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PMID:24332 | Orungo virus, an orbivirus from Africa: effects of physical and chemical agents. | Orungo virus is relatively sensitive to lipid solvents and sodium deoxycholate. It is readily inactivated at 37 degrees C and above, by u.v. irradiation, BP and formalin. Orungo virus is ph 3.0 labile, but stable at ph range of 5.0-7.0. Multiplication of Orungo virus in BHK-21 cell cultures is not affected in the presence of 5-iododeoxyuridine. | Orungo virus, an orbivirus from Africa: effects of physical and chemical agents. Orungo virus is relatively sensitive to lipid solvents and sodium deoxycholate. It is readily inactivated at 37 degrees C and above, by u.v. irradiation, BP and formalin. Orungo virus is ph 3.0 labile, but stable at ph range of 5.0-7.0. Multiplication of Orungo virus in BHK-21 cell cultures is not affected in the presence of 5-iododeoxyuridine. | [
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PMID:24334 | Pulmonary function following feeding in low-birth-weight infants. | Determination of functional residual capacity, arterial gas tensions and pH, and arterial-alveolar differences was carried out in a group of nondistressed premature infants to determine if significant alterations in lung volumes or ventilation perfusion relationships sufficiently large to cause cyanosis could be detected after feeding. The only statistically significant changes observed in these parameters were immediately following feeding, a fall of .01 pH units and a fall of 6 mm Hg in arterial oxygen tension; and 15 minutes after feeding, a fall of .01 pH units and a rise in arterial PCO2 of 3 mm Hg. In addition, significant falls in peripheral blood flow were observed five minutes after feeding in association with marked elevations in peripheral vascular resistance. The mild impairment in pulmonary function in association with the more profound changes in limb blood flow are consistent with other studies in premature and full-term infants. These data suggest that cyanotic attacks following feeding may have a complex etiology and require an aggressive multisystem approach for adequate diagnosis and therapy. | Pulmonary function following feeding in low-birth-weight infants. Determination of functional residual capacity, arterial gas tensions and pH, and arterial-alveolar differences was carried out in a group of nondistressed premature infants to determine if significant alterations in lung volumes or ventilation perfusion relationships sufficiently large to cause cyanosis could be detected after feeding. The only statistically significant changes observed in these parameters were immediately following feeding, a fall of .01 pH units and a fall of 6 mm Hg in arterial oxygen tension; and 15 minutes after feeding, a fall of .01 pH units and a rise in arterial PCO2 of 3 mm Hg. In addition, significant falls in peripheral blood flow were observed five minutes after feeding in association with marked elevations in peripheral vascular resistance. The mild impairment in pulmonary function in association with the more profound changes in limb blood flow are consistent with other studies in premature and full-term infants. These data suggest that cyanotic attacks following feeding may have a complex etiology and require an aggressive multisystem approach for adequate diagnosis and therapy. | [
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PMID:24335 | Toxic epidermal necrolysis from graft-vs-host disease. Occurrence in a patient with thymic hypoplasia. | An infant with diarrhea, failure to thrive, and a seborrhea-like skin eruption was thought to have fatal familial Leiner's syndrome. Treatment with nonirradiated plasma was followed by graft-vs-host disease and fatal toxic epidermal necrolysis; thymic hypoplasia was found at autopsy. Accurate diagnosis of immunodeficiency syndromes is essential to avoid potentially harmful therapy. | Toxic epidermal necrolysis from graft-vs-host disease. Occurrence in a patient with thymic hypoplasia. An infant with diarrhea, failure to thrive, and a seborrhea-like skin eruption was thought to have fatal familial Leiner's syndrome. Treatment with nonirradiated plasma was followed by graft-vs-host disease and fatal toxic epidermal necrolysis; thymic hypoplasia was found at autopsy. Accurate diagnosis of immunodeficiency syndromes is essential to avoid potentially harmful therapy. | [
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PMID:24337 | Diagnosis and treatment of diffuse ileojejunitis. | Thirteen patients with diffuse ileojejunitis have been diagnosed and treated by us over the past ten years. The disease bears close resemblance to Crohn's disease and may represent a variant of it. No clearcut relationship to celiac sprue was observed in this group of patients. Therapeutic success was obtained in the majority of patients treated with the anti-inflammatory drugs, sulfasalazine and steroids, with four patients requiring resectional surgery, all others manageable by nonsurgical means. There was no mortality in this series of patients. | Diagnosis and treatment of diffuse ileojejunitis. Thirteen patients with diffuse ileojejunitis have been diagnosed and treated by us over the past ten years. The disease bears close resemblance to Crohn's disease and may represent a variant of it. No clearcut relationship to celiac sprue was observed in this group of patients. Therapeutic success was obtained in the majority of patients treated with the anti-inflammatory drugs, sulfasalazine and steroids, with four patients requiring resectional surgery, all others manageable by nonsurgical means. There was no mortality in this series of patients. | [
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PMID:24338 | H2 receptor antagonists. | The uses of the histamine H2 receptor antagonists (H2 antagonists) in the management of peptic ulcer disease and the Zollinger-Ellison syndrome are reviewed. Drugs included in the discussion are burimamide, metiamide and cimetidine. The secretion and the pharmacology of the H2 antagonists are described. A discussion of the clinical use of the H2 receptor antagonists in the treatment of gastric hyperacidity and hypersecretory states, and the side effects encountered is presented. Cimetidine is effective in the treatment of duodenal ulcer, and its use appears promising in Zollinger-Ellison syndrome and gastric ulceration. | H2 receptor antagonists. The uses of the histamine H2 receptor antagonists (H2 antagonists) in the management of peptic ulcer disease and the Zollinger-Ellison syndrome are reviewed. Drugs included in the discussion are burimamide, metiamide and cimetidine. The secretion and the pharmacology of the H2 antagonists are described. A discussion of the clinical use of the H2 receptor antagonists in the treatment of gastric hyperacidity and hypersecretory states, and the side effects encountered is presented. Cimetidine is effective in the treatment of duodenal ulcer, and its use appears promising in Zollinger-Ellison syndrome and gastric ulceration. | [
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PMID:24339 | Pharmacists' knowledge and attitudes regarding monitoring the behaviors of children with learning disabilities. | The attitudes and knowledge of pharmacists in Nebraska regarding children's learning disabilities were surveyed by questionnaire. One hundred thirty-three pharmacists responded. The results were analyzed according to type of practice (community, chain and hospital) and age (20--30 years, 31--40 years, 41--50 years, and 51 years and over). The results indicate that pharmacists are being asked about learning disabilities. Overall, 53% of the pharmacists responding had been asked about learning disabilities by parents, physicians, teachers or others. No significant interactions were found between the type of practice and age, and no significant main effect for type of practice was found in the ANOVA results for knowledge scores or attitude scores. Even though there were no differences owing to age in pharmacists' knowledge of learning disabilities, age was found to have a significant effect on attitudes toward increased pharmacist involvement in monitoring behaviors of learning-disabled children; the younger groups had a more positive attitude. The pharmacist seems to have the knowledge and attitudes required to become more actively involved in monitoring the medications and behaviors of children with learning disabilities. | Pharmacists' knowledge and attitudes regarding monitoring the behaviors of children with learning disabilities. The attitudes and knowledge of pharmacists in Nebraska regarding children's learning disabilities were surveyed by questionnaire. One hundred thirty-three pharmacists responded. The results were analyzed according to type of practice (community, chain and hospital) and age (20--30 years, 31--40 years, 41--50 years, and 51 years and over). The results indicate that pharmacists are being asked about learning disabilities. Overall, 53% of the pharmacists responding had been asked about learning disabilities by parents, physicians, teachers or others. No significant interactions were found between the type of practice and age, and no significant main effect for type of practice was found in the ANOVA results for knowledge scores or attitude scores. Even though there were no differences owing to age in pharmacists' knowledge of learning disabilities, age was found to have a significant effect on attitudes toward increased pharmacist involvement in monitoring behaviors of learning-disabled children; the younger groups had a more positive attitude. The pharmacist seems to have the knowledge and attitudes required to become more actively involved in monitoring the medications and behaviors of children with learning disabilities. | [
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PMID:24341 | Development of neurohumoral control of fetal, neonatal, and adult cardiovascular functions. | Neurohumoral control of fetal, neonatal, and adult cardiovascular functions have been reviewed. Resting fetal heart rate remains fairly constant but neonatal heart rate declines progressively, reaching adult levels within six to eight weeks; systemic arterial pressure rises while pulmonary pressure falls to adult levels within the first week after birth. Sympathetic and parasympathetic control of circulatory functions matures at different rates during fetal and neonatal development; the sympathetic system becomes active earlier in fetal life than does the parasympathetic system. After birth, the parasympathetic tone of the resting heart rate rises to adult levels while adrenergic tone decreases. Despite changing autonomic activities, resting heart rate is set at given levels through alterations in intrinsic control. In the fetus, peripheral circulation is under neurohumoral tone of increasing magnitude; after birth, neurohumoral tone declines progressively, reaching levels comparable to those of adult nonpregnant sheep. Fetal cardiovascular response to neurotransmitters increases with age because of maturation of the effector system. The pulmonary bed responds primarily to acetylcholine whereas the systemic circulation responds to norepinephrine. After birth, the neonatal cardiovascular system becomes four to five times more sensitive to the action of neurotransmitters mainly because of closure of vascular shunts and elimination of umbilicoplacental circulation. In the neonate and adult, the pulmonary vascular bed loses its reactivity to neurotransmitters. | Development of neurohumoral control of fetal, neonatal, and adult cardiovascular functions. Neurohumoral control of fetal, neonatal, and adult cardiovascular functions have been reviewed. Resting fetal heart rate remains fairly constant but neonatal heart rate declines progressively, reaching adult levels within six to eight weeks; systemic arterial pressure rises while pulmonary pressure falls to adult levels within the first week after birth. Sympathetic and parasympathetic control of circulatory functions matures at different rates during fetal and neonatal development; the sympathetic system becomes active earlier in fetal life than does the parasympathetic system. After birth, the parasympathetic tone of the resting heart rate rises to adult levels while adrenergic tone decreases. Despite changing autonomic activities, resting heart rate is set at given levels through alterations in intrinsic control. In the fetus, peripheral circulation is under neurohumoral tone of increasing magnitude; after birth, neurohumoral tone declines progressively, reaching levels comparable to those of adult nonpregnant sheep. Fetal cardiovascular response to neurotransmitters increases with age because of maturation of the effector system. The pulmonary bed responds primarily to acetylcholine whereas the systemic circulation responds to norepinephrine. After birth, the neonatal cardiovascular system becomes four to five times more sensitive to the action of neurotransmitters mainly because of closure of vascular shunts and elimination of umbilicoplacental circulation. In the neonate and adult, the pulmonary vascular bed loses its reactivity to neurotransmitters. | [
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PMID:24342 | Immune complexes of hepatitis B surface antigen in the pathogenesis of periarteritis nodosa. A study of seven necropsy cases. | In 7 unselected necropsy cases of clinically diagnosed periarteritis nodosa, the detection of hepatitis B surface antigen (HBsAg) and hepatitis B core antigen (HBcAg) in the cytoplasm and nuclei of hepatocytes indicated an ongoing infection with hepititis B virus (HBV). In all these cases histologic changes found in the liver varied from "minimal" to chronic aggressive hepatitis. In all the cases, deposits of HBsAg, immunoglobulins, beta1C-globulin and C1q were detected in vascular lesions. That these deposits could represent HBsAg-anti-HBs immune complexes was supported by demonstrating their strong binding of guinea pig complement and by the successful elution of all HBsAg and part of the immunoglobulin from these deposits by treatment with buffers known to dissociate antigen-antibody bonds but not with phosphate-buffered saline, pH 7.6 (PBS). Glomerulonephritis associated with these immune complexes was found in 6 cases. The presence of larger masses of HBsAg immune complexes, chiefly in recent insudative and fibrinoid vascular lesions, their lesser amounts in lesions undergoing involution, and their absence from healed lesions strongly suggest that these complexes play a primary role in the pathogenesis of acute vascular damage in periarteritis nodosa. | Immune complexes of hepatitis B surface antigen in the pathogenesis of periarteritis nodosa. A study of seven necropsy cases. In 7 unselected necropsy cases of clinically diagnosed periarteritis nodosa, the detection of hepatitis B surface antigen (HBsAg) and hepatitis B core antigen (HBcAg) in the cytoplasm and nuclei of hepatocytes indicated an ongoing infection with hepititis B virus (HBV). In all these cases histologic changes found in the liver varied from "minimal" to chronic aggressive hepatitis. In all the cases, deposits of HBsAg, immunoglobulins, beta1C-globulin and C1q were detected in vascular lesions. That these deposits could represent HBsAg-anti-HBs immune complexes was supported by demonstrating their strong binding of guinea pig complement and by the successful elution of all HBsAg and part of the immunoglobulin from these deposits by treatment with buffers known to dissociate antigen-antibody bonds but not with phosphate-buffered saline, pH 7.6 (PBS). Glomerulonephritis associated with these immune complexes was found in 6 cases. The presence of larger masses of HBsAg immune complexes, chiefly in recent insudative and fibrinoid vascular lesions, their lesser amounts in lesions undergoing involution, and their absence from healed lesions strongly suggest that these complexes play a primary role in the pathogenesis of acute vascular damage in periarteritis nodosa. | [
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PMID:24343 | Tumor induction in host-versus-graft disease. I. Clinical and pathologic features. | Perinatal C57BL/1 mice given injections of (SJL/J X C57BL/1)F1 spleen cells developed a highly lethal runting syndrome, designated host-versus-graft disease (HVGD). The mortality was related to the dosage of F1 cells. Acute pathologic changes resembled those occurring in parent leads to F1 graft-versus-host disease (GVHD), except for more pronounced plasmacytosis. Mice suffering from HVGD recovered clinically with no sequelae except for a slight increase in the incidence of lymphomas over control mice. Such mice were hyperreactive to F1 cells utilized to initiate the original HVGD syndrome. Most of the tumors developed in those animals receiving the initial injection of F1 spleen cells within 24 hours of birth. Tumor incidence was unrelated to the clinical severity of HVGD. By contrast, GVHD in the same strain combination resulted in a much higher incidence of lymphomas in a much shorter time. Parental strain cells were detectable in the F1 hosts up to the time of tumor development. HVGD has a low tumor induction potential; GVHD has a high tumor induction potential. | Tumor induction in host-versus-graft disease. I. Clinical and pathologic features. Perinatal C57BL/1 mice given injections of (SJL/J X C57BL/1)F1 spleen cells developed a highly lethal runting syndrome, designated host-versus-graft disease (HVGD). The mortality was related to the dosage of F1 cells. Acute pathologic changes resembled those occurring in parent leads to F1 graft-versus-host disease (GVHD), except for more pronounced plasmacytosis. Mice suffering from HVGD recovered clinically with no sequelae except for a slight increase in the incidence of lymphomas over control mice. Such mice were hyperreactive to F1 cells utilized to initiate the original HVGD syndrome. Most of the tumors developed in those animals receiving the initial injection of F1 spleen cells within 24 hours of birth. Tumor incidence was unrelated to the clinical severity of HVGD. By contrast, GVHD in the same strain combination resulted in a much higher incidence of lymphomas in a much shorter time. Parental strain cells were detectable in the F1 hosts up to the time of tumor development. HVGD has a low tumor induction potential; GVHD has a high tumor induction potential. | [
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PMID:24345 | Association of increased pHi with calcium ion release in skeletal muscle. | The pH difference across the cell membrane of frog sartorius muscle cells was measured with the distribution of 5,5-dimethyl-2,4-oxazolidine-dione (DMO) as the marker. Depolarization of the muscles to values at or below the contraction threshold caused by elevating external potassium up to approximately 20 mM resulted in an internal alkalinization. The change was smaller with superthreshold depolarization (20--30 mM [K+]). The alkalinization was blocked by agents that block calcium release from the sarcoplasmic reticulum (procaine and dantrolene sodium). Other agents that cause calcium release (caffeine, theophylline, and quinine) were found to give alkalinization when tested at concentrations just below the contracture threshold. Increased acidification of the extracellular medium was associated with the internal alkalinization. The data were interpreted as indicating the presence of a calcium-stimulated H+ and/or OH- ion transport system in the muscle membrane. | Association of increased pHi with calcium ion release in skeletal muscle. The pH difference across the cell membrane of frog sartorius muscle cells was measured with the distribution of 5,5-dimethyl-2,4-oxazolidine-dione (DMO) as the marker. Depolarization of the muscles to values at or below the contraction threshold caused by elevating external potassium up to approximately 20 mM resulted in an internal alkalinization. The change was smaller with superthreshold depolarization (20--30 mM [K+]). The alkalinization was blocked by agents that block calcium release from the sarcoplasmic reticulum (procaine and dantrolene sodium). Other agents that cause calcium release (caffeine, theophylline, and quinine) were found to give alkalinization when tested at concentrations just below the contracture threshold. Increased acidification of the extracellular medium was associated with the internal alkalinization. The data were interpreted as indicating the presence of a calcium-stimulated H+ and/or OH- ion transport system in the muscle membrane. | [
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PMID:24346 | Na+ transport by mammalian stomach. | Gastric mucosas from newborn pigs (0--20 days) and rabbits (0--20 days) were used for in vitro investigation of active Na+ transport during resting (no HCl secretion) conditions. As measured with 22Na+, these tissues actively absorb Na+ from the mucosal-to serosal (m-t-s) bathing solution during both open-circuit and short-circuit current (Is) conditions. In the nonsecreting state, net Na+ transport accounts for 40--60% of Isc. The remaining current is provided by net s-to-m flux of Cl-. Amiloride (2-5 X 10(-5) M) in the mucosal solution abolishes this active Na+ transport by inhibiting m-to-s fluxes of Na+ (JNams). In vivo-in vitro experiments showed that active Na+ transport is a normal function of the resting mammalian stomach. Decreasing pH of the mucosal solution below pH 5 reversibly causes decreased current-generating capability of the tissue. Pretreatment of the tissue with amiloride abolishes this pH effect. The implication is that the low pH affects the Na+-entry step into cells. "Titration curves" of current vs. pH had an apparent pK approximately 4.0. Ouabain and K+-free solutions both cause decreases in active Na+ and Cl- current. Calculations indicate that a shunt may account for only a small (less than 30%) percentage of total transepithelial conductance. | Na+ transport by mammalian stomach. Gastric mucosas from newborn pigs (0--20 days) and rabbits (0--20 days) were used for in vitro investigation of active Na+ transport during resting (no HCl secretion) conditions. As measured with 22Na+, these tissues actively absorb Na+ from the mucosal-to serosal (m-t-s) bathing solution during both open-circuit and short-circuit current (Is) conditions. In the nonsecreting state, net Na+ transport accounts for 40--60% of Isc. The remaining current is provided by net s-to-m flux of Cl-. Amiloride (2-5 X 10(-5) M) in the mucosal solution abolishes this active Na+ transport by inhibiting m-to-s fluxes of Na+ (JNams). In vivo-in vitro experiments showed that active Na+ transport is a normal function of the resting mammalian stomach. Decreasing pH of the mucosal solution below pH 5 reversibly causes decreased current-generating capability of the tissue. Pretreatment of the tissue with amiloride abolishes this pH effect. The implication is that the low pH affects the Na+-entry step into cells. "Titration curves" of current vs. pH had an apparent pK approximately 4.0. Ouabain and K+-free solutions both cause decreases in active Na+ and Cl- current. Calculations indicate that a shunt may account for only a small (less than 30%) percentage of total transepithelial conductance. | [
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PMID:24347 | Norepinephrine: hormone and neurotransmitter in man. | To determine whether norepinephrine could subserve a hormonal as well as a neurotransmitter function, norepinephrine was infused for 60 min into each of five normal young men in doses of 0.1, 0.5, 1.0, 2.5, and 5.0 microgram/min. After infusion, the plasma norepinephrine concentration fell with a mean (+/-SD) half-time of 2.4 +/- 0.7 min. The mean (+/-SD) norepinephrine metabolic clearance rate was 3,070 +/- 200 ml/min. The calculated basal plasma norepinephrine production rate was 0.7 microgram/min. The blood pressure and circulating glycerol, acetoacetate, beta-hydroxybutyrate, and glucose (increased) and the heart rate and circulating insulin, lactate, pyruvate, and alanine (decreased) exhibited highly significant parabolic relationships with the steady-state plasma norepinephrine concentrations. However, norepinephrine levels in excess of 1,800 pg/ml were required to produce hemodynamic and/or metabolic effects. Thus, under usual conditions, the biologic actions of norepinephrine can be attributed only to its sympathetic neurotransmitter function. Plasma norepinephrine concentrations do at times exceed 1,800 pg/ml during exercise and during major acute illness. Thus, under conditions of stress, norepinephrine may subserve a hormonal, as well as a neurotransmitter, function. | Norepinephrine: hormone and neurotransmitter in man. To determine whether norepinephrine could subserve a hormonal as well as a neurotransmitter function, norepinephrine was infused for 60 min into each of five normal young men in doses of 0.1, 0.5, 1.0, 2.5, and 5.0 microgram/min. After infusion, the plasma norepinephrine concentration fell with a mean (+/-SD) half-time of 2.4 +/- 0.7 min. The mean (+/-SD) norepinephrine metabolic clearance rate was 3,070 +/- 200 ml/min. The calculated basal plasma norepinephrine production rate was 0.7 microgram/min. The blood pressure and circulating glycerol, acetoacetate, beta-hydroxybutyrate, and glucose (increased) and the heart rate and circulating insulin, lactate, pyruvate, and alanine (decreased) exhibited highly significant parabolic relationships with the steady-state plasma norepinephrine concentrations. However, norepinephrine levels in excess of 1,800 pg/ml were required to produce hemodynamic and/or metabolic effects. Thus, under usual conditions, the biologic actions of norepinephrine can be attributed only to its sympathetic neurotransmitter function. Plasma norepinephrine concentrations do at times exceed 1,800 pg/ml during exercise and during major acute illness. Thus, under conditions of stress, norepinephrine may subserve a hormonal, as well as a neurotransmitter, function. | [
