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PMID:13654 | Studies of glucose turnover and renal function in an unusual case of hereditary fructose intolerance. | Examination of glucose kinetics, pancreatic alpha and beta cell function, plasma lipids, urinary acidification and calcium excretion has been undertaken in a patient with hereditary fructose intolerance. This case was unusual as it was associated with insulin-requiring diabetes, type IV hyperlipemia, hypercalciuria and renal calculi. He also demonstrated the previously described fructose-induced defect of urine acidification. Glucagon and C-peptide assays showed that the pancreatic alpha cells were stimulated by fructose and that the beta cells did not respond to fructose. It is not known whether the latter was due to his diabetes or to the lack of a beta cell response to this sugar. Primed 14C-glucose infusions were used for the first time to study nonsteady state glucose kinetics in man. They showed that, 24 hours after the last insulin injection and under basal conditions, the glucose concentrations increased because glucose production exceeded glucose utilization. However, after the administration of sorbitol the plasma glucose concentration decreased because glucose production decreased. After the administration of sorbitol there was no change in the metabolic clearance of glucose. This reflects the lack of a peripheral insulin effect and is consistent with the lack of any measurable C-peptide. Glucose utilization also decreased, but this decrease was less than the decrease in glucose production. Because the metabolic clearance of glucose remained unchanged, it was concluded that the change in glucose utilization was solely due to the decrease in glucose concentration. The absence of C-peptide in the plasma indicated that changes in glucose turnover were not related to any changes in endogenous plasma insulin. Furthermore, the plasma glucagon concentration increased and, hence, changes in this hormone could not account for the decrease in glucose production. Therefore, it was concluded that the sorbitol-induced decline in glucose production was due to a direct effect on hepatic metabolism. | Studies of glucose turnover and renal function in an unusual case of hereditary fructose intolerance. Examination of glucose kinetics, pancreatic alpha and beta cell function, plasma lipids, urinary acidification and calcium excretion has been undertaken in a patient with hereditary fructose intolerance. This case was unusual as it was associated with insulin-requiring diabetes, type IV hyperlipemia, hypercalciuria and renal calculi. He also demonstrated the previously described fructose-induced defect of urine acidification. Glucagon and C-peptide assays showed that the pancreatic alpha cells were stimulated by fructose and that the beta cells did not respond to fructose. It is not known whether the latter was due to his diabetes or to the lack of a beta cell response to this sugar. Primed 14C-glucose infusions were used for the first time to study nonsteady state glucose kinetics in man. They showed that, 24 hours after the last insulin injection and under basal conditions, the glucose concentrations increased because glucose production exceeded glucose utilization. However, after the administration of sorbitol the plasma glucose concentration decreased because glucose production decreased. After the administration of sorbitol there was no change in the metabolic clearance of glucose. This reflects the lack of a peripheral insulin effect and is consistent with the lack of any measurable C-peptide. Glucose utilization also decreased, but this decrease was less than the decrease in glucose production. Because the metabolic clearance of glucose remained unchanged, it was concluded that the change in glucose utilization was solely due to the decrease in glucose concentration. The absence of C-peptide in the plasma indicated that changes in glucose turnover were not related to any changes in endogenous plasma insulin. Furthermore, the plasma glucagon concentration increased and, hence, changes in this hormone could not account for the decrease in glucose production. Therefore, it was concluded that the sorbitol-induced decline in glucose production was due to a direct effect on hepatic metabolism. | [
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PMID:13656 | Evaluation of success in treatment of threatening premature labor by betamimetic drugs. | For prevention of premature labor the betasympathicomimetic drugs buphenin, fenoterol, and ritodrine were administered to 135 patients intravenously for 1 week and afterward orally up to the end of week 37 of gestation. No satisfactory evaluation of success could be obtained by analyzing only the time gained by the treatment. Two supplemental parameters were needed. In addition to the previously introduced Tocolysis Index (TI) which describes the obstetric situation at the beginning of the treatment, a Prolongation Index (PI) was defined transforming the lag time gained by the treatment into a relative measure. By means of this PI, extremely differing cases could be compared, though the gestational ages at the start of treatment and the lag times between admission and delivery showed wide variation. In addition, satisfactory statistical correlations between the PI and the clinical signs as well as the neonatal outcome could be computed, indicating the PI to be a suitable parameter for the estimation of success. | Evaluation of success in treatment of threatening premature labor by betamimetic drugs. For prevention of premature labor the betasympathicomimetic drugs buphenin, fenoterol, and ritodrine were administered to 135 patients intravenously for 1 week and afterward orally up to the end of week 37 of gestation. No satisfactory evaluation of success could be obtained by analyzing only the time gained by the treatment. Two supplemental parameters were needed. In addition to the previously introduced Tocolysis Index (TI) which describes the obstetric situation at the beginning of the treatment, a Prolongation Index (PI) was defined transforming the lag time gained by the treatment into a relative measure. By means of this PI, extremely differing cases could be compared, though the gestational ages at the start of treatment and the lag times between admission and delivery showed wide variation. In addition, satisfactory statistical correlations between the PI and the clinical signs as well as the neonatal outcome could be computed, indicating the PI to be a suitable parameter for the estimation of success. | [
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|
PMID:13657 | Fetal breathing and adaptation to maternal hemorrhage in the sheep. | The effect of maternal hemorrhage in chronic preparations was studied on fetal lambs in the last month of gestation. Fourteen to 20 per cent of maternal blood was estimated to have been removed within 30 minutes, which resulted in a drop of 30 per cent of mean maternal arterial pressure. A fetal bradycardia started 28 +/- 13 minutes after the beginning of maternal hemorrhage. It lasted 30 +/- 15 minutes and was concomitant with a rise in fetal arterial pressure. It was followed by a long-lasting fetal tachycardia of 130 +/- 38 minutes and was corrected only by reinfusion of blood to the mother. The fetal blood gases demonstrated a mild asphyxia with a persistent metabolic acidemia until reinfusion of blood to the mother. Maternal and fetal plasma cortisol levels rose significantly at the end of the hemorrhage. Tracheal fluid flow did not change. Fetal breathing recorded 20 hours before and 24 hours after the experiment did not show consistent changes, but during fetal bradycardia there was no fetal breathing. Recent clinical investigations in this field have been made in the human fetus to estimate standards of fetal well being. These peculiar animal experiments do not show any significant improvement by recording fetal breathing over the recording of prelabor fetal heart rate. | Fetal breathing and adaptation to maternal hemorrhage in the sheep. The effect of maternal hemorrhage in chronic preparations was studied on fetal lambs in the last month of gestation. Fourteen to 20 per cent of maternal blood was estimated to have been removed within 30 minutes, which resulted in a drop of 30 per cent of mean maternal arterial pressure. A fetal bradycardia started 28 +/- 13 minutes after the beginning of maternal hemorrhage. It lasted 30 +/- 15 minutes and was concomitant with a rise in fetal arterial pressure. It was followed by a long-lasting fetal tachycardia of 130 +/- 38 minutes and was corrected only by reinfusion of blood to the mother. The fetal blood gases demonstrated a mild asphyxia with a persistent metabolic acidemia until reinfusion of blood to the mother. Maternal and fetal plasma cortisol levels rose significantly at the end of the hemorrhage. Tracheal fluid flow did not change. Fetal breathing recorded 20 hours before and 24 hours after the experiment did not show consistent changes, but during fetal bradycardia there was no fetal breathing. Recent clinical investigations in this field have been made in the human fetus to estimate standards of fetal well being. These peculiar animal experiments do not show any significant improvement by recording fetal breathing over the recording of prelabor fetal heart rate. | [
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|
PMID:13658 | Secretory surface area and phosphatase activity of frog gastric mucosa. | Gastric mucosae were isolated from the European frog, R. temporaria, and after 5-7 h, stimulated with histamine (10(-4) M) and theophylline (5 mM). Acid secretion increased about fourfold, and this was accompanied by a threefold increase in secretory surface of the oxyntic cells, as determined in electron micrographs with conventional morphometric techniques. At the same time phosphatase activity of the secretory surface increased. Other experiments showed that the latter was due to an acid phosphatase, with pH optimum near 3. It appears that the increase in surface phosphatase with stimulation can be attributed to a diminished local pH and not to the increase in surface area demonstrated in this study. | Secretory surface area and phosphatase activity of frog gastric mucosa. Gastric mucosae were isolated from the European frog, R. temporaria, and after 5-7 h, stimulated with histamine (10(-4) M) and theophylline (5 mM). Acid secretion increased about fourfold, and this was accompanied by a threefold increase in secretory surface of the oxyntic cells, as determined in electron micrographs with conventional morphometric techniques. At the same time phosphatase activity of the secretory surface increased. Other experiments showed that the latter was due to an acid phosphatase, with pH optimum near 3. It appears that the increase in surface phosphatase with stimulation can be attributed to a diminished local pH and not to the increase in surface area demonstrated in this study. | [
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|
PMID:13659 | HCO3 transport in rat jejunum: relationship to NaCl and H2O transport in vivo. | HCO3- absorption and its association with Na+ absorption has been studied in the rat jejunum in vivo, using a single-pass perfusion technique. The method of disequilibrium pH, the only valid way of demonstrating jejunal H+ secretion, was used to distinguish between an HCO3- pump and H+ secretion as the mechanism of HCO3- absorption. HCO3- stimulated Na+ absorption; Na+ deletion inhibited HCO3 absorption, decreased luminal acidification, and decreased the level of luminal PCO2. These results confirmed an Na+:H+ cation exchange, the possible mechanism of which is discussed in terms of results using other tissues. Na+-dependent HCO3-absorption made up a larger part of total HCO3-absorption as the luminal HCO3-concentrations diminished, although the precise degree of Na+-dependency could not be determined because of the unstirred layer effect. The mechanism of Na+-independent HCO3-absorption was not established, but it was not affected by PD, Cl-, or H2O movements. Glucose-stimulated and HCO3-stimulated Na+ absorption were less than additive. The physiological importance of HCO3-stimulated Na+ absorption in the acidic postprandial jejunum is probably due entirely to the effect of free CO2 in the lumen. | HCO3 transport in rat jejunum: relationship to NaCl and H2O transport in vivo. HCO3- absorption and its association with Na+ absorption has been studied in the rat jejunum in vivo, using a single-pass perfusion technique. The method of disequilibrium pH, the only valid way of demonstrating jejunal H+ secretion, was used to distinguish between an HCO3- pump and H+ secretion as the mechanism of HCO3- absorption. HCO3- stimulated Na+ absorption; Na+ deletion inhibited HCO3 absorption, decreased luminal acidification, and decreased the level of luminal PCO2. These results confirmed an Na+:H+ cation exchange, the possible mechanism of which is discussed in terms of results using other tissues. Na+-dependent HCO3-absorption made up a larger part of total HCO3-absorption as the luminal HCO3-concentrations diminished, although the precise degree of Na+-dependency could not be determined because of the unstirred layer effect. The mechanism of Na+-independent HCO3-absorption was not established, but it was not affected by PD, Cl-, or H2O movements. Glucose-stimulated and HCO3-stimulated Na+ absorption were less than additive. The physiological importance of HCO3-stimulated Na+ absorption in the acidic postprandial jejunum is probably due entirely to the effect of free CO2 in the lumen. | [
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|
PMID:13660 | Vitamin K1 intestinal absorption in vivo: influence of luminal contents on transport. | Intestinal absorption of [3H]phylloquinone was investigated in the unanesthetized rat by the use of a technique of recirculating perfused isolated intestinal segments. Apparent saturation kinetics were found as the concentration of the vitamin in the perfusate was increased in a stepwise fashion from 15 nM to 300 muM. Alkalinization of the perfusate or the addition of 2.5 mM linoleic acid to the perfusate caused a significant (P less than 0.05) decrease in the absorption rate of phylloquinone. Modifications in the perfusate concentration of sodium taurocholate, the substitution of a nonionic detergent (Pluronic F-68) for sodium taurocholate, the addition of medium- and long-chain saturated fatty acids, or the addition of vitamins K2 and K3 to the perfusate did not alter the absorption rate of the vitamin. Decreasing the thickness of the unstirred water layer by increasing the perfusion rate caused a significant increase in phylloquinone absorption rate. In vivo absorption of vitamin K1 appears to be mediated by an energy requiring saturable transport mechanism. The composition of the perfusate, its pH, and its rate of flow are all important determinants of vitamin K1 absorption rate. | Vitamin K1 intestinal absorption in vivo: influence of luminal contents on transport. Intestinal absorption of [3H]phylloquinone was investigated in the unanesthetized rat by the use of a technique of recirculating perfused isolated intestinal segments. Apparent saturation kinetics were found as the concentration of the vitamin in the perfusate was increased in a stepwise fashion from 15 nM to 300 muM. Alkalinization of the perfusate or the addition of 2.5 mM linoleic acid to the perfusate caused a significant (P less than 0.05) decrease in the absorption rate of phylloquinone. Modifications in the perfusate concentration of sodium taurocholate, the substitution of a nonionic detergent (Pluronic F-68) for sodium taurocholate, the addition of medium- and long-chain saturated fatty acids, or the addition of vitamins K2 and K3 to the perfusate did not alter the absorption rate of the vitamin. Decreasing the thickness of the unstirred water layer by increasing the perfusion rate caused a significant increase in phylloquinone absorption rate. In vivo absorption of vitamin K1 appears to be mediated by an energy requiring saturable transport mechanism. The composition of the perfusate, its pH, and its rate of flow are all important determinants of vitamin K1 absorption rate. | [
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|
PMID:13661 | Fluoride absorption from the rat urinary bladder: a pH-dependent event. | Urinary bladder absorption of stable and radiofluoride was studied as a function of pH in anesthetized rats to further evaluate the influence of pH gradients on fluoride transport. Buffered pH values and stable fluoride concentrations ranged from 1.85 to 7.90 and from 0.012 to 8.81 mM, respectively. [14c]inulin served as a marker for solute concentration changes due to water migration or dilution. The results indicate that bladder fluoride absorption is inversely related to pH over the 1.85-5.50 range. Mean, 15-min radiofluoride absorption values of 70% at pH 1,85, 37% at pH 3.95, and 5% at pH 5.50 WERE OBSERVED. These fractional absorption values were not significantly influenced by carrier fluoride concentration, the buffers used, or the presence of urine. Above pH 5.50, pH-independent absorption occurs to a slight extent. The results are consistent with a first-order absorptive process which occurs by the nonionic diffusion of hydrogen fluoride. | Fluoride absorption from the rat urinary bladder: a pH-dependent event. Urinary bladder absorption of stable and radiofluoride was studied as a function of pH in anesthetized rats to further evaluate the influence of pH gradients on fluoride transport. Buffered pH values and stable fluoride concentrations ranged from 1.85 to 7.90 and from 0.012 to 8.81 mM, respectively. [14c]inulin served as a marker for solute concentration changes due to water migration or dilution. The results indicate that bladder fluoride absorption is inversely related to pH over the 1.85-5.50 range. Mean, 15-min radiofluoride absorption values of 70% at pH 1,85, 37% at pH 3.95, and 5% at pH 5.50 WERE OBSERVED. These fractional absorption values were not significantly influenced by carrier fluoride concentration, the buffers used, or the presence of urine. Above pH 5.50, pH-independent absorption occurs to a slight extent. The results are consistent with a first-order absorptive process which occurs by the nonionic diffusion of hydrogen fluoride. | [
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|
PMID:13662 | Effect of autonomic blockade on the hemodynamic findings in acute cardiac tamponade. | Twenty-three closed-chest, alpha-chloralose-anesthetized, volume-expanded, alpha- and beta-adrenergic-blockaded dogs with rate fixed by atrial pacing had 30-90 ml of saline at 37 degrees C infused into the pericardial sac a) with vagus intact, b) after vagotomy, and c) with vagus intact but with systolic pressure augmented with a balloon. A significant reduction in left ventricular (LV) systolic pressure (SP), and cardiac output (CO) occurred at a pericardial volume of 30-60 ml, when LV end-diastolic (ED) and right atrial (RA) pressures were not increased. Whereas the percentage decline of CO, LVSP, maximum negative and maximum positive dP/dt was greater in group A (vagus intact) than in group B (vagus cut), significant residual depressed performance was demonstrated only in group B. In four paced, atropinized, beta-blockaded dogs, response to tamponade was similar to that in intact dogs; vagotomy at 90 ml in these dogs resulted in a fall in CO, a rise of LVSP and a significant elevation in LVED and RA pressures. Thus, in the early phases of cardiac tamponade a sympathetic neurohumoral response supports cardiac performance while the vagus nerve exerts a myocardial protective effect. Vagal afferents appear to modulate this response. | Effect of autonomic blockade on the hemodynamic findings in acute cardiac tamponade. Twenty-three closed-chest, alpha-chloralose-anesthetized, volume-expanded, alpha- and beta-adrenergic-blockaded dogs with rate fixed by atrial pacing had 30-90 ml of saline at 37 degrees C infused into the pericardial sac a) with vagus intact, b) after vagotomy, and c) with vagus intact but with systolic pressure augmented with a balloon. A significant reduction in left ventricular (LV) systolic pressure (SP), and cardiac output (CO) occurred at a pericardial volume of 30-60 ml, when LV end-diastolic (ED) and right atrial (RA) pressures were not increased. Whereas the percentage decline of CO, LVSP, maximum negative and maximum positive dP/dt was greater in group A (vagus intact) than in group B (vagus cut), significant residual depressed performance was demonstrated only in group B. In four paced, atropinized, beta-blockaded dogs, response to tamponade was similar to that in intact dogs; vagotomy at 90 ml in these dogs resulted in a fall in CO, a rise of LVSP and a significant elevation in LVED and RA pressures. Thus, in the early phases of cardiac tamponade a sympathetic neurohumoral response supports cardiac performance while the vagus nerve exerts a myocardial protective effect. Vagal afferents appear to modulate this response. | [
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|
PMID:13663 | Action of histamine and H1 and H2 blockers on the cardiopulmonary circulation. | Systemic and pulmonary hemodynamic responses to histamine were investigated inchronically instrumented unanesthetized nonpregnant ewes. Histamine was administered intravenously and into the pulmonary artery. The effects of the same doses of histamine were assessed following H1 and H2 receptor blockade. The effects ocular changes were also monitored. Results indicate that intravenous histamine produces tachycardia, systemic hypotension, pulmonary hypertension, and reduced cardiac output. The pulmonary response could be modified significantly by pentobarbital anesthesia. When injected directly into the pulmonary artery histamine failed to elicit any circulatory response. Blockade of H1 and H2 receptors, as well as autonomic ganglia, resulted in a comparable attentuation of the histamine circulatory response. It is concluded that a) central hemodynamic responses do not seem to be mediated through specific H1 and H2 receptors; b) histamine-induced pulmonary vasoconstriction can be reversed by pentobarbital anesthesia, and c) the absence of circulatory response to intrapulmonary histamine administration suggests that whatever receptors that may exist in the pulmonary vascular bed are not necessary for the central hemodynamic effects. | Action of histamine and H1 and H2 blockers on the cardiopulmonary circulation. Systemic and pulmonary hemodynamic responses to histamine were investigated inchronically instrumented unanesthetized nonpregnant ewes. Histamine was administered intravenously and into the pulmonary artery. The effects of the same doses of histamine were assessed following H1 and H2 receptor blockade. The effects ocular changes were also monitored. Results indicate that intravenous histamine produces tachycardia, systemic hypotension, pulmonary hypertension, and reduced cardiac output. The pulmonary response could be modified significantly by pentobarbital anesthesia. When injected directly into the pulmonary artery histamine failed to elicit any circulatory response. Blockade of H1 and H2 receptors, as well as autonomic ganglia, resulted in a comparable attentuation of the histamine circulatory response. It is concluded that a) central hemodynamic responses do not seem to be mediated through specific H1 and H2 receptors; b) histamine-induced pulmonary vasoconstriction can be reversed by pentobarbital anesthesia, and c) the absence of circulatory response to intrapulmonary histamine administration suggests that whatever receptors that may exist in the pulmonary vascular bed are not necessary for the central hemodynamic effects. | [
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|
PMID:13664 | Physiologic effects of normal-or low-oxygen-affinity red cells in hypoxic baboons. | Baboons were bled one-third their red cell mass and were given homologous transfusions of red blood cells to restore the red cell volume. One group of baboons received red blood cells with a normal 2,3-diphosphoglycerate 2,3-DPG) level and normal affinity for oxygen, and in this group the 2,3-DPG level after transfusion was normal. The other group received red blood cells with a 160% of normal 2,3-DPG level and decreased affinity for oxygen, and in this group the 2,3-DPG level after transfusion was 125% of normal. In both groups of baboons, the inspired oxygen concentration was lowered and arterial PO2 tension was maintained at 55-60 mmHg for 2 h after transfusion. During the hypoxic state, systemic oxygen extraction was similar in the two groups, whereas oxygen saturation was lower in the high 2,3-DPG group than in the control animals. Cardiac output was significantly reduced 30 min after the arterial PO2 was restored to normal. These data indicate that red blood cells with decreased affinity for oxygen maintained satisfactory oxygen delivery to tissue during hypoxia. | Physiologic effects of normal-or low-oxygen-affinity red cells in hypoxic baboons. Baboons were bled one-third their red cell mass and were given homologous transfusions of red blood cells to restore the red cell volume. One group of baboons received red blood cells with a normal 2,3-diphosphoglycerate 2,3-DPG) level and normal affinity for oxygen, and in this group the 2,3-DPG level after transfusion was normal. The other group received red blood cells with a 160% of normal 2,3-DPG level and decreased affinity for oxygen, and in this group the 2,3-DPG level after transfusion was 125% of normal. In both groups of baboons, the inspired oxygen concentration was lowered and arterial PO2 tension was maintained at 55-60 mmHg for 2 h after transfusion. During the hypoxic state, systemic oxygen extraction was similar in the two groups, whereas oxygen saturation was lower in the high 2,3-DPG group than in the control animals. Cardiac output was significantly reduced 30 min after the arterial PO2 was restored to normal. These data indicate that red blood cells with decreased affinity for oxygen maintained satisfactory oxygen delivery to tissue during hypoxia. | [
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|
PMID:13665 | Acid-base balance and plasma composition in the aestivating lungfish (Protopterus). | Upon entering into aestivation, Protopterus aethiopicus develops a respiratory acidosis. A slow compensatory increase in plasma bicarbonate suffices only to partially restore arterial pH toward normal. The cessation of water intake from the start of aestivation results in hemoconcentration and marked oliguria. The concentrations of most plasma constituents continue to increase progressively, and the electrolyte ratios change. The increase in urea concentration is disproportionately high for the degree of dehydration and constitutes an increasing fraction of total plasma osmolality. Acid-base and electrolyte balance do not reach a new equilibrium within 1 yr in the cocoon. | Acid-base balance and plasma composition in the aestivating lungfish (Protopterus). Upon entering into aestivation, Protopterus aethiopicus develops a respiratory acidosis. A slow compensatory increase in plasma bicarbonate suffices only to partially restore arterial pH toward normal. The cessation of water intake from the start of aestivation results in hemoconcentration and marked oliguria. The concentrations of most plasma constituents continue to increase progressively, and the electrolyte ratios change. The increase in urea concentration is disproportionately high for the degree of dehydration and constitutes an increasing fraction of total plasma osmolality. Acid-base and electrolyte balance do not reach a new equilibrium within 1 yr in the cocoon. | [
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|
PMID:13668 | Secretory state of gastric mucosa and resistance to injury by exogenous acid. | The capacity of the stomach to resist the effects of highly acid solutions was assessed by comparing the effects of such solutions on spontaneously secreting, stimulated, and inhibited gastric mucosae of rabbits in vivo and frogs in vitro. Exposure of unstimulated resting mucosa to HC1, 120 mM, for 60 minutes produced superficial erosions in all rabbits, whereas such lesions were observed in only one of ten animals stimulated with histamine. Metiamide obviated the protective effect of histamine against ulcerations even though it did not reduce H+ secretion to zero. Exposure of inhibited isolated frog fundic mucosa to HC1 resulted in significant deterioration of electrical parameters, suggesting impairment of active transport processes and increased tissue permeability. These data are consistent with the hypothesis that actively secreting gastric mucosae from two species resist injury to exogenous acid more effectively than do resting or inhibited tissues, perhaps in part as a result of a greater alkaline tide. | Secretory state of gastric mucosa and resistance to injury by exogenous acid. The capacity of the stomach to resist the effects of highly acid solutions was assessed by comparing the effects of such solutions on spontaneously secreting, stimulated, and inhibited gastric mucosae of rabbits in vivo and frogs in vitro. Exposure of unstimulated resting mucosa to HC1, 120 mM, for 60 minutes produced superficial erosions in all rabbits, whereas such lesions were observed in only one of ten animals stimulated with histamine. Metiamide obviated the protective effect of histamine against ulcerations even though it did not reduce H+ secretion to zero. Exposure of inhibited isolated frog fundic mucosa to HC1 resulted in significant deterioration of electrical parameters, suggesting impairment of active transport processes and increased tissue permeability. These data are consistent with the hypothesis that actively secreting gastric mucosae from two species resist injury to exogenous acid more effectively than do resting or inhibited tissues, perhaps in part as a result of a greater alkaline tide. | [
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|
