.99.3] to the portion of the respiratory chain containing the energy-coupling site. The findings demonstrate unequivocally that inefficient coupling of NADH oxidation to active transport cannot be due to the presence of inverted vesicles. In addition, they provide further support for specific localization of the energy-coupling site.

pubmed23n0001_347

Protection of lethally irradiated mice with allogeneic fetal liver cells: influence of irradiation dose on immunologic reconstitution.

After lethal irradiation long-lived, immunologically vigorous C3Hf mice were produced by treatment with syngeneic fetal liver cells or syngeneic newborn or adult spleen cells. Treatment of lethally irradiated mice with syngeneic or allogeneic newborn thymus cells or allogeneic newborn or adult spleen cells regularly led to fatal secondary disease or graft-versus-host reactions. Treatment of the lethally irradiated mice with fetal liver cells regularly yielded long-lived, immunologically vigorous chimeras. The introduction of the fetal liver cells into the irradiated mice appeared to be followed by development of immunological tolerance of the donor cells. The findings suggest that T-cells at an early stage of differentiation are more susceptible to tolerance induction than are T-lymphocytes at later stages of differentiation. These investigations turned up a perplexing paradox which suggests that high doses of irradiation may injure the thymic stroma, rendering it less capable of supporting certain T-cell populations in the peripheral lymphoid tissue. Alternatively, the higher and not the lower dose of irradiation may have eliminated a host cell not readily derived from fetal liver precursors which represents an important helper cell in certain cell-mediated immune functions, e.g., graft-versus-host reactions, but which is not important in others, e.g., allograft rejections. The higher dose of lethal irradiation did not permit development or maintenance of a population of spleen cells that could initiate graft-versus-host reactions but did permit the development of a population of donor cells capable of achieving vigorous allograft rejection. These observations contribute to understanding of some of the persisting immunodeficiencies that are observed in man after fatal irradiation and bone marrow transplantation. These results should suggest better approaches to more effective cellular engineering for correction of immunodeficiency diseases and for treatment of immunodeficiency diseases and of leukemias and malignancies of man.

Protection of lethally irradiated mice with allogeneic fetal liver cells: influence of irradiation dose on immunologic reconstitution. After lethal irradiation long-lived, immunologically vigorous C3Hf mice were produced by treatment with syngeneic fetal liver cells or syngeneic newborn or adult spleen cells. Treatment of lethally irradiated mice with syngeneic or allogeneic newborn thymus cells or allogeneic newborn or adult spleen cells regularly led to fatal secondary disease or graft-versus-host reactions. Treatment of the lethally irradiated mice with fetal liver cells regularly yielded long-lived, immunologically vigorous chimeras. The introduction of the fetal liver cells into the irradiated mice appeared to be followed by development of immunological tolerance of the donor cells. The findings suggest that T-cells at an early stage of differentiation are more susceptible to tolerance induction than are T-lymphocytes at later stages of differentiation. These investigations turned up a perplexing paradox which suggests that high doses of irradiation may injure the thymic stroma, rendering it less capable of supporting certain T-cell populations in the peripheral lymphoid tissue. Alternatively, the higher and not the lower dose of irradiation may have eliminated a host cell not readily derived from fetal liver precursors which represents an important helper cell in certain cell-mediated immune functions, e.g., graft-versus-host reactions, but which is not important in others, e.g., allograft rejections. The higher dose of lethal irradiation did not permit development or maintenance of a population of spleen cells that could initiate graft-versus-host reactions but did permit the development of a population of donor cells capable of achieving vigorous allograft rejection. These observations contribute to understanding of some of the persisting immunodeficiencies that are observed in man after fatal irradiation and bone marrow transplantation. These results should suggest better approaches to more effective cellular engineering for correction of immunodeficiency diseases and for treatment of immunodeficiency diseases and of leukemias and malignancies of man.

pubmed23n0001_348

Mechanism of action of penicillin: triggering of the pneumococcal autolytic enzyme by inhibitors of cell wall synthesis.

During penicillin treatment of an autolysin defective mutant pneumococcus we have observed three novel phenomena: (i) Growth of the mutant cultures is inhibited by the same concentrations of penicillin that induce lysis in the wild type. (ii) Mutant bacteria treated with the minimum growth inhibitory concentration of penicillin will lyse upon the addition of wild-type autolysin to the growth medium. Chloramphenicol and other inhibitors of protein synthesis protect the cells against lysis by exogenous enzyme. Sensitivity of the cells to exogenous autolysin requires treatment with penicillin or other inhibitors of cell wall synthesis (e.g., D-cycloserine or fosfonomycin) since exogenous autolysin alone has no effect on bacterial growth. (iii) Treatment with penicillin (or other inhibitors of cell wall synthesis) causes the escape into the medium of a choline-containing macromolecule that has properties suggesting that it contains pneumococcal lipoteichoic acid (Forssman antigen). Each one of these three phenomena (growth inhibition, sensitization to exogenous autolysin, and leakage of lipoteichoic acid) shows the same dose response as that of the penicillin-induced lysis of wild-type pneumococci. On the basis of these findings we propose a new hypothesis for the mechanism of penicillin-induced lysis of bacteria. It is suggested that inhibition of cell wall synthesis by any means triggers bacterial autolytic enzymes by destabilizing the endogenous complex of an autolysin inhibitor (lipoteichoic acid) and autolytic enzyme. Escape of lipoteichoic acid-like material to the growth medium is a consequence of this labilization. Chloramphenicol protects bacteria against penicillin-induced lysis by interfering with the activity of the autolytic enzyme, rather than by depleting the concentration of the enzyme at the cell surface.

Mechanism of action of penicillin: triggering of the pneumococcal autolytic enzyme by inhibitors of cell wall synthesis. During penicillin treatment of an autolysin defective mutant pneumococcus we have observed three novel phenomena: (i) Growth of the mutant cultures is inhibited by the same concentrations of penicillin that induce lysis in the wild type. (ii) Mutant bacteria treated with the minimum growth inhibitory concentration of penicillin will lyse upon the addition of wild-type autolysin to the growth medium. Chloramphenicol and other inhibitors of protein synthesis protect the cells against lysis by exogenous enzyme. Sensitivity of the cells to exogenous autolysin requires treatment with penicillin or other inhibitors of cell wall synthesis (e.g., D-cycloserine or fosfonomycin) since exogenous autolysin alone has no effect on bacterial growth. (iii) Treatment with penicillin (or other inhibitors of cell wall synthesis) causes the escape into the medium of a choline-containing macromolecule that has properties suggesting that it contains pneumococcal lipoteichoic acid (Forssman antigen). Each one of these three phenomena (growth inhibition, sensitization to exogenous autolysin, and leakage of lipoteichoic acid) shows the same dose response as that of the penicillin-induced lysis of wild-type pneumococci. On the basis of these findings we propose a new hypothesis for the mechanism of penicillin-induced lysis of bacteria. It is suggested that inhibition of cell wall synthesis by any means triggers bacterial autolytic enzymes by destabilizing the endogenous complex of an autolysin inhibitor (lipoteichoic acid) and autolytic enzyme. Escape of lipoteichoic acid-like material to the growth medium is a consequence of this labilization. Chloramphenicol protects bacteria against penicillin-induced lysis by interfering with the activity of the autolytic enzyme, rather than by depleting the concentration of the enzyme at the cell surface.

pubmed23n0001_349

Identification of NADPH-thioredoxin reductase system in Euglena gracillis.

Euglena gracilis contains a protein system which can utilize the reducing power of NADPH in the ribonucleotide reductase-catalyzed reduction of CTP. The proteins required for this reaction are a flavoprotien with a molecular weight of approximately 185,000 which is functionally similar to thioredoxin reductase (NADPH), EC 1.6.4.5, and another protein (Protein I) whose function in the reaction is unknown. This new protein does not appear to contain a prosthetic group and has a molecular weight of approximately 240,000. In addition, the ribonucleotide reductase active in the Euglena NADPH-thioredoxin reductase system is more complex than the protein reported in a previous publication [(1974) j. Biol. Chem. 249, 4428-4434]. The enzyme preparation described in this report contains four different types of polypeptide chains which may complex to form the active enzyme.

Identification of NADPH-thioredoxin reductase system in Euglena gracillis. Euglena gracilis contains a protein system which can utilize the reducing power of NADPH in the ribonucleotide reductase-catalyzed reduction of CTP. The proteins required for this reaction are a flavoprotien with a molecular weight of approximately 185,000 which is functionally similar to thioredoxin reductase (NADPH), EC 1.6.4.5, and another protein (Protein I) whose function in the reaction is unknown. This new protein does not appear to contain a prosthetic group and has a molecular weight of approximately 240,000. In addition, the ribonucleotide reductase active in the Euglena NADPH-thioredoxin reductase system is more complex than the protein reported in a previous publication [(1974) j. Biol. Chem. 249, 4428-4434]. The enzyme preparation described in this report contains four different types of polypeptide chains which may complex to form the active enzyme.

pubmed23n0001_350

Purification of folate binding factor in normal umbilical cord serum.

Human umbilical cord serum was found to contain both free folate and folate complexed to a high-molecular weight factor. The complexed folate was bound to a very high affinity binder and was present in concentrations equivalent to as much as 60 ng of 5-methyltetrahydrofolic acid per ml of serum. Acidification of the serum caused disassociation of the folate-binder complex. Released folates were separated from binder by Sephadex gel filtration, zonal centrifugation through sucrose gradients, or adsorption onto activated charcoal. The separated binding factor, either saturated or unsaturated with folate, had a molecular weight of about 40,000 on Sephadex G-200 chromatography. Binding of [3H]pteroylglutamic acid was rapid and, as in the original endogenous folate-binder complex, was essentially irreversible at neutral pH. The affinity and specificity of the binder were examined by competition experiments using [3H]pteroylglutamic acid and nonradioactive folate derivatives. Oxidized folates were bound in preference to reduced derivatives, but only three to four times more unlabeled 5-methyltetrahydrofolic acid than pteroylglutamic acid was required to produce an equal level of competition. The strong affinity for 5-methyltetrahydrofolic acid, the main serum folate, suggests that the binder could be part of the mechanism by which the fetus concentrates maternally supplied folate for its growth and development.

Purification of folate binding factor in normal umbilical cord serum. Human umbilical cord serum was found to contain both free folate and folate complexed to a high-molecular weight factor. The complexed folate was bound to a very high affinity binder and was present in concentrations equivalent to as much as 60 ng of 5-methyltetrahydrofolic acid per ml of serum. Acidification of the serum caused disassociation of the folate-binder complex. Released folates were separated from binder by Sephadex gel filtration, zonal centrifugation through sucrose gradients, or adsorption onto activated charcoal. The separated binding factor, either saturated or unsaturated with folate, had a molecular weight of about 40,000 on Sephadex G-200 chromatography. Binding of [3H]pteroylglutamic acid was rapid and, as in the original endogenous folate-binder complex, was essentially irreversible at neutral pH. The affinity and specificity of the binder were examined by competition experiments using [3H]pteroylglutamic acid and nonradioactive folate derivatives. Oxidized folates were bound in preference to reduced derivatives, but only three to four times more unlabeled 5-methyltetrahydrofolic acid than pteroylglutamic acid was required to produce an equal level of competition. The strong affinity for 5-methyltetrahydrofolic acid, the main serum folate, suggests that the binder could be part of the mechanism by which the fetus concentrates maternally supplied folate for its growth and development.

pubmed23n0001_351

Intramolecular arsanilazotyrosine-248-Zn complex of carboxypeptidase A: a monitor of multiple conformational states in solution.

The red azoTyr-248-Zn complex of arnilazocarboxypeptidase, previously used to demonstrate differences in conformation of the enzyme in crystals and in solution, has now provided means to detect multiple conformations of the enzyme in solution by stopped-flow pH and temperature jump experiments. These studies identify two distinct processes. Er + H+ in equilibrium Ey (I), is the extremely rapid, Kfast about 10(5) sec-1, pH dependent dissociation of the metal complex. Ey in equilibrium Ey' (II), is much slower, Kslow about 5 sec-1, pH independent interconversion of two distinct populations of protein molecules, Ey and Ey', in which the yellow azo-Tyr-248 is different conformations. These two conformations can be differentiated readily by stopped-flow pH-jump experiments, since I is three to four orders of magnitude faster than II. Mathematical expressions derived from this mechanism accurately predict all observations over the pH range from 6.0 to 8.5. In a previous stopped-flow pH-jump experiment, Lipcomb and coworkers [Quiocho, F. A., McMurray, C. H. & Lipcomb, W. H. (1972), Proc. Nat. Acad. Sci. USA 69, 2850-2854] recognized only a single process with a rate constant of about 6 sec-1, but not the major, very rapid rate observed here. The failure to detect this fast process led to the postulation of a number of explanations intended to account for the detection of only a single, slow rate. The present observations show that the premise for those conjectures is not valid. The azoprobe reveals the existence of rapidly interconvertible substructures of carboxypeptidase A, and the results support the view that in solution, enzymes can adopt multiple, readily interconvertible and related conformations which could then either facilitate or impede catalysis. In crystals, rearrangement of molecular structure could be severely impaired or restricted, and crystallization might single out either active or inactive conformations. In the latter case, such crystals would have greatly reduced activities and markedly altered catalytic behavior, as is observed for carboxypeptidase A. In combination with detailed kinetic analysis of crystals, conformational analysis in solution should be a valuable guide to discern enzyme mechanisms and select crystals for x-ray structure analysis.