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PMID:24348 | Acetazolamide and renal ammoniagenesis. | The effect of acetazolamide on ammonia-producing enzyme systems was determined in vitro at concentrations comparable to those which have been shown to abolish ammonium excretion in vivo. No change in the activity of glutaminase or gamma-glutamyl transpeptidase could be observed at concentrations up to 0.2 mM acetazolamide, and concentrations up to 1 mM were without effect on D-glutamyltransferase activity. Therefore, the effect of acetazolamide to abolish ammonium excretion cannot be explained by an action of the drug to inhibit ammoniagenesis. | Acetazolamide and renal ammoniagenesis. The effect of acetazolamide on ammonia-producing enzyme systems was determined in vitro at concentrations comparable to those which have been shown to abolish ammonium excretion in vivo. No change in the activity of glutaminase or gamma-glutamyl transpeptidase could be observed at concentrations up to 0.2 mM acetazolamide, and concentrations up to 1 mM were without effect on D-glutamyltransferase activity. Therefore, the effect of acetazolamide to abolish ammonium excretion cannot be explained by an action of the drug to inhibit ammoniagenesis. | [
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PMID:24349 | Cerebral blood flow in birds: effect of hypoxia. | The effect of hypoxia on cerebral blood flow in ducks was investigated by the rate at which arterially injected xenon-133 was cleared from the duck's brain. A two-component clearance curve resulted, which we have attributed to flow through the grey and white matter. Decreasing the arterial oxygen tension (PaO2) to 75 mmHg had no effect on cerebral blood flow. However, decreasing the PaO2 below 75 mmHg significantly increased blood flow to the fast-clearing compartment. The greatest increase in blood flow was seen when the arterial PO2 was below 50 mmHg. At an arterial PO2 of 30 mmHg, the cerebral blood flow to the fast-clearing compartment was increased more than 600% above the normoxic level. The magnitude of this increase is much greater in the duck than has been reported for mammals at roughly equivalent arterial oxygen tensions. The ability of avian cerebral blood flow to increase at moderate levels of hypoxia, plus the magnitude of the increase, may play a role in the exceptional tolerance of birds to hypoxia. | Cerebral blood flow in birds: effect of hypoxia. The effect of hypoxia on cerebral blood flow in ducks was investigated by the rate at which arterially injected xenon-133 was cleared from the duck's brain. A two-component clearance curve resulted, which we have attributed to flow through the grey and white matter. Decreasing the arterial oxygen tension (PaO2) to 75 mmHg had no effect on cerebral blood flow. However, decreasing the PaO2 below 75 mmHg significantly increased blood flow to the fast-clearing compartment. The greatest increase in blood flow was seen when the arterial PO2 was below 50 mmHg. At an arterial PO2 of 30 mmHg, the cerebral blood flow to the fast-clearing compartment was increased more than 600% above the normoxic level. The magnitude of this increase is much greater in the duck than has been reported for mammals at roughly equivalent arterial oxygen tensions. The ability of avian cerebral blood flow to increase at moderate levels of hypoxia, plus the magnitude of the increase, may play a role in the exceptional tolerance of birds to hypoxia. | [
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PMID:24350 | Altered hemodynamic responses to acute hypoxemia in spontaneously hypertensive rats. | Conscious spontaneously hypertensive rats (SHR), 5--7 wk old, were studied hemodynamically by the direct Fick procedure to determine whether high total peripheral resistance (TPR) coexisted with increased oxygen consumption (QO2) at an early stage of hypertension development. Since under resting conditions cardiac output in SHR was not significantly different from normotensive controls, the elevated arterial pressure and QO2 were associated with increased TPR. Arterial hypoxemia was induced to reduce oxygen availability and to assess whether increased TPR in SHR could be reversed by this procedure. During hypoxemia, normotensive controls (WKY) responded with increased cardiac output and decreased arterial pressure and TPR. In contrast, arterial pressure and cardiac output fell in SHR; and the increased TPR persisted. QO2 fell in hypoxemic SHR demonstrating that the relationship between total body oxygen consumption and cardiac output was abnormal in young SHR, and that increased TPR in SHR was not dependent on resting levels of QO2 or oxygen availability. Although QO2 was elevated in SHR compared to age-matched WKY, this condition was not essential for maintained elevated vascular resistance. | Altered hemodynamic responses to acute hypoxemia in spontaneously hypertensive rats. Conscious spontaneously hypertensive rats (SHR), 5--7 wk old, were studied hemodynamically by the direct Fick procedure to determine whether high total peripheral resistance (TPR) coexisted with increased oxygen consumption (QO2) at an early stage of hypertension development. Since under resting conditions cardiac output in SHR was not significantly different from normotensive controls, the elevated arterial pressure and QO2 were associated with increased TPR. Arterial hypoxemia was induced to reduce oxygen availability and to assess whether increased TPR in SHR could be reversed by this procedure. During hypoxemia, normotensive controls (WKY) responded with increased cardiac output and decreased arterial pressure and TPR. In contrast, arterial pressure and cardiac output fell in SHR; and the increased TPR persisted. QO2 fell in hypoxemic SHR demonstrating that the relationship between total body oxygen consumption and cardiac output was abnormal in young SHR, and that increased TPR in SHR was not dependent on resting levels of QO2 or oxygen availability. Although QO2 was elevated in SHR compared to age-matched WKY, this condition was not essential for maintained elevated vascular resistance. | [
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PMID:24351 | Controlling the highest lactate dehydrogenase activity known in nature. | In the shipjack, Euthynnus pelamis, white muscle appears to possess a powerful anaerobic capacity as well as a significant carbohydrate based aerobic potential. Lactate dehydrogenase occurs at higher activities than found thus far anywhere else in nature and clearly functions in anaerobic glycolysis. Alpha-glycerophosphate dehydrogenase also occurs in unusually high activities and appears to play a role in aerobic glycolysis. Regulation of these two reactions is accomplished by temperature, pH, and creatine phosphate levels. High temperature, low pH, and low creatine phosphate levels all appear to favor lactate dehydrogenase over alpha-glycerophosphate dehydrogenase; low temperature, high pH, and high creatine-phosphate levels, all expected during the quiescent state in this species, and when metabolism in aerobic, all favor alpha-glycerophosphate dehydrogenase activity. | Controlling the highest lactate dehydrogenase activity known in nature. In the shipjack, Euthynnus pelamis, white muscle appears to possess a powerful anaerobic capacity as well as a significant carbohydrate based aerobic potential. Lactate dehydrogenase occurs at higher activities than found thus far anywhere else in nature and clearly functions in anaerobic glycolysis. Alpha-glycerophosphate dehydrogenase also occurs in unusually high activities and appears to play a role in aerobic glycolysis. Regulation of these two reactions is accomplished by temperature, pH, and creatine phosphate levels. High temperature, low pH, and low creatine phosphate levels all appear to favor lactate dehydrogenase over alpha-glycerophosphate dehydrogenase; low temperature, high pH, and high creatine-phosphate levels, all expected during the quiescent state in this species, and when metabolism in aerobic, all favor alpha-glycerophosphate dehydrogenase activity. | [
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PMID:24353 | Gastric mucosal defense mechanisms: effects of salicylate and histamine. | Some of the recent concepts about the gastric mucosal defense mechanisms against damage by luminal acid and the effects of histamine and salicylate on these mechanisms are reviewed. The mucosal barrier to acid appears to consist of at least two physiologic components: a permeability mechanism and a metabolic mechanism related to cellular bicarbonate production as a result of acid secretion. In the absence of salicylate, histamine appears to exert some protection by affecting both mechanisms, but in the presence of salicylate, histamine's protective effect is limited to altering mucosal permeability. The actions of salicylate on the gastric mucosa are complex, related in part to the concentration of salicylate and the pH of the luminal fluid. The damaging effects of salicylate appear to be related more to the concentration of acid in the lumen than to the lipid solubility of the drug. Salicylate increases permeability regardless of pH; the increase is initially selective for cations and subsequently becomes nonselective, involving both cations and anions. Although both low and high concentrations of salicylate increase mucosal permeability to hydrogen ions, only high concentrations of salicylate affect cellular bicarbonate production. | Gastric mucosal defense mechanisms: effects of salicylate and histamine. Some of the recent concepts about the gastric mucosal defense mechanisms against damage by luminal acid and the effects of histamine and salicylate on these mechanisms are reviewed. The mucosal barrier to acid appears to consist of at least two physiologic components: a permeability mechanism and a metabolic mechanism related to cellular bicarbonate production as a result of acid secretion. In the absence of salicylate, histamine appears to exert some protection by affecting both mechanisms, but in the presence of salicylate, histamine's protective effect is limited to altering mucosal permeability. The actions of salicylate on the gastric mucosa are complex, related in part to the concentration of salicylate and the pH of the luminal fluid. The damaging effects of salicylate appear to be related more to the concentration of acid in the lumen than to the lipid solubility of the drug. Salicylate increases permeability regardless of pH; the increase is initially selective for cations and subsequently becomes nonselective, involving both cations and anions. Although both low and high concentrations of salicylate increase mucosal permeability to hydrogen ions, only high concentrations of salicylate affect cellular bicarbonate production. | [
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PMID:24354 | The neurohumoral response to trauma. | The bodily response to injury is an integrated and protective mechanism involving all systems, and hence it should be considered as a whole. The nervous system, especially the psychic center of the cerebral cortex, seems to initiate these responses. In the present study the neurohumoral response to trauma was assessed in experimental animals and clinical subjects. Immediately after trauma acetylcholine levels increased, suggesting enhanced activity of the central nervous system. This was followed by predominant activity of the sympathetic nervous system, as evidenced by increased catecholamine levels. During the later postoperative period, histamine levels also increased. There appears to be a good correlation between increased histamine content and fibroblastic proliferation in the traumatized area. These observations suggest that levels of the neurohumors increase in a definite sequence after trauma, facilitating the process of repair, regeneration, and normal convalescence. Therefore, trauma should be minimized during repair and the patient should receive proper sedation and rest during the pre- and postoperative periods, since each quantum of response depends upon the amount of sensory input that occurs at the time of trauma and in the post-traumatic period. | The neurohumoral response to trauma. The bodily response to injury is an integrated and protective mechanism involving all systems, and hence it should be considered as a whole. The nervous system, especially the psychic center of the cerebral cortex, seems to initiate these responses. In the present study the neurohumoral response to trauma was assessed in experimental animals and clinical subjects. Immediately after trauma acetylcholine levels increased, suggesting enhanced activity of the central nervous system. This was followed by predominant activity of the sympathetic nervous system, as evidenced by increased catecholamine levels. During the later postoperative period, histamine levels also increased. There appears to be a good correlation between increased histamine content and fibroblastic proliferation in the traumatized area. These observations suggest that levels of the neurohumors increase in a definite sequence after trauma, facilitating the process of repair, regeneration, and normal convalescence. Therefore, trauma should be minimized during repair and the patient should receive proper sedation and rest during the pre- and postoperative periods, since each quantum of response depends upon the amount of sensory input that occurs at the time of trauma and in the post-traumatic period. | [
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PMID:24356 | [Comparative investigations concerning the influence of pilocarpine and aceclidine on the glucose consumption of surviving ocular tissues (author's transl)]. | Conservative glaucoma therapy repeated during a whole lifetime raises the question of medicamentuous tolerance. Recently, cultivation of normal lens epithelia and of iris tissue has made possible determination of metabolic functions with and without application of different medicaments. Preceding experiments revealed a diminution in glucose consumption by these tissues after addition of pilocarpine. Comparative studies show 0.25 g/100 ml medium to be maximum concentration of aceclidine allowing cellular survival. With this dilution, the glucose metabolism of lens epithelia was much more sensitive to aceclidine than to pilocarpine. This was less true for iris tissue. Accordingly, the lens of an aceclidine-treated patient requires careful observation. | [Comparative investigations concerning the influence of pilocarpine and aceclidine on the glucose consumption of surviving ocular tissues (author's transl)]. Conservative glaucoma therapy repeated during a whole lifetime raises the question of medicamentuous tolerance. Recently, cultivation of normal lens epithelia and of iris tissue has made possible determination of metabolic functions with and without application of different medicaments. Preceding experiments revealed a diminution in glucose consumption by these tissues after addition of pilocarpine. Comparative studies show 0.25 g/100 ml medium to be maximum concentration of aceclidine allowing cellular survival. With this dilution, the glucose metabolism of lens epithelia was much more sensitive to aceclidine than to pilocarpine. This was less true for iris tissue. Accordingly, the lens of an aceclidine-treated patient requires careful observation. | [
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PMID:24357 | Plasma lorazepam levels. A study following single dose administration of 2 and 4 mg by different routes. | Plasma lorazepam levels were studied following single doses of 2 mg and 4 mg in patients and volunteers. There was a slightly more rapid uptake of drug following deep intramuscular injection than when taken by mouth. Plasma levels declined rapidly following intravenous injection (t1/2 of approximately 2-3 hours). Two hours after giving the drug the plasma levels were similar irrespective of the route of administration. The second phase decline was very slow and one third of the peak concentration was still in the blood at 24 hours. | Plasma lorazepam levels. A study following single dose administration of 2 and 4 mg by different routes. Plasma lorazepam levels were studied following single doses of 2 mg and 4 mg in patients and volunteers. There was a slightly more rapid uptake of drug following deep intramuscular injection than when taken by mouth. Plasma levels declined rapidly following intravenous injection (t1/2 of approximately 2-3 hours). Two hours after giving the drug the plasma levels were similar irrespective of the route of administration. The second phase decline was very slow and one third of the peak concentration was still in the blood at 24 hours. | [
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PMID:24376 | [Metabolism of glucides and polyols and disorders of their utilization]. | The first part of this report is devoted to a description of the metabolic pathways of the principle carbohydrates and polyols. The second part is concerned with the utilisation of these energy-providing substrates, when used in parenteral alimentation. Notions emerge of "total clearance" and of coefficient of utilisation in relation to the rate of administration for each of them. Finally, the third part deals with disturbances of carbolydate and polyol metabolism during stress and diabetes and the possiblilities of the development of hyperlactatemia according to the substances used or the pathological circumstances encountered. A table attempts to summarise the potential complications in comparison with all the substrates used in parenteral alimentation. | [Metabolism of glucides and polyols and disorders of their utilization]. The first part of this report is devoted to a description of the metabolic pathways of the principle carbohydrates and polyols. The second part is concerned with the utilisation of these energy-providing substrates, when used in parenteral alimentation. Notions emerge of "total clearance" and of coefficient of utilisation in relation to the rate of administration for each of them. Finally, the third part deals with disturbances of carbolydate and polyol metabolism during stress and diabetes and the possiblilities of the development of hyperlactatemia according to the substances used or the pathological circumstances encountered. A table attempts to summarise the potential complications in comparison with all the substrates used in parenteral alimentation. | [
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PMID:24377 | [Glucide intolerance and its pathogenic mechanisms during parenteral feeding]. | Hyperglycaemia during parenteral alimentation occurs either as a result of an error in the supplies provided or as a result of diminished carbohydrate tolerance. The circumstances surrounding the development of carbohydrate intolerance are essentially : severe infections, major catabolic states, renal insufficiency, extensive burns, pancreatic problems and diabetes. From a pathogenic standpoint, there are two dominant elements : disturbances in hepatic gluconeogenesis and changes in insulin secretion and in resistance to insulin. The physiopathology is dominated by the risk of hyperosmolarity. Hypoglycaemia occurs most frequently as the result of a manit fest error : too sudded interruption of carbohydrate supplies or two high dosage of exogenous insulin. | [Glucide intolerance and its pathogenic mechanisms during parenteral feeding]. Hyperglycaemia during parenteral alimentation occurs either as a result of an error in the supplies provided or as a result of diminished carbohydrate tolerance. The circumstances surrounding the development of carbohydrate intolerance are essentially : severe infections, major catabolic states, renal insufficiency, extensive burns, pancreatic problems and diabetes. From a pathogenic standpoint, there are two dominant elements : disturbances in hepatic gluconeogenesis and changes in insulin secretion and in resistance to insulin. The physiopathology is dominated by the risk of hyperosmolarity. Hypoglycaemia occurs most frequently as the result of a manit fest error : too sudded interruption of carbohydrate supplies or two high dosage of exogenous insulin. | [
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PMID:24375 | Neonatal gastric pH. | The pH of gastric juice, obtained 3 to 4 minutes after birth in 158 unselected neonates, varied between 7.5 and 8.5 in 8 meconium-containing specimens and ranged from 1.4 to 7.8 in 150 meconium-free samples. In mature infants of the latter group, pH was (1) significantly lower after vaginal delivery than after cesarean section; (2) tended to be lower after section preceded by labor than after elective section; and (3) was lowest after precipitate delivery. In premature infants, pH was above 7 regardless of mode of delivery. Three was no correlation between neonatal gastric pH and pH of simultaneously obtained maternal gastric juice, pH of amniotic fluid, pH of cord blood duration of rupture of membranes, birth weight, or Apgar score. It was concluded that the mature human fetus produces gastric acidity in response to stresses associated with labor and vaginal delivery. The possibility of a low gastric pH and the resultant pulmonary damage if aspirated must be considered in the initial care of thew newborn with poor muscle tone or reflex activity as well as in the anesthetic management of neonates undergoing emergency surgery. | Neonatal gastric pH. The pH of gastric juice, obtained 3 to 4 minutes after birth in 158 unselected neonates, varied between 7.5 and 8.5 in 8 meconium-containing specimens and ranged from 1.4 to 7.8 in 150 meconium-free samples. In mature infants of the latter group, pH was (1) significantly lower after vaginal delivery than after cesarean section; (2) tended to be lower after section preceded by labor than after elective section; and (3) was lowest after precipitate delivery. In premature infants, pH was above 7 regardless of mode of delivery. Three was no correlation between neonatal gastric pH and pH of simultaneously obtained maternal gastric juice, pH of amniotic fluid, pH of cord blood duration of rupture of membranes, birth weight, or Apgar score. It was concluded that the mature human fetus produces gastric acidity in response to stresses associated with labor and vaginal delivery. The possibility of a low gastric pH and the resultant pulmonary damage if aspirated must be considered in the initial care of thew newborn with poor muscle tone or reflex activity as well as in the anesthetic management of neonates undergoing emergency surgery. | [
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PMID:24380 | [Circulating lipids in prolonged postoperative parenteral feeding: their changes depending on the nature of energy intake]. | Repeated estimations of circulating total lipids and various fractions were made during the postoperative period in patients receiving parenteral nutrition for more than ten days. The patients were paired in relation to the underlying pathology and divided into two group : the first receiving energy supplements in the form of carbohydrates alone, whilst the second received part of this supply in the form of lipid emulsions (Trive 1000). Total lipids and various plasma fractions, with the exception of free fatty acids, increased progressively during the postoperative period, regardless of the inital values and independently of the lipid content of the nutrition fluids given. There was no significant difference between those patients who received lipid emulsions and those who did not. Free fatty acids remained at levels slightly greater than normal in the group which did not receive lipids. They were significantly higher in the group given lipid emulsions, though it was not possible to precisely define their orgin. | [Circulating lipids in prolonged postoperative parenteral feeding: their changes depending on the nature of energy intake]. Repeated estimations of circulating total lipids and various fractions were made during the postoperative period in patients receiving parenteral nutrition for more than ten days. The patients were paired in relation to the underlying pathology and divided into two group : the first receiving energy supplements in the form of carbohydrates alone, whilst the second received part of this supply in the form of lipid emulsions (Trive 1000). Total lipids and various plasma fractions, with the exception of free fatty acids, increased progressively during the postoperative period, regardless of the inital values and independently of the lipid content of the nutrition fluids given. There was no significant difference between those patients who received lipid emulsions and those who did not. Free fatty acids remained at levels slightly greater than normal in the group which did not receive lipids. They were significantly higher in the group given lipid emulsions, though it was not possible to precisely define their orgin. | [
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PMID:24381 | [Fat overload during intake of lipids]. | A review of the recent literature indicates that fat overload during the infusion of lipid emulsions remains rare, at least clinically. Nevertheless, two pictures may be seen, the physiopathology of which is discussed:--overload syndrome, with an association essentially of general and digestive signs, related to the accumulation of lipids in a certain number of parenchymatous tissues;--fat embolism. Problem may be seen at any time after administration of the solution is begun. Youth seems to predominate amongst the favourising factors. Prevention is possible if rules of caution concerning dose and the rate of administration are respected, combined with biological surveillance involving at laboratory studies at least once a week. | [Fat overload during intake of lipids]. A review of the recent literature indicates that fat overload during the infusion of lipid emulsions remains rare, at least clinically. Nevertheless, two pictures may be seen, the physiopathology of which is discussed:--overload syndrome, with an association essentially of general and digestive signs, related to the accumulation of lipids in a certain number of parenchymatous tissues;--fat embolism. Problem may be seen at any time after administration of the solution is begun. Youth seems to predominate amongst the favourising factors. Prevention is possible if rules of caution concerning dose and the rate of administration are respected, combined with biological surveillance involving at laboratory studies at least once a week. | [