PMID:13669 | Effects of chronic hypoxia on canine gastric secretion. | Interdigestive gastirc acid output and acid secretory response to histamine, insulin and pentagastrin were studied in five dogs before and after creation of a right to left extracardiac vascular shunt. The arterial PO2 decreased from 81.5 +/- 5.5 mm Hg to 39.8 +/- 6.0 mm Hg postoperatively with no change in arterial pH or PaCO2. Basal (interdigestive) acid output increased from 1.19 +/- 0.26 mEq/hr to 4.97 +/- 0.81 mEq/hr postoperatively. Acid secretory response to insulin was increased after the induction of chronic hypoxia. Increased sensitivity to pentagastric was also observed although parietal cell mass (maximum histamine-stimulated output) was unchanged postoperatively. Changes in acid secretory response and PaO2 were present at one week and were sustained through twelve weeks after shunting. Chronic hypoxia resulted in gastric acid hypersecretion secondary to enhanced sensitivity of the parietal cell to a combination of neural and humoral stimuli. | Effects of chronic hypoxia on canine gastric secretion. Interdigestive gastirc acid output and acid secretory response to histamine, insulin and pentagastrin were studied in five dogs before and after creation of a right to left extracardiac vascular shunt. The arterial PO2 decreased from 81.5 +/- 5.5 mm Hg to 39.8 +/- 6.0 mm Hg postoperatively with no change in arterial pH or PaCO2. Basal (interdigestive) acid output increased from 1.19 +/- 0.26 mEq/hr to 4.97 +/- 0.81 mEq/hr postoperatively. Acid secretory response to insulin was increased after the induction of chronic hypoxia. Increased sensitivity to pentagastric was also observed although parietal cell mass (maximum histamine-stimulated output) was unchanged postoperatively. Changes in acid secretory response and PaO2 were present at one week and were sustained through twelve weeks after shunting. Chronic hypoxia resulted in gastric acid hypersecretion secondary to enhanced sensitivity of the parietal cell to a combination of neural and humoral stimuli. | [
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PMID:13673 | The redox state of the glutathione in the bovine corneal epithelium. | In the corneal epithelium the levels of the oxidized and reduced glutathione, the oxidized and reduced triphosphopyridine nucleotide, the glucose-6-phosphate and the 6-phosphogluconate were investigated. The in vivo steady state levels were defined by preparation procedures and by the energy state of the adenosine phosphate system. The ratios of the levels of the metabolites involved suggested that the reactions of the glucose-6-phosphate dehydrogenase and of the glutathione reductase operate dependent on the redox state of the triphosphopyridine nucleotides. | The redox state of the glutathione in the bovine corneal epithelium. In the corneal epithelium the levels of the oxidized and reduced glutathione, the oxidized and reduced triphosphopyridine nucleotide, the glucose-6-phosphate and the 6-phosphogluconate were investigated. The in vivo steady state levels were defined by preparation procedures and by the energy state of the adenosine phosphate system. The ratios of the levels of the metabolites involved suggested that the reactions of the glucose-6-phosphate dehydrogenase and of the glutathione reductase operate dependent on the redox state of the triphosphopyridine nucleotides. | [
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PMID:13677 | Acid-base balance and blood gases changes and "lactate excess" in acute respiratory alkalosis during general anaesthesia. | In 40 young males aged 18-20 years operated on for inguinal hernioplasty acute respiratory alkalosis was obtained in the 45th minute of general anaesthesia. The values of basic acid-base balance parameters, blood gases, pyruvate and lactate levels and "lactate excess" were determined before and after hyperventilation. Shifts in the concentrations of hydrogen and bicarbonate ions were found which are both typical of acute respiratory alkalosis. No changes were observed in the oxygenation of capillary blood and the values of "lactate excess" were normal which rules out tissue hypoxia during acute respiratory alkalosis. Passive hyperventilation being a less dangerous alternative of hypoventilation is a frequent occurrence during general anaesthesia and it causes transient respiratory alkalosis. | Acid-base balance and blood gases changes and "lactate excess" in acute respiratory alkalosis during general anaesthesia. In 40 young males aged 18-20 years operated on for inguinal hernioplasty acute respiratory alkalosis was obtained in the 45th minute of general anaesthesia. The values of basic acid-base balance parameters, blood gases, pyruvate and lactate levels and "lactate excess" were determined before and after hyperventilation. Shifts in the concentrations of hydrogen and bicarbonate ions were found which are both typical of acute respiratory alkalosis. No changes were observed in the oxygenation of capillary blood and the values of "lactate excess" were normal which rules out tissue hypoxia during acute respiratory alkalosis. Passive hyperventilation being a less dangerous alternative of hypoventilation is a frequent occurrence during general anaesthesia and it causes transient respiratory alkalosis. | [
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|
PMID:13678 | The effect of adrenergic receptor blocking agents on arrhythmia caused by trauma. | Experiments on animals have shown that blunt trauma to the anterior chest wall causes arrhythmia and conduction disturbances. In most cases nodal rhythm and ST segment elevation were observed. Microscopic examination disclosed myocardial damage in the vicinity of the atrioventricular node. Administration of alpha or beta adrenergic receptor blocking agents immediately before trauma failed to prevent arrhythmia and conduction disturbances, on the contrary, they became even more pronounced. | The effect of adrenergic receptor blocking agents on arrhythmia caused by trauma. Experiments on animals have shown that blunt trauma to the anterior chest wall causes arrhythmia and conduction disturbances. In most cases nodal rhythm and ST segment elevation were observed. Microscopic examination disclosed myocardial damage in the vicinity of the atrioventricular node. Administration of alpha or beta adrenergic receptor blocking agents immediately before trauma failed to prevent arrhythmia and conduction disturbances, on the contrary, they became even more pronounced. | [
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|
PMID:13682 | General anesthetics and regional hypoxic pulmonary vasoconstriction. | Administration of N2O, fluroxene and isoflurane to the left lower lobe (LLL) of dogs anesthetized with pentobarbital was previously shown to inhibit LLL hypoxic pulmonary vasoconstriction (HPV). Using the same experimental model, the present study examined the effect of whole-lung administration of N2O, fluroxene, isoflurane, halothane, and enflurane on left-lower-lobe HPV. Selective ventilation of the LLL with N2 alone caused blood flow to the lobe to decrease 53.3 +/- 3.0 per cent. Responses to LLL hypoxia were remeasured during administration of inhalation anesthetics at 1 and 2 MAC to both the LLL and the rest of the lung. Isoflurane and fluroxene progressively inhibited and at 2 MAC halved lobar HPV. N2O (one third MAC) caused slight but significant inhibition, while halothane and enflurane caused slight and nonsignificant changes in lobar HPV. These effects of whole-lung administration of anesthetics on HPV were almost identical to those obtained when the administration was confined to the test lobe alone. It is concluded that N2O, isoflurane, and fluroxene locally inhibit regional HPV and via this mechanism increase total venous admixture, while halothane and enflurane do not have this effect. | General anesthetics and regional hypoxic pulmonary vasoconstriction. Administration of N2O, fluroxene and isoflurane to the left lower lobe (LLL) of dogs anesthetized with pentobarbital was previously shown to inhibit LLL hypoxic pulmonary vasoconstriction (HPV). Using the same experimental model, the present study examined the effect of whole-lung administration of N2O, fluroxene, isoflurane, halothane, and enflurane on left-lower-lobe HPV. Selective ventilation of the LLL with N2 alone caused blood flow to the lobe to decrease 53.3 +/- 3.0 per cent. Responses to LLL hypoxia were remeasured during administration of inhalation anesthetics at 1 and 2 MAC to both the LLL and the rest of the lung. Isoflurane and fluroxene progressively inhibited and at 2 MAC halved lobar HPV. N2O (one third MAC) caused slight but significant inhibition, while halothane and enflurane caused slight and nonsignificant changes in lobar HPV. These effects of whole-lung administration of anesthetics on HPV were almost identical to those obtained when the administration was confined to the test lobe alone. It is concluded that N2O, isoflurane, and fluroxene locally inhibit regional HPV and via this mechanism increase total venous admixture, while halothane and enflurane do not have this effect. | [
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PMID:13683 | Control of breathing using an extracorporeal membrane lung. | Various amounts of carbon dioxide were removed through an extracorporeal membrane lung in spontaneously breathing lambs. The decrease in alveolar ventilation was proportional to the fraction of total carbon dioxide removed by the membrane lung. When extracorporeal CO2 removal approximated CO2 production (VCO2), alveolar ventilation almost ceased. Pulmonary ventilation can be controlled by extracorporeal carbon dioxide removal. | Control of breathing using an extracorporeal membrane lung. Various amounts of carbon dioxide were removed through an extracorporeal membrane lung in spontaneously breathing lambs. The decrease in alveolar ventilation was proportional to the fraction of total carbon dioxide removed by the membrane lung. When extracorporeal CO2 removal approximated CO2 production (VCO2), alveolar ventilation almost ceased. Pulmonary ventilation can be controlled by extracorporeal carbon dioxide removal. | [
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|
PMID:13684 | Kinetic analysis of the AH8165-receptor interaction at the mammalian neuromuscular junction. | The effect of a new neuromuscular blocking agent, AH8165, on carbachol-induced end-plate depolarization was measured in isolated guinea pig lumbrical muscles. The results showed that the kinetics were competitive (parallel shift of dose-response curves to the right and a slope of Schild plot of 1.09 +/- 0.07) and the AH8165-receptor dissociation constant was estimated as 0.337 +/- 0.008 muM. Once the dissociation constant was known, the fraction of receptors occupied by any given concentration of AH8165 could be determined. This fractional receptor occupancy was then compared with the indirect twitch response in an isolated guinea pig nerve-lumbrical muscle preparation. These measurements of the effect of AH8165 on the margin of safety of neuromuscular transmission gave values comparable to those obtained with d-tubocurarine, i.e., the twitch remained normal until 75-80 per cent of the receptors were blocked and was abolished when 90-95 per cent of the receptors were occluded. Thus, the neuromuscular blocking action of AH8165 appears to be consistent with a simple postsynaptic competitive interaction with the transmitter. | Kinetic analysis of the AH8165-receptor interaction at the mammalian neuromuscular junction. The effect of a new neuromuscular blocking agent, AH8165, on carbachol-induced end-plate depolarization was measured in isolated guinea pig lumbrical muscles. The results showed that the kinetics were competitive (parallel shift of dose-response curves to the right and a slope of Schild plot of 1.09 +/- 0.07) and the AH8165-receptor dissociation constant was estimated as 0.337 +/- 0.008 muM. Once the dissociation constant was known, the fraction of receptors occupied by any given concentration of AH8165 could be determined. This fractional receptor occupancy was then compared with the indirect twitch response in an isolated guinea pig nerve-lumbrical muscle preparation. These measurements of the effect of AH8165 on the margin of safety of neuromuscular transmission gave values comparable to those obtained with d-tubocurarine, i.e., the twitch remained normal until 75-80 per cent of the receptors were blocked and was abolished when 90-95 per cent of the receptors were occluded. Thus, the neuromuscular blocking action of AH8165 appears to be consistent with a simple postsynaptic competitive interaction with the transmitter. | [
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PMID:13681 | Comparison of THAM and sodium bicarbonate in resuscitation of the heart after ventricular fibrillation in dogs. | Tris (hydroxymethyl) aminomethane (tromethamine or THAM) has been suggested as an effective substitute for sodium bicarbonate (NaHCO3) in the treatment of metabolic acidosis accompanying cardiac arrest. Even though several reports on its appraisal have been published, there is still no clear agreement on its therapeutic value. A double-blind study was therefore lndertaken to compare in 36 dogs the effectiveness of 0.6 M THAM, 0.3 M THAM, and NaHCO3 (0.892 mEq/ml) to correct metabolic acidosis produced during 3 minutes of cardiac fibrillation, followed by a 3-minute period of cardiac compression. The dogs were then defibrillated and observed for 45 minutes. One group of 8 dogs was treated with 0.9 percent NaCl infusion. Compared with 0.9 percent NaCl, both THAM and NaHCO3 were equally effective in correcting metabolic acidosis (p less than 0.05). Initially, 0.6 M THAM produced a more pronounced (p less than 0.05) elevation of blood pH, but this effect was not sustained during the later postdefibrillation period. There was little difference in the effect of either of these drugs on mean aortic pressure and total peripheral vascular resistance. It is concluded that adequate ventilation and effective cardiac compression are still the chief criteria on which the final outcome of cardiac resuscitation depends. Correction of metabolic acidosis is important supportive therapy, but either THAM or NaHCO3 can be used with comparatively equivalent effect. | Comparison of THAM and sodium bicarbonate in resuscitation of the heart after ventricular fibrillation in dogs. Tris (hydroxymethyl) aminomethane (tromethamine or THAM) has been suggested as an effective substitute for sodium bicarbonate (NaHCO3) in the treatment of metabolic acidosis accompanying cardiac arrest. Even though several reports on its appraisal have been published, there is still no clear agreement on its therapeutic value. A double-blind study was therefore lndertaken to compare in 36 dogs the effectiveness of 0.6 M THAM, 0.3 M THAM, and NaHCO3 (0.892 mEq/ml) to correct metabolic acidosis produced during 3 minutes of cardiac fibrillation, followed by a 3-minute period of cardiac compression. The dogs were then defibrillated and observed for 45 minutes. One group of 8 dogs was treated with 0.9 percent NaCl infusion. Compared with 0.9 percent NaCl, both THAM and NaHCO3 were equally effective in correcting metabolic acidosis (p less than 0.05). Initially, 0.6 M THAM produced a more pronounced (p less than 0.05) elevation of blood pH, but this effect was not sustained during the later postdefibrillation period. There was little difference in the effect of either of these drugs on mean aortic pressure and total peripheral vascular resistance. It is concluded that adequate ventilation and effective cardiac compression are still the chief criteria on which the final outcome of cardiac resuscitation depends. Correction of metabolic acidosis is important supportive therapy, but either THAM or NaHCO3 can be used with comparatively equivalent effect. | [
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|
PMID:13685 | [Drugs combined with neuroleptics in anesthesia]. | The neuroleptics are characterised by the large number of pharmacological effects they develop. When they are associated with other substances, it is frequent for the latter to have with neuroleptics one or several common sites of action. At this level, they develop phenomena of drug interaction, for example, synergy with depressors of the central nervous system or, in the periphery, with alpha-adrenolytic or parasympatholytic drugs. This type of interaction is used in anesthesia to obtain more intense effects with lesser doses of each component and also avoid the toxic effects of efficacious doses of general anesthetics used alone. However, although certain interactions are useful, others may prove harmful. This is in particular the case where neuroleptic drugs are administered after ingestion of alcohol. Various mechanisms, central and peripheral, explain the marked depression of the central nervous system which results. Potentialisation of the effects of tricyclic antidepressor drugs by neuroleptics raises a difficult problem for the anesthetist, the elimination of tricyclic antidepressor drugs is slow and requires several weeks after stopping treatment. During this period, the tissue and plasma concentrations become reduced gradually. They are pharmacologically insufficient, but are potentialised by injection of neuroleptic drugs, and may become active again, and even toxic. It seems to us advisable to avoid the use of neuroleptic drugs in patients treated with tricyclic antidepressor drugs and, if necessary, to use another method of anesthesia. | [Drugs combined with neuroleptics in anesthesia]. The neuroleptics are characterised by the large number of pharmacological effects they develop. When they are associated with other substances, it is frequent for the latter to have with neuroleptics one or several common sites of action. At this level, they develop phenomena of drug interaction, for example, synergy with depressors of the central nervous system or, in the periphery, with alpha-adrenolytic or parasympatholytic drugs. This type of interaction is used in anesthesia to obtain more intense effects with lesser doses of each component and also avoid the toxic effects of efficacious doses of general anesthetics used alone. However, although certain interactions are useful, others may prove harmful. This is in particular the case where neuroleptic drugs are administered after ingestion of alcohol. Various mechanisms, central and peripheral, explain the marked depression of the central nervous system which results. Potentialisation of the effects of tricyclic antidepressor drugs by neuroleptics raises a difficult problem for the anesthetist, the elimination of tricyclic antidepressor drugs is slow and requires several weeks after stopping treatment. During this period, the tissue and plasma concentrations become reduced gradually. They are pharmacologically insufficient, but are potentialised by injection of neuroleptic drugs, and may become active again, and even toxic. It seems to us advisable to avoid the use of neuroleptic drugs in patients treated with tricyclic antidepressor drugs and, if necessary, to use another method of anesthesia. | [
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|
PMID:13686 | [The origin and success of neuroleptics]. | The authors retrace the origin and the development of the idea of neurolepsy in anesthesiology, first under the influence of Laborit, and in psychiatry, under the influence of Delay and Deniker, during the early 1950's. In these two fields, neuroleptic drugs were considered contrary to ideas and practice at that time and disturbed current ideas. | [The origin and success of neuroleptics]. The authors retrace the origin and the development of the idea of neurolepsy in anesthesiology, first under the influence of Laborit, and in psychiatry, under the influence of Delay and Deniker, during the early 1950's. In these two fields, neuroleptic drugs were considered contrary to ideas and practice at that time and disturbed current ideas. | [
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|
PMID:13688 | [Neuroplegia: results of an interdisciplinary approach to its physiopathology]. | Neuroplegia was born from a physiopathological study of states of shock and research on inhibition of the autonomic reaction to aggression. Over the last 25 years, the experimental facts have become accumulated in favour of the first theory of H. Laborit, i.e. that this so-called defence reaction defended our lives only through conservation of motor activity in the environment. When this motor activity is inefficacious or useless, the neuroendocrine reaction may lead, during the acute phase, to states of shock and to chronic psychosomatic pathology. On this general theme, inhibition of this reaction by neuroplegic drugs, among which the phenothiazine derivatives have played a very important role, has led in numerous surgical and medical disciplines, to basic research and therapeutic consequences. Thus, in anesthetics, it was at the origin of potentialised anesthesia, then neuroleptanalgesia. In general intensive care, it is used in various ways in the study and treatment of states of shock. In psychiatry, it has initiated neuro-psychopharmacology and the neuro-physiological and biochemical study of the nervous system in its response to the psycho-social environment. It has found its place in anesthesia and in obstetric pathology. But above all, has led pharmacologists to the study of metabolic and biochemical activities. The result of an interdisciplinary approach, neuroplegia has on the contrary, often been the origin of an interdisciplinary development of our physiological and physiopathological knowledge. Today this development seems to better understand its mode of action at various levels of organisation of living systems. One may thus say that neuroplegia, apart from its therapeutic interest, has been a good working instrument and led to better understanding of numerous biological disciplines. | [Neuroplegia: results of an interdisciplinary approach to its physiopathology]. Neuroplegia was born from a physiopathological study of states of shock and research on inhibition of the autonomic reaction to aggression. Over the last 25 years, the experimental facts have become accumulated in favour of the first theory of H. Laborit, i.e. that this so-called defence reaction defended our lives only through conservation of motor activity in the environment. When this motor activity is inefficacious or useless, the neuroendocrine reaction may lead, during the acute phase, to states of shock and to chronic psychosomatic pathology. On this general theme, inhibition of this reaction by neuroplegic drugs, among which the phenothiazine derivatives have played a very important role, has led in numerous surgical and medical disciplines, to basic research and therapeutic consequences. Thus, in anesthetics, it was at the origin of potentialised anesthesia, then neuroleptanalgesia. In general intensive care, it is used in various ways in the study and treatment of states of shock. In psychiatry, it has initiated neuro-psychopharmacology and the neuro-physiological and biochemical study of the nervous system in its response to the psycho-social environment. It has found its place in anesthesia and in obstetric pathology. But above all, has led pharmacologists to the study of metabolic and biochemical activities. The result of an interdisciplinary approach, neuroplegia has on the contrary, often been the origin of an interdisciplinary development of our physiological and physiopathological knowledge. Today this development seems to better understand its mode of action at various levels of organisation of living systems. One may thus say that neuroplegia, apart from its therapeutic interest, has been a good working instrument and led to better understanding of numerous biological disciplines. | [
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|
PMID:13690 | [Neurophysiologic justification of the place of neuroleptics in neuroleptanalgesia]. | We studied the action of drugs on the cortical and systemic hemodynamic responses to reticular stimulation and somatic nociceptive stimulation. Central analgesics, even in high dosage, do not suppress the awakening reaction and the attack of hypertension produced by sciatic stimulation in curarised cats. The addition of a neuroplegic, such as Droperidol or Ethrane in low dosage, abolishes completely the response to painful stimulation. The action of pure analgesic anesthesia seems to be situated, above all, at spinal level. | [Neurophysiologic justification of the place of neuroleptics in neuroleptanalgesia]. We studied the action of drugs on the cortical and systemic hemodynamic responses to reticular stimulation and somatic nociceptive stimulation. Central analgesics, even in high dosage, do not suppress the awakening reaction and the attack of hypertension produced by sciatic stimulation in curarised cats. The addition of a neuroplegic, such as Droperidol or Ethrane in low dosage, abolishes completely the response to painful stimulation. The action of pure analgesic anesthesia seems to be situated, above all, at spinal level. | [
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|
PMID:13691 | [Endocrine effects of neuroleptics]. | The neuroleptic drugs used in anesthetics belong to the group of phenothiazines or butyrophenones. The endocrine response to their intravenous administration is still ill-known and usually only concerns the association of anesthetics and neuroleptics. However, as far as the catecholamines are concerned, it is known that neuroleptic drugs do not prevent either their secretion nor their liberation but, depending on their dosage, they block the dopaminergic receptors and the alpha-receptors and induce disturbances in the metabolism of the mono-amines. The injection of neuroleptics associated with analgesics, raises the blood levels of catecholamines, does not induce a rise in ACTH and cortisol levels in the absence of stress, but does not totally prevent their rise in cases of aggression. As far as growth hormone is concerned, the effects are variable depending on the association studied. There is a rise with droperidol + pethidine or pentazocine, no change with chloroprotixene-dextromoramide. In both cases, the blood sugar rises. As far as STH, or growth hormone, free fatty acids and insulin are concerned, one may note a rise with associations containing droperidol even in the absence of any stress and stability with a mixture of chlorprotixene and dextromoramide. With none of these well known associations was there any variation either in levels of pituitary thyreo-stimulin, nor in thyroxine levels. Testosterone becomes reduced with the association of droperidol + analgesics but this effect does not seem to be specific to droperidol. These responses are frequently disturbed in case of additional stress. | [Endocrine effects of neuroleptics]. The neuroleptic drugs used in anesthetics belong to the group of phenothiazines or butyrophenones. The endocrine response to their intravenous administration is still ill-known and usually only concerns the association of anesthetics and neuroleptics. However, as far as the catecholamines are concerned, it is known that neuroleptic drugs do not prevent either their secretion nor their liberation but, depending on their dosage, they block the dopaminergic receptors and the alpha-receptors and induce disturbances in the metabolism of the mono-amines. The injection of neuroleptics associated with analgesics, raises the blood levels of catecholamines, does not induce a rise in ACTH and cortisol levels in the absence of stress, but does not totally prevent their rise in cases of aggression. As far as growth hormone is concerned, the effects are variable depending on the association studied. There is a rise with droperidol + pethidine or pentazocine, no change with chloroprotixene-dextromoramide. In both cases, the blood sugar rises. As far as STH, or growth hormone, free fatty acids and insulin are concerned, one may note a rise with associations containing droperidol even in the absence of any stress and stability with a mixture of chlorprotixene and dextromoramide. With none of these well known associations was there any variation either in levels of pituitary thyreo-stimulin, nor in thyroxine levels. Testosterone becomes reduced with the association of droperidol + analgesics but this effect does not seem to be specific to droperidol. These responses are frequently disturbed in case of additional stress. | [
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|
PMID:13692 | [Sympathetic-adrenergic and hypophysial response to different anesthesia-analgesia technics]. | The authors used techniques of pure anesthesia analgesia with Fentathienyl-Pavulon, or anesthesia and analgesia potentialised with Fentathienyl-Flunitrazepam, or Pentothal-Fentathienyl-Pavulon on various series of patients. The study was oriented on changes in the sympathetico-adrenalin pituitary response, and the use of glucose under the effect of these techniques. There was noted a fall in plasma cortisol levels, a fall in urinary excretion of catecholamines, an increase in growth hormone and better peripheral use of glucose. A comparative study was carried out with similar techniques using morphine, pentazocine or fentanyl. | [Sympathetic-adrenergic and hypophysial response to different anesthesia-analgesia technics]. The authors used techniques of pure anesthesia analgesia with Fentathienyl-Pavulon, or anesthesia and analgesia potentialised with Fentathienyl-Flunitrazepam, or Pentothal-Fentathienyl-Pavulon on various series of patients. The study was oriented on changes in the sympathetico-adrenalin pituitary response, and the use of glucose under the effect of these techniques. There was noted a fall in plasma cortisol levels, a fall in urinary excretion of catecholamines, an increase in growth hormone and better peripheral use of glucose. A comparative study was carried out with similar techniques using morphine, pentazocine or fentanyl. | [
