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PMID:3304 | Multiple changes in distal stop-flow electrolyte patterns and reduction of acid excretion induced in rabbits by angiotensin. | 1. Angiotensin has previously been shown to inhibit distal renal tubular sodium reabsorption. As a consequence of this, or independently, it might influence the distal handling of other electrolytes. We have therefore examined the effects of angiotensin on the distal reabsorption or secretion of a spectrum of electrolytes. 2. Standard bilateral stop-flow studies were done on anaesthetized, adrenalectomized rabbits, in which the effects of intravenous infusions of either 0-02-0-05 mug min-1 kg-1 or 1 mug min-1 kg-1 of angiotensin were compared with control stop-flow results. 3. The lower dose of angiotensin inhibited distal sodium, chloride, water and magnesium reabsorption, inhibited distal hydrogen secretion and stimulated distal potassium secretion. The higher dose of angiotensin produced these changes and additionally inhibited distal calcium reabsorption. Most of the observed changes were dose-related. The low dose of angiotensin did not significantly raise blood pressure but the high dose was pressor. 4. Changes in the stop-flow patterns induced by the higher dose of angiotensin were compatible with, and may help to explain, the changes it produced in urinary excretion of sodium, chloride, potassium, magnesium and calcium in clearance studies before stop-flow. Suppression of hydrogen secretion caused by both doses of angiotensin in the stop-flow studies was also reflected by reductions in acid excretion produced by these infusion rates in additional experiments performed by clearance methods in acid-loaded, conscious rabbits. 5. The results support the view that angiotensin may have an important intrarenal role, at least in rabbits. | Multiple changes in distal stop-flow electrolyte patterns and reduction of acid excretion induced in rabbits by angiotensin. 1. Angiotensin has previously been shown to inhibit distal renal tubular sodium reabsorption. As a consequence of this, or independently, it might influence the distal handling of other electrolytes. We have therefore examined the effects of angiotensin on the distal reabsorption or secretion of a spectrum of electrolytes. 2. Standard bilateral stop-flow studies were done on anaesthetized, adrenalectomized rabbits, in which the effects of intravenous infusions of either 0-02-0-05 mug min-1 kg-1 or 1 mug min-1 kg-1 of angiotensin were compared with control stop-flow results. 3. The lower dose of angiotensin inhibited distal sodium, chloride, water and magnesium reabsorption, inhibited distal hydrogen secretion and stimulated distal potassium secretion. The higher dose of angiotensin produced these changes and additionally inhibited distal calcium reabsorption. Most of the observed changes were dose-related. The low dose of angiotensin did not significantly raise blood pressure but the high dose was pressor. 4. Changes in the stop-flow patterns induced by the higher dose of angiotensin were compatible with, and may help to explain, the changes it produced in urinary excretion of sodium, chloride, potassium, magnesium and calcium in clearance studies before stop-flow. Suppression of hydrogen secretion caused by both doses of angiotensin in the stop-flow studies was also reflected by reductions in acid excretion produced by these infusion rates in additional experiments performed by clearance methods in acid-loaded, conscious rabbits. 5. The results support the view that angiotensin may have an important intrarenal role, at least in rabbits. | [
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PMID:3305 | The acute effects of respiratory and metabolic acidosis on renal function in the dog. | 1. Effective renal plasma flow, glomerular filtration rate and cardiac output were measured in osmotically loaded dogs before and during comparable acute respiratory and metabolic acidosis. 2. Urine output increased in control dogs and in animals with metabolic acidosis, but declined with respiratory acidosis. Effective renal plasma flow and glomerular filtration rate declined with respiratory and metabolic acidosis. 3. When respiratory acidosis was buffered with sodium bicarbonate, urine volume increased and glomerular filtration rate and effective renal plasma flow were unchanged; with trihydroxymethylaminomethane, urine volume increased but glomerular filtration rate and effective renal plasma flow fell. 4. When metabolic acidosis was buffered with sodium bicarbonate, urine volume increased; with trihydroxymethylaminomethane, urine volume increased but glomerular filtration rate fell. Cardiac output declined only during metabolic acidosis, both buffered and unbuffered. 5. These studies demonstrate that, even with osmotic loading: (1) respiratory acidosis caused a decrease in glomerular filtration rate, effective renal plasma flow and urine volume; (2) metabolic acidosis depresses glomerular filtration rate and effective renal plasma flow but does not change urine volume even though cardiac output falls; (3) sodium bicarbonate is mor effective than trihydroxymethylaminomethane in preserving renal function during respiratory and metabolic acidosis. | The acute effects of respiratory and metabolic acidosis on renal function in the dog. 1. Effective renal plasma flow, glomerular filtration rate and cardiac output were measured in osmotically loaded dogs before and during comparable acute respiratory and metabolic acidosis. 2. Urine output increased in control dogs and in animals with metabolic acidosis, but declined with respiratory acidosis. Effective renal plasma flow and glomerular filtration rate declined with respiratory and metabolic acidosis. 3. When respiratory acidosis was buffered with sodium bicarbonate, urine volume increased and glomerular filtration rate and effective renal plasma flow were unchanged; with trihydroxymethylaminomethane, urine volume increased but glomerular filtration rate and effective renal plasma flow fell. 4. When metabolic acidosis was buffered with sodium bicarbonate, urine volume increased; with trihydroxymethylaminomethane, urine volume increased but glomerular filtration rate fell. Cardiac output declined only during metabolic acidosis, both buffered and unbuffered. 5. These studies demonstrate that, even with osmotic loading: (1) respiratory acidosis caused a decrease in glomerular filtration rate, effective renal plasma flow and urine volume; (2) metabolic acidosis depresses glomerular filtration rate and effective renal plasma flow but does not change urine volume even though cardiac output falls; (3) sodium bicarbonate is mor effective than trihydroxymethylaminomethane in preserving renal function during respiratory and metabolic acidosis. | [