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PMID:24382 | [Disorders of hemostasis during intake of lipids]. | Lipid emulsions based upon soya triglycerides and widely used in parenteral nutrition. Haematological surveillance of the patients, and in particular the study of coagulation, shows slight changes in platelet adhesiveness and a transient hypercoagulability state proportional to the concentration of the lipid emulsion. Nevertheless, these findings must be interpreted with the greatest caution, inasmuch as the laboratory tests involved are technically delicate. The results obtained may be contradictory according to the protocol used. Finally, in patients receiving intensive therapy, there are multiple factors (stress, prolonge;d bedrest, hypercatabolism) which may alter coagulation quite independently of the prescription of lipids. | [Disorders of hemostasis during intake of lipids]. Lipid emulsions based upon soya triglycerides and widely used in parenteral nutrition. Haematological surveillance of the patients, and in particular the study of coagulation, shows slight changes in platelet adhesiveness and a transient hypercoagulability state proportional to the concentration of the lipid emulsion. Nevertheless, these findings must be interpreted with the greatest caution, inasmuch as the laboratory tests involved are technically delicate. The results obtained may be contradictory according to the protocol used. Finally, in patients receiving intensive therapy, there are multiple factors (stress, prolonge;d bedrest, hypercatabolism) which may alter coagulation quite independently of the prescription of lipids. | [
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PMID:24383 | [Blood coagulation during prolonged postoperative parenteral feeding containing lipids]. | Blood coagulation was studied in two groups of fifteen patients of the same type during parenteral nutrition lasting for more then ten days and given during the postoperative period. The first batch of patients, taken as a control group, received parenteral nutrition with carbohydrates and proteins, whilst the second group received lipids in addition. Study of blood coagulation, carried out every three days, involved : thromboelastogen on whole blood and plasma, platelet count, platelet adhesiveness and the estimation of factors II, V, VII and X, fibrinogen, plasminogen and fibrinogen break-down products. In the patients not receiving lipids, the normal alterations seen during the postoperative period occurred : normalisation in plasminogen levels and in F.B.P., and an increase in the number of platelets and in fibrinogen level. The presence of lipids in the nutritional fluids resulted in changes in the same direction, but the increase in platelet count was slighter and fibrinogen level, after a slight increase, returned to initial values on the 20th day of parenteral nutrition. Fibrinogen was the only factor which showed any significant difference between the two groups. | [Blood coagulation during prolonged postoperative parenteral feeding containing lipids]. Blood coagulation was studied in two groups of fifteen patients of the same type during parenteral nutrition lasting for more then ten days and given during the postoperative period. The first batch of patients, taken as a control group, received parenteral nutrition with carbohydrates and proteins, whilst the second group received lipids in addition. Study of blood coagulation, carried out every three days, involved : thromboelastogen on whole blood and plasma, platelet count, platelet adhesiveness and the estimation of factors II, V, VII and X, fibrinogen, plasminogen and fibrinogen break-down products. In the patients not receiving lipids, the normal alterations seen during the postoperative period occurred : normalisation in plasminogen levels and in F.B.P., and an increase in the number of platelets and in fibrinogen level. The presence of lipids in the nutritional fluids resulted in changes in the same direction, but the increase in platelet count was slighter and fibrinogen level, after a slight increase, returned to initial values on the 20th day of parenteral nutrition. Fibrinogen was the only factor which showed any significant difference between the two groups. | [
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PMID:24384 | [Cholestatic icterus during parenteral feeding in the newborn ano infants]. | The authors observed six cases of cholestatis jaundice occuring during parenteral alimentation in the newborn and the infant. In 4 cases, local factors (duodenal atresia, necrosing enteritis of prematurity) or general factors (mucoviscidosis) probably played an essential role. In two cases, the parenteral alimentation seemed to be a main factor, but in one case with excessive and imbalanced supplies of amino acids. Three patients survived, with regression of the jaundice in 15 days to three weeks following the interruption of parenteral alimentation. | [Cholestatic icterus during parenteral feeding in the newborn ano infants]. The authors observed six cases of cholestatis jaundice occuring during parenteral alimentation in the newborn and the infant. In 4 cases, local factors (duodenal atresia, necrosing enteritis of prematurity) or general factors (mucoviscidosis) probably played an essential role. In two cases, the parenteral alimentation seemed to be a main factor, but in one case with excessive and imbalanced supplies of amino acids. Three patients survived, with regression of the jaundice in 15 days to three weeks following the interruption of parenteral alimentation. | [
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PMID:24385 | [Prevention of metabolic complications in prolonged parenteral feeding]. | Modern technics in parenteral nutrition can prevent metabolic and infectious complications. Supplying a patient with a daily ration according to his needs in the form of either hyper or normo-nutrition serves as the basis for this prevention. All that is required for success is a deep venous approach and regular biological and clinical surveillance of the patient. The authors conclusions are based on a study of 1.800 patients. | [Prevention of metabolic complications in prolonged parenteral feeding]. Modern technics in parenteral nutrition can prevent metabolic and infectious complications. Supplying a patient with a daily ration according to his needs in the form of either hyper or normo-nutrition serves as the basis for this prevention. All that is required for success is a deep venous approach and regular biological and clinical surveillance of the patient. The authors conclusions are based on a study of 1.800 patients. | [
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PMID:24386 | [Metabolic disorders in experimental hepatic insufficiency and parenteral feeding]. | Anhepatia is a complete experimental model for the study of metabolic disturbances associated with hepatic insufficiency. The authors report their experience based upon 16 cases of total hepatectomy in the dog. The animals received different types of hepatic assistance : group 1 (2 dogs), carbohydrate assistance (glucose 7 g/kg/24 h) ; group II (2 dogs), carbohydrate and haemostatic assistance : homologous plasma 50 ml/kg body weight per 24 hours previously stored at - 80 degrees C ; group III (2 dogs) same nutrition as in group II, combined with amino acids : leucine 53, 3 mg, isoleucine 46, 6 mg ; valine 53,3 mg alpha-ketonic acid (lg/kg/24 h) ; group IV (10 dogs, same nutrition as in group III, combined with supplements in neuro transmitter precursors : 6 dopa 1,66 mg, 5 hydroxy-tryptophane 2 mg and choline 16, 66 mg/kg/24h. Metabolic disturbances and their course were analysed in detail. Results : (mean survival) - group I : 24 +/- 4 hours; group II : 36 +/- 9 hours; group III : 42 +/- 4 hours; group IV : 90 +/- 9 hours. The average survival differed significantly for groups III and IV (p less than 0,01) in comparison with control groups I and II. The replacement of essential hepatic functions such as blood glucose regulation, coagulation, detoxication and the provision of neurotransmitter precursors made possible 4 days survival after total hepatectomy. | [Metabolic disorders in experimental hepatic insufficiency and parenteral feeding]. Anhepatia is a complete experimental model for the study of metabolic disturbances associated with hepatic insufficiency. The authors report their experience based upon 16 cases of total hepatectomy in the dog. The animals received different types of hepatic assistance : group 1 (2 dogs), carbohydrate assistance (glucose 7 g/kg/24 h) ; group II (2 dogs), carbohydrate and haemostatic assistance : homologous plasma 50 ml/kg body weight per 24 hours previously stored at - 80 degrees C ; group III (2 dogs) same nutrition as in group II, combined with amino acids : leucine 53, 3 mg, isoleucine 46, 6 mg ; valine 53,3 mg alpha-ketonic acid (lg/kg/24 h) ; group IV (10 dogs, same nutrition as in group III, combined with supplements in neuro transmitter precursors : 6 dopa 1,66 mg, 5 hydroxy-tryptophane 2 mg and choline 16, 66 mg/kg/24h. Metabolic disturbances and their course were analysed in detail. Results : (mean survival) - group I : 24 +/- 4 hours; group II : 36 +/- 9 hours; group III : 42 +/- 4 hours; group IV : 90 +/- 9 hours. The average survival differed significantly for groups III and IV (p less than 0,01) in comparison with control groups I and II. The replacement of essential hepatic functions such as blood glucose regulation, coagulation, detoxication and the provision of neurotransmitter precursors made possible 4 days survival after total hepatectomy. | [
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PMID:24387 | [Three cases of coma during parenteral alimentation]. | Three cases of coma occuring during parenteral alimentation are presented. The common factor was that they occurred during the phase of realimentation in patients with severely impaired nutrition. In two cases out of three, the coma marked the beginning of the absolute or relative anabolic period. They were studied clinically, biologically and electrically. In one case only, major hypophosphataemia was discovered (2mg/l). Of the three patients, one only died 4 days after the onset of coma, whilst clinical and biological characteristics had returned to normal. | [Three cases of coma during parenteral alimentation]. Three cases of coma occuring during parenteral alimentation are presented. The common factor was that they occurred during the phase of realimentation in patients with severely impaired nutrition. In two cases out of three, the coma marked the beginning of the absolute or relative anabolic period. They were studied clinically, biologically and electrically. In one case only, major hypophosphataemia was discovered (2mg/l). Of the three patients, one only died 4 days after the onset of coma, whilst clinical and biological characteristics had returned to normal. | [
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PMID:24388 | [Deficiencies in trace elements during parenteral alimentation]. | During recent years, techniques of parenteral alimentation have seen the development of complications due to deficiencies in trace elements. The main deficiency states described involve iron, copper and zinc. In addition to these three trace elements, manganèse and cobalt are included in this study. The metabolism, physiological role, needs in parenteral alimentation and clinical and metabolic effects of possible deficiency are described for each of these elements. | [Deficiencies in trace elements during parenteral alimentation]. During recent years, techniques of parenteral alimentation have seen the development of complications due to deficiencies in trace elements. The main deficiency states described involve iron, copper and zinc. In addition to these three trace elements, manganèse and cobalt are included in this study. The metabolism, physiological role, needs in parenteral alimentation and clinical and metabolic effects of possible deficiency are described for each of these elements. | [
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PMID:24389 | [Routine comparison of trace element deficiencies during parenteral alimentation]. | In 50 patients aged between 3 and 84 years treated in a multidisciplinary Intensive Care Unit and receiving parenteral alimentation, deficiency in certain trace elements or electrolytes (Cu++, Zn++, Mn++, Co++, PO-4, Mg++) was prevented or treated by the administration of a glucose solution (MB 147 G) enriched in trace elements. The aim of the present study was to demonstrate, on the basis of assay of serum levels of the trace elements involved, with the exception of Mn and Co, the effectiveness of treatment. Reference values were determined on the one hand in healthy individuals for normal figures and secondly on subjects included in the study, already on parenteral alimentation for several days, before treatment with MB 147 G, in order to demonstrate the existence of a deficiency (patient control values). In the case of PO--4, however, the patient control values concerned at one and the same time subjects in the study before treatment with MB 147 G and other patients receiving parenteral alimentation who were not part of the trial. MB 147 G solution was presented in units of 500 ml associated with glucose of varying concentrations (15 p. 100, 30 p. 100, 50 p. 100). The average daily amount administered, over a period of 236 days, was 3 unites per 24 hours, corresponding to an intake of copper of 3.78 mg, 3.90 mg of zinc, 0.20 mg of manganese, 0.24 mg of cobalt, 363 mg of magnesium, 240 mg of calcium and 15 mEz of phosphates. The results show that levels of copper, zinc, magnesium and phosphates were low during parenteral alimentation. The administration of MB 147 G resulted in a significant increase in these values, without there being any evidence of accumulation. | [Routine comparison of trace element deficiencies during parenteral alimentation]. In 50 patients aged between 3 and 84 years treated in a multidisciplinary Intensive Care Unit and receiving parenteral alimentation, deficiency in certain trace elements or electrolytes (Cu++, Zn++, Mn++, Co++, PO-4, Mg++) was prevented or treated by the administration of a glucose solution (MB 147 G) enriched in trace elements. The aim of the present study was to demonstrate, on the basis of assay of serum levels of the trace elements involved, with the exception of Mn and Co, the effectiveness of treatment. Reference values were determined on the one hand in healthy individuals for normal figures and secondly on subjects included in the study, already on parenteral alimentation for several days, before treatment with MB 147 G, in order to demonstrate the existence of a deficiency (patient control values). In the case of PO--4, however, the patient control values concerned at one and the same time subjects in the study before treatment with MB 147 G and other patients receiving parenteral alimentation who were not part of the trial. MB 147 G solution was presented in units of 500 ml associated with glucose of varying concentrations (15 p. 100, 30 p. 100, 50 p. 100). The average daily amount administered, over a period of 236 days, was 3 unites per 24 hours, corresponding to an intake of copper of 3.78 mg, 3.90 mg of zinc, 0.20 mg of manganese, 0.24 mg of cobalt, 363 mg of magnesium, 240 mg of calcium and 15 mEz of phosphates. The results show that levels of copper, zinc, magnesium and phosphates were low during parenteral alimentation. The administration of MB 147 G resulted in a significant increase in these values, without there being any evidence of accumulation. | [
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PMID:24390 | [Hydrogen ion metabolism and parenteral alimentation]. | The necessity of using solutions and emulsions in parenteral nutrition of solutions directly utilisable by the cell has led to the manufactur of substances with a composition and physico-chemical nature which greatly alters H+ ions balance. Theoretical calculation and various experimental protocois have shown that the production of H+ ions may be multiplied by 3 or 4. This explains the complications of metabolic acidosis seen in association with prolonged parenteral nutrition. There are two main sources : lipid emulsions and amino-acid solutions. One litre of lipid emulsion may provide 37 to 60 mEq of H+ ions. According to their formula, amino acid solutions may consume, or more often provide, large amounts of H+ ions (up to 20 mEq/gram of nitrogen). These physiopathological concepts should be taken into consideration in defining the compositions of fluids for parenteral nutrition, in particular when renal function is impaired. | [Hydrogen ion metabolism and parenteral alimentation]. The necessity of using solutions and emulsions in parenteral nutrition of solutions directly utilisable by the cell has led to the manufactur of substances with a composition and physico-chemical nature which greatly alters H+ ions balance. Theoretical calculation and various experimental protocois have shown that the production of H+ ions may be multiplied by 3 or 4. This explains the complications of metabolic acidosis seen in association with prolonged parenteral nutrition. There are two main sources : lipid emulsions and amino-acid solutions. One litre of lipid emulsion may provide 37 to 60 mEq of H+ ions. According to their formula, amino acid solutions may consume, or more often provide, large amounts of H+ ions (up to 20 mEq/gram of nitrogen). These physiopathological concepts should be taken into consideration in defining the compositions of fluids for parenteral nutrition, in particular when renal function is impaired. | [
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PMID:24391 | [Metabolic problems associated with ethanol administration]. | After reviewing the effects of ethanol on different metabolic processes, the author defines the doses of this alcohol which may be used in parenteral alimentation, its advantages and disadvantages. The latter in general outweigh the advantages and limit its use in parenteral nutrition. | [Metabolic problems associated with ethanol administration]. After reviewing the effects of ethanol on different metabolic processes, the author defines the doses of this alcohol which may be used in parenteral alimentation, its advantages and disadvantages. The latter in general outweigh the advantages and limit its use in parenteral nutrition. | [
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PMID:24392 | [Inherited abnormalities in amino acid metabolism]. | Inherited abnormalities in amino-acid metabolism, which up to a few years ago remained curiosities in the domain of the specialist, are now becoming almost of everyday significance. Improved knowledge, as a result of the combined efforts of paediatricians, genetecists and biochemists, has led to substantial progress, both clinically (diagnosis and treatment) as well as in the understanding of intermediary metabolisms and enzyme reactions. This "jigsay puzzle", the pieces of which fall gradually into place, has already resulted in spectacular results, the best example of which is that of phenylketonuria. In the past this disease inevitably led to increasing brain damage and has now become a purely biochemical disorder wince early diagnosis and appropriate diet enable the young infant to become a normal child and remain a healthy adult. These data are illustrated here. | [Inherited abnormalities in amino acid metabolism]. Inherited abnormalities in amino-acid metabolism, which up to a few years ago remained curiosities in the domain of the specialist, are now becoming almost of everyday significance. Improved knowledge, as a result of the combined efforts of paediatricians, genetecists and biochemists, has led to substantial progress, both clinically (diagnosis and treatment) as well as in the understanding of intermediary metabolisms and enzyme reactions. This "jigsay puzzle", the pieces of which fall gradually into place, has already resulted in spectacular results, the best example of which is that of phenylketonuria. In the past this disease inevitably led to increasing brain damage and has now become a purely biochemical disorder wince early diagnosis and appropriate diet enable the young infant to become a normal child and remain a healthy adult. These data are illustrated here. | [
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PMID:24393 | [Biological and hormonal balance during oral nutrition, exclusive perenteral and mixed nutrition in children]. | Exclusive parenteral nutrition and mixed feeding provided by the infusion of a solution of amino-acids, glucose and trace elements by a drip method, result in changes in biological and hormonal balance which have been compared with balance in the child on normal oral feeding. The 24-hour cortisol cycle tends to be attenuated. Blood glucose and, above all, blood insulin levels show major variations. Plasma free fatty acid levels, stable in exclusive parenteral nutrition, in general vary with mixed nutrition. On the basis of these findings, a constant flow should be ensured by use of a pump during exclusive parenteral nutrition. In mixed feeding, meals should be frequent and energy supplies given in the form of slowly absorbed carbohydrates. | [Biological and hormonal balance during oral nutrition, exclusive perenteral and mixed nutrition in children]. Exclusive parenteral nutrition and mixed feeding provided by the infusion of a solution of amino-acids, glucose and trace elements by a drip method, result in changes in biological and hormonal balance which have been compared with balance in the child on normal oral feeding. The 24-hour cortisol cycle tends to be attenuated. Blood glucose and, above all, blood insulin levels show major variations. Plasma free fatty acid levels, stable in exclusive parenteral nutrition, in general vary with mixed nutrition. On the basis of these findings, a constant flow should be ensured by use of a pump during exclusive parenteral nutrition. In mixed feeding, meals should be frequent and energy supplies given in the form of slowly absorbed carbohydrates. | [
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PMID:24394 | [Clinical signs and etiological aspects of metabolic encephalopathies excluding liver encephalopathy and realimentation syndrome]. | The clinical picture of metabolic encephalopathies has no aetiological specificity. It combines disturbances in conscious level dominated by disorientation and time and space and disturbances in motor activity, in particular tremor and asterixis. For each of the aetiologies studied, the following are considered: the circumstances of onset, the clinical and laboratory picture, the physiopathology and the treatment. From a diagnostic standpoint, particular emphasis should be placed upon the circumstances of onset which alone give any indication. The majority of these encephalopathies are caused by a lack of respect for simple rules in parenteral alimentation or by deficiencies. It is thus essentially an iatrogenic pathology. Treatment should be above all preventive. | [Clinical signs and etiological aspects of metabolic encephalopathies excluding liver encephalopathy and realimentation syndrome]. The clinical picture of metabolic encephalopathies has no aetiological specificity. It combines disturbances in conscious level dominated by disorientation and time and space and disturbances in motor activity, in particular tremor and asterixis. For each of the aetiologies studied, the following are considered: the circumstances of onset, the clinical and laboratory picture, the physiopathology and the treatment. From a diagnostic standpoint, particular emphasis should be placed upon the circumstances of onset which alone give any indication. The majority of these encephalopathies are caused by a lack of respect for simple rules in parenteral alimentation or by deficiencies. It is thus essentially an iatrogenic pathology. Treatment should be above all preventive. | [
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] |
PMID:24395 | [Amino acid disturbances in liver failure]. | We have attempted to trace the development of the concept which stresses that the encephalopathy often seen with hepatic failure may be the result of derangement in amino acid metabolism in patients with failing livers. We have also proposed that the infusion of solutions which may normalize these plasma amino acids and decrease the amount of toxic aromatic amino acids within the circulation may help in the amelioration of the symptoms of hepatic encephalopathy. We believe that in our studies to date, the use of this solution has been a significant advantage in the care of these extremely ill patients. We would hope that when the solution becomes generally available, the provision of such solutions to such patients will result in the salvage of some lives lost at the present time because of hepatic failure and encephalopathy. | [Amino acid disturbances in liver failure]. We have attempted to trace the development of the concept which stresses that the encephalopathy often seen with hepatic failure may be the result of derangement in amino acid metabolism in patients with failing livers. We have also proposed that the infusion of solutions which may normalize these plasma amino acids and decrease the amount of toxic aromatic amino acids within the circulation may help in the amelioration of the symptoms of hepatic encephalopathy. We believe that in our studies to date, the use of this solution has been a significant advantage in the care of these extremely ill patients. We would hope that when the solution becomes generally available, the provision of such solutions to such patients will result in the salvage of some lives lost at the present time because of hepatic failure and encephalopathy. | [