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PMID:13693 | [Limits of pure analgesic anesthesia]. | Before dealing with this subject, the author believes that it is useful to give a definition of this technique and justify it. He then recalls the results which he obtained with various analgesic drugs, few products permit this technique, either because of too low analgesic power, or because of the appearance in high dosage, of troublesome or dangerous side-effects. At present, in this type of anesthetic, one may use either morphine or fentanyl or fentathienyl. Analgesic anesthesia with morphine. This technique presents advantages and disadvantages. The author has no personal experience of high doses of morphine and can only report that of other authors who have studied this problem. Anesthesia analgesia with fentanyl and fentathienyl. The author believes that this technique presents advantages over the previous technique, he has personally studied the clinical pharmacology of high doses of fentanyl and he believes that this drug used by the intravenous route, at a dose of 0.05 mg/kg associated with a curare derivative, possesses favourable cardiovascular and respiratory metabolic effects during the surgical operation, but one must recognize that +/-20 p. 100 of cases, fentanyl associated only with curare is not sufficient to give a stable anesthetic. At present, the author has replaced, in his technique, fentanyl by fentathienyl which is a morphine derivative 6 to 7 times more powerful. He found with this drug most of the properties of high doses of fentanyl but the analgesia obtained is deeper, one does not find more than 20 p. 100 of resistant cases. However, the author noted the fairly frequent appearance of tachycardia for which there was no valid explanation. In conclusion. The author believes that in the reproaches which are made to analgesic anesthesia, some are justified, others are less so. It requires prolonged post-operative supervision, but the latter would be advisable for any other type of anesthesia. It dose not always permit one, in its pure form, to obtain stable anesthesia, but this is less and less true with the appearance of more and more powerful derivatives. The use of analgesic anesthesia as a routine technique is only possible in the form of sequential analgesic anesthesia, a technique which uses an antimorphine drug to awaken the patient. In this field, various new products are presented are discussed. | [Limits of pure analgesic anesthesia]. Before dealing with this subject, the author believes that it is useful to give a definition of this technique and justify it. He then recalls the results which he obtained with various analgesic drugs, few products permit this technique, either because of too low analgesic power, or because of the appearance in high dosage, of troublesome or dangerous side-effects. At present, in this type of anesthetic, one may use either morphine or fentanyl or fentathienyl. Analgesic anesthesia with morphine. This technique presents advantages and disadvantages. The author has no personal experience of high doses of morphine and can only report that of other authors who have studied this problem. Anesthesia analgesia with fentanyl and fentathienyl. The author believes that this technique presents advantages over the previous technique, he has personally studied the clinical pharmacology of high doses of fentanyl and he believes that this drug used by the intravenous route, at a dose of 0.05 mg/kg associated with a curare derivative, possesses favourable cardiovascular and respiratory metabolic effects during the surgical operation, but one must recognize that +/-20 p. 100 of cases, fentanyl associated only with curare is not sufficient to give a stable anesthetic. At present, the author has replaced, in his technique, fentanyl by fentathienyl which is a morphine derivative 6 to 7 times more powerful. He found with this drug most of the properties of high doses of fentanyl but the analgesia obtained is deeper, one does not find more than 20 p. 100 of resistant cases. However, the author noted the fairly frequent appearance of tachycardia for which there was no valid explanation. In conclusion. The author believes that in the reproaches which are made to analgesic anesthesia, some are justified, others are less so. It requires prolonged post-operative supervision, but the latter would be advisable for any other type of anesthesia. It dose not always permit one, in its pure form, to obtain stable anesthesia, but this is less and less true with the appearance of more and more powerful derivatives. The use of analgesic anesthesia as a routine technique is only possible in the form of sequential analgesic anesthesia, a technique which uses an antimorphine drug to awaken the patient. In this field, various new products are presented are discussed. | [
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|
PMID:13695 | [Definition of the antineurotic, antipsychotic, neuroplegic and neuroleptic properties of psychotropic substances used in anesthesia and resuscitation]. | There is a great deal of evidences (pharmacological, experimental and clinical, therapeutic, biological, biochemical, metabolic, toxicological and neurophysiological) which permits one to characterise among the psychotropic substances, the antineurotic or antipsychotic properties of certain psycholeptic drugs. They authorize also the differentiation in the sub-group of "antipsychotics" of substances with a dominant neuroplegic or neuroleptic activity and others, such as lithium, which do not have this activity. This revision of the terminology avoids the confusion maintained by the use of terms "tranquillisers" and "neuroleptics" in the classification of psychotropic drugs. | [Definition of the antineurotic, antipsychotic, neuroplegic and neuroleptic properties of psychotropic substances used in anesthesia and resuscitation]. There is a great deal of evidences (pharmacological, experimental and clinical, therapeutic, biological, biochemical, metabolic, toxicological and neurophysiological) which permits one to characterise among the psychotropic substances, the antineurotic or antipsychotic properties of certain psycholeptic drugs. They authorize also the differentiation in the sub-group of "antipsychotics" of substances with a dominant neuroplegic or neuroleptic activity and others, such as lithium, which do not have this activity. This revision of the terminology avoids the confusion maintained by the use of terms "tranquillisers" and "neuroleptics" in the classification of psychotropic drugs. | [
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|
PMID:13696 | Bone marrow transplantation with intensive combination chemotherapy/radiation therapy (SCARI) in acute leukemia. | Fifteen patients with acute leukemia resistant to standard chemotherapy were treated by bone marrow transplantation from HLA-matched siblings after conditioning with a new combination chemotherapy/radiation therapy regimen--SCARI. SCARI consists of 5 days of high-dose cytosine arabinoside and 6-thioguanine followed by 3 days of daunorubicin. After a rest period, cyclophosphamide and total-body irradiation are given sequentially. This regimen had acceptable morbidity. Median survival was 169 days. Overall survival and disease-free survival was 27% at over 11 months. Relapse rate was 13% of the entire group and 30% by actuarial projection. Relapses were late and initially extramedullary. Deaths from causes other than leukemia occurred early secondary to fungal infection and late secondary to interstitial pneumonia (frequently cytomegalovirus). Graft-versus-host disease and graft rejection were not causes of mortality. In these patients conditioned with SCARI, leukemic recurrences were infrequent but infectious complications were a major hazard. | Bone marrow transplantation with intensive combination chemotherapy/radiation therapy (SCARI) in acute leukemia. Fifteen patients with acute leukemia resistant to standard chemotherapy were treated by bone marrow transplantation from HLA-matched siblings after conditioning with a new combination chemotherapy/radiation therapy regimen--SCARI. SCARI consists of 5 days of high-dose cytosine arabinoside and 6-thioguanine followed by 3 days of daunorubicin. After a rest period, cyclophosphamide and total-body irradiation are given sequentially. This regimen had acceptable morbidity. Median survival was 169 days. Overall survival and disease-free survival was 27% at over 11 months. Relapse rate was 13% of the entire group and 30% by actuarial projection. Relapses were late and initially extramedullary. Deaths from causes other than leukemia occurred early secondary to fungal infection and late secondary to interstitial pneumonia (frequently cytomegalovirus). Graft-versus-host disease and graft rejection were not causes of mortality. In these patients conditioned with SCARI, leukemic recurrences were infrequent but infectious complications were a major hazard. | [
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|
PMID:13697 | Effect of sera from immunologically altered mice on phytohemagglutinin-induced mouse lymphocyte transformation. | The DNA-synthetic response of mouse spleen cells to phytohemagglutinin in vitro is depressed by the addition of normal mouse serum. The effects of sera from mice that had been modified immunologically were examined. Increased depressive activity was found in sera from nude mice and from mice recently treated with anti-thymocyte globulin, but no change was found in sera from thymectomized, irradiated, bone marrow-reconstituted mice, nor from lethally or sublethally irradiated or cyclophosphamide-treated mice. Base line DNA synthesis was, however, less inhibited by sera from irradiated or cyclophosphamide-treated mice. Increased depressive activity was found in sera from mice with graft-versus-host reaction and from mice injected with Salmonella enteritidis 11RX, in which macrophage stimulation occurred. Increased depressive activity was also found in the sera of mice injected with sheep erythrocytes. T cells do not appear to be a source of depressive activity but B cells, macrophages and nonlymphoid cells may be sources. | Effect of sera from immunologically altered mice on phytohemagglutinin-induced mouse lymphocyte transformation. The DNA-synthetic response of mouse spleen cells to phytohemagglutinin in vitro is depressed by the addition of normal mouse serum. The effects of sera from mice that had been modified immunologically were examined. Increased depressive activity was found in sera from nude mice and from mice recently treated with anti-thymocyte globulin, but no change was found in sera from thymectomized, irradiated, bone marrow-reconstituted mice, nor from lethally or sublethally irradiated or cyclophosphamide-treated mice. Base line DNA synthesis was, however, less inhibited by sera from irradiated or cyclophosphamide-treated mice. Increased depressive activity was found in sera from mice with graft-versus-host reaction and from mice injected with Salmonella enteritidis 11RX, in which macrophage stimulation occurred. Increased depressive activity was also found in the sera of mice injected with sheep erythrocytes. T cells do not appear to be a source of depressive activity but B cells, macrophages and nonlymphoid cells may be sources. | [
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|
PMID:13699 | [Feeding and growth of elvers of anguilla Anguilla L. (eel-like teleosten fish) experimentally reared at various temperatures in the laboratory]. | In France, there are few researches carried out on the eel's elvers rearing. So, we have undertaken to do, in a first time, a study of their alimentation and growth, in relation to the temperature, at laboratory. The structure of european eels' elvers population, drawn in its natural environment, shows a polymodal frequency distribution of individual weights. The temperature has a direct effect on general metabolism of animals and consequently on their growth. For temperatures included between 10 degrees C and 25 degrees C, any increase of this factor induced an individual activity more important. The quantity of ingested food and its rapidity of utilization, the precocity of alimentary behaviour, are in relation with temperature. The best growth is obtained at 25 degrees C. It seems that this temperature permits to have a very low mortality. From 10 degrees C and below, growth is inhibited, the individual activity and the alimentary taking are very low or non-existent. However, if temperature is elevated, from 11,5 degrees C, growth can start off again, animals are very active and an alimentary behavious is observed. At least, rearing of the european eels' elvers is possible with a salt water (salinity : 32 p. 1000); this system permits suppress in totality fungi desease (saprolegnia). | [Feeding and growth of elvers of anguilla Anguilla L. (eel-like teleosten fish) experimentally reared at various temperatures in the laboratory]. In France, there are few researches carried out on the eel's elvers rearing. So, we have undertaken to do, in a first time, a study of their alimentation and growth, in relation to the temperature, at laboratory. The structure of european eels' elvers population, drawn in its natural environment, shows a polymodal frequency distribution of individual weights. The temperature has a direct effect on general metabolism of animals and consequently on their growth. For temperatures included between 10 degrees C and 25 degrees C, any increase of this factor induced an individual activity more important. The quantity of ingested food and its rapidity of utilization, the precocity of alimentary behaviour, are in relation with temperature. The best growth is obtained at 25 degrees C. It seems that this temperature permits to have a very low mortality. From 10 degrees C and below, growth is inhibited, the individual activity and the alimentary taking are very low or non-existent. However, if temperature is elevated, from 11,5 degrees C, growth can start off again, animals are very active and an alimentary behavious is observed. At least, rearing of the european eels' elvers is possible with a salt water (salinity : 32 p. 1000); this system permits suppress in totality fungi desease (saprolegnia). | [
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|
PMID:13702 | Measurement of ionised calcium in body fluids-a review. | This paper reviews the techniques available to the clinical biochemist for measuring ionised calcium concentrations in biological fluids with particular reference to serum. At present ionised calcium may be measured colorimetrically, using tetramethyl murexide, or potentiometrically, using a calcium ion-selective electrode. These techniques compare favourably in terms of precision with existing methods for measuring total calcium. Advantages of measuring ionised calcium in preference to total calcium are (a) that there is no effect of venous occlusion or change of posture on the ionised fraction of the effect on total calcium, (b) that ionised calcium is the physiologically active form of the element, and (c) that the ionised calcium concentration is a more reliable indicator of the calcium status of patients in certain clinical conditions. The main problem in the measurement of ionised calcium is the marked dependence of the ionised fraction of the pH of the sample. Extreme care must be taken to avoid loss of CO2 or build-up of acid during the handling of the blood sample. | Measurement of ionised calcium in body fluids-a review. This paper reviews the techniques available to the clinical biochemist for measuring ionised calcium concentrations in biological fluids with particular reference to serum. At present ionised calcium may be measured colorimetrically, using tetramethyl murexide, or potentiometrically, using a calcium ion-selective electrode. These techniques compare favourably in terms of precision with existing methods for measuring total calcium. Advantages of measuring ionised calcium in preference to total calcium are (a) that there is no effect of venous occlusion or change of posture on the ionised fraction of the effect on total calcium, (b) that ionised calcium is the physiologically active form of the element, and (c) that the ionised calcium concentration is a more reliable indicator of the calcium status of patients in certain clinical conditions. The main problem in the measurement of ionised calcium is the marked dependence of the ionised fraction of the pH of the sample. Extreme care must be taken to avoid loss of CO2 or build-up of acid during the handling of the blood sample. | [
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|
PMID:13704 | Comparative in vitro activity of piribenicillin, ticarcillin, and carbenicillin against Pseudomonas aeruginosa. | Pirbenicillin, a semisynthetic penicillin, showed greater in vitro activity against 68 recent isolates of Pseudomonas aeruginosa than did ticarcillin or carbenicillin. The median minimum inhibitory concentration of each of these three compounds, respectively, was 3.1, 12.5, and 25 mug/ml when a 10(-4) dilution of an overnight culture (about 10(5) colony-forming units [CFU]/ml) was used as inoculum, but these differences were less striking when larger inocula were used: at 10(7) CFU/ml these values were 6.25, 12.5, and 50 mug/ml, and at 10(8) to 10(9) CFU/ml these values were 50, 50, and 100 mug/ml. All three compounds showed greater inhibitory activity at pH 6 than at pH 8. This pH effect was greatest with pirbenicillin, for 6.25 mug of pirbenicillin per ml inhibited 7, 11, and 57% of the strains at pH 6, 7, and 8, respectively; these values were 4, 4, and 11% with ticarcillin and 0, 0, and 7% with carbenicillin. At sufficient inhibitory concentrations, the rates of bacterial killing of the three compounds were similar. The observed differences in anti-pseudomonad activity were not due to differences in stability to pseudomonad beta-lactamases, but all three compounds were more stable than were cefazolin, cephaloridine, and benzylpenicillin. | Comparative in vitro activity of piribenicillin, ticarcillin, and carbenicillin against Pseudomonas aeruginosa. Pirbenicillin, a semisynthetic penicillin, showed greater in vitro activity against 68 recent isolates of Pseudomonas aeruginosa than did ticarcillin or carbenicillin. The median minimum inhibitory concentration of each of these three compounds, respectively, was 3.1, 12.5, and 25 mug/ml when a 10(-4) dilution of an overnight culture (about 10(5) colony-forming units [CFU]/ml) was used as inoculum, but these differences were less striking when larger inocula were used: at 10(7) CFU/ml these values were 6.25, 12.5, and 50 mug/ml, and at 10(8) to 10(9) CFU/ml these values were 50, 50, and 100 mug/ml. All three compounds showed greater inhibitory activity at pH 6 than at pH 8. This pH effect was greatest with pirbenicillin, for 6.25 mug of pirbenicillin per ml inhibited 7, 11, and 57% of the strains at pH 6, 7, and 8, respectively; these values were 4, 4, and 11% with ticarcillin and 0, 0, and 7% with carbenicillin. At sufficient inhibitory concentrations, the rates of bacterial killing of the three compounds were similar. The observed differences in anti-pseudomonad activity were not due to differences in stability to pseudomonad beta-lactamases, but all three compounds were more stable than were cefazolin, cephaloridine, and benzylpenicillin. | [
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|
PMID:13705 | Beta-Lactamase activity in strains of Bacteroides melaninogenicus and Bacteroides oralis. | beta-Lactamase from strains of Bacteroides melaninogenicus and Bacteroides oralis hydrolyzed penicillin more rapidly than ampicillin or carbenicillin. Cephalothin and a chromogenic cephalosporin (87/312) were also hydrolyzed by the enzyme. Activity was found only in beta-lactam-resistant strains, but there was considerable variation in activity among strains having the same minimal inhibitory concentrations of antibiotic. beta-Lactamase activity was cell bound and appeared to be tightly associated with the cell envelope since detergents were required to elute this activity. | Beta-Lactamase activity in strains of Bacteroides melaninogenicus and Bacteroides oralis. beta-Lactamase from strains of Bacteroides melaninogenicus and Bacteroides oralis hydrolyzed penicillin more rapidly than ampicillin or carbenicillin. Cephalothin and a chromogenic cephalosporin (87/312) were also hydrolyzed by the enzyme. Activity was found only in beta-lactam-resistant strains, but there was considerable variation in activity among strains having the same minimal inhibitory concentrations of antibiotic. beta-Lactamase activity was cell bound and appeared to be tightly associated with the cell envelope since detergents were required to elute this activity. | [
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|
PMID:13706 | Purification and properties of an NADP+-specific isocitrate dehydrogenase from Rhizobium meliloti. | An NADP+-specific isocitrate dehydrogenase has been purified and characterized from Rhizobium meliloti. The enzyme showed Mn++ or Mg++ requirement. The apparent Km values were 2.00 x 10(-5) M and 1.51 x 10(-5) M for DL-isocitrate and NADP+, respectively. The enzyme was inhibited by ATP, to a lesser extent by ADP and AMP. alpha-Ketoglutarate also inhibited the enzyme activity. Oxalacetate and glyoxylate together inhibited the enzyme activity. The inhibition was competitive. Studies with thiol inhibitors suggested that the enzyme contained a sulfhydryl group at or near the active site. The enzyme has an approximate molecular weight of 60,000. Fluorescence studies suggested that the enzyme contained tryptophan. | Purification and properties of an NADP+-specific isocitrate dehydrogenase from Rhizobium meliloti. An NADP+-specific isocitrate dehydrogenase has been purified and characterized from Rhizobium meliloti. The enzyme showed Mn++ or Mg++ requirement. The apparent Km values were 2.00 x 10(-5) M and 1.51 x 10(-5) M for DL-isocitrate and NADP+, respectively. The enzyme was inhibited by ATP, to a lesser extent by ADP and AMP. alpha-Ketoglutarate also inhibited the enzyme activity. Oxalacetate and glyoxylate together inhibited the enzyme activity. The inhibition was competitive. Studies with thiol inhibitors suggested that the enzyme contained a sulfhydryl group at or near the active site. The enzyme has an approximate molecular weight of 60,000. Fluorescence studies suggested that the enzyme contained tryptophan. | [
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|
PMID:13703 | Significance of gamma-glutamyl transferase (GGT) activity measurements in alcohol-induced hepatic injury. | Experimental evidence is presented that the determination of gamma-glutamyl transferase (GGT) activity in serum is useful in the assessment of alcohol-induced liver disease and for demonstrating to patients the toxic effects of their drinking habits on the liver. Serial measurements of GGT activity in serum have also proved valuable in monitoring the progress of therapy as well as alleged abstention from alcohol in the known alcoholic. | Significance of gamma-glutamyl transferase (GGT) activity measurements in alcohol-induced hepatic injury. Experimental evidence is presented that the determination of gamma-glutamyl transferase (GGT) activity in serum is useful in the assessment of alcohol-induced liver disease and for demonstrating to patients the toxic effects of their drinking habits on the liver. Serial measurements of GGT activity in serum have also proved valuable in monitoring the progress of therapy as well as alleged abstention from alcohol in the known alcoholic. | [
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|
PMID:13707 | Mating reaction in Saccharomyces cerevisiae. X. Agglutinability-inactivating factor: a factor which destroys sexual agglutinability of a mating-type cells. | From cells of Saccharomyces cerevisiae a factor has been extracted that destroys the agglutinability of a mating-type cells specifically. It was found in the cell extracts of diploid and tetraploid strains as well as haploid strains of a and alpha mating types. It is heat-labile and the molecular weight is about 50,000. It is adsorbed by neither a cells nor alpha cells. Its biological activity is dependent on the incubation temperature and the pH, and is completely inhibited by phenylmethylsulfonyl fluoride, a potent inhibitor of the serine proteases. All the results described in this paper indicate that this factor is a proteolytic enzyme. | Mating reaction in Saccharomyces cerevisiae. X. Agglutinability-inactivating factor: a factor which destroys sexual agglutinability of a mating-type cells. From cells of Saccharomyces cerevisiae a factor has been extracted that destroys the agglutinability of a mating-type cells specifically. It was found in the cell extracts of diploid and tetraploid strains as well as haploid strains of a and alpha mating types. It is heat-labile and the molecular weight is about 50,000. It is adsorbed by neither a cells nor alpha cells. Its biological activity is dependent on the incubation temperature and the pH, and is completely inhibited by phenylmethylsulfonyl fluoride, a potent inhibitor of the serine proteases. All the results described in this paper indicate that this factor is a proteolytic enzyme. | [
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PMID:13709 | Hydrolysis of lactose by immobilized microorganisms. | Cells of Lactobacillus bulgaricus, Escherichia coli, and Kluyveromyces (Saccharomyces) lactis immobilized in polyacrylamide gel beads retained 27 to 61% of the beta-galactosidase activity of intact cells. Optimum temperature and pH and thermostability of these microbial beta-galactosidases were negligibly affected by the immobilization. Km values of beta-galactosidase in immobilized cells of L. bulgaricus, E. coli, and K. lactis toward lactose were 4.2, 5.4, and 30 mM, respectively. Neither inhibition nor activation of beta-galactosidase in immobilized L. bulgaricus and E. coli appeared in the presence of galactose, but remarkable inhibition by galactose was detected in the case of the enzyme of immobilized K. lactis. Glucose inhibited noncompetitively the activity of three species of immobilized microbial cells. These kinetic properties were almost the same as those of free beta-galactosidase extracted from individual microorganisms. The activity of immobilized K. lactis was fairly stable during repeated runs, but those of E. coli and L. bulgaricus decreased gradually. These immobilized microbial cells, when introduced into skim milk, demonstrated high activity for converting lactose to monosaccharides. The flavor of skim milk was hardly affected by treatment with these immobilized cells, although the degree of sweetness was raised considerably. | Hydrolysis of lactose by immobilized microorganisms. Cells of Lactobacillus bulgaricus, Escherichia coli, and Kluyveromyces (Saccharomyces) lactis immobilized in polyacrylamide gel beads retained 27 to 61% of the beta-galactosidase activity of intact cells. Optimum temperature and pH and thermostability of these microbial beta-galactosidases were negligibly affected by the immobilization. Km values of beta-galactosidase in immobilized cells of L. bulgaricus, E. coli, and K. lactis toward lactose were 4.2, 5.4, and 30 mM, respectively. Neither inhibition nor activation of beta-galactosidase in immobilized L. bulgaricus and E. coli appeared in the presence of galactose, but remarkable inhibition by galactose was detected in the case of the enzyme of immobilized K. lactis. Glucose inhibited noncompetitively the activity of three species of immobilized microbial cells. These kinetic properties were almost the same as those of free beta-galactosidase extracted from individual microorganisms. The activity of immobilized K. lactis was fairly stable during repeated runs, but those of E. coli and L. bulgaricus decreased gradually. These immobilized microbial cells, when introduced into skim milk, demonstrated high activity for converting lactose to monosaccharides. The flavor of skim milk was hardly affected by treatment with these immobilized cells, although the degree of sweetness was raised considerably. | [
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PMID:13710 | Deconjugation of bile acids by intestinal lactobacilli. | Lactobacillus species normally found in the intestinal tract of humans varied in the ability to deconjugate bile acids, whereas laboratory strains of Lactobacillus acidophilus deconjugated both glycocholate and taurocholate. All isolates of L. acidophilus from human feces deconjugated taurocholate, whereas only one of six deconjugated glycocholate. None of 13 isolates identified as L. casei deconjugated taurocholate, whereas 9 deconjugated glycocholate. The deconjugating system of L. acidophilus appeared to be constitutive, required low oxidation-reduction potential, and was most active at pH 6. No degradation beyond deconjugation was detected. | Deconjugation of bile acids by intestinal lactobacilli. Lactobacillus species normally found in the intestinal tract of humans varied in the ability to deconjugate bile acids, whereas laboratory strains of Lactobacillus acidophilus deconjugated both glycocholate and taurocholate. All isolates of L. acidophilus from human feces deconjugated taurocholate, whereas only one of six deconjugated glycocholate. None of 13 isolates identified as L. casei deconjugated taurocholate, whereas 9 deconjugated glycocholate. The deconjugating system of L. acidophilus appeared to be constitutive, required low oxidation-reduction potential, and was most active at pH 6. No degradation beyond deconjugation was detected. | [
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|
PMID:13711 | Aggregation of poliovirus and reovirus by dilution in water. | Poliovirus and reovirus were found to aggregate into clumps of up to several hundred particles when diluted 10-fold into distilled water from a stock preparation of minimal aggregation in 0.05 M phosphate buffer, pH 7.2, plus 22 to 30% sucrose. Reovirus was also found to aggregate when diluted into phosphate-buffered saline. The aggregation was concentration dependent and did not occur when either virus was diluted into water 100-fold or greater. The aggregation of poliovirus was reversible by further addition of saline and produced a dispersed preparation of virus. Reovirus aggregation was not reversible. Both viruses aggregated when diluted into buffers at pH 5 and 3, and poliovirus aggregated at pH 6, and this aggregation of both viruses was reversible when returned to pH 7. Aggregation did not occur at alkaline pH values. Aggregation at low pH could be caused aggregation of either virus at pH 7. Calcium ions, however, were found to aggregate both viruses at a concentration of 0.01 M. | Aggregation of poliovirus and reovirus by dilution in water. Poliovirus and reovirus were found to aggregate into clumps of up to several hundred particles when diluted 10-fold into distilled water from a stock preparation of minimal aggregation in 0.05 M phosphate buffer, pH 7.2, plus 22 to 30% sucrose. Reovirus was also found to aggregate when diluted into phosphate-buffered saline. The aggregation was concentration dependent and did not occur when either virus was diluted into water 100-fold or greater. The aggregation of poliovirus was reversible by further addition of saline and produced a dispersed preparation of virus. Reovirus aggregation was not reversible. Both viruses aggregated when diluted into buffers at pH 5 and 3, and poliovirus aggregated at pH 6, and this aggregation of both viruses was reversible when returned to pH 7. Aggregation did not occur at alkaline pH values. Aggregation at low pH could be caused aggregation of either virus at pH 7. Calcium ions, however, were found to aggregate both viruses at a concentration of 0.01 M. | [