Intramolecular arsanilazotyrosine-248-Zn complex of carboxypeptidase A: a monitor of multiple conformational states in solution. The red azoTyr-248-Zn complex of arnilazocarboxypeptidase, previously used to demonstrate differences in conformation of the enzyme in crystals and in solution, has now provided means to detect multiple conformations of the enzyme in solution by stopped-flow pH and temperature jump experiments. These studies identify two distinct processes. Er + H+ in equilibrium Ey (I), is the extremely rapid, Kfast about 10(5) sec-1, pH dependent dissociation of the metal complex. Ey in equilibrium Ey' (II), is much slower, Kslow about 5 sec-1, pH independent interconversion of two distinct populations of protein molecules, Ey and Ey', in which the yellow azo-Tyr-248 is different conformations. These two conformations can be differentiated readily by stopped-flow pH-jump experiments, since I is three to four orders of magnitude faster than II. Mathematical expressions derived from this mechanism accurately predict all observations over the pH range from 6.0 to 8.5. In a previous stopped-flow pH-jump experiment, Lipcomb and coworkers [Quiocho, F. A., McMurray, C. H. & Lipcomb, W. H. (1972), Proc. Nat. Acad. Sci. USA 69, 2850-2854] recognized only a single process with a rate constant of about 6 sec-1, but not the major, very rapid rate observed here. The failure to detect this fast process led to the postulation of a number of explanations intended to account for the detection of only a single, slow rate. The present observations show that the premise for those conjectures is not valid. The azoprobe reveals the existence of rapidly interconvertible substructures of carboxypeptidase A, and the results support the view that in solution, enzymes can adopt multiple, readily interconvertible and related conformations which could then either facilitate or impede catalysis. In crystals, rearrangement of molecular structure could be severely impaired or restricted, and crystallization might single out either active or inactive conformations. In the latter case, such crystals would have greatly reduced activities and markedly altered catalytic behavior, as is observed for carboxypeptidase A. In combination with detailed kinetic analysis of crystals, conformational analysis in solution should be a valuable guide to discern enzyme mechanisms and select crystals for x-ray structure analysis.

pubmed23n0001_352

Presence of norepinephrine and related enzymes in isolated brain microvessels.

Norepinephrine and the enzymes involved in its synthesis and degradation were found to be associated with isolated brain microvessels. The significance of these results are discussed with respect to adrenergic innervation of the cerebral microvessels and thereby neural regulation of the cerebral microcirculation.

Presence of norepinephrine and related enzymes in isolated brain microvessels. Norepinephrine and the enzymes involved in its synthesis and degradation were found to be associated with isolated brain microvessels. The significance of these results are discussed with respect to adrenergic innervation of the cerebral microvessels and thereby neural regulation of the cerebral microcirculation.

pubmed23n0001_353

Disorders of micturition in bacterial prostatitis.

Measurements of urinary flow rate were performed in 16 patients with established prostatitis before and after a course of antimicrobial therapy. Before treatment the maximum flow rates were poor with abnormal flow curves and significant improvement in voiding characteristics were observed with treatment (P less than 0.01). A preliminary electrophysiological (EMG) study of sphincter activity suggested that the obstruction to the flow of urine was at least in part due to failure of the external sphincter to relax during micturition. Although the total number of cases in this series was small the study showed that prostatitis was associated with a disorder of micturition which correlated with the other clinical features of the disease and could be objectively evaluated. Eradication of infection restored normal conditions in the lower urinary tract.

Disorders of micturition in bacterial prostatitis. Measurements of urinary flow rate were performed in 16 patients with established prostatitis before and after a course of antimicrobial therapy. Before treatment the maximum flow rates were poor with abnormal flow curves and significant improvement in voiding characteristics were observed with treatment (P less than 0.01). A preliminary electrophysiological (EMG) study of sphincter activity suggested that the obstruction to the flow of urine was at least in part due to failure of the external sphincter to relax during micturition. Although the total number of cases in this series was small the study showed that prostatitis was associated with a disorder of micturition which correlated with the other clinical features of the disease and could be objectively evaluated. Eradication of infection restored normal conditions in the lower urinary tract.

pubmed23n0001_354

Nonenzymatic reactivation of des-acetyl citrate lyase by acetyl adenylate. First example of enzyme activation by chemotrophic modification.

The experimental conditions of nonenzymatic reactivation of des-acetyl citrate lyase from K. aerogenes were studied. It was found that at pH 8.5 0.2 MM acetyl AMP causes a fast reactivation of the enzyme. The pH dependence of activity and the Km for citrate are very similar for both native and reactivated enzyme.

Nonenzymatic reactivation of des-acetyl citrate lyase by acetyl adenylate. First example of enzyme activation by chemotrophic modification. The experimental conditions of nonenzymatic reactivation of des-acetyl citrate lyase from K. aerogenes were studied. It was found that at pH 8.5 0.2 MM acetyl AMP causes a fast reactivation of the enzyme. The pH dependence of activity and the Km for citrate are very similar for both native and reactivated enzyme.

pubmed23n0001_355

Relationship between bovine serum albumin structure and its chemical equilibria with hydrogen and 5-hydroxytryptamine ions.

From the analysis of titration curves with hydrogen and 5-HT ions, it was found that the electrostatic interaction parameter of protein macroion and the number sites of 5-HT fixing were smaller over an acid and base pH range as compared to their values at neutral pH. Our data were interpreted by the conformational changes which can be induced when BSA is exposed to denaturation by acid and alkali pH.

Relationship between bovine serum albumin structure and its chemical equilibria with hydrogen and 5-hydroxytryptamine ions. From the analysis of titration curves with hydrogen and 5-HT ions, it was found that the electrostatic interaction parameter of protein macroion and the number sites of 5-HT fixing were smaller over an acid and base pH range as compared to their values at neutral pH. Our data were interpreted by the conformational changes which can be induced when BSA is exposed to denaturation by acid and alkali pH.

pubmed23n0001_356

Some psysicochemical properties of mitochondrial and cell sap alanine aminotransferase from the rat CNS.

The authors describe different properties of brain mitochondrial and cell sap alanine aminotransferase. They showed that the mitochondrial enzyme was inhibited by maleate, chlorides, acetate and phosphate with a high ionic strength (over 1.8), that its pH optimum lay between 7.5 and 8.5, that it was thermolabile at over 40 degrees C and that it was salted out from solutions with ammonium sulphate at 0.6--0.8 saturation. The activity of the cell sap enzyme was inhibited by phosphate at an ionic strength of only 0.12, less markedly by maleate and not at all by chlorides and acetate; its pH optimum was about 8, it was thermostable up to 60 degrees C and was precipitated from ammonium sulphate solution at between 0.35 and 0.6 saturation. The authors conclude from their results that two different alanine aminotransferase enzymes are present in the CNS.

Some psysicochemical properties of mitochondrial and cell sap alanine aminotransferase from the rat CNS. The authors describe different properties of brain mitochondrial and cell sap alanine aminotransferase. They showed that the mitochondrial enzyme was inhibited by maleate, chlorides, acetate and phosphate with a high ionic strength (over 1.8), that its pH optimum lay between 7.5 and 8.5, that it was thermolabile at over 40 degrees C and that it was salted out from solutions with ammonium sulphate at 0.6--0.8 saturation. The activity of the cell sap enzyme was inhibited by phosphate at an ionic strength of only 0.12, less markedly by maleate and not at all by chlorides and acetate; its pH optimum was about 8, it was thermostable up to 60 degrees C and was precipitated from ammonium sulphate solution at between 0.35 and 0.6 saturation. The authors conclude from their results that two different alanine aminotransferase enzymes are present in the CNS.

pubmed23n0001_357

[Postvacccinal encephalomyelitis after protective antirabic innoculation, combined with polioencephalomyelitis (lyssa?)].

By means of two cases of postvaccinal encephalomyelitis following antirabies vaccination has been demonstrated the meaning of these complications. In one of these cases has been presumed a simultaneous illness of quiet rabies. The difficulty of an aetiological diagnostic by the morphological effigy of the encephalomyelitids has been referred. An improvement of the dispensaire-system and of the diagnostic of complications following antirabies vaccination is postulated.

[Postvacccinal encephalomyelitis after protective antirabic innoculation, combined with polioencephalomyelitis (lyssa?)]. By means of two cases of postvaccinal encephalomyelitis following antirabies vaccination has been demonstrated the meaning of these complications. In one of these cases has been presumed a simultaneous illness of quiet rabies. The difficulty of an aetiological diagnostic by the morphological effigy of the encephalomyelitids has been referred. An improvement of the dispensaire-system and of the diagnostic of complications following antirabies vaccination is postulated.

pubmed23n0001_358

The effect of L-dopa on young patients with simple schizophrenia, treated with neuroleptic drugs: a double-blind cross-over trial with Madopar and placebo.

Thirteen out of 18 young out-patients with simple schizophrenia under neuroleptic treatment completed a double-blind cross-over trial with Madopar [L-Dopa + benserazid (a peripheral decarboxylase inhibitor)] and placebo. Nine patients were given 900 mg L-Dopa + 225 mg benserazid daily, 1 patient received 600 mg L-Dopa + 150 mg benserazid, and 3 patients, 300 mg L-Dopa + 75 mg benserazid. In these doses, L-Dopa was effective against emotional withdrawal, blunted affect, tendency to isolation and apathy, without inducing or aggravating productive, accessory symptoms. The activity score, according to the specific activity-withdrawal scale, was significantly increased (P less than 0.05), whereas the total BPRS score (Brief Psychiatric Rating Scale) was slightly, but significantly reduced (P less than 0.05). In cases where L-Dopa had to be limited to 600 and 300 mg daily, a tendency to anxiety, distortion of thinking, and a sense of unreality were observed, depending on the dose of L-Dopa. In no case were gastrointestinal, cardiovascular or neurological side-effects observed.

The effect of L-dopa on young patients with simple schizophrenia, treated with neuroleptic drugs: a double-blind cross-over trial with Madopar and placebo. Thirteen out of 18 young out-patients with simple schizophrenia under neuroleptic treatment completed a double-blind cross-over trial with Madopar [L-Dopa + benserazid (a peripheral decarboxylase inhibitor)] and placebo. Nine patients were given 900 mg L-Dopa + 225 mg benserazid daily, 1 patient received 600 mg L-Dopa + 150 mg benserazid, and 3 patients, 300 mg L-Dopa + 75 mg benserazid. In these doses, L-Dopa was effective against emotional withdrawal, blunted affect, tendency to isolation and apathy, without inducing or aggravating productive, accessory symptoms. The activity score, according to the specific activity-withdrawal scale, was significantly increased (P less than 0.05), whereas the total BPRS score (Brief Psychiatric Rating Scale) was slightly, but significantly reduced (P less than 0.05). In cases where L-Dopa had to be limited to 600 and 300 mg daily, a tendency to anxiety, distortion of thinking, and a sense of unreality were observed, depending on the dose of L-Dopa. In no case were gastrointestinal, cardiovascular or neurological side-effects observed.

pubmed23n0001_359

The specificity of binding of the narcotic agonist etorphine in synaptic membranes of rat brain in vivo.

When 3H-etorphine was administered to rats in a pharmacologically effective dose (0.75 mug/kg intracisternally), the labeled drug was concentrated in synaptic membrane fractions isolated from the brains of rats killed 10 min after etorphine injection. Pretreatment of the animals with the narcotic antagonists naloxone, diprenorphine or l-cyclorphan, blocked the pharmacological responses to etorphine and reduced 3H-etorphine binding in the membrane fractions. The differences between 3H-etorphine bound in synaptic membranes of rats treated with d-cyclorphan (inactive isomer) and l-cyclorphan (active antagonist) were in the same range as the reductions in etorphine binding in antagonist-treated rats, indicating that stereospecific and pharmacologically-specific binding sites in synaptic membranes in vivo were of the same magnitude: about 0.04 pmol/g brain.