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PMID:24397 | [Comas of realimentation]. | Parenteral realimentation in the patient with severely impaired nutrition is sometimes associated with the development of a metabolic encephalopathy with coma. The onset of this complication is sudden. The coma is deep, accompanied by signs of neuromuscular hyperexcitability and very marked hyperventilation. It generally regresses rapidly, without sequellae. The onset of such a realimentation coma should be feared in the presence of a certain combination of conditions : severely impaired nutrition, parenteral alimentation with a high level of nitrogen and calories, transfer to the anabolic phase (perfect carbohydrate utilisation, fall in blood phosphate levels with hypophosphaturia, sometimes very marked positivisation of nitrogen balance) and, sometimes, mild premonitory clinical signs. The relationship between this type of complication and hypophosphoraemia is quite definite, but the fall in serum phosphates would not appear to be directly responsible for the coma. The exact mechanism is not known. In order to avoid such complications, great caution should be observed in both the quality as well as the quantity of the intake, as well as in the clinical and laboratory surveillance of the malnourished patient undergoing realimentation. | [Comas of realimentation]. Parenteral realimentation in the patient with severely impaired nutrition is sometimes associated with the development of a metabolic encephalopathy with coma. The onset of this complication is sudden. The coma is deep, accompanied by signs of neuromuscular hyperexcitability and very marked hyperventilation. It generally regresses rapidly, without sequellae. The onset of such a realimentation coma should be feared in the presence of a certain combination of conditions : severely impaired nutrition, parenteral alimentation with a high level of nitrogen and calories, transfer to the anabolic phase (perfect carbohydrate utilisation, fall in blood phosphate levels with hypophosphaturia, sometimes very marked positivisation of nitrogen balance) and, sometimes, mild premonitory clinical signs. The relationship between this type of complication and hypophosphoraemia is quite definite, but the fall in serum phosphates would not appear to be directly responsible for the coma. The exact mechanism is not known. In order to avoid such complications, great caution should be observed in both the quality as well as the quantity of the intake, as well as in the clinical and laboratory surveillance of the malnourished patient undergoing realimentation. | [
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PMID:24398 | [Electroencephalographic study of functional metabolic encephalopathies and comas during parenteral alimentation]. | This review deals with the common E.E.G. characteristics of dysmetabolic encephalopathies and described the particular features of hepatic, respiratory and renal encephalopathies, as well as those resulting from a disturbance in carbohydrate, water and electrolyte metabolism. These now classical data are compared with the principal electroclinical appearances seen during parenteral alimentation : slow, ample wases, non-reactive, associated with a calm coma; overall depression of basal rhythm, with excessive myogram activity and corresponding to paroxysms of muscular hypertonia seen during the coma. | [Electroencephalographic study of functional metabolic encephalopathies and comas during parenteral alimentation]. This review deals with the common E.E.G. characteristics of dysmetabolic encephalopathies and described the particular features of hepatic, respiratory and renal encephalopathies, as well as those resulting from a disturbance in carbohydrate, water and electrolyte metabolism. These now classical data are compared with the principal electroclinical appearances seen during parenteral alimentation : slow, ample wases, non-reactive, associated with a calm coma; overall depression of basal rhythm, with excessive myogram activity and corresponding to paroxysms of muscular hypertonia seen during the coma. | [
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] |
PMID:24399 | Effect of aflatoxin on the humoral and cell-mediated immune systems of the chicken. | Chickens fed 2.5 microgram of aflatoxin/g of diet from 2 to 4 weeks of age or from hatching to 4 weeks were deficient in cell-mediated immunity, as measured at 4 weeks of age by the graft-versus-host reaction. Delayed-type hypersensitive skin reactions to tuberculin were also reduced in chickens given dietary aflatoxin from hatching to 7 weeks of age. Humoral immunity, as measured by the ability of 4-week-old chicks to produce natural agglutinins to rabbit red blood cells, was not significantly altered by dietary aflatoxin. A significant decrease in concentrations of serum immunoglobulins (Ig) IgG and IgA, but no IgM, however, did occur in chicks given dietary aflatoxin from hatching to 4 weeks or between 2 and 4 weeks of age. Aflatoxin consumption from 0 to 2 weeks of age produced no marked effect on either cell-mediated or humoral immunity in 4-week-old chicks. | Effect of aflatoxin on the humoral and cell-mediated immune systems of the chicken. Chickens fed 2.5 microgram of aflatoxin/g of diet from 2 to 4 weeks of age or from hatching to 4 weeks were deficient in cell-mediated immunity, as measured at 4 weeks of age by the graft-versus-host reaction. Delayed-type hypersensitive skin reactions to tuberculin were also reduced in chickens given dietary aflatoxin from hatching to 7 weeks of age. Humoral immunity, as measured by the ability of 4-week-old chicks to produce natural agglutinins to rabbit red blood cells, was not significantly altered by dietary aflatoxin. A significant decrease in concentrations of serum immunoglobulins (Ig) IgG and IgA, but no IgM, however, did occur in chicks given dietary aflatoxin from hatching to 4 weeks or between 2 and 4 weeks of age. Aflatoxin consumption from 0 to 2 weeks of age produced no marked effect on either cell-mediated or humoral immunity in 4-week-old chicks. | [
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PMID:24401 | Less dialysis-induced morbidity and vascular instability with bicarbonate in dialysate. | We devised three protocols to test the postulate that increased morbidity during high-efficiency dialysis with large-surface-area units (LS) might be due in part to the increased flux of bicarbonate out and acetate into the patient inherent is LS dialysis. The first protocol showed that with LS-acetate dialysis there was a marked fall in plasma bicarbonate and Pco2 during the first 3 to 4 h, followed by a rapid rise in bicarbonate above normal and return to control in Pco2. With LS-bicarbonate dialysis, these oscillations were largely eliminated. A second double-blind protocol showed that central nervous system-type symptoms noted during and after LS-acetate dialysis were reduced significantly by switching to LS-bicarbonate dialysis. The third protocol showed that with LS-carbonate the tolerable rate of ultrafiltration could be increased 67% compared with LS-acetate dialysis. | Less dialysis-induced morbidity and vascular instability with bicarbonate in dialysate. We devised three protocols to test the postulate that increased morbidity during high-efficiency dialysis with large-surface-area units (LS) might be due in part to the increased flux of bicarbonate out and acetate into the patient inherent is LS dialysis. The first protocol showed that with LS-acetate dialysis there was a marked fall in plasma bicarbonate and Pco2 during the first 3 to 4 h, followed by a rapid rise in bicarbonate above normal and return to control in Pco2. With LS-bicarbonate dialysis, these oscillations were largely eliminated. A second double-blind protocol showed that central nervous system-type symptoms noted during and after LS-acetate dialysis were reduced significantly by switching to LS-bicarbonate dialysis. The third protocol showed that with LS-carbonate the tolerable rate of ultrafiltration could be increased 67% compared with LS-acetate dialysis. | [
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PMID:24402 | Metabolic acidosis after hyperalimentation with casein hydrolysate. Occurrence in a starved patient. | A 29-year-old woman with short bowel syndrome and prolonged starvation developed hyperchloremic metabolic acidosis after initiation of hyoeralimentation with a casein hydrolysate solution. The acidosis was not due to bicarbonate loss but was associated with diminished ability of the kidney to increase urinary acid excretion, particularly titratable acidity. Supplemental parenteral bicarbonate administration was necessary for two weeks until urinary acid excretion rose to normal. | Metabolic acidosis after hyperalimentation with casein hydrolysate. Occurrence in a starved patient. A 29-year-old woman with short bowel syndrome and prolonged starvation developed hyperchloremic metabolic acidosis after initiation of hyoeralimentation with a casein hydrolysate solution. The acidosis was not due to bicarbonate loss but was associated with diminished ability of the kidney to increase urinary acid excretion, particularly titratable acidity. Supplemental parenteral bicarbonate administration was necessary for two weeks until urinary acid excretion rose to normal. | [
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PMID:24403 | Costs of educating child health associates. | Child health associates can be fully prepared to competently diagnose and treat 90% of ambulatory pediatric patients five years after completion of high school at a cost of approximately one-fourth the expense of educating a pediatrician to perform the same tasks and functions. A rational health planning policy should include the extensive utilization of child health associates as primary health care providers for children. | Costs of educating child health associates. Child health associates can be fully prepared to competently diagnose and treat 90% of ambulatory pediatric patients five years after completion of high school at a cost of approximately one-fourth the expense of educating a pediatrician to perform the same tasks and functions. A rational health planning policy should include the extensive utilization of child health associates as primary health care providers for children. | [
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PMID:24405 | A chemiluminescent effect encountered in the liquid scintillation counting of 32P phosphate. | During the course of a kinetic enzyme study of bisphosphoglyceromutase (EC 2.7.5.4.) a chemiluminescent effect was encountered in the liquid scintillation counting of the beta-emission from 32Pi. A method is described for overcoming the effect, thereby permitting the accurate determination of the radioactive compounds. It is important to check for chemiluminescent effects early in the development of new procedures that involve liquid scintillation counting. | A chemiluminescent effect encountered in the liquid scintillation counting of 32P phosphate. During the course of a kinetic enzyme study of bisphosphoglyceromutase (EC 2.7.5.4.) a chemiluminescent effect was encountered in the liquid scintillation counting of the beta-emission from 32Pi. A method is described for overcoming the effect, thereby permitting the accurate determination of the radioactive compounds. It is important to check for chemiluminescent effects early in the development of new procedures that involve liquid scintillation counting. | [
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PMID:24406 | The origin of the acidosis in hyperlactataemia. | Anaerobic glycolysis produces lactate, ATP, and water but there is no net change in the number of hydrogen ions: it does not produce lactic acid. The acidosis usually associated with hyperlactataemia is caused by hydrolysis of the ATP, with release of hydrogen ions. By contrast, ATP turnover by aerobic mechanisms is not acidifying because the released hydrogen ion is reutilised as more ATP is formed. Gluconeogenesis from lactate does not utilise hydrogen ions directly--in fact, it produces them. The associated net H+ utilisation is caused by the aerobic generation of the ATP and GTP required to drive glycolysis in reverse. It is suggested that only by understanding these important biochemical facts can the clinician found his diagnosis and treatment on a firm, rational basis. | The origin of the acidosis in hyperlactataemia. Anaerobic glycolysis produces lactate, ATP, and water but there is no net change in the number of hydrogen ions: it does not produce lactic acid. The acidosis usually associated with hyperlactataemia is caused by hydrolysis of the ATP, with release of hydrogen ions. By contrast, ATP turnover by aerobic mechanisms is not acidifying because the released hydrogen ion is reutilised as more ATP is formed. Gluconeogenesis from lactate does not utilise hydrogen ions directly--in fact, it produces them. The associated net H+ utilisation is caused by the aerobic generation of the ATP and GTP required to drive glycolysis in reverse. It is suggested that only by understanding these important biochemical facts can the clinician found his diagnosis and treatment on a firm, rational basis. | [
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PMID:24408 | [Study of E. coli penicillin amidase. The pH dependence of the equilibrium constant of ampicillin enzymatic hydrolysis]. | The equilibrium parameters of the hydrolysis of ampicillin catalysed by penicillin amidase were determined within the pH range of 4.5 to 5.5. The values of the ionization constants of the carboxy group of D-(-)-ALPHA-AMINOPHENYLACETIC ACID (PK1=1.80) and amino group of 6-aminopenicillanic acid (pK2=4.60) were estimated and pH-dependence of the effective free energy of ampicillin hydrolysis was calculated. It was shown that the thermodynamic optimum of ampicillin synthesis was at 3.20 (the value of the effective free energy under the experimental conditions was 3.27 kcal/mole). The value of the "true", pH-independent free energy of hydrolysis (deltasigma) of the amide bond in the ampicillin molecule was determined to be equal to 9.72 kcal/mole. The thermodynamic parameters of ampicillin and benzylpenicillin hydrolysis were compared. The amino group in the alpha-position of phenylacetic acid was shown to have a significant effect on the values of "true" free energy of hydrolysis of the penicillin amide bond and free ionization energy in the system. | [Study of E. coli penicillin amidase. The pH dependence of the equilibrium constant of ampicillin enzymatic hydrolysis]. The equilibrium parameters of the hydrolysis of ampicillin catalysed by penicillin amidase were determined within the pH range of 4.5 to 5.5. The values of the ionization constants of the carboxy group of D-(-)-ALPHA-AMINOPHENYLACETIC ACID (PK1=1.80) and amino group of 6-aminopenicillanic acid (pK2=4.60) were estimated and pH-dependence of the effective free energy of ampicillin hydrolysis was calculated. It was shown that the thermodynamic optimum of ampicillin synthesis was at 3.20 (the value of the effective free energy under the experimental conditions was 3.27 kcal/mole). The value of the "true", pH-independent free energy of hydrolysis (deltasigma) of the amide bond in the ampicillin molecule was determined to be equal to 9.72 kcal/mole. The thermodynamic parameters of ampicillin and benzylpenicillin hydrolysis were compared. The amino group in the alpha-position of phenylacetic acid was shown to have a significant effect on the values of "true" free energy of hydrolysis of the penicillin amide bond and free ionization energy in the system. | [
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PMID:24409 | [Use of the penicillanic acid formation reaction for the analysis of penicillins. The quantitative determination of oxacillin]. | A procedure for quantitative determination of oxacillin by means of spectrophotometry is described. The procedure is based on the capacity of penicillins to transform on heating in acid buffer solutions containing copper ions into respective penicillenic acids having specific absorption characteristics in the UV-spectrum region. The amount of oxacillin was estimated by the optical density of the solutions at a wave length of 335 nm corresponding to the absorption maximum of the penicillenic acid of oxacillin. The optimal conditions for the reaction of the penicillenic acid formation, i.e. pH of the buffer solution and the level of the copper ions in it, time and temperature of heating were determined. The data on the comparison of the specificity and reproducibility of the new method with the currently used ones for determination of oxacillin are presented. | [Use of the penicillanic acid formation reaction for the analysis of penicillins. The quantitative determination of oxacillin]. A procedure for quantitative determination of oxacillin by means of spectrophotometry is described. The procedure is based on the capacity of penicillins to transform on heating in acid buffer solutions containing copper ions into respective penicillenic acids having specific absorption characteristics in the UV-spectrum region. The amount of oxacillin was estimated by the optical density of the solutions at a wave length of 335 nm corresponding to the absorption maximum of the penicillenic acid of oxacillin. The optimal conditions for the reaction of the penicillenic acid formation, i.e. pH of the buffer solution and the level of the copper ions in it, time and temperature of heating were determined. The data on the comparison of the specificity and reproducibility of the new method with the currently used ones for determination of oxacillin are presented. | [
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PMID:24410 | Stability and blood level determinations of cefaclor, a new oral cephalosporin antibiotic. | Cefaclor solutions in pH 2.5 and 4.5 buffers contained at least 90% of their initial activity after 72 h at 4 degrees C. Samples in pH 6.0, 7.0, and 8.0 buffers contained 70, 46, and 34%, respectively, of their initial activity after 72 h at 4 degrees C. After 72 h at 25 degrees C, samples prepared with pH 2.5, 4.5, 6.0, 7.0, and 8.0 buffers contained 95, 69, 16, 5, and 3%, respectively, of their initial activity. After 72 h at 37 degrees C, cefaclor solutions in pH 2.5 buffer contained 80% of the initial activity, whereas samples prepared in pH 4.5, 6.0, 7.0, and 8.0 buffers contained less than 20%. Laboratory-prepared plasma and serum samples showed an 8% loss in activity when incubated for 6 h at 4 degrees C, a 51% loss when incubated for 6 h at 25 degrees C, and a 48% loss when incubated for 2 h at 37 degrees C. Clinical samples demonstrated a similar stability pattern. Degradation rates for cefaclor in commercially prepared serum increased from 4- to 10-fold in comparison to rates obtained when samples were made in human serum freshly prepared in our laboratory. Consequently, serum standards should be made in freshly prepared human serum. | Stability and blood level determinations of cefaclor, a new oral cephalosporin antibiotic. Cefaclor solutions in pH 2.5 and 4.5 buffers contained at least 90% of their initial activity after 72 h at 4 degrees C. Samples in pH 6.0, 7.0, and 8.0 buffers contained 70, 46, and 34%, respectively, of their initial activity after 72 h at 4 degrees C. After 72 h at 25 degrees C, samples prepared with pH 2.5, 4.5, 6.0, 7.0, and 8.0 buffers contained 95, 69, 16, 5, and 3%, respectively, of their initial activity. After 72 h at 37 degrees C, cefaclor solutions in pH 2.5 buffer contained 80% of the initial activity, whereas samples prepared in pH 4.5, 6.0, 7.0, and 8.0 buffers contained less than 20%. Laboratory-prepared plasma and serum samples showed an 8% loss in activity when incubated for 6 h at 4 degrees C, a 51% loss when incubated for 6 h at 25 degrees C, and a 48% loss when incubated for 2 h at 37 degrees C. Clinical samples demonstrated a similar stability pattern. Degradation rates for cefaclor in commercially prepared serum increased from 4- to 10-fold in comparison to rates obtained when samples were made in human serum freshly prepared in our laboratory. Consequently, serum standards should be made in freshly prepared human serum. | [
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PMID:24425 | Mal perforans and spina bifida occulta. | A 26-year-old man had mal perforans pedis as the principal manifestation of spina bifida occulta with an associated intradural lipoma. Two lumbosacral dimples and a tuft of hair overlying the spina bifida led to the diagnosis. We include a review of the causes of mal perforans and a review of spina bifida occulta and associated abnormalities. | Mal perforans and spina bifida occulta. A 26-year-old man had mal perforans pedis as the principal manifestation of spina bifida occulta with an associated intradural lipoma. Two lumbosacral dimples and a tuft of hair overlying the spina bifida led to the diagnosis. We include a review of the causes of mal perforans and a review of spina bifida occulta and associated abnormalities. | [
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PMID:24426 | Alclofenac and D-penicillamine. Comparative trial in rheumatoid arthritis. | Forty-six patients with rheumatoid arthritis, 22 receiving D-penicillamine and 2j alclofenac, took part in a 6-month single-bind external observer trial to compare the efficacy and toxicity of these drugs in the treatment of severe rheumatoid arthritis. Both drugs were active and similar in their efficacy at 6 months as judged by clinical and laboratory measurements. Penicillamine was active therapeutically by 3 months, one month before alclofenac. 9 patients, 8 on alclofenac and one on D-penicillamine, had to stop treatment because of lack of effect or toxic effects. Skin rashes within the first week of treatment were a major problem with alclofenac and led to 6 withdrawals. | Alclofenac and D-penicillamine. Comparative trial in rheumatoid arthritis. Forty-six patients with rheumatoid arthritis, 22 receiving D-penicillamine and 2j alclofenac, took part in a 6-month single-bind external observer trial to compare the efficacy and toxicity of these drugs in the treatment of severe rheumatoid arthritis. Both drugs were active and similar in their efficacy at 6 months as judged by clinical and laboratory measurements. Penicillamine was active therapeutically by 3 months, one month before alclofenac. 9 patients, 8 on alclofenac and one on D-penicillamine, had to stop treatment because of lack of effect or toxic effects. Skin rashes within the first week of treatment were a major problem with alclofenac and led to 6 withdrawals. | [
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PMID:24428 | Evaluation of physician assistants in rural primary care. | The advent of physician assistants raises questions regarding their appropriate utilization and monitoring. To determine practice patterns of physician assistants in settings independent of training programs, we studied 14 primary care practices in the rural southeast. Detailed observations, including appropriateness of medical care, were made on 788 outpatient-provider encounters. Physician assistants handled minor medical problems well and 61% of the patients observed in these practices fitted this category. Three practice patterns were observed: all patients were seen by the assistant initially, followed by the physician; patients managed concurrently by physician and assistant were not preselected; and patients with specific problems were assigned to the assistant. Properly managed, each of these patterns yielded competent care. Using these observations, proposed models of management and audit are presented for each practice pattern. | Evaluation of physician assistants in rural primary care. The advent of physician assistants raises questions regarding their appropriate utilization and monitoring. To determine practice patterns of physician assistants in settings independent of training programs, we studied 14 primary care practices in the rural southeast. Detailed observations, including appropriateness of medical care, were made on 788 outpatient-provider encounters. Physician assistants handled minor medical problems well and 61% of the patients observed in these practices fitted this category. Three practice patterns were observed: all patients were seen by the assistant initially, followed by the physician; patients managed concurrently by physician and assistant were not preselected; and patients with specific problems were assigned to the assistant. Properly managed, each of these patterns yielded competent care. Using these observations, proposed models of management and audit are presented for each practice pattern. | [
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PMID:24429 | Allergic angiitis and granulomatosis. Prolonged remission induced by combined prednisone--azathioprine therapy. | Two patients with acute, rapidly progressive generalized vasculitis initially had symptoms of asthma. Progressive increase in severity of asthma was followed by systemic disease, including pulmonary infiltrative disease, mononeuritis multiplex, and abdominal pain. Examination of the tissues demonstrated vasculitis with eosinophilia, and clinically both cases appeared in a near terminal state. High-dose prednisone did not induce a remission. In particular, the lesions of mononeuritis multiplex progressed after initiation of high-dose prednisone. The addition of azathioprine to the regimen was followed by a gradual and then complete remission of clinical and laboratory abnormalities, except for some residual nerve damage and asthma of varying severity in the two patients. These two patients, whose cases are classified as the allergic granulomatosis variant of polyarteritis nodosa, have had a remission of seven and almost two years, respectively, after combined prednisone-azathioprine therapy. | Allergic angiitis and granulomatosis. Prolonged remission induced by combined prednisone--azathioprine therapy. Two patients with acute, rapidly progressive generalized vasculitis initially had symptoms of asthma. Progressive increase in severity of asthma was followed by systemic disease, including pulmonary infiltrative disease, mononeuritis multiplex, and abdominal pain. Examination of the tissues demonstrated vasculitis with eosinophilia, and clinically both cases appeared in a near terminal state. High-dose prednisone did not induce a remission. In particular, the lesions of mononeuritis multiplex progressed after initiation of high-dose prednisone. The addition of azathioprine to the regimen was followed by a gradual and then complete remission of clinical and laboratory abnormalities, except for some residual nerve damage and asthma of varying severity in the two patients. These two patients, whose cases are classified as the allergic granulomatosis variant of polyarteritis nodosa, have had a remission of seven and almost two years, respectively, after combined prednisone-azathioprine therapy. | [