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PMID:3306 | The haemodynamic effects of metabolic acidosis in the rat. | 1. The effect of metabolic acidosis of 4-6 h duration on cardiac output, blood pressure, heart rate, and hepatic and renal blood flow has been studied in the rat. 2. In anaesthetized rats, blood pressure and heart rate fell linearly with blood pH in both sham-operated and nephrectomized rats. There was no significant difference between the two groups in the effect of acidosis on either variable. 3. Cardiac output showed a significant fall with increasing acidosis in the conscious rat. 4. Estimated hepatic blood flow in conscious rats showed a significant positive correlation with blood pH in both sham-operated and nephrectomized animals. There was no significant difference in estimated hepatic blood flow between the two groups of animals at any blood pH. 5. In conscious rats, increasing acidosis caused a progressive decrease in estimated renal blood flow. 6. It is concluded that the increase in the previously described apparent renal contribution to lactate removal in the acidotic rat cannot be explained by any circulatory effect mediated by the kidney. The possible relevance of the findings to lactate homeostasis is discussed. | The haemodynamic effects of metabolic acidosis in the rat. 1. The effect of metabolic acidosis of 4-6 h duration on cardiac output, blood pressure, heart rate, and hepatic and renal blood flow has been studied in the rat. 2. In anaesthetized rats, blood pressure and heart rate fell linearly with blood pH in both sham-operated and nephrectomized rats. There was no significant difference between the two groups in the effect of acidosis on either variable. 3. Cardiac output showed a significant fall with increasing acidosis in the conscious rat. 4. Estimated hepatic blood flow in conscious rats showed a significant positive correlation with blood pH in both sham-operated and nephrectomized animals. There was no significant difference in estimated hepatic blood flow between the two groups of animals at any blood pH. 5. In conscious rats, increasing acidosis caused a progressive decrease in estimated renal blood flow. 6. It is concluded that the increase in the previously described apparent renal contribution to lactate removal in the acidotic rat cannot be explained by any circulatory effect mediated by the kidney. The possible relevance of the findings to lactate homeostasis is discussed. | [
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PMID:3307 | The effect of acidosis on lactate removal by the perfused rat kidney. | 1. The isolated perfused kidneys of fed rats in normal acid-base status showed a constant rate of lactate removal from the perfusate between 5 and 90 min of perfusion at a perfusate pH of 7-4-7-5. 2. Lactate removal by kidneys of rats in normal acid-base status was stimulated within 30 min by a reduction in perfusate pH to 7-1-7-2, but depressed when perfusate pH was reduced further. 3. Kidneys taken from rats previously made acidotic and perfused with media of various pH values showed a progressive fall in the rate of lactate removal during the perfusion. 4. Glucose output by the kidneys of rats in normal acid-base status perfused with lactate as substrate was not affected by an alteration in perfusate pH. The kidneys of acidotic rats generally showed an increased rate of glucose output compared with those of control rats. | The effect of acidosis on lactate removal by the perfused rat kidney. 1. The isolated perfused kidneys of fed rats in normal acid-base status showed a constant rate of lactate removal from the perfusate between 5 and 90 min of perfusion at a perfusate pH of 7-4-7-5. 2. Lactate removal by kidneys of rats in normal acid-base status was stimulated within 30 min by a reduction in perfusate pH to 7-1-7-2, but depressed when perfusate pH was reduced further. 3. Kidneys taken from rats previously made acidotic and perfused with media of various pH values showed a progressive fall in the rate of lactate removal during the perfusion. 4. Glucose output by the kidneys of rats in normal acid-base status perfused with lactate as substrate was not affected by an alteration in perfusate pH. The kidneys of acidotic rats generally showed an increased rate of glucose output compared with those of control rats. | [
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PMID:3386 | Elastin--proteoglycan interaction. Conformational changes of alpha-elastin induced by the interaction. | The interaction between alpha-elastin and a connective tissue proteoglycan was followed by optical density measurements and circular dichroism spectroscopy. It was found that interaction takes place at pH values below the isoelectric point of elastin with the formation of a complex coacervate. CD spectra demonstrated conformational changes of alpha-elastin caused by the interaction and resulting in an increase in the content of helical structure. This finding suggests the possibility of the involvement of proteoglycans in the molecular organization of elastin. | Elastin--proteoglycan interaction. Conformational changes of alpha-elastin induced by the interaction. The interaction between alpha-elastin and a connective tissue proteoglycan was followed by optical density measurements and circular dichroism spectroscopy. It was found that interaction takes place at pH values below the isoelectric point of elastin with the formation of a complex coacervate. CD spectra demonstrated conformational changes of alpha-elastin caused by the interaction and resulting in an increase in the content of helical structure. This finding suggests the possibility of the involvement of proteoglycans in the molecular organization of elastin. | [
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PMID:3387 | Bilateral cryptorchidism in a bull. | Clinical and pathological observations were made on a case of bilateral cryptorchidism in a bull. Sexual libido could not be assessed because the bull was housed alone. The location of the intraabdominal testes indicated that surgical castration would necessitate a flank laparotomy incision. | Bilateral cryptorchidism in a bull. Clinical and pathological observations were made on a case of bilateral cryptorchidism in a bull. Sexual libido could not be assessed because the bull was housed alone. The location of the intraabdominal testes indicated that surgical castration would necessitate a flank laparotomy incision. | [
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PMID:3393 | Treatment of tardive dyskinesia. | The pathogenesis of tardive dyskinesia is distinct from and may be functionally opposite to that of parkinsonism. The former is thought to be related to central nervous system dopaminergic hyperactivity, while the latter is known to be related to dopamine deficiency. An effective schema for the treatment of tardive dyskinesia includes avoiding antiparkinsonian medication and prescribing deanol, an acetylcholine precursor, while continuing or increasing phenothiazine dosages. | Treatment of tardive dyskinesia. The pathogenesis of tardive dyskinesia is distinct from and may be functionally opposite to that of parkinsonism. The former is thought to be related to central nervous system dopaminergic hyperactivity, while the latter is known to be related to dopamine deficiency. An effective schema for the treatment of tardive dyskinesia includes avoiding antiparkinsonian medication and prescribing deanol, an acetylcholine precursor, while continuing or increasing phenothiazine dosages. | [
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PMID:3394 | Epidemiology of tardive dyskinesia Part I. | We have performed an epidemiological study concerning tardive dyskinesia on a sample of 332 chronic schizophrenic patients (142 males and 190 females, mean age 48.6 years, mean duration of neuroleptic treatment 14.5 years). We could conclude that the age of patients at the time of assessment procedures is the most important variable. The prevalence of tardive dyskinesia was significantly higher in the older population. The significance of an insidious beginning of the illness might be only secondary to the highly significant role of the age. Other factors, such as sex, type of schizophrenia, initial syndrome, present psychic state, organic syndromes and neuroleptic-induced extrapyramidal syndrome, do not seen to play a role in the prevalence of tardive dyskinesia. | Epidemiology of tardive dyskinesia Part I. We have performed an epidemiological study concerning tardive dyskinesia on a sample of 332 chronic schizophrenic patients (142 males and 190 females, mean age 48.6 years, mean duration of neuroleptic treatment 14.5 years). We could conclude that the age of patients at the time of assessment procedures is the most important variable. The prevalence of tardive dyskinesia was significantly higher in the older population. The significance of an insidious beginning of the illness might be only secondary to the highly significant role of the age. Other factors, such as sex, type of schizophrenia, initial syndrome, present psychic state, organic syndromes and neuroleptic-induced extrapyramidal syndrome, do not seen to play a role in the prevalence of tardive dyskinesia. | [