The specificity of binding of the narcotic agonist etorphine in synaptic membranes of rat brain in vivo. When 3H-etorphine was administered to rats in a pharmacologically effective dose (0.75 mug/kg intracisternally), the labeled drug was concentrated in synaptic membrane fractions isolated from the brains of rats killed 10 min after etorphine injection. Pretreatment of the animals with the narcotic antagonists naloxone, diprenorphine or l-cyclorphan, blocked the pharmacological responses to etorphine and reduced 3H-etorphine binding in the membrane fractions. The differences between 3H-etorphine bound in synaptic membranes of rats treated with d-cyclorphan (inactive isomer) and l-cyclorphan (active antagonist) were in the same range as the reductions in etorphine binding in antagonist-treated rats, indicating that stereospecific and pharmacologically-specific binding sites in synaptic membranes in vivo were of the same magnitude: about 0.04 pmol/g brain.

pubmed23n0001_360

The cholinergic system and nociception in the primate: interactions with morphine.

In Experiment 1 the shock titration task was used to evaluate the antinoceptive properties of 5 different classes of cholinergic compounds in the rhesus monkey. Only scopolamine and high doses of physostigmine were effective in elevating the shock threshold. The apparent antinociceptive effect of physostigmine, however, was difficult to separate from its nonspecific behavioral depressant effect and was probably not related to an increase in cholinergic tone. Experiment 2 examined the interaction of morphine with arecoline, scopolamine and physostigmine. Only scopolamine (0.05 and 0.1 mg/kg) and high doses of physostigmine (0.1 mg/kg) interacted with morphine in the shock titration paradigm. The multiplicative interaction of morphine with scopolamine was confirmed in Experiment 3 over a wider range of doses. It was concluded that morphine and the cholinergic compounds produce antinociceptive effects through different mechanisms of the pain system.

The cholinergic system and nociception in the primate: interactions with morphine. In Experiment 1 the shock titration task was used to evaluate the antinoceptive properties of 5 different classes of cholinergic compounds in the rhesus monkey. Only scopolamine and high doses of physostigmine were effective in elevating the shock threshold. The apparent antinociceptive effect of physostigmine, however, was difficult to separate from its nonspecific behavioral depressant effect and was probably not related to an increase in cholinergic tone. Experiment 2 examined the interaction of morphine with arecoline, scopolamine and physostigmine. Only scopolamine (0.05 and 0.1 mg/kg) and high doses of physostigmine (0.1 mg/kg) interacted with morphine in the shock titration paradigm. The multiplicative interaction of morphine with scopolamine was confirmed in Experiment 3 over a wider range of doses. It was concluded that morphine and the cholinergic compounds produce antinociceptive effects through different mechanisms of the pain system.

pubmed23n0001_361

Effects of thienodiazepine derivatives on human sleep as compared to those of benzodiazepine derivatives.

The effects of new thienodiazepine derivatives, such as clotiazepam and Y-7131, on normal human sleep were investigated on 5 subjects and compared to those of benzodiazepine derivatives, such as diazepam and nitrazepam. REM sleep was significantly decreased only with 2 mg of Y-7131 and rebound elevation of REM sleep did not follow in recovery 1 and 2 nights. By using partial differential REM deprivation which was designed by us, there was also no rebound elevation of REM sleep noted in recovery 2 nights following 2 mg of Y-7131 medication. REM sleep was not suppressed with 15 mg of clotiazepam, 6 mg of diazepam and 10 mg of nitrazepam when compared to the baseline night. With regard to NREM sleep, stage 2 was significantly increased with 15 mg of clotiazepam and 10 mg of nitrazepam, but stage SWS was significantly decreased with 10 mg of nitrazepam.

Effects of thienodiazepine derivatives on human sleep as compared to those of benzodiazepine derivatives. The effects of new thienodiazepine derivatives, such as clotiazepam and Y-7131, on normal human sleep were investigated on 5 subjects and compared to those of benzodiazepine derivatives, such as diazepam and nitrazepam. REM sleep was significantly decreased only with 2 mg of Y-7131 and rebound elevation of REM sleep did not follow in recovery 1 and 2 nights. By using partial differential REM deprivation which was designed by us, there was also no rebound elevation of REM sleep noted in recovery 2 nights following 2 mg of Y-7131 medication. REM sleep was not suppressed with 15 mg of clotiazepam, 6 mg of diazepam and 10 mg of nitrazepam when compared to the baseline night. With regard to NREM sleep, stage 2 was significantly increased with 15 mg of clotiazepam and 10 mg of nitrazepam, but stage SWS was significantly decreased with 10 mg of nitrazepam.

pubmed23n0001_362

Effect of two weeks' treatment with thioridazine, chlorpromazine, sulpiride and bromazepam, alone or in combination with alcohol, on learning and memory in man.

Forty paid healthy male students participated in two subacute experiments of 6 weeks each. In the first trial 20 of them received bromazepam, thioridazine, and placebo double blind cross over for 2 weeks each, and in the second trial the active agents administered to the other 20 participants were chlorpromazine and sulpiride. The tests used were paired associate learning with nonsense syllables and digit memory span. Before testing the subjects took either an alcoholic or a nonalcoholic bitter drink. As in the previous study from this laboratory, alcohol was found to impair learning capacity. Of the drugs used only bromazepam impaired learning significantly, and the combined effect of alcohol and bromazepam on learning capacity was very deleterious. The adrenolytic effect of drugs did not correlate with their effect on learning. Caution is necessary when prescribing bromazepam for active outpatients at least in doses used in this study.

Effect of two weeks' treatment with thioridazine, chlorpromazine, sulpiride and bromazepam, alone or in combination with alcohol, on learning and memory in man. Forty paid healthy male students participated in two subacute experiments of 6 weeks each. In the first trial 20 of them received bromazepam, thioridazine, and placebo double blind cross over for 2 weeks each, and in the second trial the active agents administered to the other 20 participants were chlorpromazine and sulpiride. The tests used were paired associate learning with nonsense syllables and digit memory span. Before testing the subjects took either an alcoholic or a nonalcoholic bitter drink. As in the previous study from this laboratory, alcohol was found to impair learning capacity. Of the drugs used only bromazepam impaired learning significantly, and the combined effect of alcohol and bromazepam on learning capacity was very deleterious. The adrenolytic effect of drugs did not correlate with their effect on learning. Caution is necessary when prescribing bromazepam for active outpatients at least in doses used in this study.

pubmed23n0001_363

Effects of chronic exposure to stressors on avoidance-escape behavior and on brain norepinephrine.

A single exposure to a severe stressor (either cold swim or inescapable shock) impairs subsequent performance in a shuttle avoidance-escape task (1), a deficit attributed to reduction in brain noradrenergic activity produced by these stressors. In the present paper, two experiments are described which examine how repeated exposure to such stressors affects (a) shuttle avoidance-escape performance (Experiment 1), and (b) aspects of brain norepinephrine metabolism (Experiment 2). Experiment 1 showed that, whereas subjects receiving the single exposure to cold swim or shock showed a large avoidance-escape deficit, subjects that received repeated exposure to these stressors for 14 days performed similarly to the control group that received no stressor. Experiment 2 showed that, whereas subjects that received one session of the inescapable shock stressor showed a lower level of norepinephrine in hypothalamus and cortex than did subjects that received no shock, subjects that received repeated exposure to inescapable shock or cold swim showed neurochemical "habituation." Subjects that received repeated shock showed elevated tyrosine hydroxylase activity and no depletion of norepinephrine level, and both repeated shock and cold swim caused a decrease in uptake of 3H-norepinephrine by slices of cortex in vitro. Thus, it is concluded that the behavioral and neurochemical changes that were observed after the stressful conditions studied are consistent with the hypothesis that changes in avoidance-escape responding following exposure to these stressful events are due to changes in brain noradrenergic activity.

Effects of chronic exposure to stressors on avoidance-escape behavior and on brain norepinephrine. A single exposure to a severe stressor (either cold swim or inescapable shock) impairs subsequent performance in a shuttle avoidance-escape task (1), a deficit attributed to reduction in brain noradrenergic activity produced by these stressors. In the present paper, two experiments are described which examine how repeated exposure to such stressors affects (a) shuttle avoidance-escape performance (Experiment 1), and (b) aspects of brain norepinephrine metabolism (Experiment 2). Experiment 1 showed that, whereas subjects receiving the single exposure to cold swim or shock showed a large avoidance-escape deficit, subjects that received repeated exposure to these stressors for 14 days performed similarly to the control group that received no stressor. Experiment 2 showed that, whereas subjects that received one session of the inescapable shock stressor showed a lower level of norepinephrine in hypothalamus and cortex than did subjects that received no shock, subjects that received repeated exposure to inescapable shock or cold swim showed neurochemical "habituation." Subjects that received repeated shock showed elevated tyrosine hydroxylase activity and no depletion of norepinephrine level, and both repeated shock and cold swim caused a decrease in uptake of 3H-norepinephrine by slices of cortex in vitro. Thus, it is concluded that the behavioral and neurochemical changes that were observed after the stressful conditions studied are consistent with the hypothesis that changes in avoidance-escape responding following exposure to these stressful events are due to changes in brain noradrenergic activity.

pubmed23n0001_364

Evaluation of combined pharmacological and psychotherapeutic treatment in patients with functional abdominal disorders.

78 patients suffering from various functional abdominal complaints have been trated in a 2 x 2 double-blind design: (a) psychotherapy with Ro 5-3350 (TH/Ro); (b) psychotherapy with placebo (TH/P); (c) Ro 5-3350 without psychotherapy (NIH/Ro); (d) placebo without psychotherapy (NTH/P). Results show that a considerable amount of improvement cannot be ascribed to the two critical factors or the interaction of both, but are due to unspecific influences in the course of treatment. Some of the results concerning the combination of TH and the psychotropic drug pose interesting questions for further research and bare implications for double-blind trials of psychotropic drugs. The results suggest that possibly properties of any psychotropic drug have to be related to a doctor-patient relationship within which the personal problems of the patient are dealt with. In order to evaluate such properties, special methodological precautions have to be taken. These will be briefly discussed.

Evaluation of combined pharmacological and psychotherapeutic treatment in patients with functional abdominal disorders. 78 patients suffering from various functional abdominal complaints have been trated in a 2 x 2 double-blind design: (a) psychotherapy with Ro 5-3350 (TH/Ro); (b) psychotherapy with placebo (TH/P); (c) Ro 5-3350 without psychotherapy (NIH/Ro); (d) placebo without psychotherapy (NTH/P). Results show that a considerable amount of improvement cannot be ascribed to the two critical factors or the interaction of both, but are due to unspecific influences in the course of treatment. Some of the results concerning the combination of TH and the psychotropic drug pose interesting questions for further research and bare implications for double-blind trials of psychotropic drugs. The results suggest that possibly properties of any psychotropic drug have to be related to a doctor-patient relationship within which the personal problems of the patient are dealt with. In order to evaluate such properties, special methodological precautions have to be taken. These will be briefly discussed.

pubmed23n0001_365

Role of cardiovascular and ionic changes in pathogenesis and prevention of isoprenaline-induced cardiac necrosis.

Blood pressure, heart rate, oxygen uptake, and blood values of PO2, PCO2, and pH were studied in unanesthetized rats for 8 hours. After a cardiotoxic dose of 20 mg/kg isoprenaline, s.c., blood pressure fell from 117 to 72 mm Hg, heart rate accelerated from 326 to 497 beats/minute, and cardiac work diminished by about 15%. Metabolic rate increased by about 80%, blood values of PO2 rose, and those of PCO2 fell somewhat, whereas blood pH dropped from 7.48 to 7.38, indicating metabolic acidosis. Propranolol (40 mg/kg, i.p.) and verapamil (50 mg/kg, i.p.), both of which almost completely prevented isoprenaline-induced cardiac necroses, inhibited the chronotropic and calorigenic actions of isoprenaline by about 50%. While propranolol inhibited the depressor effect of isoprenaline completely, verapamil enhanced it: blood pressure fell to 46 mm Hg. Isoprenaline-induced fall of blood pH was not prevented by either propranolol or verapamil. Decrease of blood pH and cardionecrotisation were enhanced when isoprenaline was given together with 4.8 g/kg ethanol, p.o. In conclusion, hemodynamic actions of isoprenaline, especially hypotension, seem to be nonessential for the production of cardiac necroses. Strong acidification can aggravate the cardiotoxicity of isoprenaline.