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PMID:24430 | [Selective local hyperthermy of tumor tissue. Homogenized energy supply also to deep-seated tissues by high-performance decametric wave coil section plus dual system raster motion (author's transl)]. | Based on results of selective tumor tissue hyperacidification and whole-body hyperthermy the paper deals with the advantages of local hyperthermy and gives a review of the various methods of energy supply for hyperthermy in living tissue. It follows a presentation of physical and biophysical fundamentals on selective local hyperthermy of tumor tissue by radiation, by r. f. rotational magnetic fields and by r. f. capacitor fields. Using a gelatine phantom for measuring the energy supply with the various modes of local hyperthermy it is then shown that it is solely the divergent rotational magnetic field which allows to substantially reduce the inhomogeneity of energy supply also in z-direction (increasing depth in body tissue) by introducing a relative raster motion of the applicator in x-y-direction (plane parallel to body surface). It is only the introduction of the x-y raster principle combined with a described dual system array (CMT Selectotherm System) which permits the high and rather homogeneous supply of thermal energy also to deep-seated tumor tissue in the patient without concomitant critical superheating of tissues near the skin. Finally the paper gives a theoretical derivation of the temperature profile for selective local hyperthermy of tumor tissue. With due consideration of heat dissipation by convection and conduction, this endeavour shows rather promising aspects for the therapeutic efficiency obtainable in each case of treatment. | [Selective local hyperthermy of tumor tissue. Homogenized energy supply also to deep-seated tissues by high-performance decametric wave coil section plus dual system raster motion (author's transl)]. Based on results of selective tumor tissue hyperacidification and whole-body hyperthermy the paper deals with the advantages of local hyperthermy and gives a review of the various methods of energy supply for hyperthermy in living tissue. It follows a presentation of physical and biophysical fundamentals on selective local hyperthermy of tumor tissue by radiation, by r. f. rotational magnetic fields and by r. f. capacitor fields. Using a gelatine phantom for measuring the energy supply with the various modes of local hyperthermy it is then shown that it is solely the divergent rotational magnetic field which allows to substantially reduce the inhomogeneity of energy supply also in z-direction (increasing depth in body tissue) by introducing a relative raster motion of the applicator in x-y-direction (plane parallel to body surface). It is only the introduction of the x-y raster principle combined with a described dual system array (CMT Selectotherm System) which permits the high and rather homogeneous supply of thermal energy also to deep-seated tumor tissue in the patient without concomitant critical superheating of tissues near the skin. Finally the paper gives a theoretical derivation of the temperature profile for selective local hyperthermy of tumor tissue. With due consideration of heat dissipation by convection and conduction, this endeavour shows rather promising aspects for the therapeutic efficiency obtainable in each case of treatment. | [
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PMID:24431 | Anxiety, arousal, and autonomic habituation. | The interaction of anxiety and autonomic activation as a factor in the development and persistency of pathological anxiety was investigated with the aid of self-rating procedures and a habituation experiment. The state of activation was varied systematically in 40 normal subjects by various experimental conditions and by the administration of a tranquilizer. The degree of anxiety and activation were able to be differentiated in the investigated range of mean attentiveness. Anxious expectancy is perceived in particular as subjective anxiety. Fatigue and sedation, on the other hand, demonstrate subjective and autonomic desactivation. Corresponding differences can be demonstrated for the anxiolytic and sedative effects of tranquilizers. The time course of habituation is a more exact indicator than the amplitude of the orienting response. Cognitively provoked apprehensiveness, thus, appears to be qualitatively different as compared to psychoautonomically caused anxieties of psychiatric disorders. | Anxiety, arousal, and autonomic habituation. The interaction of anxiety and autonomic activation as a factor in the development and persistency of pathological anxiety was investigated with the aid of self-rating procedures and a habituation experiment. The state of activation was varied systematically in 40 normal subjects by various experimental conditions and by the administration of a tranquilizer. The degree of anxiety and activation were able to be differentiated in the investigated range of mean attentiveness. Anxious expectancy is perceived in particular as subjective anxiety. Fatigue and sedation, on the other hand, demonstrate subjective and autonomic desactivation. Corresponding differences can be demonstrated for the anxiolytic and sedative effects of tranquilizers. The time course of habituation is a more exact indicator than the amplitude of the orienting response. Cognitively provoked apprehensiveness, thus, appears to be qualitatively different as compared to psychoautonomically caused anxieties of psychiatric disorders. | [
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PMID:24432 | Purification of human diploid fibroblast interferon by immobilized neuraminidase. | An efficient method for the purification of human fibroblast interferon (IF) based on binding via the N-acetyl neuraminic acid (N-ANA) residue of the IF molecules to the immobilized neuraminidase has been developed. Binding of IF occurred at pH 4.5 and elution of the bound activity was effected at pH 9.5. Specific activity of IF in the alkaline eluate was increased by a factor in excess of 200 and the IF thus recovered had probably retained most of the N-ANA moieties. | Purification of human diploid fibroblast interferon by immobilized neuraminidase. An efficient method for the purification of human fibroblast interferon (IF) based on binding via the N-acetyl neuraminic acid (N-ANA) residue of the IF molecules to the immobilized neuraminidase has been developed. Binding of IF occurred at pH 4.5 and elution of the bound activity was effected at pH 9.5. Specific activity of IF in the alkaline eluate was increased by a factor in excess of 200 and the IF thus recovered had probably retained most of the N-ANA moieties. | [
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PMID:24433 | [State of the microcirculation in periarteritis nodosa based on morphological and biomicroscopic data]. | The status of microcirculation in periarteritis nodosa was studied by impregnation method (serous membranes) and biomicroscopy (eyeball conjunctiva). Pathological and adaptative changes in terminal vessels were found. It is suggested that the involvement of microcirculation ways in this disease is systemic and that morphological changes in microvessels are comparable to biomicroscopic changes. | [State of the microcirculation in periarteritis nodosa based on morphological and biomicroscopic data]. The status of microcirculation in periarteritis nodosa was studied by impregnation method (serous membranes) and biomicroscopy (eyeball conjunctiva). Pathological and adaptative changes in terminal vessels were found. It is suggested that the involvement of microcirculation ways in this disease is systemic and that morphological changes in microvessels are comparable to biomicroscopic changes. | [
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PMID:24436 | Oxidative phosphorylation. Halide-dependent and halide-independent effects of triorganotin and trioganolead compounds on mitochondrial functions. | 1. Each of five triorganotin and five triorganolead compounds was shown to perturb mithochondrial functions in three different ways. One is dependent and two are independent of Cl- in the medium. 2. Structure-activity relationships for the three interactions are described, and compounds suitable as tools for the separate study of each process are defined. 3. In a Cl- -containing medium trimethyltin, triethyltin, trimethyl-lead, triethyl-lead and tri-n-propyl-lead all produce the same maximum rate of ATP hydrolysis and O2 uptake; this rate is much less than that produced by uncoupling agents such as 2,4-dinitrophenol. 4. Increase in ATP hydrolysis and O2 uptake are measures on energy ultilization when triogranotin and triorganolead compounds bring about an exchange of external C1- for intramitochondrial OH- ions. Possible rate-limiting steps in this process are discussed. 5. In a C1- -containing medium ATP synthesis linked to the oxidation of beta-hydroxybutyrate or reduced cytochrone c is less inhibited by triethyltin or triethyl-lead than is ATP synthesis linked to the oxidation of succinate, pyruvate or L-glutamate. 6. The inhibition of ATP synthesis linked to the oxidation of both beta-hydroxybutyrate and reduced cytochrome c consists of two processes: one is a limited uncoupling and is C1- -dependent and the other is a C1- -independent inhibition of the energy-conservation system. 7. The different sensitivities to inhibition by triethyltin of mitochondrial functions involving the oxidation of beta-hydroxybutyrate and succinate are compared and discussed. | Oxidative phosphorylation. Halide-dependent and halide-independent effects of triorganotin and trioganolead compounds on mitochondrial functions. 1. Each of five triorganotin and five triorganolead compounds was shown to perturb mithochondrial functions in three different ways. One is dependent and two are independent of Cl- in the medium. 2. Structure-activity relationships for the three interactions are described, and compounds suitable as tools for the separate study of each process are defined. 3. In a Cl- -containing medium trimethyltin, triethyltin, trimethyl-lead, triethyl-lead and tri-n-propyl-lead all produce the same maximum rate of ATP hydrolysis and O2 uptake; this rate is much less than that produced by uncoupling agents such as 2,4-dinitrophenol. 4. Increase in ATP hydrolysis and O2 uptake are measures on energy ultilization when triogranotin and triorganolead compounds bring about an exchange of external C1- for intramitochondrial OH- ions. Possible rate-limiting steps in this process are discussed. 5. In a C1- -containing medium ATP synthesis linked to the oxidation of beta-hydroxybutyrate or reduced cytochrone c is less inhibited by triethyltin or triethyl-lead than is ATP synthesis linked to the oxidation of succinate, pyruvate or L-glutamate. 6. The inhibition of ATP synthesis linked to the oxidation of both beta-hydroxybutyrate and reduced cytochrome c consists of two processes: one is a limited uncoupling and is C1- -dependent and the other is a C1- -independent inhibition of the energy-conservation system. 7. The different sensitivities to inhibition by triethyltin of mitochondrial functions involving the oxidation of beta-hydroxybutyrate and succinate are compared and discussed. | [
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PMID:24437 | Demonstration of the phosphorylation of acetyl-coenzyme A carboxylase within intact rat epididymal fat-cells. | Intact rat epididymal fat-cells were incubated with 32Pi and the intracellular proteins separated by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. One of the phosphorylated proteins has the same RF value as [14C]biotin-labelled acetyl-CoA carboxylase purified from fat-cells and is specifically precipitated after incubation with antiserum raised against acetyl-CoA carboxylase. No significant changes in the extent of phosphorylation of acetyl-CoA carboxylase were detected after exposure of the cells to insulin. | Demonstration of the phosphorylation of acetyl-coenzyme A carboxylase within intact rat epididymal fat-cells. Intact rat epididymal fat-cells were incubated with 32Pi and the intracellular proteins separated by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. One of the phosphorylated proteins has the same RF value as [14C]biotin-labelled acetyl-CoA carboxylase purified from fat-cells and is specifically precipitated after incubation with antiserum raised against acetyl-CoA carboxylase. No significant changes in the extent of phosphorylation of acetyl-CoA carboxylase were detected after exposure of the cells to insulin. | [
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PMID:24438 | Isolation of proteoglycans from human articular cartilage. | Proteoglycans were extracted from normal human articular cartilage of various ages with 4M-guanidinium chloride and were purified and characterized by using preformed linear CsCl density gradients. With advancing age, there was a decrease in high-density proteoglycans of low protein/uronic acid weight ratio and an increase in the proportion of lower-density proteoglycans, richer in keratan sulphate and protein. Proteoglycans of each age were also shown to disaggregate in 4M-guanidinium chloride and at low pH and to reaggregate in the presence of hyaluronic acid and/or low-density fractions. Osteoarthrotic-cartilage extracts had an increased content of higher-density proteoglycans compared with normal cartilage of the same age, and results also suggested that these were not mechanical or enzymic degradation products, but were possibly proteoglycans of an immature nature. | Isolation of proteoglycans from human articular cartilage. Proteoglycans were extracted from normal human articular cartilage of various ages with 4M-guanidinium chloride and were purified and characterized by using preformed linear CsCl density gradients. With advancing age, there was a decrease in high-density proteoglycans of low protein/uronic acid weight ratio and an increase in the proportion of lower-density proteoglycans, richer in keratan sulphate and protein. Proteoglycans of each age were also shown to disaggregate in 4M-guanidinium chloride and at low pH and to reaggregate in the presence of hyaluronic acid and/or low-density fractions. Osteoarthrotic-cartilage extracts had an increased content of higher-density proteoglycans compared with normal cartilage of the same age, and results also suggested that these were not mechanical or enzymic degradation products, but were possibly proteoglycans of an immature nature. | [
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PMID:24439 | Activation of immobilized acetyl-Coenzyme A carboxylase. | Partially purified acetyl-CoA carboxylase was covalently bound to a Sepharose 4B matrix. Although aggregation was thus prevented, the enzymic activity was stimulated by citrate and isocitrate. | Activation of immobilized acetyl-Coenzyme A carboxylase. Partially purified acetyl-CoA carboxylase was covalently bound to a Sepharose 4B matrix. Although aggregation was thus prevented, the enzymic activity was stimulated by citrate and isocitrate. | [
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PMID:24440 | Synthesis of the native copper(II)-transport site of human serum albumin and its copper(II)-binding properties. | A derivative of the native-sequence tripeptide of the specific Cu(II)-transport site of human serum albumin, L-aspartyl-L-alanyl-L-histidine N-methylamide, was synthesized, and its binding to Cu(II) was examined to determine the influence of the side-chain groups on the Cu(II) binding. The equilibria involved in the Cu(II)-L-aspartyl-L-alanyl-L-histidine N-methylamide system were investigated by analytical potentiometry. Three complex species were found in the pH range 4-10. The same species were identified in both the visible and circular-dichroism spectra. The main species present in the physiological pH range is shown to have the same ligands around the square-planar Cu(II) ion as those reported for albumin and tripeptides diglycyl-L-histidine and its N-methylamide derivative. The results obtained from competition experiments showed that this tripeptide has a higher affinity towards Cu(II) than has albumin itself. The overall findings are compared with those from albumin. At neutral pH the side chains do not play any important role in the Cu(II) binding, but at low pH the beta-carboxyl group of the N-terminal aspartic residue becomes important. A possible competition site on albumin for Cu(II) at low pH is discussed. | Synthesis of the native copper(II)-transport site of human serum albumin and its copper(II)-binding properties. A derivative of the native-sequence tripeptide of the specific Cu(II)-transport site of human serum albumin, L-aspartyl-L-alanyl-L-histidine N-methylamide, was synthesized, and its binding to Cu(II) was examined to determine the influence of the side-chain groups on the Cu(II) binding. The equilibria involved in the Cu(II)-L-aspartyl-L-alanyl-L-histidine N-methylamide system were investigated by analytical potentiometry. Three complex species were found in the pH range 4-10. The same species were identified in both the visible and circular-dichroism spectra. The main species present in the physiological pH range is shown to have the same ligands around the square-planar Cu(II) ion as those reported for albumin and tripeptides diglycyl-L-histidine and its N-methylamide derivative. The results obtained from competition experiments showed that this tripeptide has a higher affinity towards Cu(II) than has albumin itself. The overall findings are compared with those from albumin. At neutral pH the side chains do not play any important role in the Cu(II) binding, but at low pH the beta-carboxyl group of the N-terminal aspartic residue becomes important. A possible competition site on albumin for Cu(II) at low pH is discussed. | [
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PMID:24441 | The binding of copper ions to copper-free bovine superoxide dismutase. Kinetic aspects. | The kinetics of reconstitution of bovine superoxide dismutase from Cu2+ and the copper-free enzyme have been studied by activity, u.v.-absorption, electron-paramagnetic-resonance and pulsed-nuclear-magnetic-resonance measurements. The process appears to be first-order up to 80% completion in most conditions, and is pH-dependent, with an apparent pK of 6.5. U.v.-absorption and solvent proton relaxation rate measurements show that fast binding of Cu2+ occurs, and the initial ligands are likely to be, at least in part, those of the native active site. The recovery of the native activity and spectroscopic properties is a slow process with activation energies of 92 kJ/mol at pH 5.3 and 8.4kJ/mol at pH 8.1 and can be described as a rearrangement of the site around the bound metal. The rate of this process is lower in partially recombined protein samples, probably because of intersubunit interactions. | The binding of copper ions to copper-free bovine superoxide dismutase. Kinetic aspects. The kinetics of reconstitution of bovine superoxide dismutase from Cu2+ and the copper-free enzyme have been studied by activity, u.v.-absorption, electron-paramagnetic-resonance and pulsed-nuclear-magnetic-resonance measurements. The process appears to be first-order up to 80% completion in most conditions, and is pH-dependent, with an apparent pK of 6.5. U.v.-absorption and solvent proton relaxation rate measurements show that fast binding of Cu2+ occurs, and the initial ligands are likely to be, at least in part, those of the native active site. The recovery of the native activity and spectroscopic properties is a slow process with activation energies of 92 kJ/mol at pH 5.3 and 8.4kJ/mol at pH 8.1 and can be described as a rearrangement of the site around the bound metal. The rate of this process is lower in partially recombined protein samples, probably because of intersubunit interactions. | [
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PMID:24442 | Partial purification and characterization of a pyruvate dehydrogenase-complex-inactivating enzyme from rat liver. | An enzyme inactivating the pyruvate dehydrogenase complex (inactivase) was purified about 8000-fold from rat liver by differential centrifugation, acid extraction of a lysosomerich 25000 g pellet, acetone fractionation, and adsorption on calcium phosphate gel. By exclusion chromatography on Sephadex G-100 a molecular weight of 21 000 was estimated. The purified enzyme was most stable at pH 5.8 in potassium phosphate buffer, and at pH 4.5 in McIlvaine buffer. At high dilutions the enzyme was very labile and was remarkably stabilized by high salt concentrations. Enzyme activity is inhibited by native rat blood serum, iodoacetamide and leupeptin, but not by phenylmethanesulphonyl fluoride, suggesting that it belongs to the class of thiol proteinases. Among various enzymes tested, only 2-oxoglutarate dehydrogenase was attacked by the inactivase to a similar extent to the pyruvate dehydrogenase complex. Studies on the inactivation mechanism indicate that although the overall reaction is completely lost after treatment with inactivase, each individual step of the multienzyme complex retains full catalytic activity. As judged from sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, the transacetylase subunit appears to be degraded into several smaller fractions. | Partial purification and characterization of a pyruvate dehydrogenase-complex-inactivating enzyme from rat liver. An enzyme inactivating the pyruvate dehydrogenase complex (inactivase) was purified about 8000-fold from rat liver by differential centrifugation, acid extraction of a lysosomerich 25000 g pellet, acetone fractionation, and adsorption on calcium phosphate gel. By exclusion chromatography on Sephadex G-100 a molecular weight of 21 000 was estimated. The purified enzyme was most stable at pH 5.8 in potassium phosphate buffer, and at pH 4.5 in McIlvaine buffer. At high dilutions the enzyme was very labile and was remarkably stabilized by high salt concentrations. Enzyme activity is inhibited by native rat blood serum, iodoacetamide and leupeptin, but not by phenylmethanesulphonyl fluoride, suggesting that it belongs to the class of thiol proteinases. Among various enzymes tested, only 2-oxoglutarate dehydrogenase was attacked by the inactivase to a similar extent to the pyruvate dehydrogenase complex. Studies on the inactivation mechanism indicate that although the overall reaction is completely lost after treatment with inactivase, each individual step of the multienzyme complex retains full catalytic activity. As judged from sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, the transacetylase subunit appears to be degraded into several smaller fractions. | [
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PMID:24443 | Fatty acid synthesis in the regenerating liver of the rat. | 1. Synthesis de novo of fatty acids in the rat liver, measured per g wet wt. of tissue, was increased by a factor of about two, between 1 and 4 days after partial hepatectomy, compared with rates in sham-operated control rat livers. 2. There were no associated changes in the rates of liver cholesterol synthesis or of adipose-tissue fatty acid synthesis in rats after partial hepatectomy, compared with rates in sham-operated rats. 3. In regenerating livers, perfused under three different conditions, there was no alteration in the capacity for fatty acid synthesis compared with that of control rats. 4. The increased synthesis of fatty acids in regenerating liver was associated with insignificant increases in plasma concentrations of tricylglycerols and free fatty acids, with a decrease in content of liver glycogen, and with no change in hepatic activity of acetyl-CoA carboxylase. 5. The accelerated rate of synthesis of fatty adids in regenerating liver appears not to be due to any intrinsic alteration in hepatic capacity for fatty acid synthesis, but it may be caused by the continuous action on liver of unidentified circulating factors. | Fatty acid synthesis in the regenerating liver of the rat. 1. Synthesis de novo of fatty acids in the rat liver, measured per g wet wt. of tissue, was increased by a factor of about two, between 1 and 4 days after partial hepatectomy, compared with rates in sham-operated control rat livers. 2. There were no associated changes in the rates of liver cholesterol synthesis or of adipose-tissue fatty acid synthesis in rats after partial hepatectomy, compared with rates in sham-operated rats. 3. In regenerating livers, perfused under three different conditions, there was no alteration in the capacity for fatty acid synthesis compared with that of control rats. 4. The increased synthesis of fatty acids in regenerating liver was associated with insignificant increases in plasma concentrations of tricylglycerols and free fatty acids, with a decrease in content of liver glycogen, and with no change in hepatic activity of acetyl-CoA carboxylase. 5. The accelerated rate of synthesis of fatty adids in regenerating liver appears not to be due to any intrinsic alteration in hepatic capacity for fatty acid synthesis, but it may be caused by the continuous action on liver of unidentified circulating factors. | [
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PMID:24454 | The extraction of antiepileptic drugs. Model study on partition coefficients. | To compare the extraction behaviour of hexane, toluene, chloroform, 1,2-dichloroethane, diethylether, ethyl acetate and acetone towards phenobarbital, phenytoin, primidone and carbamazepine, partition coefficients of these drugs between the above solvents and water (saturated with ammonium sulphate when working with acetone) were measured at different pH-conditions. Acetone rendered the highest partition coefficients for all drugs. Ethyl acetate was second in the range, except for carbamazepine, which showed a specific affinity for the chlorinated hydrocarbons. | The extraction of antiepileptic drugs. Model study on partition coefficients. To compare the extraction behaviour of hexane, toluene, chloroform, 1,2-dichloroethane, diethylether, ethyl acetate and acetone towards phenobarbital, phenytoin, primidone and carbamazepine, partition coefficients of these drugs between the above solvents and water (saturated with ammonium sulphate when working with acetone) were measured at different pH-conditions. Acetone rendered the highest partition coefficients for all drugs. Ethyl acetate was second in the range, except for carbamazepine, which showed a specific affinity for the chlorinated hydrocarbons. | [