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PMID:13712 | Ionic bonding, the mechanism of viral uptake by shellfish mucus. | An investigation was conducted to determine the processes involved in the contamination of shellfish by viruses. Results of binding-release studies show that the process involves the attachment of viruses to mucus secreted, and then ingested, by shellfish during feeding. Analysis of the mucus-virus bond involved selective degradation of the mucus and use of chemical agents to block carboxyl and sulfate groups on the mucus. Results obtained indicate that the attachment of virus to mucus is primarily ionic and involves the binding of viral particles to sulfate radicals on the mucopolysaccharide moiety of shellfish mucus. | Ionic bonding, the mechanism of viral uptake by shellfish mucus. An investigation was conducted to determine the processes involved in the contamination of shellfish by viruses. Results of binding-release studies show that the process involves the attachment of viruses to mucus secreted, and then ingested, by shellfish during feeding. Analysis of the mucus-virus bond involved selective degradation of the mucus and use of chemical agents to block carboxyl and sulfate groups on the mucus. Results obtained indicate that the attachment of virus to mucus is primarily ionic and involves the binding of viral particles to sulfate radicals on the mucopolysaccharide moiety of shellfish mucus. | [
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PMID:13713 | Fermentation of L-aspartate by a saccharolytic strain of Bacteroides melaninogenicus. | Resting cells of Bacteroides melaninogenicus fermented L-[14C]aspartate as a single substrate. The 14C-labeled products included succinate, acetate, CO2, oxaloacetate, formate, malate, glycine, alanine, and fumarate in the relative percentages 68, 15, 9.9, 2.7, 1.8, 1.0, 0.7, 0.5, and 0.06, respectively, based on the total counts per minute of the L-[14C]aspartate fermented. Ammonia was produced in high amounts, indicating that 96% of the L-aspartate fermented was deaminated. These data suggest that L-aspartate is mainly being reduced through a number of intermediate reactions involving enzymes of the tricarboxylic acid cycle to succinate. L-[14C]asparagine was also fermented by resting cells of B. melaninogenicus to form L-aspartate, which was subsequently, but less actively, fermented. | Fermentation of L-aspartate by a saccharolytic strain of Bacteroides melaninogenicus. Resting cells of Bacteroides melaninogenicus fermented L-[14C]aspartate as a single substrate. The 14C-labeled products included succinate, acetate, CO2, oxaloacetate, formate, malate, glycine, alanine, and fumarate in the relative percentages 68, 15, 9.9, 2.7, 1.8, 1.0, 0.7, 0.5, and 0.06, respectively, based on the total counts per minute of the L-[14C]aspartate fermented. Ammonia was produced in high amounts, indicating that 96% of the L-aspartate fermented was deaminated. These data suggest that L-aspartate is mainly being reduced through a number of intermediate reactions involving enzymes of the tricarboxylic acid cycle to succinate. L-[14C]asparagine was also fermented by resting cells of B. melaninogenicus to form L-aspartate, which was subsequently, but less actively, fermented. | [
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|
PMID:13714 | Sanitation in self-service automatic washers. | The potential for microbial transfer in self-service laundry washing machines was investigated by obtaining swab samples from the interior surfaces of commercial machines and wash water samples before and after disinfectant treatment. Three disinfectants (chlorine, a quaternary ammonium product, and a phenolic disinfectant) were used. Four self-service laundry facilities were sampled, with 10 replications of the procedure for each treatment at each location. Although washers were set on a warmwater setting, the wash water temperatures ranged from 24 to 51 degrees C. The quaternary ammonium product seemed most effective, averaging a 97% microbial kill; chlorine was the second most effective, with a 58% kill, and the phenolic disinfectant was least effective, with only a 25% kill. The efficacies of the chlorine and phenolic disinfectants were reduced at low water temperatures commonly experienced in self-service laundries. Interfamily cross-contamination in self-service facilities is a potential public health problem, which is aggravated by environmental conditions, such as water temperature and the practices of the previous users of the equipment. Procedural changes in laundering are recommended, including the use of a disinfectant to maintain adequate levels of sanitation. | Sanitation in self-service automatic washers. The potential for microbial transfer in self-service laundry washing machines was investigated by obtaining swab samples from the interior surfaces of commercial machines and wash water samples before and after disinfectant treatment. Three disinfectants (chlorine, a quaternary ammonium product, and a phenolic disinfectant) were used. Four self-service laundry facilities were sampled, with 10 replications of the procedure for each treatment at each location. Although washers were set on a warmwater setting, the wash water temperatures ranged from 24 to 51 degrees C. The quaternary ammonium product seemed most effective, averaging a 97% microbial kill; chlorine was the second most effective, with a 58% kill, and the phenolic disinfectant was least effective, with only a 25% kill. The efficacies of the chlorine and phenolic disinfectants were reduced at low water temperatures commonly experienced in self-service laundries. Interfamily cross-contamination in self-service facilities is a potential public health problem, which is aggravated by environmental conditions, such as water temperature and the practices of the previous users of the equipment. Procedural changes in laundering are recommended, including the use of a disinfectant to maintain adequate levels of sanitation. | [
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|
PMID:13739 | Solar uticaria. Photoallergen in a patient's serum. | A 45-year-old man had solar urticaria that was activated by visible light. Passive transfer of the reactivity with the patient's serum to the skin of normal recipients was accomplished. Results of reverse passive transfer studies were negative. The patient developed an urticarial wheal at the site of injection of his own serum that had been previously exposed to light in vitro. The experimental data suggested that his condition was attributable to an allergic response. Systemic administration of reserpine was of some therapeutic value, and increasing exposure to natural sunlight was associated with substantial in crease in his tolerance to sunlight. Unfortunately, the possible loss of reactivity that may occur in the natural course of the disease makes substantiation of the therapeutic effects difficult. | Solar uticaria. Photoallergen in a patient's serum. A 45-year-old man had solar urticaria that was activated by visible light. Passive transfer of the reactivity with the patient's serum to the skin of normal recipients was accomplished. Results of reverse passive transfer studies were negative. The patient developed an urticarial wheal at the site of injection of his own serum that had been previously exposed to light in vitro. The experimental data suggested that his condition was attributable to an allergic response. Systemic administration of reserpine was of some therapeutic value, and increasing exposure to natural sunlight was associated with substantial in crease in his tolerance to sunlight. Unfortunately, the possible loss of reactivity that may occur in the natural course of the disease makes substantiation of the therapeutic effects difficult. | [
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|
PMID:13740 | Some aspects of the persistence and fate of acrolein herbicide in water. | Experimental data for the decay of acrolein approximated first order kinetics. The reaction continued to completion in local waters but in buffered solution (pH 5.1-8.6) an equilibrium was reached after reaction of about 92% of the acrolein. It is proposed that data presented on the effects of pH on decay of acrolein may be used as a conservative estimate of dissipation rates in water where non-target organisms are at risk. In flowing water in two channels the 8 to 10 fold discrepancy between observed and predicted rates of dissipation was attributed to major losses in volatilization and adsorption. A relatively non-volatile reaction product (which gave a positive reaction with dinitrophenylhydrazine) accumulated initially but dissipated rapidly, probably by microbiological processes, when acrolein concentrations fell below about 2 to 3 ppm. | Some aspects of the persistence and fate of acrolein herbicide in water. Experimental data for the decay of acrolein approximated first order kinetics. The reaction continued to completion in local waters but in buffered solution (pH 5.1-8.6) an equilibrium was reached after reaction of about 92% of the acrolein. It is proposed that data presented on the effects of pH on decay of acrolein may be used as a conservative estimate of dissipation rates in water where non-target organisms are at risk. In flowing water in two channels the 8 to 10 fold discrepancy between observed and predicted rates of dissipation was attributed to major losses in volatilization and adsorption. A relatively non-volatile reaction product (which gave a positive reaction with dinitrophenylhydrazine) accumulated initially but dissipated rapidly, probably by microbiological processes, when acrolein concentrations fell below about 2 to 3 ppm. | [
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|
PMID:13741 | [Bacteriological examination of sputum (author's transl)]. | The AA., emphasizing the importance of ascertain the bronchial source of microorganisms recoverable from sputum, recommend for this purpose the use of properly collected sputum specimens and the application of washing treatment for mucopurulent materials. The aim of the present study has been to verify the effectiveness of mechanical apparatus to homogenize sputum without affecting the viability of resident microorganisms and the usefulness of prior microscopical examination to establish the rate of dilute inocula for the culture. | [Bacteriological examination of sputum (author's transl)]. The AA., emphasizing the importance of ascertain the bronchial source of microorganisms recoverable from sputum, recommend for this purpose the use of properly collected sputum specimens and the application of washing treatment for mucopurulent materials. The aim of the present study has been to verify the effectiveness of mechanical apparatus to homogenize sputum without affecting the viability of resident microorganisms and the usefulness of prior microscopical examination to establish the rate of dilute inocula for the culture. | [
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|
PMID:13742 | [Study on the viscolytic activity of the sputum (author's transl)]. | Several mucolytic agents were evaluated on sputum for testing their viscolytic activity and the bacterial tollerance to each of them. Proteolytic enzymes (trypsin, pepsin, papain, pancreatin), KJ, and dithiothreitol (or its derivatives) were better tollerated by common respiratory pathogens (H. influenzae, D. pneumoniae, Klebsiella, etc.) than other mucolytic agents, as acetil-cysteine, cisteamine-HCl, tension active substances, mercaptoethanol, and others. The dithiothreitol showed also one of the strongest viscolytic effect and therefore it was selected for the routinary sputum digestion at the concentration 0.1% in PBS pH 7.2. Such a solution was added to sputum specimen in different proportions according to the macroscopic "apparent" viscosity of each specimen. However researches on the comparative viscolytic activity of all the agents hereinafter considered are still in progress. | [Study on the viscolytic activity of the sputum (author's transl)]. Several mucolytic agents were evaluated on sputum for testing their viscolytic activity and the bacterial tollerance to each of them. Proteolytic enzymes (trypsin, pepsin, papain, pancreatin), KJ, and dithiothreitol (or its derivatives) were better tollerated by common respiratory pathogens (H. influenzae, D. pneumoniae, Klebsiella, etc.) than other mucolytic agents, as acetil-cysteine, cisteamine-HCl, tension active substances, mercaptoethanol, and others. The dithiothreitol showed also one of the strongest viscolytic effect and therefore it was selected for the routinary sputum digestion at the concentration 0.1% in PBS pH 7.2. Such a solution was added to sputum specimen in different proportions according to the macroscopic "apparent" viscosity of each specimen. However researches on the comparative viscolytic activity of all the agents hereinafter considered are still in progress. | [
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PMID:13743 | [Aerobic flora examination of sputum (author's transl)]. | The routine bacteriological test of expectorate, except for mycobacteria, is usually unsatisfactory. There is a need of standardization which results in the present paper from a comparison between the data obtained by two different laboratories on the same samples. It is possible to achieve reasonable and uniform results establishing a uniformity of some procedures, namely; collection of specimens, homogenization, number and type of media, interpretation of data and so on. | [Aerobic flora examination of sputum (author's transl)]. The routine bacteriological test of expectorate, except for mycobacteria, is usually unsatisfactory. There is a need of standardization which results in the present paper from a comparison between the data obtained by two different laboratories on the same samples. It is possible to achieve reasonable and uniform results establishing a uniformity of some procedures, namely; collection of specimens, homogenization, number and type of media, interpretation of data and so on. | [
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|
PMID:13744 | [Resistance to antibiotics of bacteria involved in respiratory infections (author's transl)]. | The present status of resistance to antibiotics of bacteria involved in respiratory infections is reviewed. Schematically it can outlined as follows. Streptococcus B-haemolyticus as well as Pneumococcus did not change their sensitivity to penicillin, but some strains are now resistant to tetracycline. Streptococcus viridans, Enterococcus and H. influenzae did not change substantially their sensitivity to antibiotics. Staphylococcus aureus is the bacterial species that always poses some problems with regard to antibiotic resistance. Due to the selection of strains penicillinase-producing because of the large use of penicillin, the most part of clinical isolates of staphylococci is now resistant to penicillin. In addition an increased number of strains resistant to the other antibiotics has been registered as soon as they has been introduced in therapy. The resistant strains spread in a particularly rapid way in hospital. The introduction in therapy of penicillinase-resistant penicillins constituted a remarkable advance in therapy of staphylococcal infection. However, there is now a growing number of indications about the emergence of methicillin resistant strains of staphylococci. On the other hand it must be recalled that since 1960 a marked reduction of incidence and mortality in severe staphylococcal infections has been noted. Incidence and mortality of respiratory infections due to Gram-negative bacilli is augmented particularly in connection with a larger use of immunosuppressive and antineoplastic therapies, of particular surgical or reanimation procedures, of intensive courses of antibiotic therapy etc. Emergence of Pseudomonas, Proteus, Serratia, Providencia, etc. infections poses many difficult problems of chemotherapy since these species are scarcely sensitive to antibiotics. Carbenicillin, cephalosporins, sisomicin, tobramycin, amikacin are the more recent drugs that alone or in combination may offer some chances of success in this field. | [Resistance to antibiotics of bacteria involved in respiratory infections (author's transl)]. The present status of resistance to antibiotics of bacteria involved in respiratory infections is reviewed. Schematically it can outlined as follows. Streptococcus B-haemolyticus as well as Pneumococcus did not change their sensitivity to penicillin, but some strains are now resistant to tetracycline. Streptococcus viridans, Enterococcus and H. influenzae did not change substantially their sensitivity to antibiotics. Staphylococcus aureus is the bacterial species that always poses some problems with regard to antibiotic resistance. Due to the selection of strains penicillinase-producing because of the large use of penicillin, the most part of clinical isolates of staphylococci is now resistant to penicillin. In addition an increased number of strains resistant to the other antibiotics has been registered as soon as they has been introduced in therapy. The resistant strains spread in a particularly rapid way in hospital. The introduction in therapy of penicillinase-resistant penicillins constituted a remarkable advance in therapy of staphylococcal infection. However, there is now a growing number of indications about the emergence of methicillin resistant strains of staphylococci. On the other hand it must be recalled that since 1960 a marked reduction of incidence and mortality in severe staphylococcal infections has been noted. Incidence and mortality of respiratory infections due to Gram-negative bacilli is augmented particularly in connection with a larger use of immunosuppressive and antineoplastic therapies, of particular surgical or reanimation procedures, of intensive courses of antibiotic therapy etc. Emergence of Pseudomonas, Proteus, Serratia, Providencia, etc. infections poses many difficult problems of chemotherapy since these species are scarcely sensitive to antibiotics. Carbenicillin, cephalosporins, sisomicin, tobramycin, amikacin are the more recent drugs that alone or in combination may offer some chances of success in this field. | [
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|
PMID:13746 | Treatment of hemorrhagic gastritis by antacid. | A simple and safe method of nonsurgical treatment for the control of massive acute gastric mucosal hemorrhage is described. The procedure was developed from experimental and clinical observations that the presence of gastric hydrocloric acid played an important part in the development and perpetuation of the entity. The treatment consists of complete neutralization of gastric acid with antacid to a pH of 7. The antacid is intermittently added and aspirate at 7. In a retrospective analysis, the hemorrhage was controlled in 44 of 49 patients (89%). Five patients who continued to bleed underwent surgery (10%). Three patients had vagotomy and pyloroplasty and their bleeding ceased without recurrence. Two patients underwent partial gastrectomy, but they developed recurrent bleeding and died. One patient whose bleeding has been controlled by vagotomy and pyloroplasty died without hemorrhage 10 days after operation. Of the 44 patients whose bleeding had been controlled by antacid, 11 patients died without hemorrhage one or more weeks later. These results of 89% control of hemorrhage compare favorably with those in the literature. | Treatment of hemorrhagic gastritis by antacid. A simple and safe method of nonsurgical treatment for the control of massive acute gastric mucosal hemorrhage is described. The procedure was developed from experimental and clinical observations that the presence of gastric hydrocloric acid played an important part in the development and perpetuation of the entity. The treatment consists of complete neutralization of gastric acid with antacid to a pH of 7. The antacid is intermittently added and aspirate at 7. In a retrospective analysis, the hemorrhage was controlled in 44 of 49 patients (89%). Five patients who continued to bleed underwent surgery (10%). Three patients had vagotomy and pyloroplasty and their bleeding ceased without recurrence. Two patients underwent partial gastrectomy, but they developed recurrent bleeding and died. One patient whose bleeding has been controlled by vagotomy and pyloroplasty died without hemorrhage 10 days after operation. Of the 44 patients whose bleeding had been controlled by antacid, 11 patients died without hemorrhage one or more weeks later. These results of 89% control of hemorrhage compare favorably with those in the literature. | [
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|
PMID:13747 | Patterns of gastroesophageal reflux in health and disease. | Twenty-four pH monitoring the distal esophagus quantitates gastroesophageal reflux in a near physiologic setting by measuring the frequency and duration of acid exposure to the esophageal mucosa. Fifteen asymptomatic volunteers were studies with 24-hour pH and esophageal manometry. The normal cardia was more competent supine than in the upright position. Physiologic reflux was unaffected by age, rarely occurred during slumber, and was the rule after alimentation. One hundred symptomatic pateitns with an abnormal 24-hour pH record (2 S.D. above the mean of controls) could be divided into three patterns of pathological reflux: those who refluxed only in the upright position (9), only in the supine position (37), and in both positions (54). Upright differed from supine refluxers by excessive aerophagia causing reflux episodes by repetitive belching. Compared to controls, they had excessive post-prandial reflux, lower DES pressure, and less DES exposed to the positive pressure of the abdomen. Supine differed from upright refluxers by having a higher incidence of esophagitis and an inability to clear the esophagus of acid after a supine reflux episode. Compared to controls, they had only a lower DES pressure. Combined refluxers had a higher incidence of esophagitis than supine refluxers. Stricture (15%) was seen only in this group. They were similar to supine refluxers in their inability to clear a supine reflux episode. Compared to controls, they had a lower DES pressure and less DES exposed to the positive pressure of the abdomen. Forty of the 100 patients had an antireflux procedure (4 upright, 8 supine, 28 combined). The most severe postoperative flatus and abdominal distention was seen in the upright refluxers. It is concluded that minimal reflux is physiological. Patients with pathological reflux all have lower DES pressure. Patients with upright reflux have less of their DES exposed to the positive pressure environment of the abdomen. Patients with supine reflux have an inability to clear the esophagus of reflux acid and are prone to develop esophagitis. Patients with both upright and supine reflux have the most severe disease and are at risk in developing strictures. In patients with only upright reflux, aerophagia and delayed gastric emptying may be an important etiological factor. | Patterns of gastroesophageal reflux in health and disease. Twenty-four pH monitoring the distal esophagus quantitates gastroesophageal reflux in a near physiologic setting by measuring the frequency and duration of acid exposure to the esophageal mucosa. Fifteen asymptomatic volunteers were studies with 24-hour pH and esophageal manometry. The normal cardia was more competent supine than in the upright position. Physiologic reflux was unaffected by age, rarely occurred during slumber, and was the rule after alimentation. One hundred symptomatic pateitns with an abnormal 24-hour pH record (2 S.D. above the mean of controls) could be divided into three patterns of pathological reflux: those who refluxed only in the upright position (9), only in the supine position (37), and in both positions (54). Upright differed from supine refluxers by excessive aerophagia causing reflux episodes by repetitive belching. Compared to controls, they had excessive post-prandial reflux, lower DES pressure, and less DES exposed to the positive pressure of the abdomen. Supine differed from upright refluxers by having a higher incidence of esophagitis and an inability to clear the esophagus of acid after a supine reflux episode. Compared to controls, they had only a lower DES pressure. Combined refluxers had a higher incidence of esophagitis than supine refluxers. Stricture (15%) was seen only in this group. They were similar to supine refluxers in their inability to clear a supine reflux episode. Compared to controls, they had a lower DES pressure and less DES exposed to the positive pressure of the abdomen. Forty of the 100 patients had an antireflux procedure (4 upright, 8 supine, 28 combined). The most severe postoperative flatus and abdominal distention was seen in the upright refluxers. It is concluded that minimal reflux is physiological. Patients with pathological reflux all have lower DES pressure. Patients with upright reflux have less of their DES exposed to the positive pressure environment of the abdomen. Patients with supine reflux have an inability to clear the esophagus of reflux acid and are prone to develop esophagitis. Patients with both upright and supine reflux have the most severe disease and are at risk in developing strictures. In patients with only upright reflux, aerophagia and delayed gastric emptying may be an important etiological factor. | [
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|
PMID:13748 | Long-term follow-up of internal mammary artery myocardial implantation. | A study was made of 100 patients who had undergone internal mammary artery myocardial implantation 7 to 10 years previously. Forty-two patients had single implantation with or without a free omental graft, and 54 received double implantations. Four patients had a single internal mammary artery implant plus a single aortocoronary bypass graft. Eleven patients died at operation or within the first month, and 17 died from 1 to 7 years following operation. Two were lost to follow-up, and 15 refused follow-up angiograms. From 7 to 10 years postoperatively, angiographic studies were performed on 55 patients with 73 internal mammary artery implants. Of these 73 implants, 17 (23%) were occluded; 10 (14%) were patent but did not show myocardial filling; 15 (21%) showed myocardial blush or filling of small vessels; and 31 (42%) showed filling of a major coronary artery. The patency rate correlated well with the amount of coronary disease and slightly with the amount of symptomatic improvement. This study shows that the Vineberg operation is physiologically sound; however, the ideal candidates are those patients with coronary arteries of adequate size who could benefit more by direct perfusion. | Long-term follow-up of internal mammary artery myocardial implantation. A study was made of 100 patients who had undergone internal mammary artery myocardial implantation 7 to 10 years previously. Forty-two patients had single implantation with or without a free omental graft, and 54 received double implantations. Four patients had a single internal mammary artery implant plus a single aortocoronary bypass graft. Eleven patients died at operation or within the first month, and 17 died from 1 to 7 years following operation. Two were lost to follow-up, and 15 refused follow-up angiograms. From 7 to 10 years postoperatively, angiographic studies were performed on 55 patients with 73 internal mammary artery implants. Of these 73 implants, 17 (23%) were occluded; 10 (14%) were patent but did not show myocardial filling; 15 (21%) showed myocardial blush or filling of small vessels; and 31 (42%) showed filling of a major coronary artery. The patency rate correlated well with the amount of coronary disease and slightly with the amount of symptomatic improvement. This study shows that the Vineberg operation is physiologically sound; however, the ideal candidates are those patients with coronary arteries of adequate size who could benefit more by direct perfusion. | [
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|