Role of cardiovascular and ionic changes in pathogenesis and prevention of isoprenaline-induced cardiac necrosis. Blood pressure, heart rate, oxygen uptake, and blood values of PO2, PCO2, and pH were studied in unanesthetized rats for 8 hours. After a cardiotoxic dose of 20 mg/kg isoprenaline, s.c., blood pressure fell from 117 to 72 mm Hg, heart rate accelerated from 326 to 497 beats/minute, and cardiac work diminished by about 15%. Metabolic rate increased by about 80%, blood values of PO2 rose, and those of PCO2 fell somewhat, whereas blood pH dropped from 7.48 to 7.38, indicating metabolic acidosis. Propranolol (40 mg/kg, i.p.) and verapamil (50 mg/kg, i.p.), both of which almost completely prevented isoprenaline-induced cardiac necroses, inhibited the chronotropic and calorigenic actions of isoprenaline by about 50%. While propranolol inhibited the depressor effect of isoprenaline completely, verapamil enhanced it: blood pressure fell to 46 mm Hg. Isoprenaline-induced fall of blood pH was not prevented by either propranolol or verapamil. Decrease of blood pH and cardionecrotisation were enhanced when isoprenaline was given together with 4.8 g/kg ethanol, p.o. In conclusion, hemodynamic actions of isoprenaline, especially hypotension, seem to be nonessential for the production of cardiac necroses. Strong acidification can aggravate the cardiotoxicity of isoprenaline.

pubmed23n0001_366

Alterations in norepinephrine pattern in the damaged myocardium in the rat.

In the albino rat, the evolvement of myocardial necrosis induced by a single injection of ISO was accompanied by a fall in total NE. Pretreatment with propranolol and pargyline protected against ISO-induced necrosis and myocardial hypertrophy, but did not influence the ISO-induced depletion of NE stores. The depletion of NE stores is not due to impairment in synthesis or increased intraneuronal metabolism of NE since, in ISO-treated rats, neither cardiac tyrosine hydroxylase activity nor MAO activity was altered. The decrease in endogenous NE is not due to a defect in the storage of NE. The ability of myocardium to take up and store NE returned to normal within 48 hours, whereas endogenous levels returned to normal within 5 days, even in the presence of demonstrable necrosis. Thus, there is lack of correlation between chemical and morphological changes, since catecholamine depletion occurred in the absence of morphologically demonstrable tissue injury, and the function of the adrenergic neuron returns to normal in the presence of demonstrable necrosis.

Alterations in norepinephrine pattern in the damaged myocardium in the rat. In the albino rat, the evolvement of myocardial necrosis induced by a single injection of ISO was accompanied by a fall in total NE. Pretreatment with propranolol and pargyline protected against ISO-induced necrosis and myocardial hypertrophy, but did not influence the ISO-induced depletion of NE stores. The depletion of NE stores is not due to impairment in synthesis or increased intraneuronal metabolism of NE since, in ISO-treated rats, neither cardiac tyrosine hydroxylase activity nor MAO activity was altered. The decrease in endogenous NE is not due to a defect in the storage of NE. The ability of myocardium to take up and store NE returned to normal within 48 hours, whereas endogenous levels returned to normal within 5 days, even in the presence of demonstrable necrosis. Thus, there is lack of correlation between chemical and morphological changes, since catecholamine depletion occurred in the absence of morphologically demonstrable tissue injury, and the function of the adrenergic neuron returns to normal in the presence of demonstrable necrosis.

pubmed23n0001_367

Prevention of myocardial Ca overload and necrotization by Mg and K salts or acidosis.

The crucial point in the pathogenesis of isoproterenol-induced myocardial necrotization is an abundant intracellular Ca accumulation leading to high energy phosphate exhaustion. Accordingly, in the early stage of the isoproterenol-induced necrotization process, the onset of ATP and creatine phosphate breakdown strictly parallels the acute Ca gain. In this type of necrosis, the Mg losses from the myocardium appear as a concomitant phenomenon. The hearts can be protected against the deleterious Ca overload and necrotization by increasing the plasma concentration of Mg, K, or H ions in order to counterbalance Ca according to the ration (see article). On the other hand, if Mg, K, or H ion concentrations are too low, isoproterenol-induced Ca uptake and myocardial lesions are potentiated.

Prevention of myocardial Ca overload and necrotization by Mg and K salts or acidosis. The crucial point in the pathogenesis of isoproterenol-induced myocardial necrotization is an abundant intracellular Ca accumulation leading to high energy phosphate exhaustion. Accordingly, in the early stage of the isoproterenol-induced necrotization process, the onset of ATP and creatine phosphate breakdown strictly parallels the acute Ca gain. In this type of necrosis, the Mg losses from the myocardium appear as a concomitant phenomenon. The hearts can be protected against the deleterious Ca overload and necrotization by increasing the plasma concentration of Mg, K, or H ions in order to counterbalance Ca according to the ration (see article). On the other hand, if Mg, K, or H ion concentrations are too low, isoproterenol-induced Ca uptake and myocardial lesions are potentiated.

pubmed23n0001_368

Explanation of the stimulation of microsomal N-demethylation reactions by soluble supernatant fraction.

Addition of the cell soluble supernatant fraction to an assay medium containing NADPH generating system, mixed function oxidase substrate and microsomes, resulted in a stimulation of drug metabolism ranging from 12-75%. This stimulation was observed only when the supply of DADPH generating system (isocitric dehydrogenase or glucose 6 phosphate dehydrogenase) was insufficient, leading to a NADPH oxidation rate which was greater than the rate of reduction of NADP+ during the oxidation of a drug. Hence, under our assay conditions, the soluble supernate (SS) is only providing sufficient NADPH generator, and possibly relieving inhibition by the generated NADP+. Finally, microsomal lipid peroxidation measurements under these same conditions indicate negligible to no peroxidation activity in the absence of SS.

Explanation of the stimulation of microsomal N-demethylation reactions by soluble supernatant fraction. Addition of the cell soluble supernatant fraction to an assay medium containing NADPH generating system, mixed function oxidase substrate and microsomes, resulted in a stimulation of drug metabolism ranging from 12-75%. This stimulation was observed only when the supply of DADPH generating system (isocitric dehydrogenase or glucose 6 phosphate dehydrogenase) was insufficient, leading to a NADPH oxidation rate which was greater than the rate of reduction of NADP+ during the oxidation of a drug. Hence, under our assay conditions, the soluble supernate (SS) is only providing sufficient NADPH generator, and possibly relieving inhibition by the generated NADP+. Finally, microsomal lipid peroxidation measurements under these same conditions indicate negligible to no peroxidation activity in the absence of SS.

pubmed23n0001_369

An attempt to correlated analgesia to changes in brain neuromediators in rats.

The analgesic activity of delta9THC, morphine and sodium salicylate was studied concomitantly with changes in brain stem levels of 5HT, 5HIAA, DA and NA. The results show that a correlation exists between analgesia and changes in the serotonergic system of the brain stem. Furthermore morphine sulfate was found to increase the DA concentration of the brain stem while delta9THC increased NA levels. We conclude that serotonergic system may be of major importance in analgesia while simultaneous changes in this system and/or the DA and NA systems may lead to a more pronounced analgesic activity.

An attempt to correlated analgesia to changes in brain neuromediators in rats. The analgesic activity of delta9THC, morphine and sodium salicylate was studied concomitantly with changes in brain stem levels of 5HT, 5HIAA, DA and NA. The results show that a correlation exists between analgesia and changes in the serotonergic system of the brain stem. Furthermore morphine sulfate was found to increase the DA concentration of the brain stem while delta9THC increased NA levels. We conclude that serotonergic system may be of major importance in analgesia while simultaneous changes in this system and/or the DA and NA systems may lead to a more pronounced analgesic activity.

pubmed23n0001_370

Colorimetric determination of 5-aminosalicylic acid and its N-acetylated metabolite on urine and feces.

A simple and convenient colorimetric method is described for the quantitative determination of 5-aminosalicylic acid (5-ASA) and N-acetyl-5-ASA in urine and feces after oral administration of salicylazosulfa-pyridine (SASP), the drug of choice in the treatment of ulcerative colitis. N-acetyl-5-ASA is extracted directly from the acidified biological specimen, deacetylated, and the liberated 5-ASA subjected to a modified Bratton-Marshall reaction. The 5-ASA present in the specimen must be acetylated with acetic anhydride prior to extraction. The violet colored product of the Bratton-Marshall reaction has a lambdamax of 560 nm and conforms to Beer's law over the concentration range of 0-70 umg/ml. Average recoveries (+/- S.D., N = 6) OF 5-ASA added to rat and human urine and rat fecal homogenates were 91.6 +/- 4.9%, 102 +/- 6.0%, and 71.0 +/- 4.8%, respectively. Interference by SASP and its sulfapyridine metabolities is negligible. As demonstrated, the colorimetric method is of sufficient sensitivity for application in most metabolic and pharmacokinetic studies conducted with SASP in laboratory animals and man.

Colorimetric determination of 5-aminosalicylic acid and its N-acetylated metabolite on urine and feces. A simple and convenient colorimetric method is described for the quantitative determination of 5-aminosalicylic acid (5-ASA) and N-acetyl-5-ASA in urine and feces after oral administration of salicylazosulfa-pyridine (SASP), the drug of choice in the treatment of ulcerative colitis. N-acetyl-5-ASA is extracted directly from the acidified biological specimen, deacetylated, and the liberated 5-ASA subjected to a modified Bratton-Marshall reaction. The 5-ASA present in the specimen must be acetylated with acetic anhydride prior to extraction. The violet colored product of the Bratton-Marshall reaction has a lambdamax of 560 nm and conforms to Beer's law over the concentration range of 0-70 umg/ml. Average recoveries (+/- S.D., N = 6) OF 5-ASA added to rat and human urine and rat fecal homogenates were 91.6 +/- 4.9%, 102 +/- 6.0%, and 71.0 +/- 4.8%, respectively. Interference by SASP and its sulfapyridine metabolities is negligible. As demonstrated, the colorimetric method is of sufficient sensitivity for application in most metabolic and pharmacokinetic studies conducted with SASP in laboratory animals and man.

pubmed23n0001_371

[Pressures, blood gases, pH, lactate and pyruvate concentrations in the portal venous blood in patients after laparotomy during the first 9 days (author's transl)].

In 24 patients (16 women, 8 men, mean age 60 years), who underwent abdominal operations and who had a uncomplicated postoperative course, a teflon-tube was inserted into the portal vein at the end of laparotomy and remained there for maximal 9 days. The mean values of the portal venous pressure as well as of the portal-central venous pressure gradient are unchanged during the period of all 9 days and are between 7,4 and 7,9 mmHg, between 5,6 and 6,9 mmHg respectively. The arterio-portal venous 0(2)-content difference shows in the mean no fluctuations and is about 2 Vol.%. On the first postoperative day the lactic and pyruvic acid concentrations in the artery and portal vein are moderately, but significantly elevated and decrease to normal values until day 7. - 9.

[Pressures, blood gases, pH, lactate and pyruvate concentrations in the portal venous blood in patients after laparotomy during the first 9 days (author's transl)]. In 24 patients (16 women, 8 men, mean age 60 years), who underwent abdominal operations and who had a uncomplicated postoperative course, a teflon-tube was inserted into the portal vein at the end of laparotomy and remained there for maximal 9 days. The mean values of the portal venous pressure as well as of the portal-central venous pressure gradient are unchanged during the period of all 9 days and are between 7,4 and 7,9 mmHg, between 5,6 and 6,9 mmHg respectively. The arterio-portal venous 0(2)-content difference shows in the mean no fluctuations and is about 2 Vol.%. On the first postoperative day the lactic and pyruvic acid concentrations in the artery and portal vein are moderately, but significantly elevated and decrease to normal values until day 7. - 9.

pubmed23n0001_372

Biochemical and biophysical changes in guinea pigs after acute head injury.

Animal experiments were set up mainly to derive additional diagnostic data from the study of biochemical changes after acute head injury. In standardized experiments guinea pigs were subjected in groups of 20 to three identical head injuries, each of either 1.0 J or 1.5 J intensity. The trauma was likely to result in a concussion or contusion syndrome similar to that found in man; 40 animals served as controls. During the 60 min after injury observation and measurement of body functions did not reveal signs of a shock-like condition or hypoxaemia in the traumatized animals compared with control animals. Superficial anaesthesia probably did not influence the findings. Temperature and respiration were altered significantly in all the animals receiving head injuries. Blood gas analysis showed a decrease of standard bicarbonate only after the 1.5 J injury but even though hypoxaemia was not present 2,3-diphosphoglycerate values and P50 increased, compared with the control animals. The fall of plasma lipid concentrations reported probably had to be seen as a sympathomimetic effect of the minor (1.0 J) trauma. Of special significance was the increased activity of malate dehydrogenase and aldolase, found only in the blood of severely traumatized animals, as this could serve as an early diagnostic aid for evaluating head injuries.