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PMID:24458 | Intracellular localization of hepatic propionyl-CoA carboxylase and methylmalonyl-CoA mutase in humans and normal and vitamin B12 deficient rats. | The intracellular localization of the enzymes in the vitamin B12 dependent pathway which involves the oxidation of propionate was studied in rat liver obtained from normal and vitamin B12 deficient rats as well as from man. The subcellular site of propionyl CoA carboxylase and the vitamin B12 dependent-methylmalonyl CoA mutase were determined. All of the activity of these two enzymes was demonstrated to be in the mitochondria and those enzymes were shown to be loosely bound to the inner membrane-matrix portion of the mitochondria. Vitamin B12 deficiency did not alter the subcellular localization. Finally, a rapid enzymatic assay for methylmalonyl-CoA mutase was described. | Intracellular localization of hepatic propionyl-CoA carboxylase and methylmalonyl-CoA mutase in humans and normal and vitamin B12 deficient rats. The intracellular localization of the enzymes in the vitamin B12 dependent pathway which involves the oxidation of propionate was studied in rat liver obtained from normal and vitamin B12 deficient rats as well as from man. The subcellular site of propionyl CoA carboxylase and the vitamin B12 dependent-methylmalonyl CoA mutase were determined. All of the activity of these two enzymes was demonstrated to be in the mitochondria and those enzymes were shown to be loosely bound to the inner membrane-matrix portion of the mitochondria. Vitamin B12 deficiency did not alter the subcellular localization. Finally, a rapid enzymatic assay for methylmalonyl-CoA mutase was described. | [
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PMID:24459 | Metabolic effects of beta-sympathomimetic drugs and dexamethasone in normal and diabetic pregnancy. | Marked hyperglycaemia and hyperinsulinaemia were observed in two normal women in premature labour treated with an intravenous infusion of a beta-sympathomimetic drug and intramuscular dexamethasone injections. Similar therapy in a chemical diabetic patient caused diabetic ketoacidosis, while treatment of an insulin dependent diabetic with dexamethasone alone resulted in a major increase in her insulin requirements. It is suggested that the diabetogenic effects of beta-sympathomimetic drugs and dexamethasone may be additive and that regular plasma glucose estimations should be made when they are used, especially in patients with impaired glucose tolerance. | Metabolic effects of beta-sympathomimetic drugs and dexamethasone in normal and diabetic pregnancy. Marked hyperglycaemia and hyperinsulinaemia were observed in two normal women in premature labour treated with an intravenous infusion of a beta-sympathomimetic drug and intramuscular dexamethasone injections. Similar therapy in a chemical diabetic patient caused diabetic ketoacidosis, while treatment of an insulin dependent diabetic with dexamethasone alone resulted in a major increase in her insulin requirements. It is suggested that the diabetogenic effects of beta-sympathomimetic drugs and dexamethasone may be additive and that regular plasma glucose estimations should be made when they are used, especially in patients with impaired glucose tolerance. | [
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PMID:24460 | Renal tubular function in normal pregnancy. | Maternal plasma and urine concentrations of beta2- microglobulin and gamma-glutamyl transpeptidase (EC:2.3.2.2) were measured serially in normal pregnancy. These parameters remained constant throughout and can therefore be used to evaluate tubular function in pregnancies complicated by renal disorders. | Renal tubular function in normal pregnancy. Maternal plasma and urine concentrations of beta2- microglobulin and gamma-glutamyl transpeptidase (EC:2.3.2.2) were measured serially in normal pregnancy. These parameters remained constant throughout and can therefore be used to evaluate tubular function in pregnancies complicated by renal disorders. | [
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] |
PMID:24462 | Electron spin resonance and nuclear relaxation studies on spin-labeled glutamate dehydrogenase. | The reaction of glutamate dehydrogenase with two different stable nitroxides (spin labels) is reported. The two compounds contain a carbonyl and an iodoacetamide group as their reactive parts. The carbonyl compound inactivates the enzyme by the formation of a 1:1 covalent complex after NaBH4 reduction of an intermediate Schiff's base. Evidence indicates that the enzyme is modified at lysine-126 in the active site. The electron spin resonance (ESR) spectrum of spin-labeled enzyme indicates a high degree of immobilization of the nitroxide. The binding of reduced coenzyme NADPH is reflected by a change (immobilization) of the ESR spectrum. Nuclear relaxation of bound substrate, oxidized coenzyme, and inhibitor by the paramagnetic group is observed. This shows the existence of a binding site for these compounds close to the active site. The distances of selected protons of the binding ligands to the nitroxide are calculated. The iodoacetamide spin label reacts with several groups, one of which is not a sulfhydryl. The reaction of this particular group causes inactivation of the enzyme. Protection against this inactivation could be achieved with certain ligands. Only enzyme that was spin labeled without such protection caused paramagnetic relaxation of bound substrate and coenzyme. | Electron spin resonance and nuclear relaxation studies on spin-labeled glutamate dehydrogenase. The reaction of glutamate dehydrogenase with two different stable nitroxides (spin labels) is reported. The two compounds contain a carbonyl and an iodoacetamide group as their reactive parts. The carbonyl compound inactivates the enzyme by the formation of a 1:1 covalent complex after NaBH4 reduction of an intermediate Schiff's base. Evidence indicates that the enzyme is modified at lysine-126 in the active site. The electron spin resonance (ESR) spectrum of spin-labeled enzyme indicates a high degree of immobilization of the nitroxide. The binding of reduced coenzyme NADPH is reflected by a change (immobilization) of the ESR spectrum. Nuclear relaxation of bound substrate, oxidized coenzyme, and inhibitor by the paramagnetic group is observed. This shows the existence of a binding site for these compounds close to the active site. The distances of selected protons of the binding ligands to the nitroxide are calculated. The iodoacetamide spin label reacts with several groups, one of which is not a sulfhydryl. The reaction of this particular group causes inactivation of the enzyme. Protection against this inactivation could be achieved with certain ligands. Only enzyme that was spin labeled without such protection caused paramagnetic relaxation of bound substrate and coenzyme. | [
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PMID:24463 | Raman spectra of heme a, cytochrome oxidase-ligand complexes, and alkaline denatured oxidase. | We report 441.6 nm excitation resonance Raman spectra of oxidized and reduced monomeric heme a-imidazole, cytochrome oxidase-exogenous ligand complexes in various redox states, and alkaline denatured oxidase. These data show that, in reduced oxidase, the cytochrome a3 Raman spectrum has bands at 215, 364, 1230, and 1670 cm-1 not observed in the cytochrome a spectrum. The appearance of these bands in the reduced cytochrome a3 spectrum is due to interactions between the heme a of cytochrome a3 and its protein environment and not to intrinsic properties of heme a. These interactions are pH sensitive and strongly influence the vibrational spectra of both heme a groups. We assign the 1670-cm-1 band to the heme a formyl substituent and propose that the intensity of the 1670 cm-1 is high for reduced cytochrome a3 because the C==O lies in the porphyrin plane and is very weak for oxidized and reduced cytochrome a, oxidized cytochrome a3, and oxidized and reduced heme a-imidazole because the C==O lies out of the plane. We suggest that movement of the C==O in and out of the plane explains the ligand induced spectral shift in the optical absorption spectrum of reduced cytochrome a3. Finally, we confirm the observation of Adar & Yonetani (private communication) that, under laser illumination, resting oxidase is photoreactive. | Raman spectra of heme a, cytochrome oxidase-ligand complexes, and alkaline denatured oxidase. We report 441.6 nm excitation resonance Raman spectra of oxidized and reduced monomeric heme a-imidazole, cytochrome oxidase-exogenous ligand complexes in various redox states, and alkaline denatured oxidase. These data show that, in reduced oxidase, the cytochrome a3 Raman spectrum has bands at 215, 364, 1230, and 1670 cm-1 not observed in the cytochrome a spectrum. The appearance of these bands in the reduced cytochrome a3 spectrum is due to interactions between the heme a of cytochrome a3 and its protein environment and not to intrinsic properties of heme a. These interactions are pH sensitive and strongly influence the vibrational spectra of both heme a groups. We assign the 1670-cm-1 band to the heme a formyl substituent and propose that the intensity of the 1670 cm-1 is high for reduced cytochrome a3 because the C==O lies in the porphyrin plane and is very weak for oxidized and reduced cytochrome a, oxidized cytochrome a3, and oxidized and reduced heme a-imidazole because the C==O lies out of the plane. We suggest that movement of the C==O in and out of the plane explains the ligand induced spectral shift in the optical absorption spectrum of reduced cytochrome a3. Finally, we confirm the observation of Adar & Yonetani (private communication) that, under laser illumination, resting oxidase is photoreactive. | [
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PMID:24464 | Use of immobilized light-harvesting chlorophyll a/b protein to study the stoichiometry of its self-association. | D. J. Davis & E. L. Gross (1976) Biochim. Biophys. Acta 449, 554-564 previously observed that the light-harvesting chlorophyll a/b protein or chlorophyll protein complex II self-associated as determined by ultracentrifugation. We have determined the stoichiometry of complex formation by immobilizing the monomer on ethylenediamine-Sepharose 4B and determing the ability of immobilized protein to bind the free protein. The amount of soluble protein bound to the immobilized protein increased as the concentration of soluble protein increased. The binding was maximal between pH 7 and 8. The maximum binding was three molecules bound per one molecule of protein immobilized. These results indicate that a tetramer is the intrinsic structural unit of the light-harvesting chlorophyll a/b protein in the chloroplast membrane. Upon complex formation, the chlorophyll fluorescence was decreased without any spectral change. The maximum binding was approximately doubled upon addition of 0.5 mM CaCl2 whereas 5 mM NaCl had no effect. Addition of CaCl2 had no effect on the fluorescence of the monomer. The light-harvesting chlorophyll a/b protein can be isolated from a sodium lauryl sulfate extract of chloroplasts by affinity chromatography using the immobilized light-harvesting chlorophyll a/b protein. | Use of immobilized light-harvesting chlorophyll a/b protein to study the stoichiometry of its self-association. D. J. Davis & E. L. Gross (1976) Biochim. Biophys. Acta 449, 554-564 previously observed that the light-harvesting chlorophyll a/b protein or chlorophyll protein complex II self-associated as determined by ultracentrifugation. We have determined the stoichiometry of complex formation by immobilizing the monomer on ethylenediamine-Sepharose 4B and determing the ability of immobilized protein to bind the free protein. The amount of soluble protein bound to the immobilized protein increased as the concentration of soluble protein increased. The binding was maximal between pH 7 and 8. The maximum binding was three molecules bound per one molecule of protein immobilized. These results indicate that a tetramer is the intrinsic structural unit of the light-harvesting chlorophyll a/b protein in the chloroplast membrane. Upon complex formation, the chlorophyll fluorescence was decreased without any spectral change. The maximum binding was approximately doubled upon addition of 0.5 mM CaCl2 whereas 5 mM NaCl had no effect. Addition of CaCl2 had no effect on the fluorescence of the monomer. The light-harvesting chlorophyll a/b protein can be isolated from a sodium lauryl sulfate extract of chloroplasts by affinity chromatography using the immobilized light-harvesting chlorophyll a/b protein. | [
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PMID:24465 | Kinetic analysis of the acid-alkaline conversion of horseradish peroxidases. | The nature of the acid-alkaline conversion of horseradish peroxidases was studied by measuring four rate constants in reactions, E + H+ (k1) in equilibrium (k2) EH+ and E + H2O (k3) in equilibrium (k4) EH+ + OH-, where EH+ and E denote the acid and alkaline forms of the enzymes. The values of k1, (k2 + k3), and k4 were obtained by measuring the relaxation rates of the acid leads to alkaline and alkaline leads to acid conversions by means of th pH jump method with a stopped-flow apparatus. The value of k3 could also be obtained by measuring the rate of reactions between hydrogen peroxide and peroxidases at alkaline pH. The measurements were conducted with four peroxidases having different pKa values: peroxidase A )pKa = 9.3), peroxidase C (pKa = 11.1), diacetyldeuteroperoxidase A (pKa = 7.7), and diacetyldeuteroperoxidase C (pKa = 9.1). The value of k1 was about 10(10) M-1 s-1 in the reaction of the four enzymes while k4 was quite different between the enzymes. The pKa was determined by k3 and k4 for the natural peroxidases and by k1 and k2 for the diacetyldeuteroperoxidases. The mechanism of the acid-alkaline conversion was discussed in comparison with that of metmyoglobin. | Kinetic analysis of the acid-alkaline conversion of horseradish peroxidases. The nature of the acid-alkaline conversion of horseradish peroxidases was studied by measuring four rate constants in reactions, E + H+ (k1) in equilibrium (k2) EH+ and E + H2O (k3) in equilibrium (k4) EH+ + OH-, where EH+ and E denote the acid and alkaline forms of the enzymes. The values of k1, (k2 + k3), and k4 were obtained by measuring the relaxation rates of the acid leads to alkaline and alkaline leads to acid conversions by means of th pH jump method with a stopped-flow apparatus. The value of k3 could also be obtained by measuring the rate of reactions between hydrogen peroxide and peroxidases at alkaline pH. The measurements were conducted with four peroxidases having different pKa values: peroxidase A )pKa = 9.3), peroxidase C (pKa = 11.1), diacetyldeuteroperoxidase A (pKa = 7.7), and diacetyldeuteroperoxidase C (pKa = 9.1). The value of k1 was about 10(10) M-1 s-1 in the reaction of the four enzymes while k4 was quite different between the enzymes. The pKa was determined by k3 and k4 for the natural peroxidases and by k1 and k2 for the diacetyldeuteroperoxidases. The mechanism of the acid-alkaline conversion was discussed in comparison with that of metmyoglobin. | [
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PMID:24468 | The mechanism of oxidation of reduced nicotinamide dinucleotide phosphate by submitochondrial particles from beef heart. | 1. Oxidation of NADPH by various acceptors catalyzed by submitochondrial particles and a partially purified NADH dehydrogenase from beef heart was investigated. Submitochondrial particles devoid of nicotinamide nucleotide transhydrogenase activity catalyze an oxidation of NADPH by oxygen. The partially purified NADH dehydrogenase prepared from these particles catalyzes an oxidation of NADPH by acetylpyridine-NAD. In both cases the rates of oxidation are about two orders of magnitude lower than those obtained with NADH as electron donor. 2. The kinetic characteristics of the NADPH oxidase reaction and reduction of acetylpyridine-NAD by NADPH are similar with regard to pH dependences and affinities for NADPH, indicating that both reactions involve the same binding site for NADPH. The binding of NADPH to this site appears to be rate limiting for the overall reactions. 3. At redox equilibrium NADPH and NADH reduce FMN and iron-sulphur center 1 of NADH dehydrogenase to the same extents. The rate of reduction of FMN by NADPH is at least two orders of magnitude lower than with NADH. 4. It is concluded that NADPH is a substrate of NADH dehydrogenase and that the nicotinamide nucleotide is oxidized by submitochondrial particles via the NADH--binding site of the enzyme. | The mechanism of oxidation of reduced nicotinamide dinucleotide phosphate by submitochondrial particles from beef heart. 1. Oxidation of NADPH by various acceptors catalyzed by submitochondrial particles and a partially purified NADH dehydrogenase from beef heart was investigated. Submitochondrial particles devoid of nicotinamide nucleotide transhydrogenase activity catalyze an oxidation of NADPH by oxygen. The partially purified NADH dehydrogenase prepared from these particles catalyzes an oxidation of NADPH by acetylpyridine-NAD. In both cases the rates of oxidation are about two orders of magnitude lower than those obtained with NADH as electron donor. 2. The kinetic characteristics of the NADPH oxidase reaction and reduction of acetylpyridine-NAD by NADPH are similar with regard to pH dependences and affinities for NADPH, indicating that both reactions involve the same binding site for NADPH. The binding of NADPH to this site appears to be rate limiting for the overall reactions. 3. At redox equilibrium NADPH and NADH reduce FMN and iron-sulphur center 1 of NADH dehydrogenase to the same extents. The rate of reduction of FMN by NADPH is at least two orders of magnitude lower than with NADH. 4. It is concluded that NADPH is a substrate of NADH dehydrogenase and that the nicotinamide nucleotide is oxidized by submitochondrial particles via the NADH--binding site of the enzyme. | [
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PMID:24470 | The mechanism of the control of carbon fixation by the pH in the chloroplast stroma. Studies with nitrite-mediated proton transfer across the envelope. | 1. CO2 fixation of intact spinach chloroplasts is inhibited by nitrite in a pH-dependent mode. At pH 7.3 in the medium 1 mM NaNO2 and at pH 7.9 5 mM NaNO2 were required for 50% inhibition. 2. The addition of nitrite leads to an acidificiation in the stroma. It appears that nitrite renders the envelope permeable for protons resulting in a breakdown of the pH gradient between the external space and the stroma. 3. In view of earlier results on the pH sensitivity of C02 fixation it is concluded that this pH shift in the stroma is responsible for the observed inhibition of CO2 fixation by nitrite. 4. Octanoate and to some extent also high concentrations of bicarbonate and acetate have a similar effect as nitrite in inhibiting CO2 fixation through an acidification in the stroma. 5. The levels of the intermediates of the CO2 fixation cycle were measured. A strong rise of the levels of fructose- and sedoheptulose biphosphates and a concomitant decrease of the corresponding monophosphates was observed during inhibition of CO2 fixation. It appears that the enzymatic steps of the CO2 fixation cycle responsible for the overall inhibition of CO2 fixation caused by lowering of the H+ concentration in the stroma are fructose- and sedopheptulose bisphosphatase. These two enzymes have an important function in the light regulation of CO2 fixation. | The mechanism of the control of carbon fixation by the pH in the chloroplast stroma. Studies with nitrite-mediated proton transfer across the envelope. 1. CO2 fixation of intact spinach chloroplasts is inhibited by nitrite in a pH-dependent mode. At pH 7.3 in the medium 1 mM NaNO2 and at pH 7.9 5 mM NaNO2 were required for 50% inhibition. 2. The addition of nitrite leads to an acidificiation in the stroma. It appears that nitrite renders the envelope permeable for protons resulting in a breakdown of the pH gradient between the external space and the stroma. 3. In view of earlier results on the pH sensitivity of C02 fixation it is concluded that this pH shift in the stroma is responsible for the observed inhibition of CO2 fixation by nitrite. 4. Octanoate and to some extent also high concentrations of bicarbonate and acetate have a similar effect as nitrite in inhibiting CO2 fixation through an acidification in the stroma. 5. The levels of the intermediates of the CO2 fixation cycle were measured. A strong rise of the levels of fructose- and sedoheptulose biphosphates and a concomitant decrease of the corresponding monophosphates was observed during inhibition of CO2 fixation. It appears that the enzymatic steps of the CO2 fixation cycle responsible for the overall inhibition of CO2 fixation caused by lowering of the H+ concentration in the stroma are fructose- and sedopheptulose bisphosphatase. These two enzymes have an important function in the light regulation of CO2 fixation. | [
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PMID:24471 | Dicarboxylate transport across the inner membrane of the chloroplast envelope. | The uptake of radioactively labeled dicarboxylates into the sorbitol-impermeable 3H2O space (the space surrounded by the inner envelope membrane) of spinach chloroplasts has been studied by means of silicone layer filtering centrifugation. 1. Malate, aspartate and a number of other dicarboxylates are rapidly transported across the envelope leading to an accumulation in the chloroplasts. This uptake proceeds mainly by a counterexchange with the dicarboxylates present there. 2. The dicarboxylate transport shows saturation characteristics allowing the determination of Km and V. 3. All dicarboxylates transported act as competitive inhibitors of the transport. 4. The activation energy of the transport as determined from the temperature dependency is evaluated to be 7 kcal/mol. 5. The rate of dicarboxylate transport is influenced by illumination, the countertransported molecules and the pH in the medium. These changes effect the transport velocity, whereas the corresponding Km values are not altered. 6. It is discussed whether there is more than one carrier involved in dicarboxylate transport in spinach chloroplasts. | Dicarboxylate transport across the inner membrane of the chloroplast envelope. The uptake of radioactively labeled dicarboxylates into the sorbitol-impermeable 3H2O space (the space surrounded by the inner envelope membrane) of spinach chloroplasts has been studied by means of silicone layer filtering centrifugation. 1. Malate, aspartate and a number of other dicarboxylates are rapidly transported across the envelope leading to an accumulation in the chloroplasts. This uptake proceeds mainly by a counterexchange with the dicarboxylates present there. 2. The dicarboxylate transport shows saturation characteristics allowing the determination of Km and V. 3. All dicarboxylates transported act as competitive inhibitors of the transport. 4. The activation energy of the transport as determined from the temperature dependency is evaluated to be 7 kcal/mol. 5. The rate of dicarboxylate transport is influenced by illumination, the countertransported molecules and the pH in the medium. These changes effect the transport velocity, whereas the corresponding Km values are not altered. 6. It is discussed whether there is more than one carrier involved in dicarboxylate transport in spinach chloroplasts. | [
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PMID:24472 | The permeability of the lysosomal membrane to small ions. | The permeability of the lysosomal membrane to small anions and cations was studied at 37 degrees C and pH 7.0 in a lysosomal-mitochondrial fraction isolated from the liver of untreated rats. The extent of osmotic lysis following ion influx was used as a measure of ion permeancy. In order to preserve electroneutrality, anion influx was coupled to an influx of K+ in the presence of valinomycin, and cation influx was coupled to an efflux of H+ using the protonophore 3-tert-butyl-5,2'-dichloro-4'-nitrosalicilylanilide. Lysosomal lysis was monitored by observing the loss of latency of two lysosomal hydrolases. The order of permeability of the lysosomal membrane to anions was found to be SCN- greater than I- greater than CH3COO- greater than Cl- approximately Pi greater than SO24- and that to cations Cs+ greater than K+ greater than Na+ greater than H+. These orders are largely in agreement with the lyotropic series of anions and cations. The implications of these findings for the mechanism by means of which a low intralysosomal pH is produced and maintained are discussed. | The permeability of the lysosomal membrane to small ions. The permeability of the lysosomal membrane to small anions and cations was studied at 37 degrees C and pH 7.0 in a lysosomal-mitochondrial fraction isolated from the liver of untreated rats. The extent of osmotic lysis following ion influx was used as a measure of ion permeancy. In order to preserve electroneutrality, anion influx was coupled to an influx of K+ in the presence of valinomycin, and cation influx was coupled to an efflux of H+ using the protonophore 3-tert-butyl-5,2'-dichloro-4'-nitrosalicilylanilide. Lysosomal lysis was monitored by observing the loss of latency of two lysosomal hydrolases. The order of permeability of the lysosomal membrane to anions was found to be SCN- greater than I- greater than CH3COO- greater than Cl- approximately Pi greater than SO24- and that to cations Cs+ greater than K+ greater than Na+ greater than H+. These orders are largely in agreement with the lyotropic series of anions and cations. The implications of these findings for the mechanism by means of which a low intralysosomal pH is produced and maintained are discussed. | [