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PMID:3392 | Postnatal development of the circadian rhythm of rat liver tyrosine aminotransferase activity. | The rhythm of tyrosine aminotransferase (TAT) activity in 2-day old rats is characterized by a maximum at the beginning and a further one at the end of light-time. In 7-day old animals, the rhythm is much less pronounced than in 2-day old pups. At the 21st day of life, the rats already exhibit the rhythmic pattern of the adults, although the absolute values are still somewhat below those of the adults. The TAT rhythm in neonates is obviously generated by periodic variations in cyclic AMP-dependent release of TAT from polysomes. | Postnatal development of the circadian rhythm of rat liver tyrosine aminotransferase activity. The rhythm of tyrosine aminotransferase (TAT) activity in 2-day old rats is characterized by a maximum at the beginning and a further one at the end of light-time. In 7-day old animals, the rhythm is much less pronounced than in 2-day old pups. At the 21st day of life, the rats already exhibit the rhythmic pattern of the adults, although the absolute values are still somewhat below those of the adults. The TAT rhythm in neonates is obviously generated by periodic variations in cyclic AMP-dependent release of TAT from polysomes. | [
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PMID:3395 | The influence of dehydrocholate on hepatic uptake and biliary excretion of 3H-taurocholate and 3H-ouabain. | The hepatic uptake and biliary excretion of 3H-taurocholate and 3H-ouabain was studied in the rat during saline (control) and dehydrocholate infusions. Dehydrocholate (140 mumol/hr) did not influence the plasma disappearance nor the biliary excretion of taurocholate after a single iv injection (37 mumol/kg). Bile production in the dehydrocholate experiment was increased 2- to 3-fold compared with controls. The biliary transport maximum for exogenously administered taurocholate was determined by constant infusion to be 135.0 +/- 3.0 mumol/hr (22 mumol/min/g of liver). Concomitant infusions of 140 mumol of dehydrocholate per hr did not alter the maximal taurocholate output. The effects of the two bile salts on bile flow were additive. Dehydrocholate (140 mumol/hr) reduced the biliary excretion of 3H-ouabain (0.8 mumol/kg) and elevated the secondary slow component of the plasma disappearance of the cardiac glycoside. The hepatic levels of ouabain were increased compared with controls. It is concluded that dehydrocholate interferes with ouabain transport at the canalicular level but not with primary hepatic uptake. Taurocholate (140 mumol/hr) failed to influence the total biliary output of ouabain. These differences and the lack of interaction between dehydrocholate and taurocholate suggest a hepatic transporting pathway for taurocholate which differs from that for taurocholate which differs from that for dehydrocholate and/or its metabolites. | The influence of dehydrocholate on hepatic uptake and biliary excretion of 3H-taurocholate and 3H-ouabain. The hepatic uptake and biliary excretion of 3H-taurocholate and 3H-ouabain was studied in the rat during saline (control) and dehydrocholate infusions. Dehydrocholate (140 mumol/hr) did not influence the plasma disappearance nor the biliary excretion of taurocholate after a single iv injection (37 mumol/kg). Bile production in the dehydrocholate experiment was increased 2- to 3-fold compared with controls. The biliary transport maximum for exogenously administered taurocholate was determined by constant infusion to be 135.0 +/- 3.0 mumol/hr (22 mumol/min/g of liver). Concomitant infusions of 140 mumol of dehydrocholate per hr did not alter the maximal taurocholate output. The effects of the two bile salts on bile flow were additive. Dehydrocholate (140 mumol/hr) reduced the biliary excretion of 3H-ouabain (0.8 mumol/kg) and elevated the secondary slow component of the plasma disappearance of the cardiac glycoside. The hepatic levels of ouabain were increased compared with controls. It is concluded that dehydrocholate interferes with ouabain transport at the canalicular level but not with primary hepatic uptake. Taurocholate (140 mumol/hr) failed to influence the total biliary output of ouabain. These differences and the lack of interaction between dehydrocholate and taurocholate suggest a hepatic transporting pathway for taurocholate which differs from that for taurocholate which differs from that for dehydrocholate and/or its metabolites. | [
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PMID:3396 | Postnatal development of mixed-function oxidation as measured in microsomes from the small intestine and liver of rabbits. | The postnatal development of aminopyrine N-demethylase, aniline 4-hydroxylase, benzpyrene hydroxylase, biphenyl 4-hydroxylase, 7-ethoxycoumarin 0-deethylase activities, NADPH-cytochrome c reductase, and cytochrome P-450 was compared in microsomes from the liver and small intestine of New Zealand white rabbits. Apart from hepatic aniline hydroxylase activity, all of the xenobiotic-metabolizing enzyme activities examined had a similar pattern of development in the liver and small intestine. In both tissues the ability to metabolize xenobiotics was generally undetectable at 2 days of age and remained relatively low for the first 20 days of life. Theresfter, a rapid 2- to 5-fold increase in all the enzyme activity studied was noted, and adult values were reached or exceeded by 30 days of age. Subsequent development of xenobiotic-metabolizing enzyme activities in the small intestine, but not in the liver, exhibited a transient fall at 50 days of age before adult activities were attained after 75 days of age. The developmental pattern of cytochrome P-450 in the small intestine closely resembled that of the xenobiotic-metabolizing enzyme activities, but in the liver this correlation was less exact. | Postnatal development of mixed-function oxidation as measured in microsomes from the small intestine and liver of rabbits. The postnatal development of aminopyrine N-demethylase, aniline 4-hydroxylase, benzpyrene hydroxylase, biphenyl 4-hydroxylase, 7-ethoxycoumarin 0-deethylase activities, NADPH-cytochrome c reductase, and cytochrome P-450 was compared in microsomes from the liver and small intestine of New Zealand white rabbits. Apart from hepatic aniline hydroxylase activity, all of the xenobiotic-metabolizing enzyme activities examined had a similar pattern of development in the liver and small intestine. In both tissues the ability to metabolize xenobiotics was generally undetectable at 2 days of age and remained relatively low for the first 20 days of life. Theresfter, a rapid 2- to 5-fold increase in all the enzyme activity studied was noted, and adult values were reached or exceeded by 30 days of age. Subsequent development of xenobiotic-metabolizing enzyme activities in the small intestine, but not in the liver, exhibited a transient fall at 50 days of age before adult activities were attained after 75 days of age. The developmental pattern of cytochrome P-450 in the small intestine closely resembled that of the xenobiotic-metabolizing enzyme activities, but in the liver this correlation was less exact. | [