Biochemical and biophysical changes in guinea pigs after acute head injury. Animal experiments were set up mainly to derive additional diagnostic data from the study of biochemical changes after acute head injury. In standardized experiments guinea pigs were subjected in groups of 20 to three identical head injuries, each of either 1.0 J or 1.5 J intensity. The trauma was likely to result in a concussion or contusion syndrome similar to that found in man; 40 animals served as controls. During the 60 min after injury observation and measurement of body functions did not reveal signs of a shock-like condition or hypoxaemia in the traumatized animals compared with control animals. Superficial anaesthesia probably did not influence the findings. Temperature and respiration were altered significantly in all the animals receiving head injuries. Blood gas analysis showed a decrease of standard bicarbonate only after the 1.5 J injury but even though hypoxaemia was not present 2,3-diphosphoglycerate values and P50 increased, compared with the control animals. The fall of plasma lipid concentrations reported probably had to be seen as a sympathomimetic effect of the minor (1.0 J) trauma. Of special significance was the increased activity of malate dehydrogenase and aldolase, found only in the blood of severely traumatized animals, as this could serve as an early diagnostic aid for evaluating head injuries.

pubmed23n0001_373

Total brain ischaemia in dogs: cerebral physiological and metabolic changes after 15 minutes of circulatory arrest.

Cross-clamping of the ascending aorta in dogs for 15 min produced severe neurological deficit, observed for up to 20 h. Immediately after restoration of the circulation, the intracranial pressure in the cisterna magna increased transiently to a mean peak of 22.8 Torr (SD +/- 1.7) because of a compensatory increase in systemic arterial pressure, without a fall in cerebral perfusion pressure. The intracranial pressure returned to control values 15-30 min after ischaemia and showed no secondary rise during the 8 h of observation. The electroencephalogram became isoelectric 34 +/- 6.5 s (mean +/-SD) after circulatory occlusion, and was abnormal when it reappeared 5 h 36 min (SD +/- 2 h 4 min) after the circulation was restored. The electrical impedance of the brain increased immediately after ischaemia and returned rapidly towards pre-ischaemic values during re-perfusion. The cerebral water had not increased measurably 4 h after ischaemia. After ischaemia, the lactate concentration in the cerebrospinal fluid increased to 4.7 mequiv./1(SEM +/-0.1) and the pH decreased to 7.17 (SEM +/-0.02); both returned to control values after 3.5 h. The cerebral glucose uptake was decreased 35 min after ischaemia, cerebral oxygen uptake remained unchanged but cerebral blood flow decreased (P less than 0.05 at 90 min). Immediately after cardiac arrest, recovery was impaired more by the presence of focal abnormal brain perfusion than by intracranial hypertension.

Total brain ischaemia in dogs: cerebral physiological and metabolic changes after 15 minutes of circulatory arrest. Cross-clamping of the ascending aorta in dogs for 15 min produced severe neurological deficit, observed for up to 20 h. Immediately after restoration of the circulation, the intracranial pressure in the cisterna magna increased transiently to a mean peak of 22.8 Torr (SD +/- 1.7) because of a compensatory increase in systemic arterial pressure, without a fall in cerebral perfusion pressure. The intracranial pressure returned to control values 15-30 min after ischaemia and showed no secondary rise during the 8 h of observation. The electroencephalogram became isoelectric 34 +/- 6.5 s (mean +/-SD) after circulatory occlusion, and was abnormal when it reappeared 5 h 36 min (SD +/- 2 h 4 min) after the circulation was restored. The electrical impedance of the brain increased immediately after ischaemia and returned rapidly towards pre-ischaemic values during re-perfusion. The cerebral water had not increased measurably 4 h after ischaemia. After ischaemia, the lactate concentration in the cerebrospinal fluid increased to 4.7 mequiv./1(SEM +/-0.1) and the pH decreased to 7.17 (SEM +/-0.02); both returned to control values after 3.5 h. The cerebral glucose uptake was decreased 35 min after ischaemia, cerebral oxygen uptake remained unchanged but cerebral blood flow decreased (P less than 0.05 at 90 min). Immediately after cardiac arrest, recovery was impaired more by the presence of focal abnormal brain perfusion than by intracranial hypertension.

pubmed23n0001_374

Oxygen and carbon dioxide dissociation of duck blood.

Oxygen and CO2 dissociation of duck blood was studied in blood samples equilibrated with known gas mixtures at the bird's body temperature (41 degrees C) and analyzed in the Van Slyke manometric apparatus and in pH electrodes. At various pH values between 7.38 and 7.55 the Hill plots yielded straight and parallel lines over a wide range of O2 saturation, the Hill coefficient being 2.9. Half saturation pressure P50 at pH = 7.50 was 36 torr. The Bohr effect factor was -0.53. Buffering properties were analyzed by equilibrating blood samples with gas mixtures of different PCO2 at 41 degrees C. The buffer value for whole blood in the range of 3-7% CO2 was 19.3 mMol-L-1-pH-1, the buffer value for true plasma 22.9 mMol-L-1-pH-1. The CO2 dissociation curve constructed using the buffer values had a slope of 0.17 mMol-L-1-torr-1 in the PCO2 range from 40 to 50 torr. The CO2 content of oxygenated blood at PCO2 = 40 torr was 21.7 mMol-L-1. The Haldane effect factor at PCO2 = 35 torr equalled 0.30 mMol of combined CO2 per mMol HbO2. With the values of PO2, PCO2 and pH measured in arterial blood of undisturbed and unrestrained, resting ducks effective dissociation curves for both O2 and CO2 were constructed assuming a metabolic R.Q. of 0.8. These curves are expected to resemble closely the actual in vitro dissociation curves of resting ducks.

Oxygen and carbon dioxide dissociation of duck blood. Oxygen and CO2 dissociation of duck blood was studied in blood samples equilibrated with known gas mixtures at the bird's body temperature (41 degrees C) and analyzed in the Van Slyke manometric apparatus and in pH electrodes. At various pH values between 7.38 and 7.55 the Hill plots yielded straight and parallel lines over a wide range of O2 saturation, the Hill coefficient being 2.9. Half saturation pressure P50 at pH = 7.50 was 36 torr. The Bohr effect factor was -0.53. Buffering properties were analyzed by equilibrating blood samples with gas mixtures of different PCO2 at 41 degrees C. The buffer value for whole blood in the range of 3-7% CO2 was 19.3 mMol-L-1-pH-1, the buffer value for true plasma 22.9 mMol-L-1-pH-1. The CO2 dissociation curve constructed using the buffer values had a slope of 0.17 mMol-L-1-torr-1 in the PCO2 range from 40 to 50 torr. The CO2 content of oxygenated blood at PCO2 = 40 torr was 21.7 mMol-L-1. The Haldane effect factor at PCO2 = 35 torr equalled 0.30 mMol of combined CO2 per mMol HbO2. With the values of PO2, PCO2 and pH measured in arterial blood of undisturbed and unrestrained, resting ducks effective dissociation curves for both O2 and CO2 were constructed assuming a metabolic R.Q. of 0.8. These curves are expected to resemble closely the actual in vitro dissociation curves of resting ducks.

pubmed23n0001_375

First apparent dissociation constant of carbonic acid, pK'1, of plasma and erythrocytes.

The first apparent dissociation constant of carbonic acid, pK'1, of plasma and red cells was determined on venous blood of ten healthy, adult, male, human subjects. pH and PCO2 of plasma and red cells were analyzed electrometrically and a micromanometric method was used for the determination of total carbon dioxide content. Erythrocyte carbamino hemoglobin levels were estimated and used for the correction of erythrocyte pK'1. Each blood sample was subjected to the following regimen before centrifugation, 1) As drawn from the antecubital vein, 2) Oxygenated with a 5% CO2, O2 balance gas mixture, and 3) Reduced with a 5% CO2, N2 balance gas mixture. pK'1 of plasma and red cells are presented: (see article). The consistently larger values for red cell pK'1 than the respective plasma data may be attributed to the greater amount of carbamino hemoglobin concentration present in the erythrocytes. A simplified method for the calculation of erythrocyte bicarbonate concentration using the experimentally determined red cell pK'1 value has been formulated. The method involves the use of a regression equation relating plasma and red cell pH, the equivalence of plasma and red cell PCO2, along with the experimentally determined red cell pK'1.

First apparent dissociation constant of carbonic acid, pK'1, of plasma and erythrocytes. The first apparent dissociation constant of carbonic acid, pK'1, of plasma and red cells was determined on venous blood of ten healthy, adult, male, human subjects. pH and PCO2 of plasma and red cells were analyzed electrometrically and a micromanometric method was used for the determination of total carbon dioxide content. Erythrocyte carbamino hemoglobin levels were estimated and used for the correction of erythrocyte pK'1. Each blood sample was subjected to the following regimen before centrifugation, 1) As drawn from the antecubital vein, 2) Oxygenated with a 5% CO2, O2 balance gas mixture, and 3) Reduced with a 5% CO2, N2 balance gas mixture. pK'1 of plasma and red cells are presented: (see article). The consistently larger values for red cell pK'1 than the respective plasma data may be attributed to the greater amount of carbamino hemoglobin concentration present in the erythrocytes. A simplified method for the calculation of erythrocyte bicarbonate concentration using the experimentally determined red cell pK'1 value has been formulated. The method involves the use of a regression equation relating plasma and red cell pH, the equivalence of plasma and red cell PCO2, along with the experimentally determined red cell pK'1.

pubmed23n0001_376

The effects of changes in pH and PCO2 in blood and water on breathing in rainbow trout, Salmo gairdneri.

The effect of sustained hypercapnia on the acid-base balance and gill ventilation in rainbow trout, Salmo gairdneri, was studied. The response to an increase in PICO2 from 0.3 to 5.2 mm Hg was a five-fold increase in gill ventilation volume and a slight increase in breathing frequency. There was a concomitant rise in PACO2 and an immediate fall in pHa. If PICO2 was maintained at 5.2 mm Hg for several days, ventilation volume gradually returned to the initial, prehypercapnic level within three days. Arterial pH also returned to the initial level within 2-3 days. These results are consistent with the hypothesis that under these conditions fish regulate pH via HCO3/C1 exchange across the gills rather than by changes in ventilation and subsequent adjustment of PACO2. A reduction in environmental pH causes a reduction in pHa but only a slow gradual increase in VG. Injections of HC1 or NaHCO3 into the blood have opposite effects on pHa but both cause a marked increase in VG. It is concluded that a rise in PACO2 results in a rise in VG and that changes in pH in blood or water have little direct effect on VG in rainbow trout. Possible location for receptors involved in this reflex response are discussed.

The effects of changes in pH and PCO2 in blood and water on breathing in rainbow trout, Salmo gairdneri. The effect of sustained hypercapnia on the acid-base balance and gill ventilation in rainbow trout, Salmo gairdneri, was studied. The response to an increase in PICO2 from 0.3 to 5.2 mm Hg was a five-fold increase in gill ventilation volume and a slight increase in breathing frequency. There was a concomitant rise in PACO2 and an immediate fall in pHa. If PICO2 was maintained at 5.2 mm Hg for several days, ventilation volume gradually returned to the initial, prehypercapnic level within three days. Arterial pH also returned to the initial level within 2-3 days. These results are consistent with the hypothesis that under these conditions fish regulate pH via HCO3/C1 exchange across the gills rather than by changes in ventilation and subsequent adjustment of PACO2. A reduction in environmental pH causes a reduction in pHa but only a slow gradual increase in VG. Injections of HC1 or NaHCO3 into the blood have opposite effects on pHa but both cause a marked increase in VG. It is concluded that a rise in PACO2 results in a rise in VG and that changes in pH in blood or water have little direct effect on VG in rainbow trout. Possible location for receptors involved in this reflex response are discussed.

pubmed23n0001_377

The effect of pH upon fluoride uptake in intact enamel.

The relationship between pH and fluoride uptake in intact enamel of permanent premolars was investigated by using: (1) a sodium fluoride dentifrice, (2) a potassium fluoride + manganese chloride dentifrice, and (3) a sodium fluoride solution of the same fluoride concentration. The first part of this paper deals with the in vitro uptake of fluoride from dentifrice slurries and from sodium fluoride solutions of different pH ranging from 7.1 to 4.5. This investigation showed that there was no significant difference between the agents but that the effect of the pH was significant. The uptake of fluoride in the form of fluorapatite was more than five times larger at the lower pH level. The second part of the paper deals with the rate of fluoride uptake (increase in fluoride content) from dentifrices in the same pH range. It was shown that the three agents gave the same initial rate of fluoride uptake (about 50 parts/10(6)/min) at pH 6 and that the rate of fluoride uptake in the outer layer of the enamel was proportional to the hydrogen ion activity.