PMID:13749 | Inhibition of prostaglandin biosynthesis by non-narcotic analgesic drugs. | The existence of a relationship between inhibition of prostaglandin biosynthesis and analgesic or anti-inflammatory activity was investigated in the case of the non-narcotic analgesics glafenine, floctafenine and clometacine, in comparison to indomethacin and acetylsalicylic acid. These compounds inhibit prostaglandin biosynthesis from arachidonic acid in a guinea-pig lung homogenate as strongly as indomethacin. On its biosynthesis in rat epididymal tissue stimulated by noradrenaline, glafenine equals indomethacin inhibitory potency, whereas floctafenine and clometacine are less active. Acetylsalicylic acid is the least active in both preparations. In vivo, prostaglandin biosynthesis induced in rat peritoneal fluid by injection of acetic acid is inhibited by the 5 drugs, ranked as follows: floctafenine greater than indomethacin greater than glafenine greater than clometacine greater than acetylsalicylic acid. The pharmacological profile of glafenine, floctafenine and clometacine is characterized by a relatively strong effect on acetic acid writhing and a relatively weak effect on carrageenin oedema, U.V. erythema and adjuvant arthritis. The inhibition of prostaglandin biosynthesis seems better correlated with their analgesic activity than with their anti-inflammatory effects. The results show that prostaglandins could play an important role in the genesis of tissulary pain in animals. | Inhibition of prostaglandin biosynthesis by non-narcotic analgesic drugs. The existence of a relationship between inhibition of prostaglandin biosynthesis and analgesic or anti-inflammatory activity was investigated in the case of the non-narcotic analgesics glafenine, floctafenine and clometacine, in comparison to indomethacin and acetylsalicylic acid. These compounds inhibit prostaglandin biosynthesis from arachidonic acid in a guinea-pig lung homogenate as strongly as indomethacin. On its biosynthesis in rat epididymal tissue stimulated by noradrenaline, glafenine equals indomethacin inhibitory potency, whereas floctafenine and clometacine are less active. Acetylsalicylic acid is the least active in both preparations. In vivo, prostaglandin biosynthesis induced in rat peritoneal fluid by injection of acetic acid is inhibited by the 5 drugs, ranked as follows: floctafenine greater than indomethacin greater than glafenine greater than clometacine greater than acetylsalicylic acid. The pharmacological profile of glafenine, floctafenine and clometacine is characterized by a relatively strong effect on acetic acid writhing and a relatively weak effect on carrageenin oedema, U.V. erythema and adjuvant arthritis. The inhibition of prostaglandin biosynthesis seems better correlated with their analgesic activity than with their anti-inflammatory effects. The results show that prostaglandins could play an important role in the genesis of tissulary pain in animals. | [
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|
PMID:13750 | Selectivity of clenbuterol (NAB 365) in guinea-pig isolated tissues containing beta-adrenoceptors. | The effects of clenbuterol (ANB 365), an aminohalogen substituted phenylethanolamine, have been examined on isolated tissue preparations from guinea-pigs. Clenbuterol produced concentration (or dose)-dependent relaxations of tracheal chains, decreases in perfusion pressure of hind limb blood vessels, inhibitions of acetylcholine-induced contractions of the uterus, increases in atrial rate and inhibitions of electrically-induced contractions of the ileum. These responses were blocked by propranolol. Clenbuterol was similar in potency to isoprenaline on the trachea (carbachol-contracted), hind limb and uterus (beta2-adrenoceptors) but was significantly less potent than isoprenaline on the atria and the ileum (beta1-adrenoceptors). Clenbuterol produced marked relaxation of intrinsic tone tracheal preparations in concentrations up to 3000 times less than were required on carbachol-contracted preparations, whereas for isoprenaline the concentrations required on intrinsic tone preparations were only 55 fold less. It is concluded that clenbuterol is a beta-adrenoceptor agonist and that it is a partial agonist on the carbachol-stimulated trachea, on the atria and on hind limb blood vessels. It shows beta2-selectivity in that its potency, relative to that of isoprenaline, on the preparations containing beta2-adrenoceptors was much higher than on those with beta1-adrenoceptors. On intrinsic tone tracheal preparations it may produce an additional relaxant effect responsible for its high potency on that preparation. | Selectivity of clenbuterol (NAB 365) in guinea-pig isolated tissues containing beta-adrenoceptors. The effects of clenbuterol (ANB 365), an aminohalogen substituted phenylethanolamine, have been examined on isolated tissue preparations from guinea-pigs. Clenbuterol produced concentration (or dose)-dependent relaxations of tracheal chains, decreases in perfusion pressure of hind limb blood vessels, inhibitions of acetylcholine-induced contractions of the uterus, increases in atrial rate and inhibitions of electrically-induced contractions of the ileum. These responses were blocked by propranolol. Clenbuterol was similar in potency to isoprenaline on the trachea (carbachol-contracted), hind limb and uterus (beta2-adrenoceptors) but was significantly less potent than isoprenaline on the atria and the ileum (beta1-adrenoceptors). Clenbuterol produced marked relaxation of intrinsic tone tracheal preparations in concentrations up to 3000 times less than were required on carbachol-contracted preparations, whereas for isoprenaline the concentrations required on intrinsic tone preparations were only 55 fold less. It is concluded that clenbuterol is a beta-adrenoceptor agonist and that it is a partial agonist on the carbachol-stimulated trachea, on the atria and on hind limb blood vessels. It shows beta2-selectivity in that its potency, relative to that of isoprenaline, on the preparations containing beta2-adrenoceptors was much higher than on those with beta1-adrenoceptors. On intrinsic tone tracheal preparations it may produce an additional relaxant effect responsible for its high potency on that preparation. | [
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|
PMID:13751 | Serum corticosterone as a quantitative test of the phlogistic potency of various agents topically applied in the rat. | The application into the rat conjunctiva of various phlogistic agents, such as croton oil, mustard oil and formaldehyde, elicits an increase of serum corticosterone linearly related to the log of the applied concentrations, so that from their parallelized regression lines it is possible to calculate the phlogistic potency of each tested agent in reference to croton oil. The time kinetic of such an increase (elicited by croton oil) is compared with that of two other parameters previously adopted as indirect quantitative indices of the phlogosis: the adrenal ascorbic acid depletion and the liver tyrosine-alpha-ketoglutarate transaminase increase. Serum corticosterone is shown to be the quickest and the most sensitive of the adopted indices, even if the phlogistic potency of the tested agents and the precision of these evaluations substantially coincides whatsoever the index adopted. Finally the pathways of adrenocortical activation are investigated and it is shown that the activation may be peripherally blocked by topical application of corticosteroids (but not of local anesthetics) and centrally by hypophysectomy or parenteral administration of pentobarbital plus morphine. | Serum corticosterone as a quantitative test of the phlogistic potency of various agents topically applied in the rat. The application into the rat conjunctiva of various phlogistic agents, such as croton oil, mustard oil and formaldehyde, elicits an increase of serum corticosterone linearly related to the log of the applied concentrations, so that from their parallelized regression lines it is possible to calculate the phlogistic potency of each tested agent in reference to croton oil. The time kinetic of such an increase (elicited by croton oil) is compared with that of two other parameters previously adopted as indirect quantitative indices of the phlogosis: the adrenal ascorbic acid depletion and the liver tyrosine-alpha-ketoglutarate transaminase increase. Serum corticosterone is shown to be the quickest and the most sensitive of the adopted indices, even if the phlogistic potency of the tested agents and the precision of these evaluations substantially coincides whatsoever the index adopted. Finally the pathways of adrenocortical activation are investigated and it is shown that the activation may be peripherally blocked by topical application of corticosteroids (but not of local anesthetics) and centrally by hypophysectomy or parenteral administration of pentobarbital plus morphine. | [
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|
PMID:13753 | Photoproduction of ammonium ion from N2 in Rhodospirillum rubrum. | NH+4 excretion was undetectable in N2-fixing cultures of Rhodospirillum rubrum (S-1) and nitrogenase activity in these cultures was repressed by the addition of 10 mM NH+4 to the medium. The glutamate analog, L-methionine-DL-sulfoximine (MSX), derepressed N2 fixation even in the presence of 10 mM extracellular NH+4. When 10 mg MSX/ml was added to cultures just prior to nitrogenase induction they developed nitrogenase activity (20% of the control activities) and excreted most of their fixed N2 as NH+4. Nitrogenase activities and NH+4 production from fixed N2 were increased considerably when a combined nitrogen source, NH+4 (greater than 40 mumoles NH+4/mg cell protein in 6 days) or L-glutamate (greater than 60 mumoles NH+4/ mg cell protein in 6 days) was added to the cultures together with MSX. Biochemical analysis revealed that R. rubrum produced glutamine synthetase and glutamate synthase (NADP-dependent) but no detectable NADP-dependent glutamate dehydrogenase. The specific activity of glutamine synthetase was observed to be maximal when nitrogenase activity was also maximal. Nitrogenase and glutamine synthetase activities were repressed by NH+4 as well as by glutamate. The results demonstrate that utilization of solar energy to photoproduce large quantities of NH+4 from N2 is possible with photosynthetic bacteria by interfering with their regulatory control of N2 fixation. | Photoproduction of ammonium ion from N2 in Rhodospirillum rubrum. NH+4 excretion was undetectable in N2-fixing cultures of Rhodospirillum rubrum (S-1) and nitrogenase activity in these cultures was repressed by the addition of 10 mM NH+4 to the medium. The glutamate analog, L-methionine-DL-sulfoximine (MSX), derepressed N2 fixation even in the presence of 10 mM extracellular NH+4. When 10 mg MSX/ml was added to cultures just prior to nitrogenase induction they developed nitrogenase activity (20% of the control activities) and excreted most of their fixed N2 as NH+4. Nitrogenase activities and NH+4 production from fixed N2 were increased considerably when a combined nitrogen source, NH+4 (greater than 40 mumoles NH+4/mg cell protein in 6 days) or L-glutamate (greater than 60 mumoles NH+4/ mg cell protein in 6 days) was added to the cultures together with MSX. Biochemical analysis revealed that R. rubrum produced glutamine synthetase and glutamate synthase (NADP-dependent) but no detectable NADP-dependent glutamate dehydrogenase. The specific activity of glutamine synthetase was observed to be maximal when nitrogenase activity was also maximal. Nitrogenase and glutamine synthetase activities were repressed by NH+4 as well as by glutamate. The results demonstrate that utilization of solar energy to photoproduce large quantities of NH+4 from N2 is possible with photosynthetic bacteria by interfering with their regulatory control of N2 fixation. | [
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|
PMID:13754 | Energy coupling and respiration in Nitrosomonas europaea. | Intact cells of Nitrosomonas europaea grown in an ammonium salts medium will oxidise ammonium ions, hydroxylamine and ascorbate-TMPD; there is no oxidation of carbon monoxide, methane or methanol. The Km value for ammonia oxidation is highly pH dependent with a minimum value of 0.5 mM above pH 8.0. This suggests that free ammonia is the species crossing the cytoplasmic membrane(s). The measurement of respiration driven proton translocation indicates that there is probably only one proton translocating loop (loop 3) association with hydroxylamine oxidation. The oxidation of "endogenous" substrates is sometimes associated with more than one proton-translocating loop. These results indicate that during growth hydroxylamine oxidation is probably associated with a maximum P/O ratio of 1. | Energy coupling and respiration in Nitrosomonas europaea. Intact cells of Nitrosomonas europaea grown in an ammonium salts medium will oxidise ammonium ions, hydroxylamine and ascorbate-TMPD; there is no oxidation of carbon monoxide, methane or methanol. The Km value for ammonia oxidation is highly pH dependent with a minimum value of 0.5 mM above pH 8.0. This suggests that free ammonia is the species crossing the cytoplasmic membrane(s). The measurement of respiration driven proton translocation indicates that there is probably only one proton translocating loop (loop 3) association with hydroxylamine oxidation. The oxidation of "endogenous" substrates is sometimes associated with more than one proton-translocating loop. These results indicate that during growth hydroxylamine oxidation is probably associated with a maximum P/O ratio of 1. | [
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|
PMID:13755 | D-Alanine dehydrogenase. Its role in the utilisation of alanine isomers as growth substrates by Pseudomonas aeruginosa PA01. | Pseudomonas aeruginosa PA01 was found to utilise both the D- and L-isomers of alpha-alanine and also beta-alanine as sole sources of carbon and energy for growth. Enzymological studies of wild-type cultures and comparison with mutants deficient in growth upon one or more isomers of alanine led to the following conclusions: (i) utilisation of D-alanine involved its direct oxidation by an inducible, membrane-bound, cytochrome-linked dehydrogenase; (ii) utilisation of L-alanine required its conversion to the directly oxidisable D-form by a soluble racemase; (iii) utilisation of beta-alanine, like L-alanine, involves both the racemase and D-alanine dehydrogenase enzymes, but in addition must involve other enzymes the identity of which is still speculative; (iv) P. aeruginosa, like Escherichia coli, appears to take up D-alanine and L-alanine by means of two specific permeases. | D-Alanine dehydrogenase. Its role in the utilisation of alanine isomers as growth substrates by Pseudomonas aeruginosa PA01. Pseudomonas aeruginosa PA01 was found to utilise both the D- and L-isomers of alpha-alanine and also beta-alanine as sole sources of carbon and energy for growth. Enzymological studies of wild-type cultures and comparison with mutants deficient in growth upon one or more isomers of alanine led to the following conclusions: (i) utilisation of D-alanine involved its direct oxidation by an inducible, membrane-bound, cytochrome-linked dehydrogenase; (ii) utilisation of L-alanine required its conversion to the directly oxidisable D-form by a soluble racemase; (iii) utilisation of beta-alanine, like L-alanine, involves both the racemase and D-alanine dehydrogenase enzymes, but in addition must involve other enzymes the identity of which is still speculative; (iv) P. aeruginosa, like Escherichia coli, appears to take up D-alanine and L-alanine by means of two specific permeases. | [
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PMID:13756 | Monosaccharide transport systems in the yeast Rhodotorula glutinis. | By using D-glucose, D-xylose, D-galactose and D-fructose in the strictly aerobic yeast Rhodotorula glutinis and by comparing the half-saturation constants with inhibition constants the yeast was shown to possess a single common system for D-xylose and D-galactose (Km's and Ki's all between 0.5 and 1.1 mM) but another distinct transport system for D-fructose. The transport of D-glucose has a special position in that glucose blocks apparently allotopically all the other systems observed although it uses at least one of them for its own transport. The different character of D-glucose uptake is underlined by its relative independence of pH (its "Km" is completely pH-insensitive) in contrast with all other sugars. At low concentrations, all sugars show mutual positive cooperativity in uptake, suggesting at least two transport sites plus possibly a modifier site on the carrier. | Monosaccharide transport systems in the yeast Rhodotorula glutinis. By using D-glucose, D-xylose, D-galactose and D-fructose in the strictly aerobic yeast Rhodotorula glutinis and by comparing the half-saturation constants with inhibition constants the yeast was shown to possess a single common system for D-xylose and D-galactose (Km's and Ki's all between 0.5 and 1.1 mM) but another distinct transport system for D-fructose. The transport of D-glucose has a special position in that glucose blocks apparently allotopically all the other systems observed although it uses at least one of them for its own transport. The different character of D-glucose uptake is underlined by its relative independence of pH (its "Km" is completely pH-insensitive) in contrast with all other sugars. At low concentrations, all sugars show mutual positive cooperativity in uptake, suggesting at least two transport sites plus possibly a modifier site on the carrier. | [
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|
PMID:13757 | Energetic aspects of anaerobic growth of Aerobacter aerogenes in complex medium. | Molar growth yields for anaerobic growth of Aerobacter aerogenes in complex medium were much higher than for growth in minimal medium. In batch cultures the molar growth yield for glucose varied from 44 to 50 and YATP from 17.1 to 18.8. For glucose-limited chemostat cultures a value of 17.5 g/mole was found for Y max ATP and a value of 2.3 mmoles ATP/g dry weight h for the maintenance coeficient. Growth-dependent pH changes were used to control the addition of fresh medium, containing excess of glucose to a continuous culture. The specific growth rate and the population density were dependent on the pH difference between the inflowing medium and the culture. At a mu value of 1.44 h-1 the molar growth yield for glucose was about 70 and Y ATP about 28.5. An equation is presented, which gives the relation between theoretical and experimental Y max ATP values. | Energetic aspects of anaerobic growth of Aerobacter aerogenes in complex medium. Molar growth yields for anaerobic growth of Aerobacter aerogenes in complex medium were much higher than for growth in minimal medium. In batch cultures the molar growth yield for glucose varied from 44 to 50 and YATP from 17.1 to 18.8. For glucose-limited chemostat cultures a value of 17.5 g/mole was found for Y max ATP and a value of 2.3 mmoles ATP/g dry weight h for the maintenance coeficient. Growth-dependent pH changes were used to control the addition of fresh medium, containing excess of glucose to a continuous culture. The specific growth rate and the population density were dependent on the pH difference between the inflowing medium and the culture. At a mu value of 1.44 h-1 the molar growth yield for glucose was about 70 and Y ATP about 28.5. An equation is presented, which gives the relation between theoretical and experimental Y max ATP values. | [
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|
PMID:13758 | Proton-motive force and the motile behavior of Bacillus subtilis. | Changes in the proton-motive force cause a transient change in the motile behavior of Bacillus subtilis cells. Both an increase and a decrease in the proton-motive force caused transient tumbling. Simultaneous decrease of proton-motive force and increase of attractant concentration lessens the response toward the attractant. A simultaneous increase of proton-motive force and increase of attractant concentration prolonges the response toward attractant. A hypothesis explaining the various effects is given. | Proton-motive force and the motile behavior of Bacillus subtilis. Changes in the proton-motive force cause a transient change in the motile behavior of Bacillus subtilis cells. Both an increase and a decrease in the proton-motive force caused transient tumbling. Simultaneous decrease of proton-motive force and increase of attractant concentration lessens the response toward the attractant. A simultaneous increase of proton-motive force and increase of attractant concentration prolonges the response toward attractant. A hypothesis explaining the various effects is given. | [
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|
PMID:13759 | Ammonium uptake and metabolism by mitrogen fixing bacteria. II. Klebsiella pneumoniae. | The primary steps of N2, ammonia and nitrate metabolism in Klebsiella pneumoniae grown in a continuous culture are regulated by the kind and supply of the nitrogenous compound. Cultures growing on N2 as the only nitrogen source have high activities of nitrogenase, unadenylated glutamine synthetase and glutamate synthase and low levels of glutamate dehydrogenase. If small amounts of ammonium salts are added continuously, initially only part of it is absorbed by the organisms. After 2-3 h complete absorption of ammonia against an ammonium gradient coinciding with an increased growth rate of the bacteria is observed. The change in the extracellular ammonium level is paralleled by the intracellular glutamine concentration which in turn regulates the glutamine synthesis and an induction of glutamate dehydrogenase synthesis. Upon deadenylation these events are reversed.--Addition of dinitrophenol causes transient leakage of intracellular ammonium into the medium. | Ammonium uptake and metabolism by mitrogen fixing bacteria. II. Klebsiella pneumoniae. The primary steps of N2, ammonia and nitrate metabolism in Klebsiella pneumoniae grown in a continuous culture are regulated by the kind and supply of the nitrogenous compound. Cultures growing on N2 as the only nitrogen source have high activities of nitrogenase, unadenylated glutamine synthetase and glutamate synthase and low levels of glutamate dehydrogenase. If small amounts of ammonium salts are added continuously, initially only part of it is absorbed by the organisms. After 2-3 h complete absorption of ammonia against an ammonium gradient coinciding with an increased growth rate of the bacteria is observed. The change in the extracellular ammonium level is paralleled by the intracellular glutamine concentration which in turn regulates the glutamine synthesis and an induction of glutamate dehydrogenase synthesis. Upon deadenylation these events are reversed.--Addition of dinitrophenol causes transient leakage of intracellular ammonium into the medium. | [
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|
PMID:13760 | Ammonia assimilation in the fission yeast Schizosaccharomyces pombe 972. | Glutamine synthetase (GS) activity of Schizosaccharomyces pombe 972 was high in ammonia-limited cultures, low in phosphate- and sulphate-limited cultures and not detected in glucose-limited cultures. When ammonia was 'pulsed' into an ammonia-limited culture then GS activity decreased at a rate faster than that calculated if enzyme synthesis ceased and enzyme was diluted out by growth. Enzyme activity increased in ammonia-starved, phosphate-limited cultures and in the ammonia 'pulse' system when the added ammonia had been utilised. These increases in enzyme activity were prevented by the presence of 100 mug/ml cycloheximide. GS activity was inversely related to the intracellular concentration of glutamate. | Ammonia assimilation in the fission yeast Schizosaccharomyces pombe 972. Glutamine synthetase (GS) activity of Schizosaccharomyces pombe 972 was high in ammonia-limited cultures, low in phosphate- and sulphate-limited cultures and not detected in glucose-limited cultures. When ammonia was 'pulsed' into an ammonia-limited culture then GS activity decreased at a rate faster than that calculated if enzyme synthesis ceased and enzyme was diluted out by growth. Enzyme activity increased in ammonia-starved, phosphate-limited cultures and in the ammonia 'pulse' system when the added ammonia had been utilised. These increases in enzyme activity were prevented by the presence of 100 mug/ml cycloheximide. GS activity was inversely related to the intracellular concentration of glutamate. | [
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|
PMID:13761 | Monoamine metabolism in human brain. | Norepinephrine (NE), dopamine (DA), tyrosine hydroxylase (TH), catechol-O-methyltransferase (COMT) and monoamine oxidase (MAO) levels were measured in human brain tissue obtained at autopsy from a series of 39 patients dying of various medical and accidental causes. The nine following brain areas were studied: globus pallidus, thalamus, hypothalamus, hippocampus, substantia nigra, floor of the fourth ventricle, orbital cortex, caudate nucleus, and mammillary bodies. Enzyme activity correlated positively with age in all brain areas for MAO (with both benzylamine and tryptamine substrates) but no consistent pattern of correlation was found for COMT and TH. Mean MAO activity was significantly higher in women than men. There is increased brain MAO activity during late childhood and adolescence. These data are consistent with previous evidence suggesting that age and sex are important determinants of amine metabolism in the human central nervous system. | Monoamine metabolism in human brain. Norepinephrine (NE), dopamine (DA), tyrosine hydroxylase (TH), catechol-O-methyltransferase (COMT) and monoamine oxidase (MAO) levels were measured in human brain tissue obtained at autopsy from a series of 39 patients dying of various medical and accidental causes. The nine following brain areas were studied: globus pallidus, thalamus, hypothalamus, hippocampus, substantia nigra, floor of the fourth ventricle, orbital cortex, caudate nucleus, and mammillary bodies. Enzyme activity correlated positively with age in all brain areas for MAO (with both benzylamine and tryptamine substrates) but no consistent pattern of correlation was found for COMT and TH. Mean MAO activity was significantly higher in women than men. There is increased brain MAO activity during late childhood and adolescence. These data are consistent with previous evidence suggesting that age and sex are important determinants of amine metabolism in the human central nervous system. | [
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|
PMID:13762 | [Management of second stage of labour: observations, reflections, advices (author's transl)]. | All vaginal deliveries of the Department of Obstetrics and Gynecology of the University Basel (N = 4081) during the year 74/73 and of the University Tübingen (N = 3249) 75/74 were analysed using an IBM-system 370/135 Only alive singletons beyond the 28th week of gestation were analysed. Clinical management was quite different in the two departments; the incidence of vaginal operations (Basal 11.2%, Tübingen 12.6%), however, as well as the distribution of pH-values and Apgar-scores after 1 min were quite similar. Basel: Acidotic risk (i.e. pHUA less than 7.200) 13.5%, severe acidotic risk (i.e. pHUA less than 7.100) 1.55%, low Apgar-scores (1--3) 0.7%. Tübingen: :12.3%, 2.11%, 1.6%. 3.5% of all parturients (Basel) had duration of second stage of labour with active maternal pressure support lasting more than 30 min. In two highly selected samples differing only with regard to the occurrence of cord-entanglements at birth (N1 = 1755, N2 = 1098) the association (rank correlation method according to Kendall) between the parameters of the fetal acid-base balance and the duration of second stage of labour as well as duration of the period with "active bearing down" was studied. Without cord encirclements pH in the umbilical artery fall --0.087 and in the umbilical vein --0.115 units and with cord complications the values amounted to --0.062 (UA) and --0.120 (UV) respectively pro 60 min duration of second stage with "bearing down efforts". Analogous computations for pCO2, pO2 and HbO2 are presented. Apgar-scores in these samples showed a very loose connection with the time variables. From these data the conclusion is drawn that the indication to perform vaginal operations for termination of delivery should not primarily be governed by the factor time but rather by the whole obstetrical situation i.e. the possible fetal risk of the intervention. This holds only if maternal welfare is established and fetal well being is monitored continuously. | [Management of second stage of labour: observations, reflections, advices (author's transl)]. All vaginal deliveries of the Department of Obstetrics and Gynecology of the University Basel (N = 4081) during the year 74/73 and of the University Tübingen (N = 3249) 75/74 were analysed using an IBM-system 370/135 Only alive singletons beyond the 28th week of gestation were analysed. Clinical management was quite different in the two departments; the incidence of vaginal operations (Basal 11.2%, Tübingen 12.6%), however, as well as the distribution of pH-values and Apgar-scores after 1 min were quite similar. Basel: Acidotic risk (i.e. pHUA less than 7.200) 13.5%, severe acidotic risk (i.e. pHUA less than 7.100) 1.55%, low Apgar-scores (1--3) 0.7%. Tübingen: :12.3%, 2.11%, 1.6%. 3.5% of all parturients (Basel) had duration of second stage of labour with active maternal pressure support lasting more than 30 min. In two highly selected samples differing only with regard to the occurrence of cord-entanglements at birth (N1 = 1755, N2 = 1098) the association (rank correlation method according to Kendall) between the parameters of the fetal acid-base balance and the duration of second stage of labour as well as duration of the period with "active bearing down" was studied. Without cord encirclements pH in the umbilical artery fall --0.087 and in the umbilical vein --0.115 units and with cord complications the values amounted to --0.062 (UA) and --0.120 (UV) respectively pro 60 min duration of second stage with "bearing down efforts". Analogous computations for pCO2, pO2 and HbO2 are presented. Apgar-scores in these samples showed a very loose connection with the time variables. From these data the conclusion is drawn that the indication to perform vaginal operations for termination of delivery should not primarily be governed by the factor time but rather by the whole obstetrical situation i.e. the possible fetal risk of the intervention. This holds only if maternal welfare is established and fetal well being is monitored continuously. | [
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|