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PMID:3397 | Characterization of the hepatic microsomal mixed-function oxidase enzyme system in miniature pigs. | Hepatic microsomal protein, cytochrome P-450, UDP-glucuronyltransferase, ethylmorphine demethylase, aniline hydroxylase, and aryl hydrocarbon hydroxylase levels were measured in the 2-, 4-, 5-, 6-, and 8-month-old Hanford miniature pig. The activities or concentrations of all of the liver parameters measured had apparently reached their adult plateau level by 2 months of age. The use of the miniature pig in toxicology research programs is discussed. | Characterization of the hepatic microsomal mixed-function oxidase enzyme system in miniature pigs. Hepatic microsomal protein, cytochrome P-450, UDP-glucuronyltransferase, ethylmorphine demethylase, aniline hydroxylase, and aryl hydrocarbon hydroxylase levels were measured in the 2-, 4-, 5-, 6-, and 8-month-old Hanford miniature pig. The activities or concentrations of all of the liver parameters measured had apparently reached their adult plateau level by 2 months of age. The use of the miniature pig in toxicology research programs is discussed. | [
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PMID:3398 | Monooxygenase-catalyzed aldrin epoxidation and dihydroisodrin hydroxylation in monkey liver needle-biopsy specimens. Assay and properties. | Aldrin epoxidation and dihydroisodrin (1,8,9,10,11,11-hexachloro-2,3-7,6-endo-2,1-7,8-endo-tetracyclo [6.2.1.1(3), (6).0(2), (7)]dodec-9-ene (DHI) hydroxylation have been studied in 0.2-ml liver monooxygenase preparations. Liver biopsy specimens of rhesus (Macaca mulatta) and bonnet (M. radiata) monkeys obtained with a 1.9-mm Menghini needle were the primary enzyme sources. Dieldrin and monohydroxydihydroisodrin (DHI-OH) were the only metabolites detected by electron-capture GLC analysis of hexane extracts of incubation media. Incubation, extraction, and analysis could be done in the same vessel. Maximum rates were obtained in the presence of NADPH and O2, and both transformations were inhibited by CO. The apparent KM and Vmax (+/-SD) for epoxidation was 1.2 +/- 0.2 X 10(-5) M aldrin and 210 +/- 20 pmol of dieldrin per mg of protein per min, and the corresponding values for hydroxylation were 2.3 +/- 0.4 X 10(-5) M DHI and 150 +/- 20 pmol of DHI-OH per mg of protein per min. Aldrin epoxidation and DHI hydroxylation activities of rhesus monkey liver biopsy and rat liver preparations were evaluated after phenobarbital treatment. The assay procedures can be used in protocols in which animals serve as their own controls. | Monooxygenase-catalyzed aldrin epoxidation and dihydroisodrin hydroxylation in monkey liver needle-biopsy specimens. Assay and properties. Aldrin epoxidation and dihydroisodrin (1,8,9,10,11,11-hexachloro-2,3-7,6-endo-2,1-7,8-endo-tetracyclo [6.2.1.1(3), (6).0(2), (7)]dodec-9-ene (DHI) hydroxylation have been studied in 0.2-ml liver monooxygenase preparations. Liver biopsy specimens of rhesus (Macaca mulatta) and bonnet (M. radiata) monkeys obtained with a 1.9-mm Menghini needle were the primary enzyme sources. Dieldrin and monohydroxydihydroisodrin (DHI-OH) were the only metabolites detected by electron-capture GLC analysis of hexane extracts of incubation media. Incubation, extraction, and analysis could be done in the same vessel. Maximum rates were obtained in the presence of NADPH and O2, and both transformations were inhibited by CO. The apparent KM and Vmax (+/-SD) for epoxidation was 1.2 +/- 0.2 X 10(-5) M aldrin and 210 +/- 20 pmol of dieldrin per mg of protein per min, and the corresponding values for hydroxylation were 2.3 +/- 0.4 X 10(-5) M DHI and 150 +/- 20 pmol of DHI-OH per mg of protein per min. Aldrin epoxidation and DHI hydroxylation activities of rhesus monkey liver biopsy and rat liver preparations were evaluated after phenobarbital treatment. The assay procedures can be used in protocols in which animals serve as their own controls. | [
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PMID:3399 | In vitro metabolism of 1-phenyl-2-propanone oxime in rat liver homogenates. | 1-Phenyl-2-propanone oxime is a known in vitro metabolite of amphetamine. Further in vitro metabolism of this oxime with the 12,000g supernatant fraction from homogenized rat liver gave one major and two minor metabolites which were identified as 2-nitro-1-phenylpropane, benzyl alcohol, and 1-phenyl-2-propanone, respectively, by means of combined gas chromatography and mass spectrometry, and by comparison with authentic samples of each product. | In vitro metabolism of 1-phenyl-2-propanone oxime in rat liver homogenates. 1-Phenyl-2-propanone oxime is a known in vitro metabolite of amphetamine. Further in vitro metabolism of this oxime with the 12,000g supernatant fraction from homogenized rat liver gave one major and two minor metabolites which were identified as 2-nitro-1-phenylpropane, benzyl alcohol, and 1-phenyl-2-propanone, respectively, by means of combined gas chromatography and mass spectrometry, and by comparison with authentic samples of each product. | [
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PMID:3400 | Anaerobic release of fluoride from halothane. Relationship to the binding of halothane metabolites to hepatic cellular constituents. | Halothane has been found to undergo a reductive defluorination. This reaction requires an active cytochrome P-450 system and NADPH, and is inducible by phenobarbital and polychlorinated biphenyls but not by methylcholanthrene. The fluoride release occurs only under low O2 tension, while high O2 tension results in the oxidation of halothane to trifluoroacetic acid, inorganic bromide, and chloride. The release of the inorganic fluoride is linear up to 60 min. Because the conditions required for fluoride release and the binding of a halothane metabolite to microsomal phospholipids are similar, the defluorinated halothane molecule is assumed to be involved with this binding. However, based on the amount of fluoride released, the defluorinated halothane metabolite represents only approximately 60% of the total amount of halothane metabolite bound, which suggests that more than one metabolite may be involved in the binding. | Anaerobic release of fluoride from halothane. Relationship to the binding of halothane metabolites to hepatic cellular constituents. Halothane has been found to undergo a reductive defluorination. This reaction requires an active cytochrome P-450 system and NADPH, and is inducible by phenobarbital and polychlorinated biphenyls but not by methylcholanthrene. The fluoride release occurs only under low O2 tension, while high O2 tension results in the oxidation of halothane to trifluoroacetic acid, inorganic bromide, and chloride. The release of the inorganic fluoride is linear up to 60 min. Because the conditions required for fluoride release and the binding of a halothane metabolite to microsomal phospholipids are similar, the defluorinated halothane molecule is assumed to be involved with this binding. However, based on the amount of fluoride released, the defluorinated halothane metabolite represents only approximately 60% of the total amount of halothane metabolite bound, which suggests that more than one metabolite may be involved in the binding. | [