The effect of pH upon fluoride uptake in intact enamel. The relationship between pH and fluoride uptake in intact enamel of permanent premolars was investigated by using: (1) a sodium fluoride dentifrice, (2) a potassium fluoride + manganese chloride dentifrice, and (3) a sodium fluoride solution of the same fluoride concentration. The first part of this paper deals with the in vitro uptake of fluoride from dentifrice slurries and from sodium fluoride solutions of different pH ranging from 7.1 to 4.5. This investigation showed that there was no significant difference between the agents but that the effect of the pH was significant. The uptake of fluoride in the form of fluorapatite was more than five times larger at the lower pH level. The second part of the paper deals with the rate of fluoride uptake (increase in fluoride content) from dentifrices in the same pH range. It was shown that the three agents gave the same initial rate of fluoride uptake (about 50 parts/10(6)/min) at pH 6 and that the rate of fluoride uptake in the outer layer of the enamel was proportional to the hydrogen ion activity.

pubmed23n0001_378

Determination of inorganic pyrophosphatase in rat odontoblast layer by a radiochemical method.

The enzyme inorganic pyrophosphatase (PPiase, EC 3.6.1.1) from the odontoblastic layer of rat incisors has been studied by means of a radiochemical micromethod. The enzyme was incubated with 32P-pyrophosphate in tris-HCl buffer at 37 degrees C. The reaction was linear with time for at least 45 min, and the pH optimum was found to be 8.8, independent of the amount of pyrophosphate present. Heating the enzyme at 56 degrees C inhibited the enzyme activity rapidly, Mg2+ ions activated the enzyme by 15% at an ion concentration of 4 mM, while higher concentrations were inhibitory. Ca2+ ions and PO43-ions inhibited the enzyme at all concentrations. F- ions did not affect the PPiase at concentrations below 8 mM, whereas higher concentrations had an inhibiting effect. Urea was found to inhibit the enzyme at concentrations above 1.5 M, while EDTA was a strong inhibitor at very low concentrations. The characteristics of PPiase agree well with the properties of the enzyme nonspecific alkaline phosphatase (EC 3.1.3.1.) studied earlier.

Determination of inorganic pyrophosphatase in rat odontoblast layer by a radiochemical method. The enzyme inorganic pyrophosphatase (PPiase, EC 3.6.1.1) from the odontoblastic layer of rat incisors has been studied by means of a radiochemical micromethod. The enzyme was incubated with 32P-pyrophosphate in tris-HCl buffer at 37 degrees C. The reaction was linear with time for at least 45 min, and the pH optimum was found to be 8.8, independent of the amount of pyrophosphate present. Heating the enzyme at 56 degrees C inhibited the enzyme activity rapidly, Mg2+ ions activated the enzyme by 15% at an ion concentration of 4 mM, while higher concentrations were inhibitory. Ca2+ ions and PO43-ions inhibited the enzyme at all concentrations. F- ions did not affect the PPiase at concentrations below 8 mM, whereas higher concentrations had an inhibiting effect. Urea was found to inhibit the enzyme at concentrations above 1.5 M, while EDTA was a strong inhibitor at very low concentrations. The characteristics of PPiase agree well with the properties of the enzyme nonspecific alkaline phosphatase (EC 3.1.3.1.) studied earlier.

pubmed23n0001_379

Assessment of denture plaque pH in subjects with and without denture stomatitis.

To evaluate the "resting" pH and induced pH changes in denture plaque, soft deposits were collected from the fitting surface of the denture, pooled and suspended in water. Plaque pH was determined with microelectrode equipment before and after mouth rinsing with a sucrose solution. A characteristic level in the "resting" pH of denture plaque was found in most of 12 subjects tested. pH values below the baseline level were recorded for more than 2 h after a rinse. The pH depressions were more pronounced in maxillary than in mandibular plaque. Further, the pH minima tended to be lower in subjects with denture stomatitis than in controls. No clear relationship could be established between the "resting" pH and the concentration of Candida hyphae in denture smears or palatal inflammation.

Assessment of denture plaque pH in subjects with and without denture stomatitis. To evaluate the "resting" pH and induced pH changes in denture plaque, soft deposits were collected from the fitting surface of the denture, pooled and suspended in water. Plaque pH was determined with microelectrode equipment before and after mouth rinsing with a sucrose solution. A characteristic level in the "resting" pH of denture plaque was found in most of 12 subjects tested. pH values below the baseline level were recorded for more than 2 h after a rinse. The pH depressions were more pronounced in maxillary than in mandibular plaque. Further, the pH minima tended to be lower in subjects with denture stomatitis than in controls. No clear relationship could be established between the "resting" pH and the concentration of Candida hyphae in denture smears or palatal inflammation.

pubmed23n0001_380

Bioluminescence assay of enzymes obtained from buccal epithelium by superficial scraping.

A method is presented for the simultaneous assay of buccal enzymes by measuring reduced nicotinamide adenine dinucleotide and adenosine triphosphate with the aid of the bioluminescence of luciferase extracts. The activity of glucose-6-phosphate dehydrogenase (G6PDH) was shown earlier to be increased in homogeneous leukoplakias of the oral mucosa. Since smoking has been implicated as an etiologic factor of leukoplakia, G6PDH was measured in the normal buccal epithelium of cigarette smokers. No difference was found in the activity of G6PDH between smokers and nonsmokers when related to the activity of pyruvate kinase, which is known to be invariable in healthy and leukoplakic oral mucosa. A new compact kinetic luminescence analyzer is briefly described.

Bioluminescence assay of enzymes obtained from buccal epithelium by superficial scraping. A method is presented for the simultaneous assay of buccal enzymes by measuring reduced nicotinamide adenine dinucleotide and adenosine triphosphate with the aid of the bioluminescence of luciferase extracts. The activity of glucose-6-phosphate dehydrogenase (G6PDH) was shown earlier to be increased in homogeneous leukoplakias of the oral mucosa. Since smoking has been implicated as an etiologic factor of leukoplakia, G6PDH was measured in the normal buccal epithelium of cigarette smokers. No difference was found in the activity of G6PDH between smokers and nonsmokers when related to the activity of pyruvate kinase, which is known to be invariable in healthy and leukoplakic oral mucosa. A new compact kinetic luminescence analyzer is briefly described.

pubmed23n0001_381

Allogeneic marrow transplantation for the treatment of leukaemia. A review.

22 HL-A antigen and mixed leukocyte culture-matched sibling bone marrow transplants were attempted in patients with acute leukaemia (at the National Cancer Institute) to define the toxicities of four different immunosuppressive regimens, the complications associated with warrow engraftment and antileukaemic effect. 73% (16/22) were engrafted as indicated by a change to donor red blood cells (RBC) type, leukocyte, immunoglobulin allotype or by the speed of morrow repopulation and the occurrence of the Graft Versus Host Disease (GVHD). 12 of 16 (75%) successful engrafted patients developed GVHD. The current published results of clinical bone marrow transplantation from major centers has been reviewed and will be discussed in relationship to current clinical complications associated with bone marrow transplantation.

Allogeneic marrow transplantation for the treatment of leukaemia. A review. 22 HL-A antigen and mixed leukocyte culture-matched sibling bone marrow transplants were attempted in patients with acute leukaemia (at the National Cancer Institute) to define the toxicities of four different immunosuppressive regimens, the complications associated with warrow engraftment and antileukaemic effect. 73% (16/22) were engrafted as indicated by a change to donor red blood cells (RBC) type, leukocyte, immunoglobulin allotype or by the speed of morrow repopulation and the occurrence of the Graft Versus Host Disease (GVHD). 12 of 16 (75%) successful engrafted patients developed GVHD. The current published results of clinical bone marrow transplantation from major centers has been reviewed and will be discussed in relationship to current clinical complications associated with bone marrow transplantation.

pubmed23n0001_382

Current status of treatment of pneumonia.

Proper treatment of pneumonia is dependent upon a correct diagnosis. Pneumonia may be due to infectious agents, allergic phenomena, or chemical causes. Treatment regimens are outlined for the various types of pneumonia--pneumococcal, staphylococcal, fungal, and pneumonia due to gram-negative and anaerobic gram-negative bacilli, to Blastomyces dermatitidis, and to the parasite Pneumocystis carinii. In discussing current concepts of treatment, several well-known methods are emphasized, as well as newer developments, knowledge of which is essential for optimal treatment of pneumonia.

Current status of treatment of pneumonia. Proper treatment of pneumonia is dependent upon a correct diagnosis. Pneumonia may be due to infectious agents, allergic phenomena, or chemical causes. Treatment regimens are outlined for the various types of pneumonia--pneumococcal, staphylococcal, fungal, and pneumonia due to gram-negative and anaerobic gram-negative bacilli, to Blastomyces dermatitidis, and to the parasite Pneumocystis carinii. In discussing current concepts of treatment, several well-known methods are emphasized, as well as newer developments, knowledge of which is essential for optimal treatment of pneumonia.

pubmed23n0001_383

'Picture frame' fibres in a carrier of the trait for malignant hyperpyrexia.

A member of a family which was known to be susceptible to malignant hyperpyrexia, who was identified as a carrier by the presence of an elevated serum creatine-phosphokinase, has been investigated further. Muscle was examined biochemically, and the study included the sarcoplasmic ATPase-activity, actinomycin, Mg2+ ATPase activity, ATP, phosphocreatine and glucose-6-phosphate. In addition, the calcium uptake by the sarcoplasmic reticulum was studied. The histochemical analysis of the muscle revealed the presence of a new fibre type characterised by a dense rim of ATPase activity, which gives the impression of a 'picture-frame'. Ultramicroscopic study revealed changes in the mitochondria and areas of myofibrillar disruption with swelling of the sarcoplasmic reticulum.

'Picture frame' fibres in a carrier of the trait for malignant hyperpyrexia. A member of a family which was known to be susceptible to malignant hyperpyrexia, who was identified as a carrier by the presence of an elevated serum creatine-phosphokinase, has been investigated further. Muscle was examined biochemically, and the study included the sarcoplasmic ATPase-activity, actinomycin, Mg2+ ATPase activity, ATP, phosphocreatine and glucose-6-phosphate. In addition, the calcium uptake by the sarcoplasmic reticulum was studied. The histochemical analysis of the muscle revealed the presence of a new fibre type characterised by a dense rim of ATPase activity, which gives the impression of a 'picture-frame'. Ultramicroscopic study revealed changes in the mitochondria and areas of myofibrillar disruption with swelling of the sarcoplasmic reticulum.

pubmed23n0001_384

The GP dilemma. Recommendations and synopsis of a student conference.

The proceedings of a conference organised by students are reported. The present standing of the general practitioner and his need in different societies are equated and the obvious deficiencies are considered. Such themes as maldistribution, service and education are discussed. Resolutions derived from the conference are reported in full.

The GP dilemma. Recommendations and synopsis of a student conference. The proceedings of a conference organised by students are reported. The present standing of the general practitioner and his need in different societies are equated and the obvious deficiencies are considered. Such themes as maldistribution, service and education are discussed. Resolutions derived from the conference are reported in full.

pubmed23n0001_385

Gastric emptying of liquids after different vagotomies and pyloroplasty.

Gastric emptying of five liquid meals which differ in their physicochemical properties have been measured in control dogs and dogs that have received a Heinecke-Mikulicz pyloroplasty alone, proximal gastric vagotomy without drainage, selective gastric vagotomy and pyloroplasty and truncal vagotomy and pyloroplasty. The first two phases of emptying have been computed by the method of least squares to obtain a logarithmic-linear pattern and are expressed as relative rates: The initial post-ingestion process is characterized by beta or the average relative rate of emptying in the first ten minutes, the basic or exponential rate as beta and the change in rate from the initial to basic pattern as deltabeta. Each measure of gastric emptying was statistically analyzed to determine specific differences in rates between the operations studied. We confirmed the earlier claims that pyloroplasty alone does not change the emptying rate of liquid meals. Each measure or phase of emptying varies consistently across the operations from meal to meal tested. Initial emptying after all three vagotomies is significantly faster than control with progressive rate increases as proximal gastric vagotomy is compared with selective gastric vagotomy with pyloroplasty and with truncal vagotomy with pyloroplasty, probably indicative of gastric fundal loss of accommodation to volume distention after denervation. The basic exponential pattern of emptying is not lost after any of the operations studied. The basic rate after proximal gastric vagotomy and selective gastric vagotomy with pyloroplasty is nearly identical, slightly delayed from the control rate and significantly slower than the more rapid rate after truncal vagotomy with pyloroplasty. Possible explanations for these are discussed and imply a particular importance of the hepatic and celiac vagal fibers, sectioned only with truncal vagotomy, in the regulation of gastric emptying of liquids.