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PMID:24473 | Active groups of extracellular endo-D-galacturonanase of aspergillus niger derived from pH effect on kinetic data. | In an attempt to characterize the groups essential for the catalytic action extracellular endo-D-galacturonanase of Aspergillus niger (poly (1,4-alpha-D-galacturonide) glycanohydrolase, EC 3.2.1.15) the behaviour of the kinetic parameters as a function of pH was examined. The dependence of kcat and kcat/Km on pH suggests that two dissociable groups are involved, for which the pK values of about 3.0 and 5.0 in the free enzyme and 3.06 and 5.72 in the catalytic complex were found at 30 degrees C. These values and the value of the heat of ionization of the acidic group, deltaHi 6.48 kcal/mol, resulting from the pKa values obtained at 20 degrees C (5.91) and at 30 degrees C (5.72) suggest the participation of a carboxylate group and a protonated imidazole group of histidine in the reaction catalyzed by endo-D-galacturonanase. | Active groups of extracellular endo-D-galacturonanase of aspergillus niger derived from pH effect on kinetic data. In an attempt to characterize the groups essential for the catalytic action extracellular endo-D-galacturonanase of Aspergillus niger (poly (1,4-alpha-D-galacturonide) glycanohydrolase, EC 3.2.1.15) the behaviour of the kinetic parameters as a function of pH was examined. The dependence of kcat and kcat/Km on pH suggests that two dissociable groups are involved, for which the pK values of about 3.0 and 5.0 in the free enzyme and 3.06 and 5.72 in the catalytic complex were found at 30 degrees C. These values and the value of the heat of ionization of the acidic group, deltaHi 6.48 kcal/mol, resulting from the pKa values obtained at 20 degrees C (5.91) and at 30 degrees C (5.72) suggest the participation of a carboxylate group and a protonated imidazole group of histidine in the reaction catalyzed by endo-D-galacturonanase. | [
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PMID:24474 | Effects of melanin on tyrosine hydroxylase and phenylalanine hydroxylase. | Melanin inhibited rat liver phenylalanine hydroxylase, but activated tyrosine hydroxylase from rat brain (caudate nucleus), rat adrenal glands, and bovine adrenal medulla. Activation of tyrosine hydroxylase by melanin was demonstrated with the extensively dialyzed enzyme and in suboptimal concentrations of the substrate (tyrosine) and the cofactor (6-methyltetrahydropterin). Tyrosine hydroxylase from rat brain was activated by melanin more markedly than that from rat adrenal glands. Purified and extensively dialyzed bovine adrenal tyrosine hydroxylase had two Km values with 6-methyltetrahydropterin, depending upon its concentrations, but the melanin-activated tyrosine hydroxylase had a single Km value and showed the classical Michaelis-Menten kinetics. | Effects of melanin on tyrosine hydroxylase and phenylalanine hydroxylase. Melanin inhibited rat liver phenylalanine hydroxylase, but activated tyrosine hydroxylase from rat brain (caudate nucleus), rat adrenal glands, and bovine adrenal medulla. Activation of tyrosine hydroxylase by melanin was demonstrated with the extensively dialyzed enzyme and in suboptimal concentrations of the substrate (tyrosine) and the cofactor (6-methyltetrahydropterin). Tyrosine hydroxylase from rat brain was activated by melanin more markedly than that from rat adrenal glands. Purified and extensively dialyzed bovine adrenal tyrosine hydroxylase had two Km values with 6-methyltetrahydropterin, depending upon its concentrations, but the melanin-activated tyrosine hydroxylase had a single Km value and showed the classical Michaelis-Menten kinetics. | [
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PMID:24475 | Immobilized D-amino acid oxidase. | 1. D-Amino acid oxidase (D-amino acid: oxygen oxidoreductase (deaminating), EC 1.4.3.3) apoenzyme, holoenzyme and the enzyme-benzoate complex were active and stable when immobilized to aminoalkyl or carboxyalkyl agarose, or to cyanogen bromide-activated agarose. The immobilized enzyme-benzoate complex could be converted into the holo- and apoenzyme without being liberated from the agarose. 2. The apparent Michaelis constant and substrate specificity of the immobilized enzyme were similar to those of the free enzyme. The optimum pH of the reaction was shifted towards acidic side by 1.0-2.0 pH units from that of the free enzyme. 3. With increasing number of methylene groups of the 'spacer' from 3 to 5, molecular activity of the immobilized enzyme increased, while the apparent Miachaelis constant decreased. | Immobilized D-amino acid oxidase. 1. D-Amino acid oxidase (D-amino acid: oxygen oxidoreductase (deaminating), EC 1.4.3.3) apoenzyme, holoenzyme and the enzyme-benzoate complex were active and stable when immobilized to aminoalkyl or carboxyalkyl agarose, or to cyanogen bromide-activated agarose. The immobilized enzyme-benzoate complex could be converted into the holo- and apoenzyme without being liberated from the agarose. 2. The apparent Michaelis constant and substrate specificity of the immobilized enzyme were similar to those of the free enzyme. The optimum pH of the reaction was shifted towards acidic side by 1.0-2.0 pH units from that of the free enzyme. 3. With increasing number of methylene groups of the 'spacer' from 3 to 5, molecular activity of the immobilized enzyme increased, while the apparent Miachaelis constant decreased. | [
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PMID:24477 | The fate of extracellular glutathione in the rat. | When intravenously administered to rats, [U-14C]glycine-labelled GSSG, GSH and its analogue ophthalmic acid were rapidly removed from the blood. In perfusion studies with isolated liver, however, the compounds did not enter the liver tissue. Thus, uptake by this tissue is obviously not responsible for the removal of gamma-glutamyl tripeptides from the blood. Instead, rapid hydrolysis of the tripeptides was observed. The undegraded tripeptides were only detected in the blood immediately after administration. Within tissue the degradation product glycine accounted for all the radioactivity. After intravenous injection of the labelled tripeptides the radioactivity accumulated first in the kidney, as shown by autoradiographic studies and chemical analysis of different tissues. The hydrolysis of the gamma-glutamyl tripeptides decreased markedly after the renal arteries were clamped. These observations strongly suggest that renal tissue is the principal site of the degradation of the tripeptides. Inhibition studies and experiments with isolated renal tubules revealed that gamma-glutamyl transpeptidase catalyses the fast hydrolysis of the extracellular peptides. The results indicate that, when entering the extracellular space, glutathione and its analogues are completely hydrolysed and must be resynthesized after reuptake of the constituent amino acids. It is concluded that the degradation occurs mainly on the luminal surface of the renal brush-border membrane and that gamma-glutamyl transpeptidase is a glutathionase acting on extracellular glutathione. | The fate of extracellular glutathione in the rat. When intravenously administered to rats, [U-14C]glycine-labelled GSSG, GSH and its analogue ophthalmic acid were rapidly removed from the blood. In perfusion studies with isolated liver, however, the compounds did not enter the liver tissue. Thus, uptake by this tissue is obviously not responsible for the removal of gamma-glutamyl tripeptides from the blood. Instead, rapid hydrolysis of the tripeptides was observed. The undegraded tripeptides were only detected in the blood immediately after administration. Within tissue the degradation product glycine accounted for all the radioactivity. After intravenous injection of the labelled tripeptides the radioactivity accumulated first in the kidney, as shown by autoradiographic studies and chemical analysis of different tissues. The hydrolysis of the gamma-glutamyl tripeptides decreased markedly after the renal arteries were clamped. These observations strongly suggest that renal tissue is the principal site of the degradation of the tripeptides. Inhibition studies and experiments with isolated renal tubules revealed that gamma-glutamyl transpeptidase catalyses the fast hydrolysis of the extracellular peptides. The results indicate that, when entering the extracellular space, glutathione and its analogues are completely hydrolysed and must be resynthesized after reuptake of the constituent amino acids. It is concluded that the degradation occurs mainly on the luminal surface of the renal brush-border membrane and that gamma-glutamyl transpeptidase is a glutathionase acting on extracellular glutathione. | [
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PMID:24478 | The dimerization of ferrihaems. I. The effect of buffer ions and specific cations on deuteroferrihaem dimerization. | The dimerization of dueteroferrihaem in aqueous solution has been investigated using a parameter, named the dimerization index (Robs). This is defined as the ratio of extinction coefficients at wavelengths corresponding to Soret band maxima for the monomeric and dimeric species, respectively. For solutions containing mainly monomeric species, Robs greater than 2, whereas for solutions containing mainly dimeric species Robs less than 1. A computer programme has been applied to determine values of the dimerization constant, K, defined as: K = [dimer] [H+]/[monomer]2. Phosphate buffer anions and Tris . HCl buffer enhanced dimerization. Monovalent and divalent cations also increased dimerization, but in a specific manner. The magnitudes of their effects increased in the order K+ less than Na+ less than Li+ less than Sr2+ less than Mg2+ approximately or equal to Ca2+. Values of K were determined for several concentrations of Na+ and Sr2+. These data are interpreted in terms of a stabilization of the ferrihaem dimer by the formation of ion triplets with the added cation 'sandwiched' between carboxyl residues of the adjacent ferrihaem monomeric units. General guidelines are recommended for the choice of conditions which minimize dimerization. | The dimerization of ferrihaems. I. The effect of buffer ions and specific cations on deuteroferrihaem dimerization. The dimerization of dueteroferrihaem in aqueous solution has been investigated using a parameter, named the dimerization index (Robs). This is defined as the ratio of extinction coefficients at wavelengths corresponding to Soret band maxima for the monomeric and dimeric species, respectively. For solutions containing mainly monomeric species, Robs greater than 2, whereas for solutions containing mainly dimeric species Robs less than 1. A computer programme has been applied to determine values of the dimerization constant, K, defined as: K = [dimer] [H+]/[monomer]2. Phosphate buffer anions and Tris . HCl buffer enhanced dimerization. Monovalent and divalent cations also increased dimerization, but in a specific manner. The magnitudes of their effects increased in the order K+ less than Na+ less than Li+ less than Sr2+ less than Mg2+ approximately or equal to Ca2+. Values of K were determined for several concentrations of Na+ and Sr2+. These data are interpreted in terms of a stabilization of the ferrihaem dimer by the formation of ion triplets with the added cation 'sandwiched' between carboxyl residues of the adjacent ferrihaem monomeric units. General guidelines are recommended for the choice of conditions which minimize dimerization. | [
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PMID:24479 | The dimerization of ferrihaems. II. Equilibrium and kinetic studies of mesoferrihaem dimerization. | Spectrophotometric data have been determined for mesoferrihaem at several pH values and over a range of concentration covering four orders of magnitude. The data reveal a dimerization process according to the equation 2 monomer in equilibrium dimer + H+, analogous to earlier findings for deuteroferrihaem and protoferrihaem. The value of K (defined as K = [dimer] [H+]/[monomer]2) was found to be 6.92.10(-2). This is close to the value for deuteroferrihaem but much less than that for protoferrihaem. This is interpreted in terms of possible additional bonding between the delocalized electron systems in protoferrihaem dimers relative to those of mesoferrihaem and deuteroferrihaem. Rate constants for dimerization were determined by temperature-jump spectrophotometry. The pH dependence of the rate constants is explained in terms of two distinct pathways for the dimerization process. These involve either direct reaction between two undissociated monomer molecules or alternatively an initial acid dissociation of a monomer molecule followed by reaction between an undissociated and dissociated molecule. | The dimerization of ferrihaems. II. Equilibrium and kinetic studies of mesoferrihaem dimerization. Spectrophotometric data have been determined for mesoferrihaem at several pH values and over a range of concentration covering four orders of magnitude. The data reveal a dimerization process according to the equation 2 monomer in equilibrium dimer + H+, analogous to earlier findings for deuteroferrihaem and protoferrihaem. The value of K (defined as K = [dimer] [H+]/[monomer]2) was found to be 6.92.10(-2). This is close to the value for deuteroferrihaem but much less than that for protoferrihaem. This is interpreted in terms of possible additional bonding between the delocalized electron systems in protoferrihaem dimers relative to those of mesoferrihaem and deuteroferrihaem. Rate constants for dimerization were determined by temperature-jump spectrophotometry. The pH dependence of the rate constants is explained in terms of two distinct pathways for the dimerization process. These involve either direct reaction between two undissociated monomer molecules or alternatively an initial acid dissociation of a monomer molecule followed by reaction between an undissociated and dissociated molecule. | [
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PMID:24480 | Spin-trapping of the trichloromethyl radical produced during enzymic NADPH oxidation in the presence of carbon tetrachloride or bromotrichloromethane. | Utilizing the spin-trapping agent phenyl-t-butyl nitrone, a free radical has been detected which is produced from carbon tetrachloride or bromotrichloromethane during the enzymic oxidation of NADPH by rat liver microsomes. The presence of NADPH is obligatory for generation of the radical. The formation of the trichloromethyl radical-phenyl-t-butyl nitrone adduct is an enzymic process, as evidenced by the inhibition of its formation in systems containing heated microsomes and in systems containing p-hydroxymercuribenzoate. A computer-simulated ESR spectrum for the trichloromethyl adduct of phenyl-t-butyl nitrone can reproduce the essential features of the spectrum of the spin-trapped radical produced enzymically from CCl4. A mechanism is proposed for the formation of the trichloromethyl radical from CCl4 or BrCCl3. | Spin-trapping of the trichloromethyl radical produced during enzymic NADPH oxidation in the presence of carbon tetrachloride or bromotrichloromethane. Utilizing the spin-trapping agent phenyl-t-butyl nitrone, a free radical has been detected which is produced from carbon tetrachloride or bromotrichloromethane during the enzymic oxidation of NADPH by rat liver microsomes. The presence of NADPH is obligatory for generation of the radical. The formation of the trichloromethyl radical-phenyl-t-butyl nitrone adduct is an enzymic process, as evidenced by the inhibition of its formation in systems containing heated microsomes and in systems containing p-hydroxymercuribenzoate. A computer-simulated ESR spectrum for the trichloromethyl adduct of phenyl-t-butyl nitrone can reproduce the essential features of the spectrum of the spin-trapped radical produced enzymically from CCl4. A mechanism is proposed for the formation of the trichloromethyl radical from CCl4 or BrCCl3. | [
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PMID:24482 | Mass spectrometry of medicines. Quantitative determination by direct probe inlet system mass chromatography. | A fundamental study was performed to establish a method of quantitative analysis using mass chromatography by the direct inlet system. The samples were aspirin, phenacetin and caffeine, which are often used as cold medicines, and barbital, allobarbital, phenobarbital and phenytoin which are difficult to analyse by gas chromatography in their intact states. N-acetylsulfamine and ethyl p-aminobenzoate were used as internal standards. Direct inlet mass chromatography was performed by an on-line system of the Shimadzu LKB-9000 and the GC-MSPAC 300. The ratio of the cumulative ions of certain peaks of sample and the internal standard was studied. It was found that, whether the sample is a pure reagent or a mixture, the ratio of cumulative ions of a peak specific to the sample and of a selected peak of the internal standard is proportional to the sample size, the error being less than +/- 3.5% for aspirin, phenacetin and caffeine, and less than +/- 2.7% for barbital, allobarbital and phenobarbital. The same relationship was observed for the phenobarbital and phenytoin mixed in rat plasma, the error being less than +/- 2.0%. It can be concluded that this method is applicable to the quantiative determination of medicines in urine, body fluids and other biological samples. | Mass spectrometry of medicines. Quantitative determination by direct probe inlet system mass chromatography. A fundamental study was performed to establish a method of quantitative analysis using mass chromatography by the direct inlet system. The samples were aspirin, phenacetin and caffeine, which are often used as cold medicines, and barbital, allobarbital, phenobarbital and phenytoin which are difficult to analyse by gas chromatography in their intact states. N-acetylsulfamine and ethyl p-aminobenzoate were used as internal standards. Direct inlet mass chromatography was performed by an on-line system of the Shimadzu LKB-9000 and the GC-MSPAC 300. The ratio of the cumulative ions of certain peaks of sample and the internal standard was studied. It was found that, whether the sample is a pure reagent or a mixture, the ratio of cumulative ions of a peak specific to the sample and of a selected peak of the internal standard is proportional to the sample size, the error being less than +/- 3.5% for aspirin, phenacetin and caffeine, and less than +/- 2.7% for barbital, allobarbital and phenobarbital. The same relationship was observed for the phenobarbital and phenytoin mixed in rat plasma, the error being less than +/- 2.0%. It can be concluded that this method is applicable to the quantiative determination of medicines in urine, body fluids and other biological samples. | [
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PMID:24483 | Continuous production of NADP by immobilized Achromobacter aceris cells. | Several microorganisms having higher nicotinamide adenine dinucleotide kinase (NAD kinase, EC 2.7.1.23) activity were immobilized into polyacrylamide gel lattices. The enzyme activity field by immobilization was highest in Achromobacter aceris AKU 0120. By the incubation of the immobilized A. aceris cells at pH 4.0, the NAD kinase activity increased and the adenosine triphosphate (ATP)-degradation activity disappeared completely. Enzymatic properties of the immobilized A. aceris cells were investigated and compared with those of intact cells. The optimal pH and the optimal temperature of immobilized cells were the same as those of intact cells. Immobilized cell NAD kinase was more stable than that of intact cells. The operational half-life of immobilized cells was 20 days when the substrate solution was passed through a column packed with immobilized cells at a flow rate which gives a space velocity (SV) of 0.1 hr-1 at 37 degrees C. On the other hand, the half-life of the intact cells was only 6 hr. | Continuous production of NADP by immobilized Achromobacter aceris cells. Several microorganisms having higher nicotinamide adenine dinucleotide kinase (NAD kinase, EC 2.7.1.23) activity were immobilized into polyacrylamide gel lattices. The enzyme activity field by immobilization was highest in Achromobacter aceris AKU 0120. By the incubation of the immobilized A. aceris cells at pH 4.0, the NAD kinase activity increased and the adenosine triphosphate (ATP)-degradation activity disappeared completely. Enzymatic properties of the immobilized A. aceris cells were investigated and compared with those of intact cells. The optimal pH and the optimal temperature of immobilized cells were the same as those of intact cells. Immobilized cell NAD kinase was more stable than that of intact cells. The operational half-life of immobilized cells was 20 days when the substrate solution was passed through a column packed with immobilized cells at a flow rate which gives a space velocity (SV) of 0.1 hr-1 at 37 degrees C. On the other hand, the half-life of the intact cells was only 6 hr. | [
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PMID:24484 | Mathematical modeling of lag phases in microbial growth. | This paper describes a mathematical method of the lap phases of Saccharomyces cerevisiae that incorporates the basic concepts previously presented in a two-stage deterministic model for the growth of this organism under conditions of oxygen excess with a sugar as the growth-limiting substrate. The model structure was suggested by an extensive investigation of the causes of the lap phases of S. cerevisiae which found that, in contrast to the traditionally accepted trends, the length of the lap phase was not inoculum-size dependent. This was consistent with other previously published work which suggested that a major factor in the length of the lag phases in S. cerevisiae was the need to synthesize adequate levels of glycolytic and respiratory enzymes. These suggestions were confirmed experimentally with lag-age data. Based on this conclusion a mathematical model was developed incorporating a description of the levels of glycolytic and respiratory enzymes and their effect on the growth rate and metabolism. This model was tested experimentally and the initial results indicate that many aspects of the lag phase of this organism may be described mathematically. The experimental findings further support the concept of primary regulatory control proposed by Bijkerk and Hall. | Mathematical modeling of lag phases in microbial growth. This paper describes a mathematical method of the lap phases of Saccharomyces cerevisiae that incorporates the basic concepts previously presented in a two-stage deterministic model for the growth of this organism under conditions of oxygen excess with a sugar as the growth-limiting substrate. The model structure was suggested by an extensive investigation of the causes of the lap phases of S. cerevisiae which found that, in contrast to the traditionally accepted trends, the length of the lap phase was not inoculum-size dependent. This was consistent with other previously published work which suggested that a major factor in the length of the lag phases in S. cerevisiae was the need to synthesize adequate levels of glycolytic and respiratory enzymes. These suggestions were confirmed experimentally with lag-age data. Based on this conclusion a mathematical model was developed incorporating a description of the levels of glycolytic and respiratory enzymes and their effect on the growth rate and metabolism. This model was tested experimentally and the initial results indicate that many aspects of the lag phase of this organism may be described mathematically. The experimental findings further support the concept of primary regulatory control proposed by Bijkerk and Hall. | [
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PMID:24487 | [Effect of narcotic analgesics on excitatory transmission in the spinal cord]. | As demonstrated on nonanesthetized curare-immobilized spinal cats morphine, promedol and fentanyl failed to alter the amplitude of induced potentials in the ventro-lateral columns of the lumbar spinal cord, evoked by a single or repetitive stimulation of the cutaneous or pelvic nerves. In some experiments the same drugs inhibited the nerurons of the posterior horns of the spinal cord activated by the nociceptive stimulation of the peripheral receptors in intraarterial administration of bradykinin. It is suggested that a spinal component was involved in the action of hypnotic analgetics. | [Effect of narcotic analgesics on excitatory transmission in the spinal cord]. As demonstrated on nonanesthetized curare-immobilized spinal cats morphine, promedol and fentanyl failed to alter the amplitude of induced potentials in the ventro-lateral columns of the lumbar spinal cord, evoked by a single or repetitive stimulation of the cutaneous or pelvic nerves. In some experiments the same drugs inhibited the nerurons of the posterior horns of the spinal cord activated by the nociceptive stimulation of the peripheral receptors in intraarterial administration of bradykinin. It is suggested that a spinal component was involved in the action of hypnotic analgetics. | [