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PMID:3401 | Microsomal spectral properties and narcotic N-demethylase activity in methadone-dependent rats. | Rats were given access ad lib. to various concentrations (0.3 to 1.0 mg/ml) of methadone hydrochloride dissolved in sucrose solution. The N-demethylation of various narcotics was studied in hepatic preparations from methadone-consuming rats in order to determine if there was substrate specificity for the microsomal demethylase system. The Vmax for the N-demethylation of methadone, ethylmorphine, and meperidine was increased by 40-65%, whereas that for morphine N-demethylation was reduced to 55% of the control value. Additive or synergistic effects on microsomal cytochrome P-450 content were seen when methadone consumption was supplemented by administration of maximally inducing doses of either 3-methylcholanthrene (3-MC) or phenobarbital (PB). This suggested that there was an increase in a type of cytochrome P-450 which was independent of that induced by PB or 3-MC. The qualitative change in cytochrome P-450 reflected in the ethylisocyanide binding spectrum was also apparent after treatment with methadone, PB, or 3-MC, and the combination of methadone and PB exhibited effects that differed from PB alone. Two-substrate kinetic analysis with methadone and morphine as substrates indicated that more than one enzymic system may be involved in the N-demethylation reaction and that a common component of this N-demethylase system could not be induced with phenobarbital. However, methadone and meperidine seem to be demethylated by the same enzymic system. | Microsomal spectral properties and narcotic N-demethylase activity in methadone-dependent rats. Rats were given access ad lib. to various concentrations (0.3 to 1.0 mg/ml) of methadone hydrochloride dissolved in sucrose solution. The N-demethylation of various narcotics was studied in hepatic preparations from methadone-consuming rats in order to determine if there was substrate specificity for the microsomal demethylase system. The Vmax for the N-demethylation of methadone, ethylmorphine, and meperidine was increased by 40-65%, whereas that for morphine N-demethylation was reduced to 55% of the control value. Additive or synergistic effects on microsomal cytochrome P-450 content were seen when methadone consumption was supplemented by administration of maximally inducing doses of either 3-methylcholanthrene (3-MC) or phenobarbital (PB). This suggested that there was an increase in a type of cytochrome P-450 which was independent of that induced by PB or 3-MC. The qualitative change in cytochrome P-450 reflected in the ethylisocyanide binding spectrum was also apparent after treatment with methadone, PB, or 3-MC, and the combination of methadone and PB exhibited effects that differed from PB alone. Two-substrate kinetic analysis with methadone and morphine as substrates indicated that more than one enzymic system may be involved in the N-demethylation reaction and that a common component of this N-demethylase system could not be induced with phenobarbital. However, methadone and meperidine seem to be demethylated by the same enzymic system. | [
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PMID:3402 | Metabolism of 2-(3-chloro-4(4-chlorobenzoyl)-phenyl)-as-triazine-3,5(2H,4H)-dione by the chicken. | The metabolism of the anticoccidial 2-[3-chloro-4-(4-chlorobenzoyl)phenyl]-as-triazine-3,-5(2H,4H)-dione (CP-25,415) was investigated in the chicken. It was shown that the predominant residue present in the chicken was 2-[3-chloro-4-(alpha-hydroxy-4-chlorobenzoyl)phenyl]-as-triazine-3,5(2H,4H)-dione (CP-25,641). A gas-liquid chromatographic assay for the analysis of CP-25,641 in biological fluids and tissues was developed which was rapid, accurate, and reproducible. Results of the analytical method correlated well with radiochemical measurements and were indicative of the total drug-related residues. The half-life of CP-25,641 in tissues was approximately 32 hr except in the kidney, where the half-life was approximately 40 hr due to urine retention by the kidneys. CP-25,641 was excreted without further change. | Metabolism of 2-(3-chloro-4(4-chlorobenzoyl)-phenyl)-as-triazine-3,5(2H,4H)-dione by the chicken. The metabolism of the anticoccidial 2-[3-chloro-4-(4-chlorobenzoyl)phenyl]-as-triazine-3,-5(2H,4H)-dione (CP-25,415) was investigated in the chicken. It was shown that the predominant residue present in the chicken was 2-[3-chloro-4-(alpha-hydroxy-4-chlorobenzoyl)phenyl]-as-triazine-3,5(2H,4H)-dione (CP-25,641). A gas-liquid chromatographic assay for the analysis of CP-25,641 in biological fluids and tissues was developed which was rapid, accurate, and reproducible. Results of the analytical method correlated well with radiochemical measurements and were indicative of the total drug-related residues. The half-life of CP-25,641 in tissues was approximately 32 hr except in the kidney, where the half-life was approximately 40 hr due to urine retention by the kidneys. CP-25,641 was excreted without further change. | [
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PMID:3403 | Physiological disposition and metabolism of N-t-butylarterenol and its di-p-toluate ester (bitolterol) in the rat. | The metabolism and disposition of the bronchodilator, N-t-butylarterenol (tBA) and its di-p-toluate ester (bitolterol) were compared in the rat. Radioactivity was preferentially retained in lungs of rats compared with heart and blood after iv medication with tritium-labeled bitolterol, but was not retained in tissues after iv medication with [3H]tBA. After oral and iv medication with [3H]bitolterol, fecal radioactivity accounted for 24% of the dose and 65 and 79% of the radioactivity, respectively, was excreted in urine (0-72 hr). In comparison, urine radioactivity after oral and iv medication with [3H]tBA was 43 and 83% of the dose, respectively, and fecal radioactivity accounted for 43 or 23% of the dose, respectively (0-72 hr). Bitolterol was hydrolyzed in vitro to tBA by esterases found in various tissues including small intestine, liver, and plasma. Moreover, tBA was a substrate for catecholamine O-methyltransferase but not for monoamine oxidase. Similar metabolites were observed in urine samples of rats given either [3H]tBA or [3H]bitolterol. Urine metabolites were identified as free and conjugated forms of both tBA and 3-O-methyl-tBA. | Physiological disposition and metabolism of N-t-butylarterenol and its di-p-toluate ester (bitolterol) in the rat. The metabolism and disposition of the bronchodilator, N-t-butylarterenol (tBA) and its di-p-toluate ester (bitolterol) were compared in the rat. Radioactivity was preferentially retained in lungs of rats compared with heart and blood after iv medication with tritium-labeled bitolterol, but was not retained in tissues after iv medication with [3H]tBA. After oral and iv medication with [3H]bitolterol, fecal radioactivity accounted for 24% of the dose and 65 and 79% of the radioactivity, respectively, was excreted in urine (0-72 hr). In comparison, urine radioactivity after oral and iv medication with [3H]tBA was 43 and 83% of the dose, respectively, and fecal radioactivity accounted for 43 or 23% of the dose, respectively (0-72 hr). Bitolterol was hydrolyzed in vitro to tBA by esterases found in various tissues including small intestine, liver, and plasma. Moreover, tBA was a substrate for catecholamine O-methyltransferase but not for monoamine oxidase. Similar metabolites were observed in urine samples of rats given either [3H]tBA or [3H]bitolterol. Urine metabolites were identified as free and conjugated forms of both tBA and 3-O-methyl-tBA. | [
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PMID:3404 | Physiological disposition and metabolism of (3H)bitolterol in man and dog. | The metabolism and disposition of bitolterol, the di-p-toluate ester of N-t-butylarterenol (tBA) was studied in man after a single oral dose and in dog after intraduodenal, iv, or oral administration. The mean (+/- SE) peak plasma radioactivity in man (dose, 70 mug/kg) was 180 +/- 18 ng equivalents of [3H]bitolterol per ml or approximately 11% of the dose, whereas peak plasma radioactivity in dog (dose, 200 mug/kg) was 144 +/- 23 ng equivalents per ml or approximately 4% of the dose. For both man and dog, the time for maximum plasma level of radioactivity varied from 0.5 to 2 hr. In man, only 1% of the plasma radioactivity represented intact [3H]bitolterol 1.0 hr after medication. In the dog, radioactivity was concentrated in lung tissue after iv administration of [3H]bitolterol. Recovery of intact [3H]bitolterol in lung at 4.5 hr ranged from 26 to 46% of total tissue radioactivity after iv dosage and from 4 to 14% total tissue radioactivity after intraduodenal administration. Radioactivity recovered in human urine and feces (0-72 hr) accounted for 86 and 8.1% of the dose, respectively. Recovery of radioactivity in dog urine and feces accounted for 58 and 23% of the dose, respectively, in the same time period. Radiochromatograms of urine samples from man and dog revealed similar patterns