Gastric emptying of liquids after different vagotomies and pyloroplasty. Gastric emptying of five liquid meals which differ in their physicochemical properties have been measured in control dogs and dogs that have received a Heinecke-Mikulicz pyloroplasty alone, proximal gastric vagotomy without drainage, selective gastric vagotomy and pyloroplasty and truncal vagotomy and pyloroplasty. The first two phases of emptying have been computed by the method of least squares to obtain a logarithmic-linear pattern and are expressed as relative rates: The initial post-ingestion process is characterized by beta or the average relative rate of emptying in the first ten minutes, the basic or exponential rate as beta and the change in rate from the initial to basic pattern as deltabeta. Each measure of gastric emptying was statistically analyzed to determine specific differences in rates between the operations studied. We confirmed the earlier claims that pyloroplasty alone does not change the emptying rate of liquid meals. Each measure or phase of emptying varies consistently across the operations from meal to meal tested. Initial emptying after all three vagotomies is significantly faster than control with progressive rate increases as proximal gastric vagotomy is compared with selective gastric vagotomy with pyloroplasty and with truncal vagotomy with pyloroplasty, probably indicative of gastric fundal loss of accommodation to volume distention after denervation. The basic exponential pattern of emptying is not lost after any of the operations studied. The basic rate after proximal gastric vagotomy and selective gastric vagotomy with pyloroplasty is nearly identical, slightly delayed from the control rate and significantly slower than the more rapid rate after truncal vagotomy with pyloroplasty. Possible explanations for these are discussed and imply a particular importance of the hepatic and celiac vagal fibers, sectioned only with truncal vagotomy, in the regulation of gastric emptying of liquids.

pubmed23n0001_386

Pancreatic enzyme response with an elemental diet.

Elemental diets can maintain or slightly improve the nutritional status without a major stimulatory effect on the pancreas. Six dogs were maintained with a regular chow diet, switched to an elemental diet and, subsequently, returned to a chow diet. Cannulation of the pancreatic duct through a duodenal cutaneous fistula revealed the enzyme response to be decreased in a dog maintained with an elemental diet, with no or only a slight weight gain. Return to a regular diet resulted in a return of pancreatic enzyme response.

Pancreatic enzyme response with an elemental diet. Elemental diets can maintain or slightly improve the nutritional status without a major stimulatory effect on the pancreas. Six dogs were maintained with a regular chow diet, switched to an elemental diet and, subsequently, returned to a chow diet. Cannulation of the pancreatic duct through a duodenal cutaneous fistula revealed the enzyme response to be decreased in a dog maintained with an elemental diet, with no or only a slight weight gain. Return to a regular diet resulted in a return of pancreatic enzyme response.

pubmed23n0001_387

Gastric fibrinolysis.

Gastric juice from 15 normals, 20 patients with gastric ulcer and 14 patients with erosive haemorrhagic gastroduodenitis was investigated in respect of its activity on unheated and heated fibrin plates and its content of FDP and plasminogen or plasmin with immunochemical methods. Gastric juice from normals showed no activity on unheated and heated fibrin plates, and no FDP or plasminogen could be demonstrated. In the patients with gastric ulcer the gastric juice showed little or no fibrinolytic activity on fibrin plates except in 2, who had regurgitation of duodenal juice and neutral pH of the juice. These patients had equally high activity on heated as on unheated plates and no plasmin could be demonstrated. It was shown that this activity was not due to fibrinolysis, but to non-specific proteolytic activity (probably trypsin). The patients with erosive haemorrhage gastroduodenitis exhibited quite a different picture. The gastric juice from these patients showed extremely high activity on fibrin plates, the activity was higher on unheated than on heated plates. The activity was inhibited in vitro by addition of EACA and in vivo after administration of AMCA. The occurence of plasmic could be demonstrated directly immunologically in the gastric juice. By comparsion of plasmin and trypsin in various assays it could further be improved that the gastric juice in these cases contained plasminogen activator and plasmin. The patients with erosive haemorrhagic gastroduodenitis showed no increase in fibrinolysis in the blood, but low values for plasminogen and alpha2-M, and the serum contained FDP. These findings in the blood and gastric juice were interpreted as signs of local fibrinolysis in the stomach and duodenum. There is reason to assume that this gastric fibrinolysis contributes substantially to the bleeding tendency. The effect of administration of AMCA on fibrinolytic activity and the haemorrhage lends support to the assumption of such a mechanism.

Gastric fibrinolysis. Gastric juice from 15 normals, 20 patients with gastric ulcer and 14 patients with erosive haemorrhagic gastroduodenitis was investigated in respect of its activity on unheated and heated fibrin plates and its content of FDP and plasminogen or plasmin with immunochemical methods. Gastric juice from normals showed no activity on unheated and heated fibrin plates, and no FDP or plasminogen could be demonstrated. In the patients with gastric ulcer the gastric juice showed little or no fibrinolytic activity on fibrin plates except in 2, who had regurgitation of duodenal juice and neutral pH of the juice. These patients had equally high activity on heated as on unheated plates and no plasmin could be demonstrated. It was shown that this activity was not due to fibrinolysis, but to non-specific proteolytic activity (probably trypsin). The patients with erosive haemorrhage gastroduodenitis exhibited quite a different picture. The gastric juice from these patients showed extremely high activity on fibrin plates, the activity was higher on unheated than on heated plates. The activity was inhibited in vitro by addition of EACA and in vivo after administration of AMCA. The occurence of plasmic could be demonstrated directly immunologically in the gastric juice. By comparsion of plasmin and trypsin in various assays it could further be improved that the gastric juice in these cases contained plasminogen activator and plasmin. The patients with erosive haemorrhagic gastroduodenitis showed no increase in fibrinolysis in the blood, but low values for plasminogen and alpha2-M, and the serum contained FDP. These findings in the blood and gastric juice were interpreted as signs of local fibrinolysis in the stomach and duodenum. There is reason to assume that this gastric fibrinolysis contributes substantially to the bleeding tendency. The effect of administration of AMCA on fibrinolytic activity and the haemorrhage lends support to the assumption of such a mechanism.

pubmed23n0001_388

Acid-base parameters in the dehydrated camel.

The effect of prolonged (10 days) dehydration on acid-base parameters of camel blood was examined. The pH and PCO2 levels rose significantly in the course of dehydration. This state was comparable with compensated non-respiratory alkalosis found in other animals. The plasma sodium, and magnesium levels rose significantly also. The plasma oxygen and calcium levels declined significantly. There were no significant changes in potassium and phosphate levels. It is concluded that the changes found in acid-base status following dehydration are further evidence of water preservation mechanisms in the dehydrated camel.

Acid-base parameters in the dehydrated camel. The effect of prolonged (10 days) dehydration on acid-base parameters of camel blood was examined. The pH and PCO2 levels rose significantly in the course of dehydration. This state was comparable with compensated non-respiratory alkalosis found in other animals. The plasma sodium, and magnesium levels rose significantly also. The plasma oxygen and calcium levels declined significantly. There were no significant changes in potassium and phosphate levels. It is concluded that the changes found in acid-base status following dehydration are further evidence of water preservation mechanisms in the dehydrated camel.

pubmed23n0001_389

[Clinical experiences with untreated homologous vein grafts in reconstruction of arteries (author's transl)].

50 transplantations of homologous vein grafts in reconstruction of arteries are reported on. Vein grafts were either transplanted immediately or used after deep freezing. This procedure has proved to be effective in the replacement of arteries during our observation period of four years. Results of homologous vein transplants are similar to those of autologous transplants.

[Clinical experiences with untreated homologous vein grafts in reconstruction of arteries (author's transl)]. 50 transplantations of homologous vein grafts in reconstruction of arteries are reported on. Vein grafts were either transplanted immediately or used after deep freezing. This procedure has proved to be effective in the replacement of arteries during our observation period of four years. Results of homologous vein transplants are similar to those of autologous transplants.

pubmed23n0001_390

The effect of a simulated subarachnoid hemorrhage on cerebral blood flow in the monkey.

The hydrogen clearance method was used to measure local and total cerebral blood flow (CBF) in the rhesus monkey before and for five hours after a simulated subarachnoid hemorrhage (SAH). CBF remained stable after SAH unless SAH was associated with a fall in cerebral perfusion pressure. In addition, cerebrovascular resistance did not increase after SAH. These results suggest that vasoactive agents in fresh whole blood, and the arterial spasm they produce when added to cerebrospinal fluid (CSF), play only a limited role in the pathogenesis of ischemic encephalopathy that follows an SAH.

The effect of a simulated subarachnoid hemorrhage on cerebral blood flow in the monkey. The hydrogen clearance method was used to measure local and total cerebral blood flow (CBF) in the rhesus monkey before and for five hours after a simulated subarachnoid hemorrhage (SAH). CBF remained stable after SAH unless SAH was associated with a fall in cerebral perfusion pressure. In addition, cerebrovascular resistance did not increase after SAH. These results suggest that vasoactive agents in fresh whole blood, and the arterial spasm they produce when added to cerebrospinal fluid (CSF), play only a limited role in the pathogenesis of ischemic encephalopathy that follows an SAH.

pubmed23n0001_391

Effects of oxygen saturation and pCO2 on brain uptake of glucose analogues in rabbits.

The effect of oxygen saturation and PCO2 on brain uptake of glucose analogues was studied in rabbits. Using a modified Oldendorf technique, 14C-labeled glucose analogues with a 3H2O reference standard were introduced into the cerebral circulation via the common carotid artery, and the radioactivity of the ipsilateral cerebral cortex was counted and expressed in terms of a brain uptake index (BUI). Severe hypoxia (oxygen saturation less than or equal to 18%) resulted in approximately a 40% decrease in the BUI of 2-deoxy-D-glucose and a 45% decrease in the BUI of 3-0-methyl-D-glucose. Severe hypercapnia (PCO2 = 100 mm Hg) caused a 45% decrease in the BUI of both of these glucose analogues. Hypercapnia superimposed on severe hypoxia had no additional effect. Hypocapnia (PCO2 = 15 mm Hg) increased the BUI of 3-0-methyl-D-glucose by 35% of the control value, and this increase was extremely sensitive to competitive inhibition. When BUI values were plotted against pH rather than PCO2 for the same experiments, there was a good correlation with the calculated linear regression. These results are compared with previous findings on pathologically induced changes in brain uptake of glucose analogues, and the possible role of blood flow is considered in detail.

Effects of oxygen saturation and pCO2 on brain uptake of glucose analogues in rabbits. The effect of oxygen saturation and PCO2 on brain uptake of glucose analogues was studied in rabbits. Using a modified Oldendorf technique, 14C-labeled glucose analogues with a 3H2O reference standard were introduced into the cerebral circulation via the common carotid artery, and the radioactivity of the ipsilateral cerebral cortex was counted and expressed in terms of a brain uptake index (BUI). Severe hypoxia (oxygen saturation less than or equal to 18%) resulted in approximately a 40% decrease in the BUI of 2-deoxy-D-glucose and a 45% decrease in the BUI of 3-0-methyl-D-glucose. Severe hypercapnia (PCO2 = 100 mm Hg) caused a 45% decrease in the BUI of both of these glucose analogues. Hypercapnia superimposed on severe hypoxia had no additional effect. Hypocapnia (PCO2 = 15 mm Hg) increased the BUI of 3-0-methyl-D-glucose by 35% of the control value, and this increase was extremely sensitive to competitive inhibition. When BUI values were plotted against pH rather than PCO2 for the same experiments, there was a good correlation with the calculated linear regression. These results are compared with previous findings on pathologically induced changes in brain uptake of glucose analogues, and the possible role of blood flow is considered in detail.

pubmed23n0001_392

Engraftment of allogeneic dog bone marrow.

Resistance to allogeneic bone-marrow grafts (AR) was found to occur in many species, including the dog. The i.v. administration of silica particles suppressed AR in vivo in this species. Genetic studies provide suggestive evidence for the existence of a previously unrecognized system or systems in the canine major histocompatibility complex controlling AR.

Engraftment of allogeneic dog bone marrow. Resistance to allogeneic bone-marrow grafts (AR) was found to occur in many species, including the dog. The i.v. administration of silica particles suppressed AR in vivo in this species. Genetic studies provide suggestive evidence for the existence of a previously unrecognized system or systems in the canine major histocompatibility complex controlling AR.

pubmed23n0001_393

Lactate and pyruvate concentrations, and acid-base balance of cerebrospinal fluid in experimentally induced intracerebral and subarachnoid hemorrhage in dogs.

The effect of blood injected into either subarachnoid space or subcortical brain tissue upon lactate and pyruvate concentrations as well as acid-base balance of cerebrospinal fluid (CSF) was studied in the anesthetized dog. CSF lactate and lactate/pyruvate ratio (L/P ratio) increased progressively following the intracranial injection of blood and reached the maximum level at six hours after injection. These changes were significantly greater in animals with intracerebral hematoma than in those with subarachnoid hemorrhage (SAH). An increase in CSF lactate and L/P ratio in hemorrhagic CSF seems to be caused by two different factors. Shed blood cells per se produce lactate and pyruvate, and blood in the subarachnoid space and intracerebral hematomas cause secondary changes in brain tissue metabolism by a probable reduction of cerebral blood flow. Therefore, an increase in CSF lactate with a concomitant rise in CSF L/P ratio is a useful indicator for brain tissue hypoxia, even when CSF is hemorrhagic. The association of an increase in CSF lactate to a disproportionate decrease in CSF HCO-3 was also observed in these animals.