PMID:13763 | Antibiotics produced by Streptomyces olivaceus 142. I. Characterization of the FPG mutant and conditions of production of antibiotic WR 142-FPG. | By treating the Streptomyces olivaceus 142 strain simultaneously with ethyleneimine and UV radiation, the FPG mutant was isolated, which was characterized by the fact that in submerged cultures it produces a cytotoxic substance for fibroblasts and tumor cells and inhibits growth of pathogenic fungi. The mutant differs from other strains not only in having a different spectrum of antimicrobial activity, but also by taxonomic properties such as color of the aerial mycelium, liquefaction of gelatin, growth on cellulose, production of ammonia and nitrate reduction. An optimal culture medium and conditions of biosynthesis of the antibiotic in submerged cultures on the shaking machine and in 20-liter fermentation tanks were elaborated. The active substance was designated by the symbol WR 142-FPG. | Antibiotics produced by Streptomyces olivaceus 142. I. Characterization of the FPG mutant and conditions of production of antibiotic WR 142-FPG. By treating the Streptomyces olivaceus 142 strain simultaneously with ethyleneimine and UV radiation, the FPG mutant was isolated, which was characterized by the fact that in submerged cultures it produces a cytotoxic substance for fibroblasts and tumor cells and inhibits growth of pathogenic fungi. The mutant differs from other strains not only in having a different spectrum of antimicrobial activity, but also by taxonomic properties such as color of the aerial mycelium, liquefaction of gelatin, growth on cellulose, production of ammonia and nitrate reduction. An optimal culture medium and conditions of biosynthesis of the antibiotic in submerged cultures on the shaking machine and in 20-liter fermentation tanks were elaborated. The active substance was designated by the symbol WR 142-FPG. | [
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|
PMID:13764 | Search for new aminoguanidine derivatives with immunosuppressive and cytostatic properties. I. Reactions of amino-, nitroamino- and diaminoguanidine with acetylpyruvic acid ethyl ester. | Reactions of amino-, nitroamino- and diaminoguanidine (I, II, III) with acetylpyruvic acid ethyl ester (IV) at varying pH of the medium were studied. It was found that the nucleophilic hydrazine NH2 group of compounds I, II, and III) attacks the keto group of ester IV in position gamma or alpha. The newly synthesized compounds were submitted to biological evaluation. | Search for new aminoguanidine derivatives with immunosuppressive and cytostatic properties. I. Reactions of amino-, nitroamino- and diaminoguanidine with acetylpyruvic acid ethyl ester. Reactions of amino-, nitroamino- and diaminoguanidine (I, II, III) with acetylpyruvic acid ethyl ester (IV) at varying pH of the medium were studied. It was found that the nucleophilic hydrazine NH2 group of compounds I, II, and III) attacks the keto group of ester IV in position gamma or alpha. The newly synthesized compounds were submitted to biological evaluation. | [
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|
PMID:13765 | Sustrate-specificity of glucomylase (E.C. 3.2.1.3) exemplified by p-nitroaniline n-glucosides. | The present paper supplements the previous investigations on the mechanism of reactions catalyzed by amylolytic enzymes. The performed experiments corroborated the data on transglucosylative properties of animal glucoamylases which were found to depend on the structure and concentration of substrate. The p-nitroanilines were first introduced in this type of research enabling to find the differences of activity mechanism of animal glucoamylases and Aspergillus niger glucoamylase. In complement to the data reported by Japanese students the transglucosylative properties of glucoamylases were shown to depend not only on the enzyme origin and substrate concentration but also on the chemical composition of applied substrate and concentration of hydrogen ions in the reaction medium. | Sustrate-specificity of glucomylase (E.C. 3.2.1.3) exemplified by p-nitroaniline n-glucosides. The present paper supplements the previous investigations on the mechanism of reactions catalyzed by amylolytic enzymes. The performed experiments corroborated the data on transglucosylative properties of animal glucoamylases which were found to depend on the structure and concentration of substrate. The p-nitroanilines were first introduced in this type of research enabling to find the differences of activity mechanism of animal glucoamylases and Aspergillus niger glucoamylase. In complement to the data reported by Japanese students the transglucosylative properties of glucoamylases were shown to depend not only on the enzyme origin and substrate concentration but also on the chemical composition of applied substrate and concentration of hydrogen ions in the reaction medium. | [
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|
PMID:13766 | Effect of acid pH, salts, and temperature on the infectivity and physical integrity of enteroviruses. | At 2 degrees and 30 degrees C, enteroviruses are more stable on the acid than on the alkaline side of neutrality. In the range from pH 3 to 9, temperature is so influential that the fastest inactivation rate at 2 degrees C is slower than the slowest inactivation rate at 30 degrees C. Specific ions or salts also affect the rate of inactivation of enteroviruses. NaCl and other chloride salts enhance the inactivation of poliovirus at pH 3. NaCl is considerably less effective against poliovirus in the range of pH 4.5 to 7.0 than at pH less than 4.5. Loss of RNA infectivity of the virus particle proceeds as rapidly as the loss of infectivity of the particle itself, except at pH 3 in the presence of MgCl2. Inactivation results in alterations to the physical integrity of enteroviruses. At pH 5 and 7, RNA hydrolysis of poliovirus particles occurs; and at pH3, 5,6, and 7 the nucleic acid becomes susceptible to ribonuclease. Only virus particles inactivated at pH 3 show a sensitivity to chymotrypsin. The hemagglutinins of echovirus type 7 are destroyed during inactivation at pH 3,4,5, and 6; but at pH 6 this alteration precedes the loss of infectivity. The pH of the suspension is a primary determinant of the mechanism of virus destruction and possibly of the loss of infectivity at these temperatures. | Effect of acid pH, salts, and temperature on the infectivity and physical integrity of enteroviruses. At 2 degrees and 30 degrees C, enteroviruses are more stable on the acid than on the alkaline side of neutrality. In the range from pH 3 to 9, temperature is so influential that the fastest inactivation rate at 2 degrees C is slower than the slowest inactivation rate at 30 degrees C. Specific ions or salts also affect the rate of inactivation of enteroviruses. NaCl and other chloride salts enhance the inactivation of poliovirus at pH 3. NaCl is considerably less effective against poliovirus in the range of pH 4.5 to 7.0 than at pH less than 4.5. Loss of RNA infectivity of the virus particle proceeds as rapidly as the loss of infectivity of the particle itself, except at pH 3 in the presence of MgCl2. Inactivation results in alterations to the physical integrity of enteroviruses. At pH 5 and 7, RNA hydrolysis of poliovirus particles occurs; and at pH3, 5,6, and 7 the nucleic acid becomes susceptible to ribonuclease. Only virus particles inactivated at pH 3 show a sensitivity to chymotrypsin. The hemagglutinins of echovirus type 7 are destroyed during inactivation at pH 3,4,5, and 6; but at pH 6 this alteration precedes the loss of infectivity. The pH of the suspension is a primary determinant of the mechanism of virus destruction and possibly of the loss of infectivity at these temperatures. | [
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|
PMID:13767 | Investigation on the infection of cucumber mesophyll protoplasts with cucumber mosaic virus. | Isolated protoplasts from the first leaf mesophyll of cucumber plants have been successfully infected in vitro with cucumber mosaic virus (CMV). Virus instability before, during and after inoculation of the protoplasts resulted in low infectivities when extracts were assayed on cowpea; however, viral RNA extraction improved the bioassay technique. Attempts to optimize inoculation and incubation of protoplasts are outlined, incorporating the improved assay. | Investigation on the infection of cucumber mesophyll protoplasts with cucumber mosaic virus. Isolated protoplasts from the first leaf mesophyll of cucumber plants have been successfully infected in vitro with cucumber mosaic virus (CMV). Virus instability before, during and after inoculation of the protoplasts resulted in low infectivities when extracts were assayed on cowpea; however, viral RNA extraction improved the bioassay technique. Attempts to optimize inoculation and incubation of protoplasts are outlined, incorporating the improved assay. | [
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|
PMID:13768 | Thymidine-kinase in cytomegalovirus infected cells. | In human diploid fibroblast LEP cells infected with AD169 strain of human cytomegalovirus (CMV) a sharp increase of cytosol thymidine kinase activity was observed. The properties of the cytosol enzymes from infected and non-infected cells were compared. No significant differences between the enzymes from infected and control cells were observed in substrate specificity, pH dependence, thermostability and relative electrophoretic mobility. Human sera containing high titres of CMV complement-fixing antibodies did not neutralize the enzyme from infected cells. It is concluded from these results that the increase of cytosol thymidinekinase activity in CMV-infected cells was due to an enhancement of cellular thymidine kinase. | Thymidine-kinase in cytomegalovirus infected cells. In human diploid fibroblast LEP cells infected with AD169 strain of human cytomegalovirus (CMV) a sharp increase of cytosol thymidine kinase activity was observed. The properties of the cytosol enzymes from infected and non-infected cells were compared. No significant differences between the enzymes from infected and control cells were observed in substrate specificity, pH dependence, thermostability and relative electrophoretic mobility. Human sera containing high titres of CMV complement-fixing antibodies did not neutralize the enzyme from infected cells. It is concluded from these results that the increase of cytosol thymidinekinase activity in CMV-infected cells was due to an enhancement of cellular thymidine kinase. | [
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PMID:13769 | [Role of viral-bacterial associations in meningitis in children]. | A total of 50 section cases of meningitis in children were investigated. All patients revealed acute viral respiratory infections with generalization, including lesions of the central nervous system (CNS). In 14 children moreover meningococcal infection was diagnosed. In 30 children lesions of the CNS were due to mixed bacterial microflora. In 6 children along with acute viral respiratory infections (AVRI) mycoplasmosis was also revealed. Etiology of the process was determined on the basis of characteristic structural changes in the CNS and other organs, findings of virological and bacteriological investigations. In the majority of children the intravascular blood coagulation was observed. Waterhouse-Friderichsen's syndrome was revealed mainly in meningococcal infection. In order to ascertain the data obtained case records of 120 children, who had undergone treatment in connection with meningococcal infection, were analysed. It turned out that all 42 children, who had developed this disease independently, recovered. Combination of the disease with AVRI led to fatal outcomes in 12 cases out of 72. | [Role of viral-bacterial associations in meningitis in children]. A total of 50 section cases of meningitis in children were investigated. All patients revealed acute viral respiratory infections with generalization, including lesions of the central nervous system (CNS). In 14 children moreover meningococcal infection was diagnosed. In 30 children lesions of the CNS were due to mixed bacterial microflora. In 6 children along with acute viral respiratory infections (AVRI) mycoplasmosis was also revealed. Etiology of the process was determined on the basis of characteristic structural changes in the CNS and other organs, findings of virological and bacteriological investigations. In the majority of children the intravascular blood coagulation was observed. Waterhouse-Friderichsen's syndrome was revealed mainly in meningococcal infection. In order to ascertain the data obtained case records of 120 children, who had undergone treatment in connection with meningococcal infection, were analysed. It turned out that all 42 children, who had developed this disease independently, recovered. Combination of the disease with AVRI led to fatal outcomes in 12 cases out of 72. | [
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|
PMID:13770 | [Structural and functional characteristics of the spleen in systemic and local allogenic graft versus host reaction]. | Morphological and histochemical characteristics of the spleen in the systemic and locally induced in the spleen transplant host reaction were compared. In both forms there were observed infiltration of the organ with macrophages and pyroninophilial blasts characterized by a high level of the activity of enzymes of lysosomas and a hexosemonophosphate cycle. In the process of interaction between genetically heterogenous mononuclear cells the disturbance of enzymatic processes in them and in the adjacent cellular elements took place, and susequently they underwent necrosis. Most frequently this process was observed in lymphoid follicles. The destructive component in the spleen tissue was more pronounced in the systemic transplant against host reaction. | [Structural and functional characteristics of the spleen in systemic and local allogenic graft versus host reaction]. Morphological and histochemical characteristics of the spleen in the systemic and locally induced in the spleen transplant host reaction were compared. In both forms there were observed infiltration of the organ with macrophages and pyroninophilial blasts characterized by a high level of the activity of enzymes of lysosomas and a hexosemonophosphate cycle. In the process of interaction between genetically heterogenous mononuclear cells the disturbance of enzymatic processes in them and in the adjacent cellular elements took place, and susequently they underwent necrosis. Most frequently this process was observed in lymphoid follicles. The destructive component in the spleen tissue was more pronounced in the systemic transplant against host reaction. | [
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|
PMID:13773 | Murray Valley encephalitis virus infection in mosquitoes and domestic fowls in Queensland, 1974. | Field studies during an epidemic of Murray Valley encephalitis (MVE) led to the isolation of MVE virus from a pool of mosquitoes (Culex annulirostris) and a sentinel chicken from Charleville, south-west Queensland. A high proportion of domestic fowls at Charleville had antibody to MVE virus at the beginning of February 1974, in advance of the first case recognized in Queensland and allowing early warning from health authorities. A survey of antibody in domestic fowls in mid-1974 suggested widespread activity of MVE virus in western and east-central Queensland. Virus isolation and serological studies showed activity in south-west Queensland of three other viruses known to infect man, Ross River, Sindbis and Kunjin viruses. | Murray Valley encephalitis virus infection in mosquitoes and domestic fowls in Queensland, 1974. Field studies during an epidemic of Murray Valley encephalitis (MVE) led to the isolation of MVE virus from a pool of mosquitoes (Culex annulirostris) and a sentinel chicken from Charleville, south-west Queensland. A high proportion of domestic fowls at Charleville had antibody to MVE virus at the beginning of February 1974, in advance of the first case recognized in Queensland and allowing early warning from health authorities. A survey of antibody in domestic fowls in mid-1974 suggested widespread activity of MVE virus in western and east-central Queensland. Virus isolation and serological studies showed activity in south-west Queensland of three other viruses known to infect man, Ross River, Sindbis and Kunjin viruses. | [
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|
PMID:13774 | Clinical evaluation of a dialysate regeneration system for maintenance haemodialysis. | A commercially available sorbent-based dialysate regeneration system has been compared to conventional single-pass dialysate delivery systems for treatment periods of six weeks in 13 patients on maintenance dialysis. The results of treatment were virtually identical in comparing sorbent and conventional systems except that seven of the eight patients using 2-5 M2 dialysers for 3--4 hours thrice per week developed asymptomatic metabolic acidosis with the dialysate regeneration system. This complication was not seen in the five patients using 1-3 M2 dialysers and having a 6--7 hour treatment thrice weekly. Dialysate regeneration systems are particularly suited for use when water supplies are limited or of insufficient purity for single-pass dialysis, and when a portable artificial kidney is required. To avoid metabolic acidosis with this system, using currently available disposable cartridges, each dialysis treatment should be of at least 4-5 hours duration. | Clinical evaluation of a dialysate regeneration system for maintenance haemodialysis. A commercially available sorbent-based dialysate regeneration system has been compared to conventional single-pass dialysate delivery systems for treatment periods of six weeks in 13 patients on maintenance dialysis. The results of treatment were virtually identical in comparing sorbent and conventional systems except that seven of the eight patients using 2-5 M2 dialysers for 3--4 hours thrice per week developed asymptomatic metabolic acidosis with the dialysate regeneration system. This complication was not seen in the five patients using 1-3 M2 dialysers and having a 6--7 hour treatment thrice weekly. Dialysate regeneration systems are particularly suited for use when water supplies are limited or of insufficient purity for single-pass dialysis, and when a portable artificial kidney is required. To avoid metabolic acidosis with this system, using currently available disposable cartridges, each dialysis treatment should be of at least 4-5 hours duration. | [
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|
PMID:13775 | Effects of induced cryptorchidism in bulls. | Ninety bull calves were made cryptorchid by forcing the testes into the abdominal cavity and ablating the scrotum with elastrator rings. Normal spermatogenesis was seen following histological examination of some of the testes approximately 17 months after the operation. No effect on bull behaviour was noted in the cryptorchids although plasma testosterone levels were lower than those reported for bulls. It is possible that this method may leave some of the cryptorchids fertile. | Effects of induced cryptorchidism in bulls. Ninety bull calves were made cryptorchid by forcing the testes into the abdominal cavity and ablating the scrotum with elastrator rings. Normal spermatogenesis was seen following histological examination of some of the testes approximately 17 months after the operation. No effect on bull behaviour was noted in the cryptorchids although plasma testosterone levels were lower than those reported for bulls. It is possible that this method may leave some of the cryptorchids fertile. | [
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|
PMID:13781 | [Tubular structure and germ cell distribution of cryptorchid or normal testes in early childhood (author's transl)]. | Many recent publications have demonstrated that the cryptorchid testicle (and, to a lesser extent, the descended partner) are progressively injured from the second year of life onwards. Do these injuries occur in an organ which has been healthy up to this time or are they superimposed on a structurally abnormal testicle? In order to answer this, parts of cryptorchid testicles, of the descended partners, and of normal testicles were compared by histological examination of serial sections. Parts of four testes from children aged 4-7 months (2 specimens obtained by biopsy and 2 from autoptic material) and parts of four testes from children 1 1/2 years old (2 obtained by biopsy and 2 from autoptic material) were examined. The biopsies were fixed in Stieve's fixative. Tissue samples from clinically healthy children who had died suddenly were fixed in 4% formalin. The tissue was embedded in paraffin and sectioned serially; 6 mum sections were stained with HE. The spermatogonia in each cross-section and in each oblique section of a same tubule were counted and the counts of the latter were adjusted to a cross-section 50-60 mum in diameter. This counting technique did not alter the density of spermatogonia. The graphs present data on the density of spermatogonia through the lengths of the tubules examined and demonstrate tubular branching and blind ends. In the first year of life the cryptorchid testis and its descended partner showed repeated long sections lacking spermatogonia in the same tubule, whereas in normal testes the spermatogonia were more evenly distributed. The cryptorchid testis showed increased tubule branching in the areas examined. In the second year of life the tubules of the cryptorchid testis and its descended partner manifest areas free of germ cells, increased branching, and blind ends. The cryptorchid testis also had a tubule completely free of spermatogonia. The germ cell-free parts were always associated with a smaller tubule diameter than normal. The normal testes did not disclose increased branching or spermatogonium-free areas within similar lengths of tubules and showed an even distribution of spermatogonia. The different distribution of spermatogonia within the tubules and the increased branching of the tubules in cryptorchid testes indicate a previous disturbance of testis development. | [Tubular structure and germ cell distribution of cryptorchid or normal testes in early childhood (author's transl)]. Many recent publications have demonstrated that the cryptorchid testicle (and, to a lesser extent, the descended partner) are progressively injured from the second year of life onwards. Do these injuries occur in an organ which has been healthy up to this time or are they superimposed on a structurally abnormal testicle? In order to answer this, parts of cryptorchid testicles, of the descended partners, and of normal testicles were compared by histological examination of serial sections. Parts of four testes from children aged 4-7 months (2 specimens obtained by biopsy and 2 from autoptic material) and parts of four testes from children 1 1/2 years old (2 obtained by biopsy and 2 from autoptic material) were examined. The biopsies were fixed in Stieve's fixative. Tissue samples from clinically healthy children who had died suddenly were fixed in 4% formalin. The tissue was embedded in paraffin and sectioned serially; 6 mum sections were stained with HE. The spermatogonia in each cross-section and in each oblique section of a same tubule were counted and the counts of the latter were adjusted to a cross-section 50-60 mum in diameter. This counting technique did not alter the density of spermatogonia. The graphs present data on the density of spermatogonia through the lengths of the tubules examined and demonstrate tubular branching and blind ends. In the first year of life the cryptorchid testis and its descended partner showed repeated long sections lacking spermatogonia in the same tubule, whereas in normal testes the spermatogonia were more evenly distributed. The cryptorchid testis showed increased tubule branching in the areas examined. In the second year of life the tubules of the cryptorchid testis and its descended partner manifest areas free of germ cells, increased branching, and blind ends. The cryptorchid testis also had a tubule completely free of spermatogonia. The germ cell-free parts were always associated with a smaller tubule diameter than normal. The normal testes did not disclose increased branching or spermatogonium-free areas within similar lengths of tubules and showed an even distribution of spermatogonia. The different distribution of spermatogonia within the tubules and the increased branching of the tubules in cryptorchid testes indicate a previous disturbance of testis development. | [
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PMID:13782 | Genetics of peroxisomal enzymes in the mouse: nonlinkage of D-amino acid oxidase locus (Dao) to catalase (Cs) and L-alpha-hydroxyacid oxidase (Hao-1) loci on chromosome 2. | An electrophoretic polymorphism of the peroxisomal enzyme D-amino acid oxidase was observed in NZC strain Mus musculus. F1 (NZC X BALB/c) mice showed a codominant allcle two-banded phenotype which is inconsistent with the dimeric subunit structure reported for this enzyme in other species. The enzyme locus (Dao) was shown to segregate independently of Hao-1, encoding the peroxisomal enzyme hydroxyacid oxidase (liver on A4 isozyme). Thus Dao is not linked to previously mapped peroxisomal enzyme loci, Hao-1 and Cs, closely localized on chromosome 2. | Genetics of peroxisomal enzymes in the mouse: nonlinkage of D-amino acid oxidase locus (Dao) to catalase (Cs) and L-alpha-hydroxyacid oxidase (Hao-1) loci on chromosome 2. An electrophoretic polymorphism of the peroxisomal enzyme D-amino acid oxidase was observed in NZC strain Mus musculus. F1 (NZC X BALB/c) mice showed a codominant allcle two-banded phenotype which is inconsistent with the dimeric subunit structure reported for this enzyme in other species. The enzyme locus (Dao) was shown to segregate independently of Hao-1, encoding the peroxisomal enzyme hydroxyacid oxidase (liver on A4 isozyme). Thus Dao is not linked to previously mapped peroxisomal enzyme loci, Hao-1 and Cs, closely localized on chromosome 2. | [
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PMID:13780 | Myocardial function during acute hypoxia in the calf. | Hemodynamics and myocardial contractility were evaluated in five unanesthetized calves during acute hypocapnic and isocapnic hypoxia and during acute hypocapnic hypoxia with beta-adrenergic blockade. Both hypocapnic and isocapnic hypoxia, with mean PaO2 levels of 33.1 and 39.1 mm Hg respectively, produced a decline in stroke volume and index, while cardiac output and index were maintained at normoxic control levels by an increase in heart rate. Evaluation of myocardial contractility indices suggested an augmentation of left ventricular contractility during both hypocapnic and isocapnic hypoxia. Beta-adrenergic blockade effectively eliminated the increase in left ventricular contractility during hypocapnic hypoxia, suggesting an important role of the adrenergic nervous system in the genesis of the cardiovascular response of the calf to acute hypoxia. Right ventricular contractility indices failed to demonstrate a clear-cut augmentation of contractility during hypocapnic and isocapnic hypoxia when the concurrent increase in afterload was considered. Mean pulmonary arterial blood pressure rose significantly during hypocapnic and isocapnic hypoxia. The pulmonary pressor response to hypocapnic hypoxia was significantly augmented by beta blockade, indicating that the autonomic nervous system is capable of modifying the hypoxic pulmonary pressor response in this species. | Myocardial function during acute hypoxia in the calf. Hemodynamics and myocardial contractility were evaluated in five unanesthetized calves during acute hypocapnic and isocapnic hypoxia and during acute hypocapnic hypoxia with beta-adrenergic blockade. Both hypocapnic and isocapnic hypoxia, with mean PaO2 levels of 33.1 and 39.1 mm Hg respectively, produced a decline in stroke volume and index, while cardiac output and index were maintained at normoxic control levels by an increase in heart rate. Evaluation of myocardial contractility indices suggested an augmentation of left ventricular contractility during both hypocapnic and isocapnic hypoxia. Beta-adrenergic blockade effectively eliminated the increase in left ventricular contractility during hypocapnic hypoxia, suggesting an important role of the adrenergic nervous system in the genesis of the cardiovascular response of the calf to acute hypoxia. Right ventricular contractility indices failed to demonstrate a clear-cut augmentation of contractility during hypocapnic and isocapnic hypoxia when the concurrent increase in afterload was considered. Mean pulmonary arterial blood pressure rose significantly during hypocapnic and isocapnic hypoxia. The pulmonary pressor response to hypocapnic hypoxia was significantly augmented by beta blockade, indicating that the autonomic nervous system is capable of modifying the hypoxic pulmonary pressor response in this species. | [