of metabolites including free and conjugated forms of both tBA and the 3-O-methyl metabolite, N-t-butylmetarterenol. The major radioactive components of the feces were bitolterol and tBA. The results indicate that bitolterol is absorbed orally and retained as the intact ester in lung. The prolonged bronchodilator activity of bitolterol is due to the slow release of the ester from lung and hydrolysis to tBA, an active beta2-adrenoceptor agonist. Pharmacological activity is terminated by metabolism of tBA via conjugation or 3-O-methylation. | Physiological disposition and metabolism of (3H)bitolterol in man and dog. The metabolism and disposition of bitolterol, the di-p-toluate ester of N-t-butylarterenol (tBA) was studied in man after a single oral dose and in dog after intraduodenal, iv, or oral administration. The mean (+/- SE) peak plasma radioactivity in man (dose, 70 mug/kg) was 180 +/- 18 ng equivalents of [3H]bitolterol per ml or approximately 11% of the dose, whereas peak plasma radioactivity in dog (dose, 200 mug/kg) was 144 +/- 23 ng equivalents per ml or approximately 4% of the dose. For both man and dog, the time for maximum plasma level of radioactivity varied from 0.5 to 2 hr. In man, only 1% of the plasma radioactivity represented intact [3H]bitolterol 1.0 hr after medication. In the dog, radioactivity was concentrated in lung tissue after iv administration of [3H]bitolterol. Recovery of intact [3H]bitolterol in lung at 4.5 hr ranged from 26 to 46% of total tissue radioactivity after iv dosage and from 4 to 14% total tissue radioactivity after intraduodenal administration. Radioactivity recovered in human urine and feces (0-72 hr) accounted for 86 and 8.1% of the dose, respectively. Recovery of radioactivity in dog urine and feces accounted for 58 and 23% of the dose, respectively, in the same time period. Radiochromatograms of urine samples from man and dog revealed similar patterns of metabolites including free and conjugated forms of both tBA and the 3-O-methyl metabolite, N-t-butylmetarterenol. The major radioactive components of the feces were bitolterol and tBA. The results indicate that bitolterol is absorbed orally and retained as the intact ester in lung. The prolonged bronchodilator activity of bitolterol is due to the slow release of the ester from lung and hydrolysis to tBA, an active beta2-adrenoceptor agonist. Pharmacological activity is terminated by metabolism of tBA via conjugation or 3-O-methylation. | [
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PMID:3405 | Adriamycin metabolism in man. Evidence from urinary metabolites. | We studied the human metabolism of adriamycin by isolating and identifying urinary metabolites which retain adriamycin's specific fluorescence properties. Metabolites were extracted by adsorption to polystyrene polymeric sorbants, separated on silicic acid columns and purified by thin-layer chromatography. Structures were determined by comparative chromatography; infrared, fluorescence, and mass spectroscopy; and enzymatic and chemical degradation. Substances identified were adriamycinol, adriamycinol aglycone, adriamycin aglycone, deoxyadriamycin aglycone, deoxyadriamycinol aglycone, demethyldeoxyadriamycinol aglycone, demethyldeoxyadriamycinol aglycone 4-O-sulfate, and demethyldeoxyadriamycinol aglycone 4-O-beta-glucuronide. Other metabolites have been purified but not identified. Human metabolism of adriamycin involved carbonyl reduction, reductive glycosidic cleavage, hydrolytic glycosidic cleavage, O-demethylation, O-sulfation, and O-beta-glucuronidation. Carbonyl reduction was the major enzymatic conversion occurring in the human. | Adriamycin metabolism in man. Evidence from urinary metabolites. We studied the human metabolism of adriamycin by isolating and identifying urinary metabolites which retain adriamycin's specific fluorescence properties. Metabolites were extracted by adsorption to polystyrene polymeric sorbants, separated on silicic acid columns and purified by thin-layer chromatography. Structures were determined by comparative chromatography; infrared, fluorescence, and mass spectroscopy; and enzymatic and chemical degradation. Substances identified were adriamycinol, adriamycinol aglycone, adriamycin aglycone, deoxyadriamycin aglycone, deoxyadriamycinol aglycone, demethyldeoxyadriamycinol aglycone, demethyldeoxyadriamycinol aglycone 4-O-sulfate, and demethyldeoxyadriamycinol aglycone 4-O-beta-glucuronide. Other metabolites have been purified but not identified. Human metabolism of adriamycin involved carbonyl reduction, reductive glycosidic cleavage, hydrolytic glycosidic cleavage, O-demethylation, O-sulfation, and O-beta-glucuronidation. Carbonyl reduction was the major enzymatic conversion occurring in the human. | [
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PMID:3411 | [Termination of pregnancy and perinatal mortality (author's transl)]. | Elective induction was practised in 1875 of 10 537 deliveries (17.8%). Duration of delivery was, if anything, shortened after elective induction, compared with spontaneous delivery. There was no evidence of soft-tissue dystocias after elective induction. The Apgar score was 8-10 in 95.7% of children born after elective induction. pH of umbilical-artery blood in 95.7% of children after elective induction was greater than or equal to 7.2. There was a striking increase in the incidence of occiput posterior position with elective induction (2.7% of cases). The incidence of operative vaginal delivery was as frequent after elective as after all other forms of delivery. The incidence of section was 4.7% after elective induction, 11.5% in the entire series, intra-uterine asphyxia being an indication in 1.7%, compared with 2.5% for the total group. The frequency of operations was inversely proportional to the cervical index. Perinatal mortality was 0.53 per thousand (one case) after elective induction, 10.15 per thousand in the total group, 5.9 per thousand in those with indication for early induction. Perinatal mortality decreased from 23.0 per thousand to 7.2 per thousand from 1967 to 1974. | [Termination of pregnancy and perinatal mortality (author's transl)]. Elective induction was practised in 1875 of 10 537 deliveries (17.8%). Duration of delivery was, if anything, shortened after elective induction, compared with spontaneous delivery. There was no evidence of soft-tissue dystocias after elective induction. The Apgar score was 8-10 in 95.7% of children born after elective induction. pH of umbilical-artery blood in 95.7% of children after elective induction was greater than or equal to 7.2. There was a striking increase in the incidence of occiput posterior position with elective induction (2.7% of cases). The incidence of operative vaginal delivery was as frequent after elective as after all other forms of delivery. The incidence of section was 4.7% after elective induction, 11.5% in the entire series, intra-uterine asphyxia being an indication in 1.7%, compared with 2.5% for the total group. The frequency of operations was inversely proportional to the cervical index. Perinatal mortality was 0.53 per thousand (one case) after elective induction, 10.15 per thousand in the total group, 5.9 per thousand in those with indication for early induction. Perinatal mortality decreased from 23.0 per thousand to 7.2 per thousand from 1967 to 1974. | [