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PMID:24489 | Exertion injuries in adolescent athletes. | A series of 147 cases of exertion injuries in less than or equal to 15 years old athletes is presented. All injuries occurred during training or athletic performances without trauma and caused symptoms that prevented athletic exercises. There were 67 girls (46%) and 80 boys (54%) in the material. About 90% of them had been training for more than one year before the onset of the symptoms; 65% were interested in track and field athletics, 13% in ball games, 11% in skiing, 4% in swimming, and 3% in orienteering. The rest were interested in other sports. About 33% of the injuries were growth disturbances or osteochondroses seen also in other children. About 15% were anomalies, deformities or earlier osteochondritic changes, which caused first symptoms during the physical exercise; 50% were typical overuse injuries that may bother adult athletes, too; 43% of the injuries were localized in ankle, foot and heel, 31% in knee, 8% in back and trunk, 7% in pelvic and hip region, and the rest in other parts of the body. The injuries were generally slight, no permanent disability was noticed. Rest and conservation therapy cured most cases; operative treatment was used in only eight cases. | Exertion injuries in adolescent athletes. A series of 147 cases of exertion injuries in less than or equal to 15 years old athletes is presented. All injuries occurred during training or athletic performances without trauma and caused symptoms that prevented athletic exercises. There were 67 girls (46%) and 80 boys (54%) in the material. About 90% of them had been training for more than one year before the onset of the symptoms; 65% were interested in track and field athletics, 13% in ball games, 11% in skiing, 4% in swimming, and 3% in orienteering. The rest were interested in other sports. About 33% of the injuries were growth disturbances or osteochondroses seen also in other children. About 15% were anomalies, deformities or earlier osteochondritic changes, which caused first symptoms during the physical exercise; 50% were typical overuse injuries that may bother adult athletes, too; 43% of the injuries were localized in ankle, foot and heel, 31% in knee, 8% in back and trunk, 7% in pelvic and hip region, and the rest in other parts of the body. The injuries were generally slight, no permanent disability was noticed. Rest and conservation therapy cured most cases; operative treatment was used in only eight cases. | [
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PMID:24490 | Perfusion characteristics of preserved canine kidneys subjected to warm ischaemia. | Canine kidneys were subjected to 0, 15 or 30 min of warm ischaemia followed by 24 hours preservation by perfusion. Changes in perfusate concentration of acid radicles, lactate, free fatty acid and lactice dehydrogenase were assessed at 1 hour and 24 hours. With the exception of LDH concentration at 1 hour, no single parameter was capable of detecting kidneys which were so damaged as to be non-life supporting. | Perfusion characteristics of preserved canine kidneys subjected to warm ischaemia. Canine kidneys were subjected to 0, 15 or 30 min of warm ischaemia followed by 24 hours preservation by perfusion. Changes in perfusate concentration of acid radicles, lactate, free fatty acid and lactice dehydrogenase were assessed at 1 hour and 24 hours. With the exception of LDH concentration at 1 hour, no single parameter was capable of detecting kidneys which were so damaged as to be non-life supporting. | [
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PMID:24493 | Effect of concanavalin A on black widow spider venom activity at the neuromuscular junction: implications for mechanisms of venom action. | Concanavalin A (Con A) inhibits black widow spider venom-induced transmitter release at both tissue-cultured and adult neuromuscular junctions and also inhibits the venom-induced destruction of cultured neurites. This inhibitory action is partially or completely prevented by prior treatment with colchicine. Neither colchicine nor Con A interacts significantly with depolarization-induced transmitter release. These results are analogous to those obtained from experiments on lymphocyte surface receptor capping. They suggest that redistribution of neuronal membrane components may be a crucial step in spider venom action. This membrane redistribution appears to be modulated in neurons, as in other cell types, by microtubule-microfilament array. How such a redistribution causes increased transmitter release cannot as yet be specified. Changes in the ionic permeability of sodium and potassium were examined as likely mechanisms. Increased sodium influx (and seondary release of calcium from intracellular stores) cannot be the basis for spider venom action. Increased potassium efflux remains a possibility, but is not consistent with all of the data. Other possible mechanisms are also suggested. | Effect of concanavalin A on black widow spider venom activity at the neuromuscular junction: implications for mechanisms of venom action. Concanavalin A (Con A) inhibits black widow spider venom-induced transmitter release at both tissue-cultured and adult neuromuscular junctions and also inhibits the venom-induced destruction of cultured neurites. This inhibitory action is partially or completely prevented by prior treatment with colchicine. Neither colchicine nor Con A interacts significantly with depolarization-induced transmitter release. These results are analogous to those obtained from experiments on lymphocyte surface receptor capping. They suggest that redistribution of neuronal membrane components may be a crucial step in spider venom action. This membrane redistribution appears to be modulated in neurons, as in other cell types, by microtubule-microfilament array. How such a redistribution causes increased transmitter release cannot as yet be specified. Changes in the ionic permeability of sodium and potassium were examined as likely mechanisms. Increased sodium influx (and seondary release of calcium from intracellular stores) cannot be the basis for spider venom action. Increased potassium efflux remains a possibility, but is not consistent with all of the data. Other possible mechanisms are also suggested. | [
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PMID:24498 | Quantitative electron miscroscopic studies on the kinetics of secretory granules in G-cells. | Ultrastructural studies of secretory granules of rat antral G-cells and measurement of serum gastrin level were performed under the condition of fasting and administration of alkaline solution into the stomach. On electron micrographs, no qualitative difference was observed among those experimental groups. However, morphometrical analysis revealed significant quantitative differences. The population density of secretory granules of the rats treated once with alkali first increased and then decreased reaching that of the fasted group, while that of the repeatedly treated group remained nearly equal to the maximum value. The average sectioned surface area of secretory granules tended to decrease for 1.5 h after the stimulation but the differences was not significant among those groups. From the results obtained at present, responding to chemical stimulation such as pH changes in the antrum, it seems probable that not only exocytosis but also migration of secretory granules from supra- and/or para-nuclear portion to the basal portion of the cell occurs rapidly in G-cells and that both these processes are inhibited immediately by antral acidification. Moreover, the present results apparently indicate that under the condition of no antral acidification G-cells have a capacity of secreting gastrin for a fairly long time, such as 4--8 h, responding to adequate stimulus. These findings are strongly suggestive of the existence of a capacious pool of granules in the supra- and/or para-nuclear cytoplasm or of fairly speedy production of secretory granules in the Golgi area. | Quantitative electron miscroscopic studies on the kinetics of secretory granules in G-cells. Ultrastructural studies of secretory granules of rat antral G-cells and measurement of serum gastrin level were performed under the condition of fasting and administration of alkaline solution into the stomach. On electron micrographs, no qualitative difference was observed among those experimental groups. However, morphometrical analysis revealed significant quantitative differences. The population density of secretory granules of the rats treated once with alkali first increased and then decreased reaching that of the fasted group, while that of the repeatedly treated group remained nearly equal to the maximum value. The average sectioned surface area of secretory granules tended to decrease for 1.5 h after the stimulation but the differences was not significant among those groups. From the results obtained at present, responding to chemical stimulation such as pH changes in the antrum, it seems probable that not only exocytosis but also migration of secretory granules from supra- and/or para-nuclear portion to the basal portion of the cell occurs rapidly in G-cells and that both these processes are inhibited immediately by antral acidification. Moreover, the present results apparently indicate that under the condition of no antral acidification G-cells have a capacity of secreting gastrin for a fairly long time, such as 4--8 h, responding to adequate stimulus. These findings are strongly suggestive of the existence of a capacious pool of granules in the supra- and/or para-nuclear cytoplasm or of fairly speedy production of secretory granules in the Golgi area. | [
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PMID:24499 | Relaxin, a male hormone? Immunocytological localization of a related antigen in the boar testis. | Convincingly demonstrated by immunocytological methods in females of several mammalian species, relaxin has not yet been localized in the male. Immunocytologically, a related antigen was identified in adult normal boar testes using and anti- [NIH P-relaxin/HSA] antiserum free of anti HSA Abs. A strong reaction was observed in interstitial cells, a weaker but very clear one in Sertoli cells. NIH P-relaxin and HC1-acetone extracts of either corpora lutea from pregnant sows or boar testes inhibited the immunofluorescence of the reactive structures in the boar testes as well as in ovaries of pregnant sows. Ethanol-acetone precipitates from boar rete testis or caudal epididymal fluids inhibited the reaction of interstitial and Sertoli cells, but this inhibition in the sow was limited only to degenerative ovarian structures, probably due to an insufficient level of inhibiting antigen in these two seminal fluids, in contrast with the very high concentration of relaxin in luteal cells of pregnant sows. Specific immunofluorescence was observed neither in ectopic testes of adult monocryptorchid boars (contrary to scrotal testes in these same animals) nor in testes of prepuberal pigs. The specificity and meaning of these results are discussed. | Relaxin, a male hormone? Immunocytological localization of a related antigen in the boar testis. Convincingly demonstrated by immunocytological methods in females of several mammalian species, relaxin has not yet been localized in the male. Immunocytologically, a related antigen was identified in adult normal boar testes using and anti- [NIH P-relaxin/HSA] antiserum free of anti HSA Abs. A strong reaction was observed in interstitial cells, a weaker but very clear one in Sertoli cells. NIH P-relaxin and HC1-acetone extracts of either corpora lutea from pregnant sows or boar testes inhibited the immunofluorescence of the reactive structures in the boar testes as well as in ovaries of pregnant sows. Ethanol-acetone precipitates from boar rete testis or caudal epididymal fluids inhibited the reaction of interstitial and Sertoli cells, but this inhibition in the sow was limited only to degenerative ovarian structures, probably due to an insufficient level of inhibiting antigen in these two seminal fluids, in contrast with the very high concentration of relaxin in luteal cells of pregnant sows. Specific immunofluorescence was observed neither in ectopic testes of adult monocryptorchid boars (contrary to scrotal testes in these same animals) nor in testes of prepuberal pigs. The specificity and meaning of these results are discussed. | [
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PMID:24501 | In vitro studies on the effects on granular gland secretion in Xenopus laevis skin of stimulation and blockade of alpha and beta adrenoceptors of myoepithelial cells. | Secretion from the granular glands of Xenopus laevis skin was stimulated by alpha-adrenergic agonists, an effect which was blocked by alpha-adrenergic antagonists and inhibited by beta-adrenergic agonists, db-cAMP and diazoxide. The inhibition by isoprenaline and salbutamol, but not that by diazoxide, was blocked by a beta-adrenergic antagonist. It is concluded that the myoepithelial cells surrounding the secretory compartment of the granular glands bear alpha and beta adrenoceptors, and that the beta receptors comprise, or at least include beta2 receptors. | In vitro studies on the effects on granular gland secretion in Xenopus laevis skin of stimulation and blockade of alpha and beta adrenoceptors of myoepithelial cells. Secretion from the granular glands of Xenopus laevis skin was stimulated by alpha-adrenergic agonists, an effect which was blocked by alpha-adrenergic antagonists and inhibited by beta-adrenergic agonists, db-cAMP and diazoxide. The inhibition by isoprenaline and salbutamol, but not that by diazoxide, was blocked by a beta-adrenergic antagonist. It is concluded that the myoepithelial cells surrounding the secretory compartment of the granular glands bear alpha and beta adrenoceptors, and that the beta receptors comprise, or at least include beta2 receptors. | [
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PMID:24503 | The mutagenic effect of 1,2-dichloroethane on Salmonella typhimurium I. Activation through conjugation with glutathion in vitro. | One of the main components in the waste products from vinyl chloride industries (EDC-tar), is ethylene dichloride (1,2-dichloroethane). This compound has been tested for mutagenicity on Salmonella typhimurium TA 1535. It is concluded that 1,2-dichloroethane gives a weak direct mutagenic effect, which is enhanced by addition of the postmitochondrial liver fraction (S-9). This activation is NADPH-independent and non microsomal. It is caused by a factor in the soluble fraction (115 000 g supernatant). This activation was further enhanced by the addition of glutathione but not by the addition of L-cysteine, N-acetyl-L-cysteine or 2-mercaptoethanol. No activation was observed when glutathione was added in the presence of a totally denaturated S-9 fraction or in the absence of this fraction. Activation of 1,2-dichloroethane was also found in the presence of glutathione and glutathione S-transferase A and C but not with glutathione S-tranferase B. A synthetic conjugate S-(2-chloroethyl)-L-cysteine gave a strong direct mutagenic effect at concentrations where no effects were seen with 1,2-dichloroethane. It is thus concluded that 1,2-dichloroethane is activated by conjugation to glutathione. Another main component in EDC-tar, 1,1,2-trichloroethane, was not mutagenic under any of our experimental conditions. For comparison 1,2-dibromoethane was also tested and gave a stronger direct mutagenic effect than 1,2-dichloroethane. Like the latter 1,2-dibromoethane was also activated by a NADPH-independent process. | The mutagenic effect of 1,2-dichloroethane on Salmonella typhimurium I. Activation through conjugation with glutathion in vitro. One of the main components in the waste products from vinyl chloride industries (EDC-tar), is ethylene dichloride (1,2-dichloroethane). This compound has been tested for mutagenicity on Salmonella typhimurium TA 1535. It is concluded that 1,2-dichloroethane gives a weak direct mutagenic effect, which is enhanced by addition of the postmitochondrial liver fraction (S-9). This activation is NADPH-independent and non microsomal. It is caused by a factor in the soluble fraction (115 000 g supernatant). This activation was further enhanced by the addition of glutathione but not by the addition of L-cysteine, N-acetyl-L-cysteine or 2-mercaptoethanol. No activation was observed when glutathione was added in the presence of a totally denaturated S-9 fraction or in the absence of this fraction. Activation of 1,2-dichloroethane was also found in the presence of glutathione and glutathione S-transferase A and C but not with glutathione S-tranferase B. A synthetic conjugate S-(2-chloroethyl)-L-cysteine gave a strong direct mutagenic effect at concentrations where no effects were seen with 1,2-dichloroethane. It is thus concluded that 1,2-dichloroethane is activated by conjugation to glutathione. Another main component in EDC-tar, 1,1,2-trichloroethane, was not mutagenic under any of our experimental conditions. For comparison 1,2-dibromoethane was also tested and gave a stronger direct mutagenic effect than 1,2-dichloroethane. Like the latter 1,2-dibromoethane was also activated by a NADPH-independent process. | [
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PMID:24505 | Neurogenic vasodilation of cat cerebral arteries. | Transmural nerve stimulation (TNS) with 0.3-msec pulses between 1 and 25 Hz dilated cat cerebral artery segments in the presence of active muscle tone. Maximum vasodilatation occurred at 8 Hz. The dilator response to exogenous acetylcholine, but not to TNS, was abolished by atropine. Neither physostigmine nor hemicholinium affected the dilator response to TNS, which persisted after administration of guanethidine, phenoxybenzamine, propranolol, reserpine, and chronic sympathectomy. However, it was abolished by tetrodotoxin and cold storage. When examined histochemically, cat and rabbit cerebral arteries exhibited a rich plexiform distribution of acetylcholinesterase which was not affected appreciably by sympathetic denervation. These results suggest that vasodilation is not mediated through modification of sympathetic activity. They also indicate the existence of a nonadrenergic, possibly noncholinergic, vasodilator innervation in cat cerebral arteries. Preliminary studies suggest that the transmitter is not histamine, ATP, prostaglandins, gamma-aminobutyric acid, dopamine, or serotonin. The cat cerebral artery segments contrast with the isolated rabbit cerebral arteries which predominantly constrict in response to TNS and show a small dilator response. | Neurogenic vasodilation of cat cerebral arteries. Transmural nerve stimulation (TNS) with 0.3-msec pulses between 1 and 25 Hz dilated cat cerebral artery segments in the presence of active muscle tone. Maximum vasodilatation occurred at 8 Hz. The dilator response to exogenous acetylcholine, but not to TNS, was abolished by atropine. Neither physostigmine nor hemicholinium affected the dilator response to TNS, which persisted after administration of guanethidine, phenoxybenzamine, propranolol, reserpine, and chronic sympathectomy. However, it was abolished by tetrodotoxin and cold storage. When examined histochemically, cat and rabbit cerebral arteries exhibited a rich plexiform distribution of acetylcholinesterase which was not affected appreciably by sympathetic denervation. These results suggest that vasodilation is not mediated through modification of sympathetic activity. They also indicate the existence of a nonadrenergic, possibly noncholinergic, vasodilator innervation in cat cerebral arteries. Preliminary studies suggest that the transmitter is not histamine, ATP, prostaglandins, gamma-aminobutyric acid, dopamine, or serotonin. The cat cerebral artery segments contrast with the isolated rabbit cerebral arteries which predominantly constrict in response to TNS and show a small dilator response. | [
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PMID:24506 | An investigation of factors influencing the measurement of creatine phosphokinase activity in serum using coupled enzymatic methods. | 1. The factors influencing the measurement of creatine phosphokinase (CPK) activity in serum by coupled enzymatic methods were investigated to establish optimum conditions for this type of assay. Such a study was indicated following observations by the authors of poor performance of commerically produced reagent kits together with the failure of most of the established an well accepted methods to operate under true optimum zero order kinetics in the reaction phase state. 2. The factors invested were the effects of pH, substrate concentrations (creatine phosphate, glucose and NADP+), added auxiliary (hexokinase) and indicator (glucose-6-phosphate dehydrogenase) enzymes, dithiothreitol (DTT) as an activator and conditions of storage of substrate stability. DTT was found to be a suitable activator but not a reactivator of the reaction. The optimum concentrations of creatine phosphate, glucose and NADP+ were found to be 20.0, 20.0 and 2.0 mmol/litre, respectively. Optimum activieies of the enzymes, glucose-6-phosphate dehydrosenase and hexokinase were 1000 and 2000 units/litre, respectively. 3. The between-day precision of the method for measuring serum at pH 6.8 and 30 degrees C at three activity levels under the optimum conditions developed was excellent yielding coefficients of variation ranging from 2.0 to 2.7%. | An investigation of factors influencing the measurement of creatine phosphokinase activity in serum using coupled enzymatic methods. 1. The factors influencing the measurement of creatine phosphokinase (CPK) activity in serum by coupled enzymatic methods were investigated to establish optimum conditions for this type of assay. Such a study was indicated following observations by the authors of poor performance of commerically produced reagent kits together with the failure of most of the established an well accepted methods to operate under true optimum zero order kinetics in the reaction phase state. 2. The factors invested were the effects of pH, substrate concentrations (creatine phosphate, glucose and NADP+), added auxiliary (hexokinase) and indicator (glucose-6-phosphate dehydrogenase) enzymes, dithiothreitol (DTT) as an activator and conditions of storage of substrate stability. DTT was found to be a suitable activator but not a reactivator of the reaction. The optimum concentrations of creatine phosphate, glucose and NADP+ were found to be 20.0, 20.0 and 2.0 mmol/litre, respectively. Optimum activieies of the enzymes, glucose-6-phosphate dehydrosenase and hexokinase were 1000 and 2000 units/litre, respectively. 3. The between-day precision of the method for measuring serum at pH 6.8 and 30 degrees C at three activity levels under the optimum conditions developed was excellent yielding coefficients of variation ranging from 2.0 to 2.7%. | [
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PMID:24507 | Improved method for fractionating gamma-glutamyltransferase by electrophoresis on cellulose acetate. | I describe a method of fractionating gamma-glutamyltransferase on cellulose acetate. The tracing is obtained in parallel with that of serum protein, and the gamma-glutamyltransferase bands are characterized by correspondence with the major protein fractions. The overall pattern of the isoenzyme activity in normal sera is one of activity in the alpha1- and alpha2-globulin regions. In hepatic diseases four bands are usually present, but some more specific observations are possible, e.g., the presence of an intense beta-globulin band in occlusive icterus, intra- or extrahepatic, and a marked alpha2-globulin band in alcoholism. The potentialities of this technique as a diagnostic and prognostic aid together with its simplicity prompt me to recommend its use in the clinical laboratory. | Improved method for fractionating gamma-glutamyltransferase by electrophoresis on cellulose acetate. I describe a method of fractionating gamma-glutamyltransferase on cellulose acetate. The tracing is obtained in parallel with that of serum protein, and the gamma-glutamyltransferase bands are characterized by correspondence with the major protein fractions. The overall pattern of the isoenzyme activity in normal sera is one of activity in the alpha1- and alpha2-globulin regions. In hepatic diseases four bands are usually present, but some more specific observations are possible, e.g., the presence of an intense beta-globulin band in occlusive icterus, intra- or extrahepatic, and a marked alpha2-globulin band in alcoholism. The potentialities of this technique as a diagnostic and prognostic aid together with its simplicity prompt me to recommend its use in the clinical laboratory. | [
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PMID:24512 | Correlation between plasma diphenhydramine level and sedative and antihistamine effects. | The sedative and antihistamine effects of diphenhydramine were assessed in relation to plasma concentration after placebo, diphenhydramine 50 mg intravenously, and diphenhydramine 50 mg orally to each of 6 healthy volunteers on three separate occasions. Diphenhydramine plasma elimination t1/2 was 3.0 to 4.3 hr, volume of distribution was 188 to 336 L, and clearance was 637 to 1,014 ml/min. Systemic bioavailability of the oral preparation ranged from 0.26 to 0.60. The sedative effect of intravenous diphenhydramine differed from that of placebo only during the first 3 hr. Antihistamine effect, as measured by reduction of histamine provoked skin wheal diameter, was significantly different from that of placebo for at least 8 hr. There was a positive correlation between plasma diphenhydramine level and sedative and antihistamine effects, but wide variation in the extent and rate of change of these effects were observed between the subject. There appears to be a concentration range of 25 to 50 ng/ml, within which there is significant antihistamine effect without significant sedation. | Correlation between plasma diphenhydramine level and sedative and antihistamine effects. The sedative and antihistamine effects of diphenhydramine were assessed in relation to plasma concentration after placebo, diphenhydramine 50 mg intravenously, and diphenhydramine 50 mg orally to each of 6 healthy volunteers on three separate occasions. Diphenhydramine plasma elimination t1/2 was 3.0 to 4.3 hr, volume of distribution was 188 to 336 L, and clearance was 637 to 1,014 ml/min. Systemic bioavailability of the oral preparation ranged from 0.26 to 0.60. The sedative effect of intravenous diphenhydramine differed from that of placebo only during the first 3 hr. Antihistamine effect, as measured by reduction of histamine provoked skin wheal diameter, was significantly different from that of placebo for at least 8 hr. There was a positive correlation between plasma diphenhydramine level and sedative and antihistamine effects, but wide variation in the extent and rate of change of these effects were observed between the subject. There appears to be a concentration range of 25 to 50 ng/ml, within which there is significant antihistamine effect without significant sedation. | [
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] |
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