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PMID:13783 | The maximum activities of hexokinase, phosphorylase, phosphofructokinase, glycerol phosphate dehydrogenases, lactate dehydrogenase, octopine dehydrogenase, phosphoenolpyruvate carboxykinase, nucleoside diphosphatekinase, glutamate-oxaloacetate transaminase and arginine kinase in relation to carbohydrate utilization in muscles from marine invertebrates. | Comparison of the activities of hexokinase, phosphorylase and phosphofructokinase in muscles from marine invertebrates indicates that they can be divided into three groups. First, the activities of the three enzymes are low in coelenterate muscles, catch muscles of molluscs and muscles of echinoderms; this indicates a low rate of carbohydrate (and energy) utilization by these muscles. Secondly, high activities of phosphorylase and phosphofructokinase relative to those of hexokinase are found in, for example, lobster abdominal and scallop snap muscles; this indicates that these muscles depend largely on anaerobic degradation of glycogen for energy production. Thirdly, high activities of hexokinase are found in the radular muscles of prosobranch molluscs and the fin muscles of squids; this indicates a high capacity for glucose utilization, which is consistent with the high activities of enzymes of the tricarboxylic acid cycle in these muscles [Alp, Newsholme & Zammit (1976) Biochem. J. 154, 689-700]. 2. The activities of lactate dehydrogenase, octopine dehydrogenase, phosphoenolpyruvate carboxykinase, cytosolic and mitochondrial glycerol 3-phosphate dehydrogenase and glutamate-oxaloacetate transaminase were measured in order to provide a qualitative indication of the importance of different processes for oxidation of glycolytically formed NADH. The muscles are divided into four groups: those that have a high activity of lactate dehydrogenase relative to the activities of phosphofructokinase (e.g. crustacean muscles); those that have high activities of octopine dehydrogenase but low activities of lactate dehydrogenase (e.g. scallop snap muscle); those that have moderate activities of both lactate dehydrogenase and octopine dehydrogenase (radular muscles of prosobranchs), and those that have low activities of both lactate dehydrogenase and octopine dehydrogenase, but which possess activities of phosphoenolpyruvate carboxykinase (oyster adductor muscles). It is suggested that, under anaerobic conditions, muscles of marine invertebrates form lactate and/or octopine or succinate (or similar end product) according to the activities of the enzymes present in the muscles (see above). The muscles investigated possess low activities of cytosolic glycerol 3-phosphate dehydrogenase, which indicates that glycerol phosphate formation is quantitatively unimportant under anaerobic conditions, and low activities of mitochondrial glycerol phosphate dehydrogenase, which indicates that the glycerol phosphate cycle is unimportant in the re-oxidation of glycolytically produced NADH in these muscles under aerobic conditions. Conversely, high activities of glutamate-oxaloacetate transaminase are present in some muscles, which indicates that the malate-aspartate cycle may be important in oxidation of glycolytically produced NADH under aerobic conditions. 3. High activities of nucleoside diphosphate kinase were found in muscles that function for prolonged periods under anaerobic conditions (e.g... | The maximum activities of hexokinase, phosphorylase, phosphofructokinase, glycerol phosphate dehydrogenases, lactate dehydrogenase, octopine dehydrogenase, phosphoenolpyruvate carboxykinase, nucleoside diphosphatekinase, glutamate-oxaloacetate transaminase and arginine kinase in relation to carbohydrate utilization in muscles from marine invertebrates. Comparison of the activities of hexokinase, phosphorylase and phosphofructokinase in muscles from marine invertebrates indicates that they can be divided into three groups. First, the activities of the three enzymes are low in coelenterate muscles, catch muscles of molluscs and muscles of echinoderms; this indicates a low rate of carbohydrate (and energy) utilization by these muscles. Secondly, high activities of phosphorylase and phosphofructokinase relative to those of hexokinase are found in, for example, lobster abdominal and scallop snap muscles; this indicates that these muscles depend largely on anaerobic degradation of glycogen for energy production. Thirdly, high activities of hexokinase are found in the radular muscles of prosobranch molluscs and the fin muscles of squids; this indicates a high capacity for glucose utilization, which is consistent with the high activities of enzymes of the tricarboxylic acid cycle in these muscles [Alp, Newsholme & Zammit (1976) Biochem. J. 154, 689-700]. 2. The activities of lactate dehydrogenase, octopine dehydrogenase, phosphoenolpyruvate carboxykinase, cytosolic and mitochondrial glycerol 3-phosphate dehydrogenase and glutamate-oxaloacetate transaminase were measured in order to provide a qualitative indication of the importance of different processes for oxidation of glycolytically formed NADH. The muscles are divided into four groups: those that have a high activity of lactate dehydrogenase relative to the activities of phosphofructokinase (e.g. crustacean muscles); those that have high activities of octopine dehydrogenase but low activities of lactate dehydrogenase (e.g. scallop snap muscle); those that have moderate activities of both lactate dehydrogenase and octopine dehydrogenase (radular muscles of prosobranchs), and those that have low activities of both lactate dehydrogenase and octopine dehydrogenase, but which possess activities of phosphoenolpyruvate carboxykinase (oyster adductor muscles). It is suggested that, under anaerobic conditions, muscles of marine invertebrates form lactate and/or octopine or succinate (or similar end product) according to the activities of the enzymes present in the muscles (see above). The muscles investigated possess low activities of cytosolic glycerol 3-phosphate dehydrogenase, which indicates that glycerol phosphate formation is quantitatively unimportant under anaerobic conditions, and low activities of mitochondrial glycerol phosphate dehydrogenase, which indicates that the glycerol phosphate cycle is unimportant in the re-oxidation of glycolytically produced NADH in these muscles under aerobic conditions. Conversely, high activities of glutamate-oxaloacetate transaminase are present in some muscles, which indicates that the malate-aspartate cycle may be important in oxidation of glycolytically produced NADH under aerobic conditions. 3. High activities of nucleoside diphosphate kinase were found in muscles that function for prolonged periods under anaerobic conditions (e.g... | [
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|
PMID:13784 | Phosphate transport into brush-border membrane vesicles isolated from rat small intestine. | Uptake of Pi into brush-border membrane vesicles isolated from rat small intestine was investigated by a rapid filtration technique. The following results were obtained. 1. At pH 7.4 in the presence of a NaCl gradient across the membrane (sodium concentration in the medium higher than sodium concentration in the vesicles), phosphate was taken up by a saturable transport system, which was competitively inhibited by arsenate. Phosphate entered the same osmotically reactive space as D-glucose, which indicates that transport into the vesicles rather than binding to the membranes was determined. 2. The amount of phosphate taken up initially was increased about fourfold by lowering the pH from 7.4 to 6.0.3. When Na+ was replaced by K+, Rb+ or Cs+, the initial rate of uptake decreased at pH 7.4 but was not altered at pH 6.0.4. Experiments with different anions (SCN-,Cl-, SO42-) and with ionophores (valinomycin, monactin) showed that at pH 7.4 phosphate transport in the presence of a Na+ gradient is almost independent of the electrical potential across the vesicle membrane, whereas at pH 6.0 phosphate transport involves the transfer of negative charge. It is concluded that intestinal brush-border membranes contain a Na+/phosphate co-transport system, which catalyses under physiological conditions an electroneutral entry of Pi and Na+ into the intestinal epithelial cell. In contrast with the kidney, probably univalent phosphate and one Na+ ion instead of bivalent phosphate and two Na+ ions are transported together. | Phosphate transport into brush-border membrane vesicles isolated from rat small intestine. Uptake of Pi into brush-border membrane vesicles isolated from rat small intestine was investigated by a rapid filtration technique. The following results were obtained. 1. At pH 7.4 in the presence of a NaCl gradient across the membrane (sodium concentration in the medium higher than sodium concentration in the vesicles), phosphate was taken up by a saturable transport system, which was competitively inhibited by arsenate. Phosphate entered the same osmotically reactive space as D-glucose, which indicates that transport into the vesicles rather than binding to the membranes was determined. 2. The amount of phosphate taken up initially was increased about fourfold by lowering the pH from 7.4 to 6.0.3. When Na+ was replaced by K+, Rb+ or Cs+, the initial rate of uptake decreased at pH 7.4 but was not altered at pH 6.0.4. Experiments with different anions (SCN-,Cl-, SO42-) and with ionophores (valinomycin, monactin) showed that at pH 7.4 phosphate transport in the presence of a Na+ gradient is almost independent of the electrical potential across the vesicle membrane, whereas at pH 6.0 phosphate transport involves the transfer of negative charge. It is concluded that intestinal brush-border membranes contain a Na+/phosphate co-transport system, which catalyses under physiological conditions an electroneutral entry of Pi and Na+ into the intestinal epithelial cell. In contrast with the kidney, probably univalent phosphate and one Na+ ion instead of bivalent phosphate and two Na+ ions are transported together. | [
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|
PMID:13785 | Validation of a simple radiochemical assay measuring hydrolysis of choline-labelled microsomal phosphatidylcholine by phospholipase C. pH-dependence. | Selective hydrolysis of phosphatidylcholine species, which are selectively radioactively labelled in vivo, does not appear to interfere with a radiochemical assay for hydrolysis of microsomal phosphatidylcholine by C-type phospholipases from Bacillus cereus or Clostridium perfringens. Both phospholipases substantially hydrolysed phosphatidylcholine over the pH range 4.0-10.0. | Validation of a simple radiochemical assay measuring hydrolysis of choline-labelled microsomal phosphatidylcholine by phospholipase C. pH-dependence. Selective hydrolysis of phosphatidylcholine species, which are selectively radioactively labelled in vivo, does not appear to interfere with a radiochemical assay for hydrolysis of microsomal phosphatidylcholine by C-type phospholipases from Bacillus cereus or Clostridium perfringens. Both phospholipases substantially hydrolysed phosphatidylcholine over the pH range 4.0-10.0. | [
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|
PMID:13786 | Transannular dioxygenation of 9,10-dimethyl-1,2-benzanthracene by cytochrome P-450 oxygenase of rat liver. | 9,10-Dimethyl-1,2-benzanthracene is oxygenated by rat liver microsomal cytochrome P-450 oxygenase to its 9,10-epidioxide. This transannular 1,4-peroxide is converted further into the diol by the microsomal preparation and NADPH. These two products constitute the majority of the metabolites found under the conditions described. | Transannular dioxygenation of 9,10-dimethyl-1,2-benzanthracene by cytochrome P-450 oxygenase of rat liver. 9,10-Dimethyl-1,2-benzanthracene is oxygenated by rat liver microsomal cytochrome P-450 oxygenase to its 9,10-epidioxide. This transannular 1,4-peroxide is converted further into the diol by the microsomal preparation and NADPH. These two products constitute the majority of the metabolites found under the conditions described. | [
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|
PMID:13803 | Fatty acid synthesis in aorta. Isolation of fatty acid synthetase from chicken aorta. | Fatty acid synthesis by subcellular fractions of human aorta was studied by measuring the incorporation of either radioactive acetyl-CoA or malonyl-CoA into long chain fatty acids. The high speed supernatant fraction contained fatty acid synthetase and was capable of de novo fatty acid synthesis. The fatty acid synthetase from chicken aorta was purified 800-fold from the high speed supernatant and was judged to be 10% pure at this level. Its molecular weight was estimated to be 450,000 on the basis of agarose gel filtration chromatography, while under dissociating conditions a molecular weight of 220,000 was obtained on sodium dodecyl sulphate disc gel electrophoresis. Fatty acid synthesis was dependent on acetyl-CoA, malonyl-CoA and NADPH. The major product was free palmitic acid. In enzymatic and physical characteristics the chicken aorta fatty acid synthetase strongly resembles the synthetase isolated from chicken liver. The two enzymes cross-react immuno-chemically and this homology provides the possibility of studying the synthesis and degradation of the aorta synthetase during the development of atherosclerosis. | Fatty acid synthesis in aorta. Isolation of fatty acid synthetase from chicken aorta. Fatty acid synthesis by subcellular fractions of human aorta was studied by measuring the incorporation of either radioactive acetyl-CoA or malonyl-CoA into long chain fatty acids. The high speed supernatant fraction contained fatty acid synthetase and was capable of de novo fatty acid synthesis. The fatty acid synthetase from chicken aorta was purified 800-fold from the high speed supernatant and was judged to be 10% pure at this level. Its molecular weight was estimated to be 450,000 on the basis of agarose gel filtration chromatography, while under dissociating conditions a molecular weight of 220,000 was obtained on sodium dodecyl sulphate disc gel electrophoresis. Fatty acid synthesis was dependent on acetyl-CoA, malonyl-CoA and NADPH. The major product was free palmitic acid. In enzymatic and physical characteristics the chicken aorta fatty acid synthetase strongly resembles the synthetase isolated from chicken liver. The two enzymes cross-react immuno-chemically and this homology provides the possibility of studying the synthesis and degradation of the aorta synthetase during the development of atherosclerosis. | [
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|
PMID:13801 | Effect of altered lymphocyte function on immunologic disorders in NZB/NZW mice. | NZB/NZW F1 female mice were treated with the immunosuppresive enzyme L-asparaginar antibodies, diminished deposition of gamma-globulins in kidneys, significantly delayed the onset of proteinuria, and reduced deaths from nephritis. These effects were associated with reduction of cellular IgM antibody synthesis to both T-dependent and T-independent antigens, but the graft-versus-host reaction was not affected. After several weeks of therapy, antibodies against Asnase appeared in the circulation, the effect on antibody synthesis was lost, ANA and anti-DNA appeared, followed by proteinuria and deaths from nephritis. Therefore Asnase proved to be an effective therapy in NZB/NZW mice, but its usefulness was limited by the appearance of inactivating antibodies. | Effect of altered lymphocyte function on immunologic disorders in NZB/NZW mice. NZB/NZW F1 female mice were treated with the immunosuppresive enzyme L-asparaginar antibodies, diminished deposition of gamma-globulins in kidneys, significantly delayed the onset of proteinuria, and reduced deaths from nephritis. These effects were associated with reduction of cellular IgM antibody synthesis to both T-dependent and T-independent antigens, but the graft-versus-host reaction was not affected. After several weeks of therapy, antibodies against Asnase appeared in the circulation, the effect on antibody synthesis was lost, ANA and anti-DNA appeared, followed by proteinuria and deaths from nephritis. Therefore Asnase proved to be an effective therapy in NZB/NZW mice, but its usefulness was limited by the appearance of inactivating antibodies. | [
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|
PMID:13805 | Acute-phase reactant protein profiles: an aid to monitoring large bowel cancer by CEA and serum enzymes. | The profiles of 4 acute-phase reactant proteins (APRPs) (haptoglobin (HPT), alpha1 antitrypsin (AAT), alpha1 acid glycoprotein (AGP) and prealbumin (PALB)) have been studied during the evolution of bowel cancer. Serial measurements of these APRPs can add to the information obtained from measurements of the level of CEA and hepatic enzymes during the monitoring of postoperative patients. There is considerable stability in the profile in a given individual in health, Rises of AAT and AGP are associated with metastases. High levels of HPT may suggest involvement of the bowel wall by recurrent cancer. PALB levels tend to reflect the nutritional status. A discriminant function based on the log CEA, AAT and AGP preoperative blood levels can considerably improve on the predictive value attained using CEA levels alone. | Acute-phase reactant protein profiles: an aid to monitoring large bowel cancer by CEA and serum enzymes. The profiles of 4 acute-phase reactant proteins (APRPs) (haptoglobin (HPT), alpha1 antitrypsin (AAT), alpha1 acid glycoprotein (AGP) and prealbumin (PALB)) have been studied during the evolution of bowel cancer. Serial measurements of these APRPs can add to the information obtained from measurements of the level of CEA and hepatic enzymes during the monitoring of postoperative patients. There is considerable stability in the profile in a given individual in health, Rises of AAT and AGP are associated with metastases. High levels of HPT may suggest involvement of the bowel wall by recurrent cancer. PALB levels tend to reflect the nutritional status. A discriminant function based on the log CEA, AAT and AGP preoperative blood levels can considerably improve on the predictive value attained using CEA levels alone. | [
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|
PMID:13806 | Amino acid sequence of the Anthopleura xanthogrammica heart stimulant, anthopleurin A. | A highly potent heart stimulant, anthopleurin A, from Anthopleura xanthogrammica was shown to exist as a single polypeptide chain consisting of 49 amino acid residues. The sequence of the peptide was shown to be: Gly-Val-Ser-Cys-Leu-Cys-Asp-Ser-Asp-Gly-Pro-Ser-Val-Arg-Gly-Asn-Thr-Leu-Ser-Gly-Thr-Leu-Trp-Leu-Tyr-Pro-Ser-Gly-Cys-Pro-Ser-Gly-Trp-His-Asn-Cys-Lys-Ala-His-Gly-Pro-Thr-Ile-Gly-Trp-Cys-Cys-Lys-Gln as judged by Edman degradation of the carboxymethylcysteine derivative and the tryptic peptides obtained from the derivative. Although six carboxymethylcysteine residues were present in the polypeptide, no cysteine residues were detectable in the native protein, indicating that there are three cystine residues in anthopleurin A. | Amino acid sequence of the Anthopleura xanthogrammica heart stimulant, anthopleurin A. A highly potent heart stimulant, anthopleurin A, from Anthopleura xanthogrammica was shown to exist as a single polypeptide chain consisting of 49 amino acid residues. The sequence of the peptide was shown to be: Gly-Val-Ser-Cys-Leu-Cys-Asp-Ser-Asp-Gly-Pro-Ser-Val-Arg-Gly-Asn-Thr-Leu-Ser-Gly-Thr-Leu-Trp-Leu-Tyr-Pro-Ser-Gly-Cys-Pro-Ser-Gly-Trp-His-Asn-Cys-Lys-Ala-His-Gly-Pro-Thr-Ile-Gly-Trp-Cys-Cys-Lys-Gln as judged by Edman degradation of the carboxymethylcysteine derivative and the tryptic peptides obtained from the derivative. Although six carboxymethylcysteine residues were present in the polypeptide, no cysteine residues were detectable in the native protein, indicating that there are three cystine residues in anthopleurin A. | [
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|
PMID:13807 | Magnetic resonance studies of the binding of 13C-labeled carbon monoxide to myoglobins and hemoglobins containing modified hemes. | The effects of changes in the groups attached to the periphery of the porphyrin ring of the heme of various hemoglobin and myoglobins on the environment experienced by the ligand, carbon monoxide, have been studied by observation of the chemical shift of the bound 13CO. The results indicate that the major interaction between bound ligands and substituents around the porphyrin is that transmitted electronically from substituent to ligand. The nature of the protein environment around the ligand and the interaction between the proximal histidine (F8) and the ligand (through the iron atom) impose differences between subunits of hemoglobin and between myoglobins and hemoglobins which are largely, but not entirely, independent of these substituent effects. To assess the influence of protein structure on the chemical shifts of bound ligand, the shifts of 13CO bound to myoglobin and hemoglobins from a wide range of species have also been measured. | Magnetic resonance studies of the binding of 13C-labeled carbon monoxide to myoglobins and hemoglobins containing modified hemes. The effects of changes in the groups attached to the periphery of the porphyrin ring of the heme of various hemoglobin and myoglobins on the environment experienced by the ligand, carbon monoxide, have been studied by observation of the chemical shift of the bound 13CO. The results indicate that the major interaction between bound ligands and substituents around the porphyrin is that transmitted electronically from substituent to ligand. The nature of the protein environment around the ligand and the interaction between the proximal histidine (F8) and the ligand (through the iron atom) impose differences between subunits of hemoglobin and between myoglobins and hemoglobins which are largely, but not entirely, independent of these substituent effects. To assess the influence of protein structure on the chemical shifts of bound ligand, the shifts of 13CO bound to myoglobin and hemoglobins from a wide range of species have also been measured. | [
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|
PMID:13808 | Correlation between the exposure of aromatic chromophores at the surface of the Fc domains of immunoglobulin G and their ability to bind complement. | The recognition that certain biological effector functions associated with the Fc region of human IgG are mediated exclusively by either the Cgamma2 or Cgamma3 domains prompted a study of some of the physical properties of the isolated domains in an attempt to correlate these with functional differentiation. The degree of aromatic chromophore exposure of intact Fc and fragments corresponding to the Cgamma2 and Cgamma3 domains were determined by solvent perturbation difference spectroscopy using 20% ethylene glycol. For the monomeric Cgamma2 fragment one of the two tryptophans and all four of the tyrosines were exposed to solvent. In the pFc' fragment, which represented a dimer of two intact Cgamma3 domains, an average of 0.4 of the two tryptophans of 3.3 of the five tyrosines per chain were exposed. These data were consistent with the suggested involvement of tryptophan in complement fixation since Cgamma2 binds C1q but pFc' does not. Several fragments derived from the Cgamma3 region had previously been shown to have differing environments for their aromatic side chains from circular dichroism studies. These fragments have now been shown to exhibit different degrees of chromophore exposure to solvent. Removal of the carboxy-termimal heptapeptide from the intact, Cgamma3 domain resulted in a fragment not only showing a greater exposure of aromatic residues but also having the ability to bind Clq. Our data suggest that the structural requirements for C1Q binding may be quite commonplace within Fc, but tertiary folding limits their expression except in Cgamma2 in the native molecule. The solvent perturbation observed with Fc was somewhat lower than would have been expected from the results with the isolated domains, suggesting that interdomain interactions may result in burial of aromatic residues. | Correlation between the exposure of aromatic chromophores at the surface of the Fc domains of immunoglobulin G and their ability to bind complement. The recognition that certain biological effector functions associated with the Fc region of human IgG are mediated exclusively by either the Cgamma2 or Cgamma3 domains prompted a study of some of the physical properties of the isolated domains in an attempt to correlate these with functional differentiation. The degree of aromatic chromophore exposure of intact Fc and fragments corresponding to the Cgamma2 and Cgamma3 domains were determined by solvent perturbation difference spectroscopy using 20% ethylene glycol. For the monomeric Cgamma2 fragment one of the two tryptophans and all four of the tyrosines were exposed to solvent. In the pFc' fragment, which represented a dimer of two intact Cgamma3 domains, an average of 0.4 of the two tryptophans of 3.3 of the five tyrosines per chain were exposed. These data were consistent with the suggested involvement of tryptophan in complement fixation since Cgamma2 binds C1q but pFc' does not. Several fragments derived from the Cgamma3 region had previously been shown to have differing environments for their aromatic side chains from circular dichroism studies. These fragments have now been shown to exhibit different degrees of chromophore exposure to solvent. Removal of the carboxy-termimal heptapeptide from the intact, Cgamma3 domain resulted in a fragment not only showing a greater exposure of aromatic residues but also having the ability to bind Clq. Our data suggest that the structural requirements for C1Q binding may be quite commonplace within Fc, but tertiary folding limits their expression except in Cgamma2 in the native molecule. The solvent perturbation observed with Fc was somewhat lower than would have been expected from the results with the isolated domains, suggesting that interdomain interactions may result in burial of aromatic residues. | [
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|
PMID:13802 | [Further remarks on histochemistry applied to myodiagnosis: findings of "type predominance" (author's transl)]. | Data concerning muscular biopsies (histochemically examined) of three patients affected by Charcot-Marie-Tooth disease, neurogenic atrophy of spondilosic origin and benign congenital hypotonia, are described. The common finding was a histochemical appearence of "type predominance". This point and the possible "neurogenic" origin of benign congenital hypotonia, are discussed. | [Further remarks on histochemistry applied to myodiagnosis: findings of "type predominance" (author's transl)]. Data concerning muscular biopsies (histochemically examined) of three patients affected by Charcot-Marie-Tooth disease, neurogenic atrophy of spondilosic origin and benign congenital hypotonia, are described. The common finding was a histochemical appearence of "type predominance". This point and the possible "neurogenic" origin of benign congenital hypotonia, are discussed. | [
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|
PMID:13809 | Purification and properties of debranching enzyme from dogfish muscle. | Glycogen debranching enzyme (4-alpha-glucanotransferase amylo-1,6-glucosidase, EC 2.4.1.25 + 3.2.1.33) was purified 140-fold from dogfish muscle in a rapid, high-yield procedure that takes advantage of a strong binding of the enzyme to glycogen, and its quantitative adsorption to concanavalin A-Sepharose only when the polysaccharide is present. The final product was hrophoresis in the presence and absence of dodecyl sulfate. A molecular weight of 162,000 +/- 5000 was determined by sedimentation equilibrium analysis in good agreement with the value of 160,000 estimated by gel electrophoresis, but a low-sedimentation constant of 6.5 S suggests that the enzyme is asymmetric. The molecule appears to be made up of a single polypeptide chain with no evidence for multiple repeating sequences: it could not be dissociated into smaller fragments by dodecyl sulfate even after complete carboxymethylation; tryptic cleavage of the native protein yielded only two fragments of molecular weight 20,000 and 140,000 without loss of enzymatic activity. The amino acid composition of the enzyme is reported; no covalently bound phosphate or carbohydrate could be detected. All 32 sulfhydryl groups present were titrated with 5,5'-dithiobis(2-nitrobenzoic acid) under denaturing conditions; eight reacted readily in the native enzyme without loss of catalytic activity, while substitution of eight additional ones lowered the activity by 50%. Inactivation was greatly reduced by glycogen; the polysaccharide also influenced markedly the electrophoretic behavior of the enzyme and large filamentous aggregates were formed when solutions of both were mixed. Purified debranching enzyme releases 3 mumol of glucose min-1 mg-1 at 19 degrees C, pH 6.0, from a glycogen limit dextrin and one-tenth this amount when the native polysaccharide is used as substrate; glycogen is quantitatively degraded in the presence of phosphorylase. None of the usual sugar phosphates or nucleotide effectors of glycolysis affected enzymatic activity. No phosphorylation by either dogfish or rabbit skeletal muscle protein kinase or phosphorylase kinase could be demonstrated, nor any direct interaction with phosphorylase as measured by SH-group reactivity, enzymatic activity, or rate of phosphorylase b to a conversion. Purification of the 160,000 molecular weight M-line protein of skeletal muscle resulted in the quantitative removal of debranching enzyme, indicating that the two proteins are different. | Purification and properties of debranching enzyme from dogfish muscle. Glycogen debranching enzyme (4-alpha-glucanotransferase amylo-1,6-glucosidase, EC 2.4.1.25 + 3.2.1.33) was purified 140-fold from dogfish muscle in a rapid, high-yield procedure that takes advantage of a strong binding of the enzyme to glycogen, and its quantitative adsorption to concanavalin A-Sepharose only when the polysaccharide is present. The final product was hrophoresis in the presence and absence of dodecyl sulfate. A molecular weight of 162,000 +/- 5000 was determined by sedimentation equilibrium analysis in good agreement with the value of 160,000 estimated by gel electrophoresis, but a low-sedimentation constant of 6.5 S suggests that the enzyme is asymmetric. The molecule appears to be made up of a single polypeptide chain with no evidence for multiple repeating sequences: it could not be dissociated into smaller fragments by dodecyl sulfate even after complete carboxymethylation; tryptic cleavage of the native protein yielded only two fragments of molecular weight 20,000 and 140,000 without loss of enzymatic activity. The amino acid composition of the enzyme is reported; no covalently bound phosphate or carbohydrate could be detected. All 32 sulfhydryl groups present were titrated with 5,5'-dithiobis(2-nitrobenzoic acid) under denaturing conditions; eight reacted readily in the native enzyme without loss of catalytic activity, while substitution of eight additional ones lowered the activity by 50%. Inactivation was greatly reduced by glycogen; the polysaccharide also influenced markedly the electrophoretic behavior of the enzyme and large filamentous aggregates were formed when solutions of both were mixed. Purified debranching enzyme releases 3 mumol of glucose min-1 mg-1 at 19 degrees C, pH 6.0, from a glycogen limit dextrin and one-tenth this amount when the native polysaccharide is used as substrate; glycogen is quantitatively degraded in the presence of phosphorylase. None of the usual sugar phosphates or nucleotide effectors of glycolysis affected enzymatic activity. No phosphorylation by either dogfish or rabbit skeletal muscle protein kinase or phosphorylase kinase could be demonstrated, nor any direct interaction with phosphorylase as measured by SH-group reactivity, enzymatic activity, or rate of phosphorylase b to a conversion. Purification of the 160,000 molecular weight M-line protein of skeletal muscle resulted in the quantitative removal of debranching enzyme, indicating that the two proteins are different. | [