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PMID:3406 | Binding of organic compounds to rat liver and lung. | The binding of various radioisotopically labeled organic compounds to rat liver and lung was investigated in vitro. Pieces of rat lung and slices of rat liver were incubated at 37 degrees C under a nitrogen atmosphere in a modified Krebs-Ringer phosphate solution (pH 7.4) CONTAININg the compound to be studied. Of the neutral compounds investigated, digitoxin, digoxin and dexamethasone were highly bound to both liver and lung tissue, whereas the degree of binding of amitrole, erythritol, and ouabain was 20% or less. The weak acids which were bound to the greatest extent in both liver and lung were phenobarbital, pentobarbital, and diphenylhydantoin. Barbital was poorly bound, and there was no evidence for the binding of 5,5-dimethyloxazolidine-2,4-dione or p-aminohippuric acid in either tissue. Binding of the cardiac glycosides and the barbiturates directly paralleled their lipid solubilities. The degree of binding of neutral compounds and weak acids to lung and liver tissue did not vary greatly with concentration, even though broad concentration ranges were studied. This was also true of the weak base morphine. On the other hand, the binding to liver and lung of the organic bases nicotine, pilocarpine, d-amphetamine, lidocaine, erythromycin, and chloroquine, did vary with concentration. The quaternary ammonium compound decamethonium was bound only to liver, and this binding also varied with concentration. Two additional quaternary ammonium compounds, tetraethylammonium and N1-methylnicotinamide, were not significantly bound to either tissue. Comparisons on the basis of equal content of solids revealed that the binding of diverse organic compounds in liver is greater than or equal to that in lung. | Binding of organic compounds to rat liver and lung. The binding of various radioisotopically labeled organic compounds to rat liver and lung was investigated in vitro. Pieces of rat lung and slices of rat liver were incubated at 37 degrees C under a nitrogen atmosphere in a modified Krebs-Ringer phosphate solution (pH 7.4) CONTAININg the compound to be studied. Of the neutral compounds investigated, digitoxin, digoxin and dexamethasone were highly bound to both liver and lung tissue, whereas the degree of binding of amitrole, erythritol, and ouabain was 20% or less. The weak acids which were bound to the greatest extent in both liver and lung were phenobarbital, pentobarbital, and diphenylhydantoin. Barbital was poorly bound, and there was no evidence for the binding of 5,5-dimethyloxazolidine-2,4-dione or p-aminohippuric acid in either tissue. Binding of the cardiac glycosides and the barbiturates directly paralleled their lipid solubilities. The degree of binding of neutral compounds and weak acids to lung and liver tissue did not vary greatly with concentration, even though broad concentration ranges were studied. This was also true of the weak base morphine. On the other hand, the binding to liver and lung of the organic bases nicotine, pilocarpine, d-amphetamine, lidocaine, erythromycin, and chloroquine, did vary with concentration. The quaternary ammonium compound decamethonium was bound only to liver, and this binding also varied with concentration. Two additional quaternary ammonium compounds, tetraethylammonium and N1-methylnicotinamide, were not significantly bound to either tissue. Comparisons on the basis of equal content of solids revealed that the binding of diverse organic compounds in liver is greater than or equal to that in lung. | [
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PMID:3407 | Pharmacokinetics of digoxin in the rat. | Previous studies on the pharmacokinetics of 3H-digoxin in the rat have been based on total radioactivity in the plasma, even though the drug is extensively metabolized in this species. A comparison of total radioactivity vs. unchanged drug in rat plasma after administration of 3H-digoxin clearly showed the need to separate digoxin from its metabolites. The pharmacokinetics of digoxin were therefore examined using solvent extraction and thin-layer chromatography to isolate unchanged drug. Digoxin levels after a 1 mg/kg iv dose were measured in the plasma and urine of adult male rats in which the bile duct or the ureters had been ligated, as well as in sham-operated controls. In all cases, digoxin concentrations were best described by a two-compartment open model. Digoxin was rapidly eliminated from the plasma of controls, with a half-life of 2.5 hr, a volume of distribution of 3.6 liter/kg, and a renal clearance somewhat lower than the glomerular filtration rate. No significant change in these parameters was observed in rats with bile duct ligation. The total body clearance of 5.77 ml/min in the controls was reduced by only 10% in the bile duct-ligated rats. In animals with bilateral ureter ligation, the body clearance was reduced by 30% and the plasma half-life of digoxin was increased to 4 hr, although no significant change in the apparent volume of distribution was noted. Approximately 60% of the total body clearance was unaffected by bile duct and ureter ligations, and was assumed to be due to biotransformation. Biliary excretion was found to be important for digoxigenin bisdigitoxoside, inasmuch as rats with bile duct ligation showed elevated metabolite levels in the plasma as well as a 3-fold increase in renal excretion of the bisglycoside. | Pharmacokinetics of digoxin in the rat. Previous studies on the pharmacokinetics of 3H-digoxin in the rat have been based on total radioactivity in the plasma, even though the drug is extensively metabolized in this species. A comparison of total radioactivity vs. unchanged drug in rat plasma after administration of 3H-digoxin clearly showed the need to separate digoxin from its metabolites. The pharmacokinetics of digoxin were therefore examined using solvent extraction and thin-layer chromatography to isolate unchanged drug. Digoxin levels after a 1 mg/kg iv dose were measured in the plasma and urine of adult male rats in which the bile duct or the ureters had been ligated, as well as in sham-operated controls. In all cases, digoxin concentrations were best described by a two-compartment open model. Digoxin was rapidly eliminated from the plasma of controls, with a half-life of 2.5 hr, a volume of distribution of 3.6 liter/kg, and a renal clearance somewhat lower than the glomerular filtration rate. No significant change in these parameters was observed in rats with bile duct ligation. The total body clearance of 5.77 ml/min in the controls was reduced by only 10% in the bile duct-ligated rats. In animals with bilateral ureter ligation, the body clearance was reduced by 30% and the plasma half-life of digoxin was increased to 4 hr, although no significant change in the apparent volume of distribution was noted. Approximately 60% of the total body clearance was unaffected by bile duct and ureter ligations, and was assumed to be due to biotransformation. Biliary excretion was found to be important for digoxigenin bisdigitoxoside, inasmuch as rats with bile duct ligation showed elevated metabolite levels in the plasma as well as a 3-fold increase in renal excretion of the bisglycoside. | [
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PMID:3408 | N-acetylation of drugs. Pharmacogenetic studies in rabbits selected for their acetylator characteristics. | Studies on acetylation of sulfadiazine, isoniazid, and p-aminobenzoic acid in selected lines of slow and rapid acetylator rabbits are described. Pedigree analysis of rabbits classified as slow or rapid sulfadiazine acetylators confirmed previous studies that the rate of sulfadiazine elimination (acetylation) is genetically controlled, with rapid elimination dominant over slow elimination of the drug. Pharmacokinetic studies in rabbits of specified sulfadiazine acetylator genotypes with isoniazid and p-aminobenzoic acid show that the rate of isoniazid elimination is under the same genetic control as is sulfadiazine, whereas the rate of p-aminobenzoic acid elimination is not. A new drug acetylation polymorphism, which controls the rate of enzymatic acetylation of p-aminobenzoic acid in peripheral blood cells and which is related to the sulfadiazine acetylation polymorphism, is described. | N-acetylation of drugs. Pharmacogenetic studies in rabbits selected for their acetylator characteristics. Studies on acetylation of sulfadiazine, isoniazid, and p-aminobenzoic acid in selected lines of slow and rapid acetylator rabbits are described. Pedigree analysis of rabbits classified as slow or rapid sulfadiazine acetylators confirmed previous studies that the rate of sulfadiazine elimination (acetylation) is genetically controlled, with rapid elimination dominant over slow elimination of the drug. Pharmacokinetic studies in rabbits of specified sulfadiazine acetylator genotypes with isoniazid and p-aminobenzoic acid show that the rate of isoniazid elimination is under the same genetic control as is sulfadiazine, whereas the rate of p-aminobenzoic acid elimination is not. A new drug acetylation polymorphism, which controls the rate of enzymatic acetylation of p-aminobenzoic acid in peripheral blood cells and which is related to the sulfadiazine acetylation polymorphism, is described. | [