Lactate and pyruvate concentrations, and acid-base balance of cerebrospinal fluid in experimentally induced intracerebral and subarachnoid hemorrhage in dogs. The effect of blood injected into either subarachnoid space or subcortical brain tissue upon lactate and pyruvate concentrations as well as acid-base balance of cerebrospinal fluid (CSF) was studied in the anesthetized dog. CSF lactate and lactate/pyruvate ratio (L/P ratio) increased progressively following the intracranial injection of blood and reached the maximum level at six hours after injection. These changes were significantly greater in animals with intracerebral hematoma than in those with subarachnoid hemorrhage (SAH). An increase in CSF lactate and L/P ratio in hemorrhagic CSF seems to be caused by two different factors. Shed blood cells per se produce lactate and pyruvate, and blood in the subarachnoid space and intracerebral hematomas cause secondary changes in brain tissue metabolism by a probable reduction of cerebral blood flow. Therefore, an increase in CSF lactate with a concomitant rise in CSF L/P ratio is a useful indicator for brain tissue hypoxia, even when CSF is hemorrhagic. The association of an increase in CSF lactate to a disproportionate decrease in CSF HCO-3 was also observed in these animals.

pubmed23n0001_394

A study of variables affecting the quality of platelets stored at "room temperature".

The effect of variables associated with the donor and with methods of collecting, processing, and storing platelets on the quality of platelets kept at ambient temperature was studied. Changes in structural integrity of platelets, decrease in pH, loss of aggregability, and kinetics in vivo of platelets tagged with 51Cr were used as indicators of the tolerance of platelets to storage. A platelet concentration of less than 2.5 x 10(6) per cu mm, a temperature of storage less than 24 C, and continuous, gentle, agitation were found to be essential for satisfactory preservation of platelet integrity, function, and post-transfusion survival. Platelets from female donors tolerated storage less well than did platelets from male donors, possibly because the lower hematocrit of blood collection from females resulted in greater initial acidity of the concentrate. A number of other variables analyzed appear to be of little or no consequence for successful platelet storage.

A study of variables affecting the quality of platelets stored at "room temperature". The effect of variables associated with the donor and with methods of collecting, processing, and storing platelets on the quality of platelets kept at ambient temperature was studied. Changes in structural integrity of platelets, decrease in pH, loss of aggregability, and kinetics in vivo of platelets tagged with 51Cr were used as indicators of the tolerance of platelets to storage. A platelet concentration of less than 2.5 x 10(6) per cu mm, a temperature of storage less than 24 C, and continuous, gentle, agitation were found to be essential for satisfactory preservation of platelet integrity, function, and post-transfusion survival. Platelets from female donors tolerated storage less well than did platelets from male donors, possibly because the lower hematocrit of blood collection from females resulted in greater initial acidity of the concentrate. A number of other variables analyzed appear to be of little or no consequence for successful platelet storage.

pubmed23n0001_395

Dopamine-containing neurons of the substantia nigra and their terminals in the neostriatum.

Tne ultrastructural and fluorescence histochemical characteristics of the mature rabbit substantia nigra and neostriatum have been reviewed as a frame of reference for the developmental study. Biochemical investigations were reported on neostriatal dopamine concentrations and the relative uptake and accumulation of 3H-dopamine by this tissue from fetal to adult stages, to provide quantitative data for correlation with the fluorexcence information. The development of the neurons of the substantia nigra and their axons which project to the neostriatum has been presented from their appearance at day 14 of gestation to their maturation in early postnatal life. The initial bipolar neuroblasts, which develop in the midline of the caudal mesencephalon, are fluorescent as soon as they emerge from the ependymal zone. Their fluorescent axons, which form the nigroneostriatal pathway, reach the telencephalon at day 16 of gestation and ramify extensively in the putamen by day 20, but do not enter the caudate nucleus until several days later. Some of the early fluorescent axonal profiles in the putamen are extremely large. Electron microscopic study of theis stage suggests that the large fluorescent profiles may correspond to axonal growth cones or early synapses. A distinct substantia nigra, pars compacta and reticulata, can be recognized by fluorescence microscopy by day 20 of gestation. Electron microscopy reveals that the young neurons are multipolar with numerous developing dendrites, some of which exhibit early synaptic junctions. The subsequent maturaition of these cells and the neuropil is described. The fluorescent axons of the substantia nigra grow into the putamen and caudate nucleus in a nonuniform manner forming fluorescent islands throughout the neostriatum in late fetal life. Occasionally, minute beaded fluorescent axons are found. These profiles might correspond to some of the axons with varicosities "en passage" revealed by electron microscopy. In an attempt to identify further the dopamine-containing axon, the ultratructure of adult neostriatum incubated in 5-hydroxydopamine was reported. Axonal varicosities "en passage" containing a dense "tag" in the vesicles were found. Most of the tagged boutons did not exhibit synaptic contacts. The possible significance of these finding s as related to dopamine secretion are discussed.

Dopamine-containing neurons of the substantia nigra and their terminals in the neostriatum. Tne ultrastructural and fluorescence histochemical characteristics of the mature rabbit substantia nigra and neostriatum have been reviewed as a frame of reference for the developmental study. Biochemical investigations were reported on neostriatal dopamine concentrations and the relative uptake and accumulation of 3H-dopamine by this tissue from fetal to adult stages, to provide quantitative data for correlation with the fluorexcence information. The development of the neurons of the substantia nigra and their axons which project to the neostriatum has been presented from their appearance at day 14 of gestation to their maturation in early postnatal life. The initial bipolar neuroblasts, which develop in the midline of the caudal mesencephalon, are fluorescent as soon as they emerge from the ependymal zone. Their fluorescent axons, which form the nigroneostriatal pathway, reach the telencephalon at day 16 of gestation and ramify extensively in the putamen by day 20, but do not enter the caudate nucleus until several days later. Some of the early fluorescent axonal profiles in the putamen are extremely large. Electron microscopic study of theis stage suggests that the large fluorescent profiles may correspond to axonal growth cones or early synapses. A distinct substantia nigra, pars compacta and reticulata, can be recognized by fluorescence microscopy by day 20 of gestation. Electron microscopy reveals that the young neurons are multipolar with numerous developing dendrites, some of which exhibit early synaptic junctions. The subsequent maturaition of these cells and the neuropil is described. The fluorescent axons of the substantia nigra grow into the putamen and caudate nucleus in a nonuniform manner forming fluorescent islands throughout the neostriatum in late fetal life. Occasionally, minute beaded fluorescent axons are found. These profiles might correspond to some of the axons with varicosities "en passage" revealed by electron microscopy. In an attempt to identify further the dopamine-containing axon, the ultratructure of adult neostriatum incubated in 5-hydroxydopamine was reported. Axonal varicosities "en passage" containing a dense "tag" in the vesicles were found. Most of the tagged boutons did not exhibit synaptic contacts. The possible significance of these finding s as related to dopamine secretion are discussed.

pubmed23n0001_396

[Separation of the hormones of the adenohypophysis of rats by use of electrophoresis in polyacrylamide gel in the presence of sodium dodecyl sulfate].

A comparative study of rat adenohypophysis extract and its alcohol fractions was performed by two variants of the method of electrophoresis in polyacrylamide gel: at pH 9,5 and with the presence of sodium dodecyl sulphate at pH 7.2. With the presence of sodium dodecylsulphate four protein zones are found which in the order from the anode towards the cathode are identified as hemoglobin, somatotropin, lactotropin and albumin. It is shown that the somatotropin zone after the extract separation at pH 9.5 is inhomogeneous and consists of somatotropin and hemoglobin.

[Separation of the hormones of the adenohypophysis of rats by use of electrophoresis in polyacrylamide gel in the presence of sodium dodecyl sulfate]. A comparative study of rat adenohypophysis extract and its alcohol fractions was performed by two variants of the method of electrophoresis in polyacrylamide gel: at pH 9,5 and with the presence of sodium dodecyl sulphate at pH 7.2. With the presence of sodium dodecylsulphate four protein zones are found which in the order from the anode towards the cathode are identified as hemoglobin, somatotropin, lactotropin and albumin. It is shown that the somatotropin zone after the extract separation at pH 9.5 is inhomogeneous and consists of somatotropin and hemoglobin.

pubmed23n0001_397

[Properties of NAD-glycohydrolase of the nuclei of the liver cells of rats].

Certain properties of the rat liver cell nuclei NAD-glycohydrolase (EC 3.2.2.5) were investigated. It is established that its highest activity is at 37 degrees with activation energy equal to 9480 cal/M and with factor Q10 equal to 1.5. The enzyme pH optimum in 0.2 M tris acetate is equal to 6.5 and in 0.2 potassium phosphate - 7.5. It was shown that the enzyme manifests its strict specificity only with beta-NAD, and it hardly decomposes NADP without affecting NADH, NADPH and NMN. The apparent Km value of the enzyme with respect to NAD is established. Isonicotinic acid hydrazide, nicotinamide and to the less extent nicotinic acid inhibit the enzymatic activity of nuclei. EDTA, EGTA, p-CMB, mercaptoethanol do not cause any changes in the rat liver cells nuclei NADase activity.

[Properties of NAD-glycohydrolase of the nuclei of the liver cells of rats]. Certain properties of the rat liver cell nuclei NAD-glycohydrolase (EC 3.2.2.5) were investigated. It is established that its highest activity is at 37 degrees with activation energy equal to 9480 cal/M and with factor Q10 equal to 1.5. The enzyme pH optimum in 0.2 M tris acetate is equal to 6.5 and in 0.2 potassium phosphate - 7.5. It was shown that the enzyme manifests its strict specificity only with beta-NAD, and it hardly decomposes NADP without affecting NADH, NADPH and NMN. The apparent Km value of the enzyme with respect to NAD is established. Isonicotinic acid hydrazide, nicotinamide and to the less extent nicotinic acid inhibit the enzymatic activity of nuclei. EDTA, EGTA, p-CMB, mercaptoethanol do not cause any changes in the rat liver cells nuclei NADase activity.

pubmed23n0001_398

[Some properties of "soluble" Na+ and K+-ATPase obtained from various subcellular membrane structures of the brain by use of non-ionic detergents].

A comparative study was carried out of some properties of "soluble" Na+, K+-ATPase obtained from different subcellular membrane brain structures by means of non-ionic detergents of triton X-100 and digitonin. It is established that temperature and pH-optima of "soluble" Na+, K+-ATPase are close to these optima of the initial membrane preparations. A certain difference is observed in the dynamics of temperature and pH-dependence of Na+, K+-ATPase activity in the extracts from different subcellular structures. The stability of the preparations in storage was investigated. A conclusion is made that more stable enzyme extracts may be obtained by means of digitonin.

[Some properties of "soluble" Na+ and K+-ATPase obtained from various subcellular membrane structures of the brain by use of non-ionic detergents]. A comparative study was carried out of some properties of "soluble" Na+, K+-ATPase obtained from different subcellular membrane brain structures by means of non-ionic detergents of triton X-100 and digitonin. It is established that temperature and pH-optima of "soluble" Na+, K+-ATPase are close to these optima of the initial membrane preparations. A certain difference is observed in the dynamics of temperature and pH-dependence of Na+, K+-ATPase activity in the extracts from different subcellular structures. The stability of the preparations in storage was investigated. A conclusion is made that more stable enzyme extracts may be obtained by means of digitonin.

pubmed23n0001_399

[Properties of glutamine synthetase of the brain of rats during ontogenesis].

When investigating activity of glutamine synthetase of the enzymatic preparations isolated from the brain of rats of 0.5, 1, 3, 12 and 24-month age, no considerable differences were found in the indices of the values Km to a-glutamate and Vmax which are respectively equal to: Km (M-10(-3))=5.5; 3.5; 3.6; 3.9; 5.5; Vmax=3.1; 4.5; 5.0; 5.2 muM were found. When adding various concentrations of a-ketoglutaric acid into the incubation medium the differences are registered in the degree and character of the age changes in brain glutamine synthetase activity in comparison with this enzyme form the liver.

[Properties of glutamine synthetase of the brain of rats during ontogenesis]. When investigating activity of glutamine synthetase of the enzymatic preparations isolated from the brain of rats of 0.5, 1, 3, 12 and 24-month age, no considerable differences were found in the indices of the values Km to a-glutamate and Vmax which are respectively equal to: Km (M-10(-3))=5.5; 3.5; 3.6; 3.9; 5.5; Vmax=3.1; 4.5; 5.0; 5.2 muM were found. When adding various concentrations of a-ketoglutaric acid into the incubation medium the differences are registered in the degree and character of the age changes in brain glutamine synthetase activity in comparison with this enzyme form the liver.