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PMID:13810 | Mechanism of pigeon liver malic enzyme: kinetics, specificity, and half-site stoichiometry of the alkylation of a cysteinyl residue by the substrate-inhibitor bromopyruvate. | Malic enzyme from pigeon liver is alkylated by the substrate analogue bromopyruvate, resulting in the concomitant loss of its oxidative decarboxylase and oxalacetate decarboxylase activities, but not its ability to reduce alpha-keto acids. The inactivation of oxidative decarboxylase activity follows saturation kinetics, indicating the formation of an enzyme-bromopyruvate complex (K congruent to 8 mM) prior to alkylation. The inactivation is inhibited by metal ions and pyridine nucleotide cofactors. Protection of malic enzyme by the substrates L-malate and pyruvate and the inhibitors tartronate and oxalate requires the presence of the above cofactors, which tighten the binding of these carboxylic acids in accord with the ordered kinetic scheme (Hsu, R. Y., Lardy, H. A., and Cleland, W. W. (1967), J. Biol. Chem. 242, 5315-5322). Bromopyruvate is reduced to L-bromolactate by malic enzyme and is an effective inhibitor of L-malate and pyruvate in the overall reaction. The apparent kinetic constants (90 muM-0.8 mM) are one to two orders of magnitude lower than the half-saturation constant (K) of inactivation, indicating a similar tightening of bromopyruvate binding in the E-NADP+ (NADPH)-Mn2+ (Mg2+)-BP complexes. During alkylation, bromopyruvate interacts initially at the carboxylic acid substrate pocket of the active site, as indicated by the protective effect of substrates and the ability of this compound to form kinetically viable complexes with malic enzyme, particularly as a competitive inhibitor of pyruvate carboxylation with a Ki (90 muM) in the same order as its apparent Michaelis constant of 98 muM. Subsequent alkylation of a cysteinyl residue blocks the C-C bond cleavage step. The incorporation of radioactivity from [14C]bromopyruvate gives a half-site stoichiometry of two carboxyketomethyl residues per tetramer, indicating strong negative cooperativity between the four subunits of equal size, or alternatively the presence of structurally dissimilar active sites. | Mechanism of pigeon liver malic enzyme: kinetics, specificity, and half-site stoichiometry of the alkylation of a cysteinyl residue by the substrate-inhibitor bromopyruvate. Malic enzyme from pigeon liver is alkylated by the substrate analogue bromopyruvate, resulting in the concomitant loss of its oxidative decarboxylase and oxalacetate decarboxylase activities, but not its ability to reduce alpha-keto acids. The inactivation of oxidative decarboxylase activity follows saturation kinetics, indicating the formation of an enzyme-bromopyruvate complex (K congruent to 8 mM) prior to alkylation. The inactivation is inhibited by metal ions and pyridine nucleotide cofactors. Protection of malic enzyme by the substrates L-malate and pyruvate and the inhibitors tartronate and oxalate requires the presence of the above cofactors, which tighten the binding of these carboxylic acids in accord with the ordered kinetic scheme (Hsu, R. Y., Lardy, H. A., and Cleland, W. W. (1967), J. Biol. Chem. 242, 5315-5322). Bromopyruvate is reduced to L-bromolactate by malic enzyme and is an effective inhibitor of L-malate and pyruvate in the overall reaction. The apparent kinetic constants (90 muM-0.8 mM) are one to two orders of magnitude lower than the half-saturation constant (K) of inactivation, indicating a similar tightening of bromopyruvate binding in the E-NADP+ (NADPH)-Mn2+ (Mg2+)-BP complexes. During alkylation, bromopyruvate interacts initially at the carboxylic acid substrate pocket of the active site, as indicated by the protective effect of substrates and the ability of this compound to form kinetically viable complexes with malic enzyme, particularly as a competitive inhibitor of pyruvate carboxylation with a Ki (90 muM) in the same order as its apparent Michaelis constant of 98 muM. Subsequent alkylation of a cysteinyl residue blocks the C-C bond cleavage step. The incorporation of radioactivity from [14C]bromopyruvate gives a half-site stoichiometry of two carboxyketomethyl residues per tetramer, indicating strong negative cooperativity between the four subunits of equal size, or alternatively the presence of structurally dissimilar active sites. | [
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|
PMID:13811 | Mechanism of mitochondrial carbamoyl-phosphate synthetase: synthesis and properties of active CO2, precursor of carbamoyl phosphate. | This paper demonstrates the formation of "active CO2" (CO2-P), a precursor of carbamoyl phosphate (CP), with frog liver carbamoyl-phosphate synthetase. Absence of ammonia is essential for the demonstration by pulse incubation with H14CO3- of CO2-P. Adenosine triphosphate (ATP) and acetylglutamate are required for the synthesis of CO2-P, which is highly unstable in aqueous solutions (t1/2 = 0.75 s at 24 degrees C at neutral pH). In the absence of ammonia, CO2-P attains rapidly a steady-state level, which depends on the concentration of ATP and HCO3-. The "apparent KM'S" are approximately equal to those found for the adenosine triphosphate (ATPase) activity of the enzyme. The maximum level of CO2-P is limited by the amount of enzyme, and approximates 4 mol of intermediate/mol of enzyme. The unprotonated form of ammonia seems to be the species reacting with CO2-P to produce CP. The reaction of CO2-P and NH3 is very fast (rate constant kn = 8 x 10(4) M-1 S-1) and does not consume free ATP. Therefore, the 2 mol of ATP necessary for CP synthesis binds or reacts with the enzyme and/or CO2 prior to reaction with NH3. The reaction of CO2-P with NH3 also takes place in acetone under conditions at which the enzyme is not active, suggesting little or no assistance from enzyme catalysis or that a part of the catalytic site is "frozen" by the solvent in the active conformation. In the light of these and other findings, a new scheme is proposed for the mechanism of frog liver carbamoyl-phosphate synthetase and some considerations are made on the chemical nature of the intermediate and on the possible evolutionary significance of the reaction of CO2-P with NH3 in acetone. | Mechanism of mitochondrial carbamoyl-phosphate synthetase: synthesis and properties of active CO2, precursor of carbamoyl phosphate. This paper demonstrates the formation of "active CO2" (CO2-P), a precursor of carbamoyl phosphate (CP), with frog liver carbamoyl-phosphate synthetase. Absence of ammonia is essential for the demonstration by pulse incubation with H14CO3- of CO2-P. Adenosine triphosphate (ATP) and acetylglutamate are required for the synthesis of CO2-P, which is highly unstable in aqueous solutions (t1/2 = 0.75 s at 24 degrees C at neutral pH). In the absence of ammonia, CO2-P attains rapidly a steady-state level, which depends on the concentration of ATP and HCO3-. The "apparent KM'S" are approximately equal to those found for the adenosine triphosphate (ATPase) activity of the enzyme. The maximum level of CO2-P is limited by the amount of enzyme, and approximates 4 mol of intermediate/mol of enzyme. The unprotonated form of ammonia seems to be the species reacting with CO2-P to produce CP. The reaction of CO2-P and NH3 is very fast (rate constant kn = 8 x 10(4) M-1 S-1) and does not consume free ATP. Therefore, the 2 mol of ATP necessary for CP synthesis binds or reacts with the enzyme and/or CO2 prior to reaction with NH3. The reaction of CO2-P with NH3 also takes place in acetone under conditions at which the enzyme is not active, suggesting little or no assistance from enzyme catalysis or that a part of the catalytic site is "frozen" by the solvent in the active conformation. In the light of these and other findings, a new scheme is proposed for the mechanism of frog liver carbamoyl-phosphate synthetase and some considerations are made on the chemical nature of the intermediate and on the possible evolutionary significance of the reaction of CO2-P with NH3 in acetone. | [
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|
PMID:13812 | PH-induced changes in the reactions controlled by the low- and high-affinity Ca2+-binding sites in sarcoplasmic reticulum. | The effect of pH on the Ca2+-binding sites of high and low affinity, located respectively on the outer and inner surfaces of the sarcoplasmic reticulum membrane, was investigated using intact and leaky sarcoplasmic reticulum vesicles. With the use of intact vesicles, different pH profiles of membrane phosphorylation and rates of nucleoside triphosphate hydrolysis were obtained depending on the assay temperature, on the Ca2+ concentration, and on whether ATP or ITP was used as substrate. The different pH profiles were related to the amount of Ca2+ accumualted by the vesicles, i.e., to different degrees of saturation of the inner, low-affinity Ca2+-binding site. With the use of leaky vesicles, the saturation of the two Ca2+-binding sites could be controlled more precisely since the Ca2+ concentration on both sides of the membrane was equal to the Ca2+ concentration of the assay medium. Using leaky vesicles and measuring the rates of nucleotide hydrolysis, nucleotide-phosphate exchange and membrane phosphorylation by nucleotide as an indication of the degree of saturation of the Ca2+-binding sites, we observed that the affinity of both the high- and low-affinity sites increased three to four orders of magnitude when the pH of the assay medium was increased from 6.1 to 8.65. | PH-induced changes in the reactions controlled by the low- and high-affinity Ca2+-binding sites in sarcoplasmic reticulum. The effect of pH on the Ca2+-binding sites of high and low affinity, located respectively on the outer and inner surfaces of the sarcoplasmic reticulum membrane, was investigated using intact and leaky sarcoplasmic reticulum vesicles. With the use of intact vesicles, different pH profiles of membrane phosphorylation and rates of nucleoside triphosphate hydrolysis were obtained depending on the assay temperature, on the Ca2+ concentration, and on whether ATP or ITP was used as substrate. The different pH profiles were related to the amount of Ca2+ accumualted by the vesicles, i.e., to different degrees of saturation of the inner, low-affinity Ca2+-binding site. With the use of leaky vesicles, the saturation of the two Ca2+-binding sites could be controlled more precisely since the Ca2+ concentration on both sides of the membrane was equal to the Ca2+ concentration of the assay medium. Using leaky vesicles and measuring the rates of nucleotide hydrolysis, nucleotide-phosphate exchange and membrane phosphorylation by nucleotide as an indication of the degree of saturation of the Ca2+-binding sites, we observed that the affinity of both the high- and low-affinity sites increased three to four orders of magnitude when the pH of the assay medium was increased from 6.1 to 8.65. | [
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|
PMID:13813 | Investigation of the open ring form of nicotinamide adenine dinucleotide. | In strong alkali, nicotinamide adenine dinucleotide (NAD+) undergoes a ring opening of the nicotinamide ring. The open form of NAD+, ONAD, has two pKa values at--1.9 and 11.2 and absorbs maximally at 350 nm in its acidic form, at 372 nm in its neutral form, and at 340 nm in its aniomic form. ONAD has the chemical properties expected for a Schiff base of 2-carboxamideglutacondialdehyde (CGDA) and adenosine diphosphate ribosylamine. The decomposition of ONAD has been studied over a wide range of pH. A final product of ONAD hydrolysis is the base fluorescent compound 2-hydroxynicotinaldehyde. In the pH range 10--13, CGDA can be trapped as an intermediate, which absorbs maximally at 345 nm in its anionic form and at 320 nm in its neutral form, pKa = 2.9. The yield of 2-hydroxynicotinaldehyde from ONAD has been estimated as 95% at NaOH concentrations of 5 N and above, and is postulated to result from ring closure of CGDA. The pseudobase hydroxide ring addition adduct of NAD+, psiNAD-OH, is reversibly formed from NAD+ and is the 370-nm precursor of ONAD. | Investigation of the open ring form of nicotinamide adenine dinucleotide. In strong alkali, nicotinamide adenine dinucleotide (NAD+) undergoes a ring opening of the nicotinamide ring. The open form of NAD+, ONAD, has two pKa values at--1.9 and 11.2 and absorbs maximally at 350 nm in its acidic form, at 372 nm in its neutral form, and at 340 nm in its aniomic form. ONAD has the chemical properties expected for a Schiff base of 2-carboxamideglutacondialdehyde (CGDA) and adenosine diphosphate ribosylamine. The decomposition of ONAD has been studied over a wide range of pH. A final product of ONAD hydrolysis is the base fluorescent compound 2-hydroxynicotinaldehyde. In the pH range 10--13, CGDA can be trapped as an intermediate, which absorbs maximally at 345 nm in its anionic form and at 320 nm in its neutral form, pKa = 2.9. The yield of 2-hydroxynicotinaldehyde from ONAD has been estimated as 95% at NaOH concentrations of 5 N and above, and is postulated to result from ring closure of CGDA. The pseudobase hydroxide ring addition adduct of NAD+, psiNAD-OH, is reversibly formed from NAD+ and is the 370-nm precursor of ONAD. | [
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|
PMID:13814 | Metal site conformational states of vanadyl(IV) human serotransferrin complexes. | This study was undertaken to investigate the conformational states of the two metal sites in the human serum transferrin molecule. The 9.2 GHz electron paramagnetic resonance spectra of frozen solutions of divanadyl(IV) transferrin consist of a superposition of two sets of resonances, A and B, due to the magnetically nonequivalent binding environments of the VO2+ ion. Examination of the intensities of the A and B resonances as a function of pH from 6.0 to 10.7 reveals that they arise from two conformational states of the metal sites in which the geometrical arrangement and/or identity of one or more ligands in the first coordination sphere are different. From pH 7.5 to 9.0, the metal sites exist in A and B conformations but above pH 9.0 the A conformation. This transformation is coupled to the ionization of an apparently noncoordinating protein functional group with a pK - 10.0 +/- 0.1. Below pH 7.0, binding in the B conformation is rapidly lost, driven in part by the protonation of a functional group, possibly the anion, with a pK - 6.6 +/- 0.1. In 90% D2O, this pK is elevated to 7.8 +/- 0.1. At pH 6.0 in H2O, essentially one VO2+ ion remains bound to the protein with the metal site in the A conformation. Experiments with mixed VO2+ -Fe3+ transferrin complexes indicate that the same may be true of Fe3+. At pH 10.7, a new set of VO2+ resonances, labeled C, are observed; they possibly arise from a third conformation of the metal site. One bicarbonate or corbonate is required per VO2+ ion bound to the protein. 2.7 H+ are released per VO2+ bound in either the A or B conformations. The above results are discussed in terms of the "equivalence" and "nonequivalence" of the metal sites. | Metal site conformational states of vanadyl(IV) human serotransferrin complexes. This study was undertaken to investigate the conformational states of the two metal sites in the human serum transferrin molecule. The 9.2 GHz electron paramagnetic resonance spectra of frozen solutions of divanadyl(IV) transferrin consist of a superposition of two sets of resonances, A and B, due to the magnetically nonequivalent binding environments of the VO2+ ion. Examination of the intensities of the A and B resonances as a function of pH from 6.0 to 10.7 reveals that they arise from two conformational states of the metal sites in which the geometrical arrangement and/or identity of one or more ligands in the first coordination sphere are different. From pH 7.5 to 9.0, the metal sites exist in A and B conformations but above pH 9.0 the A conformation. This transformation is coupled to the ionization of an apparently noncoordinating protein functional group with a pK - 10.0 +/- 0.1. Below pH 7.0, binding in the B conformation is rapidly lost, driven in part by the protonation of a functional group, possibly the anion, with a pK - 6.6 +/- 0.1. In 90% D2O, this pK is elevated to 7.8 +/- 0.1. At pH 6.0 in H2O, essentially one VO2+ ion remains bound to the protein with the metal site in the A conformation. Experiments with mixed VO2+ -Fe3+ transferrin complexes indicate that the same may be true of Fe3+. At pH 10.7, a new set of VO2+ resonances, labeled C, are observed; they possibly arise from a third conformation of the metal site. One bicarbonate or corbonate is required per VO2+ ion bound to the protein. 2.7 H+ are released per VO2+ bound in either the A or B conformations. The above results are discussed in terms of the "equivalence" and "nonequivalence" of the metal sites. | [
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PMID:13815 | Isolation, characterization, and activation of the magnesium dependent endodeoxyribonuclease from Bacillus subtilis. | A major endodeoxyribonulcease was isolated from a mutant of the transformable Bacillus subtilis 168. The magnesium-dependent endonuclease was purified approximately 750-fold to electrophoretic homogeneity. The enzyme had a molecular weight of about 31 000, as determined by gel filtration and polyacrylamide gel electrophoresis. The protein appears to be composed of two subunits. The nuclease was dependent on magnesium or maganese ions for hydrolytic activity. The purified nuclease degraded DNA from several species of Bacillus, as well as Escherichia coli DNA, alkylated, depurinated, and thymine-dimer containing B. subtilis DNA, and hydroxymethyluracil-containing phage DNA. The enzyme also hydrolyzed single-stranded DNA, although native DNA was the preferred substrate. However, the nuclease was unable to degrade ribosomal RNA. The cleavage products of the DNA hydrolysis have 5'-phosphate and 3'-hydroxyl ends. The enzyme could be activated in crude extracts by heat treatment or treatment with guanidine hydrochloride. The nuclease activity was inhibited by phosphate and by high concentrations of NaCl. A possible function for this endonuclease in bacterial transformation is discussed. | Isolation, characterization, and activation of the magnesium dependent endodeoxyribonuclease from Bacillus subtilis. A major endodeoxyribonulcease was isolated from a mutant of the transformable Bacillus subtilis 168. The magnesium-dependent endonuclease was purified approximately 750-fold to electrophoretic homogeneity. The enzyme had a molecular weight of about 31 000, as determined by gel filtration and polyacrylamide gel electrophoresis. The protein appears to be composed of two subunits. The nuclease was dependent on magnesium or maganese ions for hydrolytic activity. The purified nuclease degraded DNA from several species of Bacillus, as well as Escherichia coli DNA, alkylated, depurinated, and thymine-dimer containing B. subtilis DNA, and hydroxymethyluracil-containing phage DNA. The enzyme also hydrolyzed single-stranded DNA, although native DNA was the preferred substrate. However, the nuclease was unable to degrade ribosomal RNA. The cleavage products of the DNA hydrolysis have 5'-phosphate and 3'-hydroxyl ends. The enzyme could be activated in crude extracts by heat treatment or treatment with guanidine hydrochloride. The nuclease activity was inhibited by phosphate and by high concentrations of NaCl. A possible function for this endonuclease in bacterial transformation is discussed. | [
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|
PMID:13816 | Mechanism of lysozyme catalysis: role of ground-state strain in subsite D in hen egg-white and human lysozymes. | The association constants for the binding of various saccharides to hen egg-white lysozyme and human lysozyme have been measured by fluorescence titration. Among these are the oligosaccharides GlcNAc-beta(1 leads to 4)-MurNAc-beta(1 leads to 4)-GlcNAc-beta(1 leads to 4)-GlcNAc, GlcNAc-beta(1 leads to 4)-MurNAc-beta(1 leads to 4)-GlcNAc-beta(1 leads to 4)-N-acetyl-D-xylosamine, and GlcNAc-beta(1 leads to 4-GlcNAc-beta(1 leads to 4)-MurNAc, prepared here for the first time. The binding constants for saccharides which must have N-acetylmuramic acid, N-acetyl-D-glucosamine, or N-acetyl-D-xylosamine bound in subsite D indicate that there is no strain involved in the binding of N-acetyl-D-glycosamine in this site, and that the lactyl group of N-acetylmuramic acid (rather than the hydroxymethyl group) is responsible for the apparent strain previously reported for binding at this subsite. For hen egg-white lysozyme, the dependence of saccharide binding on pH or on a saturating concentration of Gd(III) suggests that the conformation of several of the complexes are different from one another and from that proposed for a productive complex. This is supported by fluorescence difference spectra of the various hen egg-white lysozyme-saccharide complexes. Human lysozyme binds most saccharides studied more weakly than the hen egg-white enzyme, but binds GlcNAc-beta(1 leads to 4)-MurNAc-beta(1leads to 4)-GlcNAc-beta(1 leads to 4)-MurNAc more strongly. It is suggested that subsite C of the human enzyme is "looser" than the equivalent site in the hen egg enzyme, so that the rearrangement of a saccharide in this subsite in response to introduction of an N-acetylmuramic acid residue into subsite D destabilizes the saccharide complexes of human lysozyme less than it does the corresponding hen egg-white lysozyme complexes. This difference and the differences in the fluorescence difference spectra of hen egg-white lysozyme and human lysozyme are ascribed mainly to the replacement of Trp-62 in hen egg-white lysozyme by Tyr-63 in the human enzyme. The implications of our findings for the assumption of superposition and additivity of energies of binding in individual subsites, and for the estimation of the role of strain in lysozyme catalysis, are discussed. | Mechanism of lysozyme catalysis: role of ground-state strain in subsite D in hen egg-white and human lysozymes. The association constants for the binding of various saccharides to hen egg-white lysozyme and human lysozyme have been measured by fluorescence titration. Among these are the oligosaccharides GlcNAc-beta(1 leads to 4)-MurNAc-beta(1 leads to 4)-GlcNAc-beta(1 leads to 4)-GlcNAc, GlcNAc-beta(1 leads to 4)-MurNAc-beta(1 leads to 4)-GlcNAc-beta(1 leads to 4)-N-acetyl-D-xylosamine, and GlcNAc-beta(1 leads to 4-GlcNAc-beta(1 leads to 4)-MurNAc, prepared here for the first time. The binding constants for saccharides which must have N-acetylmuramic acid, N-acetyl-D-glucosamine, or N-acetyl-D-xylosamine bound in subsite D indicate that there is no strain involved in the binding of N-acetyl-D-glycosamine in this site, and that the lactyl group of N-acetylmuramic acid (rather than the hydroxymethyl group) is responsible for the apparent strain previously reported for binding at this subsite. For hen egg-white lysozyme, the dependence of saccharide binding on pH or on a saturating concentration of Gd(III) suggests that the conformation of several of the complexes are different from one another and from that proposed for a productive complex. This is supported by fluorescence difference spectra of the various hen egg-white lysozyme-saccharide complexes. Human lysozyme binds most saccharides studied more weakly than the hen egg-white enzyme, but binds GlcNAc-beta(1 leads to 4)-MurNAc-beta(1leads to 4)-GlcNAc-beta(1 leads to 4)-MurNAc more strongly. It is suggested that subsite C of the human enzyme is "looser" than the equivalent site in the hen egg enzyme, so that the rearrangement of a saccharide in this subsite in response to introduction of an N-acetylmuramic acid residue into subsite D destabilizes the saccharide complexes of human lysozyme less than it does the corresponding hen egg-white lysozyme complexes. This difference and the differences in the fluorescence difference spectra of hen egg-white lysozyme and human lysozyme are ascribed mainly to the replacement of Trp-62 in hen egg-white lysozyme by Tyr-63 in the human enzyme. The implications of our findings for the assumption of superposition and additivity of energies of binding in individual subsites, and for the estimation of the role of strain in lysozyme catalysis, are discussed. | [
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|
PMID:13817 | Energetics of the cooperative and noncooperative binding of nicotinamide adenine dinucleotide to yeast glyceraldehyde-3-phosphate dehydrogenase at pH 6.5 and pH 8.5. Equilibrium and calorimetric analysis over a range of temperature. | The binding of nicotinamide adenine dinucleotide (NAD+) to yeast glyceraldehyde-3-phosphate dehydrogenase (GPDH) has been studied at pH 6.5 and 8.5, at 5,25, and 40 degrees C, by calorimetry, fluorometry, spectrophotometry, equilibrium dialysis, and flow dialysis. As reported earlier for pH 7.3 (Velick S.F., Baggott, J.P., and Sturtevant, J.M. (1971), Biochemistry 10, 779), the binding is accompanied by enthalpy changes which become rapidly more negative as the temperature increases, with delta Cp = -500 to -750 cal deg-1 (mole of NAD+ bound)-1, and by entropy changes which also, as required by the large negative delta Cp, become rapidly more negative with increasing temperature. The binding data at pH 6.5 can be fitted on the basis of either four identical noninteracting sites, or of four sites showing a small degree of negative cooperativity. The data at pH 8.5, particularly at 40 degrees C, require the introduction of positive cooperativity, as was previously shown by Kirschner et al. (Kirschner, K., Eigen, M., Bittman, R., and Voigt, B. (1966), Proc. Natl. Acad. Sci. U.S.A. 56, 1661), and can be equally well fitted on the basis of a sequential model (Adair, G.S. (1925), J. Biol. Chem. 63, 529) or a concerted model (Monod, J., Wyman, J., and Changeux, J.P. (1965), J. Mol. Biol. 12, 88). It is proposed that the observed thermodynamic changes are largely the result of a hydrophobic effect due to a decrease in the exposure of nonpolar groups to the solvent, and of a tightening of the protein structure when the coenzyme is bound with concomitant decrease in the number of easily excitable internal degrees of freedom. | Energetics of the cooperative and noncooperative binding of nicotinamide adenine dinucleotide to yeast glyceraldehyde-3-phosphate dehydrogenase at pH 6.5 and pH 8.5. Equilibrium and calorimetric analysis over a range of temperature. The binding of nicotinamide adenine dinucleotide (NAD+) to yeast glyceraldehyde-3-phosphate dehydrogenase (GPDH) has been studied at pH 6.5 and 8.5, at 5,25, and 40 degrees C, by calorimetry, fluorometry, spectrophotometry, equilibrium dialysis, and flow dialysis. As reported earlier for pH 7.3 (Velick S.F., Baggott, J.P., and Sturtevant, J.M. (1971), Biochemistry 10, 779), the binding is accompanied by enthalpy changes which become rapidly more negative as the temperature increases, with delta Cp = -500 to -750 cal deg-1 (mole of NAD+ bound)-1, and by entropy changes which also, as required by the large negative delta Cp, become rapidly more negative with increasing temperature. The binding data at pH 6.5 can be fitted on the basis of either four identical noninteracting sites, or of four sites showing a small degree of negative cooperativity. The data at pH 8.5, particularly at 40 degrees C, require the introduction of positive cooperativity, as was previously shown by Kirschner et al. (Kirschner, K., Eigen, M., Bittman, R., and Voigt, B. (1966), Proc. Natl. Acad. Sci. U.S.A. 56, 1661), and can be equally well fitted on the basis of a sequential model (Adair, G.S. (1925), J. Biol. Chem. 63, 529) or a concerted model (Monod, J., Wyman, J., and Changeux, J.P. (1965), J. Mol. Biol. 12, 88). It is proposed that the observed thermodynamic changes are largely the result of a hydrophobic effect due to a decrease in the exposure of nonpolar groups to the solvent, and of a tightening of the protein structure when the coenzyme is bound with concomitant decrease in the number of easily excitable internal degrees of freedom. | [
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|
PMID:13818 | Interaction of acetazolamide and 4-nitrothiophenolate ion with bivalent metal ion derivatives of bovine carbonic anhydrase. | The stability and rate constants for the interaction of acetazolamide (diamox) and 4-nitrothiophenolate ion (NTP) with the bivalent Mn, Co, Ni, Cu and Cd forms of bovine carbonic anhydrase have been measured by utilizing the distinct visible spectra of each metalloenzyme-NTP adduct. Differing stabilities of the various NTP and (particularly) diamox complexes reside mainly in varying values for the dissociation rate constants (kd). Intrinsic formation rate constants (for the acid form of the enzyme reacting with the basic form of the ligand) are uniformly high (greater than or equal 2 X 10(7) M-1 s-1 at 25 degrees C). Invariance of kd with pH and a bell-shaped log K-pH profile with the Cu-enzyme adducts are features observed previously with the native enzyme. Binding of NTP with the Cu and Cd metalloenzymes is stronger than to the native form. | Interaction of acetazolamide and 4-nitrothiophenolate ion with bivalent metal ion derivatives of bovine carbonic anhydrase. The stability and rate constants for the interaction of acetazolamide (diamox) and 4-nitrothiophenolate ion (NTP) with the bivalent Mn, Co, Ni, Cu and Cd forms of bovine carbonic anhydrase have been measured by utilizing the distinct visible spectra of each metalloenzyme-NTP adduct. Differing stabilities of the various NTP and (particularly) diamox complexes reside mainly in varying values for the dissociation rate constants (kd). Intrinsic formation rate constants (for the acid form of the enzyme reacting with the basic form of the ligand) are uniformly high (greater than or equal 2 X 10(7) M-1 s-1 at 25 degrees C). Invariance of kd with pH and a bell-shaped log K-pH profile with the Cu-enzyme adducts are features observed previously with the native enzyme. Binding of NTP with the Cu and Cd metalloenzymes is stronger than to the native form. | [
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|
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