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PMID:3414 | Histidyl transfer ribonucleic acid synthetase from Salmonella typhimurium. Interaction with substrates and ATP analogues. | Structural requirements for substrate binding to histidyl-tRNA synthetase from Salmonella typhimurium have been investigated using ATP analogues. Ki values and the relative binding affinity of the enzyme for these analogues have been determined in the tRNA aminoacylation reaction. The enzyme is highly specific for ATP: no binding was found for GTP, CTP, TTP and UTP. dATP is a very poor substrate for acylation of tRNA, with a Km 40-fold higher than that of ATP. Binding of adenosine 5'-triphosphate requires interactions of the amino group of adenosine and the sugar moiety; the 2' and the 5' positions of the ribose appear to be essential for recognition; the phosphate groups enhance the binding. AMP is a noncompetitive inhibitor with ATP. The interaction of histidyl-tRNA synthetase, a dimeric enzyme, with histidine and ATP was examined by fluorescence measurements at equilibrium and by equilibrium dialysis. Binding with L-histidine is significantly tighter at pH 6 than at pH 7, while the ATP binding is independent of pH. The stoichiometry was measured at pH 6 than at pH 7, while the ATP binding is independent of pH. The stoichiometry was measured at pH 7.5 by equilibrium dialysis and is 1 mol ATP/mol enzyme and, variably, close to 2 or 1 mol histidine/mol enzyme. | Histidyl transfer ribonucleic acid synthetase from Salmonella typhimurium. Interaction with substrates and ATP analogues. Structural requirements for substrate binding to histidyl-tRNA synthetase from Salmonella typhimurium have been investigated using ATP analogues. Ki values and the relative binding affinity of the enzyme for these analogues have been determined in the tRNA aminoacylation reaction. The enzyme is highly specific for ATP: no binding was found for GTP, CTP, TTP and UTP. dATP is a very poor substrate for acylation of tRNA, with a Km 40-fold higher than that of ATP. Binding of adenosine 5'-triphosphate requires interactions of the amino group of adenosine and the sugar moiety; the 2' and the 5' positions of the ribose appear to be essential for recognition; the phosphate groups enhance the binding. AMP is a noncompetitive inhibitor with ATP. The interaction of histidyl-tRNA synthetase, a dimeric enzyme, with histidine and ATP was examined by fluorescence measurements at equilibrium and by equilibrium dialysis. Binding with L-histidine is significantly tighter at pH 6 than at pH 7, while the ATP binding is independent of pH. The stoichiometry was measured at pH 6 than at pH 7, while the ATP binding is independent of pH. The stoichiometry was measured at pH 7.5 by equilibrium dialysis and is 1 mol ATP/mol enzyme and, variably, close to 2 or 1 mol histidine/mol enzyme. | [
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PMID:3415 | [The coupling of beta1-24-corticotropin to the adenylate-cylase system in rat adipocytes. Evidence for hormone-nucleotides interaction (author's transl)]. | The general aim was to define some of the most important parameters involved in the coupling step between the synthetic analog of adrenocoricotropin hormone (beta1-24-corticotropin tetracosa peptide) and the catalytic unit of the adenylate-cyclase system of fat cells. These studies were performed with a purified plasma membrane fraction from rat adipose tissue. In this regard, some effects of ions, pH, and nucleotides (ATP nad GTP) on this hormone sensitive system were studied A simple model based on a random association process of reactants yeilded a statisfactory approximation of the kinetic data. In contract to results obtained by two other groups, which were analyzed by De Haen, no evidence was found for a regulation of the adenylate-cyclase activity by the adenosine triphosphate which was not complexed to magnesium... | [The coupling of beta1-24-corticotropin to the adenylate-cylase system in rat adipocytes. Evidence for hormone-nucleotides interaction (author's transl)]. The general aim was to define some of the most important parameters involved in the coupling step between the synthetic analog of adrenocoricotropin hormone (beta1-24-corticotropin tetracosa peptide) and the catalytic unit of the adenylate-cyclase system of fat cells. These studies were performed with a purified plasma membrane fraction from rat adipose tissue. In this regard, some effects of ions, pH, and nucleotides (ATP nad GTP) on this hormone sensitive system were studied A simple model based on a random association process of reactants yeilded a statisfactory approximation of the kinetic data. In contract to results obtained by two other groups, which were analyzed by De Haen, no evidence was found for a regulation of the adenylate-cyclase activity by the adenosine triphosphate which was not complexed to magnesium... | [
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PMID:3416 | Ion-binding to phospholipids. Interaction of calcium with phosphatidylserine. | The binding of Ca2+ to monolayers and bilayers of phosphatidylserine has been investigated as a function of pH, ionic strength (NaCl concentration) and Ca2+ concentration using surface and colloid chemical techniques. The molar ratio of lipid to bound calcium decreases to 2 as the Ca2+ concentration is increased to about 0.1 mM. At [Ca2+] greater than 0.1 mM a 1:1 complex is formed. The apparent binding constant Ka ranges from about approximately 10(6) - 10(4) l/mol depending on the Ca2+ concentration. After allowing for electrostatic effects and neighbour group interactions, the intrinsic binding constant Ki of the phosphorylserine polar group at pH 7 (I = 0.01 M), where it carries a net negative charge of one, is approximately 10(4) l/mol; consistent values for Ki were obtained using several independent approaches. Ka for Ca2+ binding decreases with increasing NaCl concentration because the monovalent cations compete with Ca2+ for the same binding site. Na+ and K+ are equally effective in displacing 45Ca2+ adsorbed to monolayers of phosphatidylserine, both with respect to the kinetics and the equilibrium of the displacement. Ka for the reaction between phosphatidylserine and monovalent cations is about 10(3)-fold smaller than that of Ca2+. An investigation of the binding of Mn2+ to phosphatidylserine by both surface chemical and nuclear magnetic resonance methods shows that this cation has a similar binding constant to that of Ca2+. The Ca2+-binding capabilities of monolayers containing only carboxyl groups (i.e. arachidic acid) and phosphodiester groups (i.e. dicetyl phosphate) have also been determined; the apparent pK for the - COOH group in monolayers is larger than or equal to 9 and that for the phosphodiester group is less than 4. Since these groups do not retain the same pK values when they are in close proximity in the phosphorylserine group, the relative contributions of the two groups to the binding of Ca2+ to phosphatidylserine is not obvious. | Ion-binding to phospholipids. Interaction of calcium with phosphatidylserine. The binding of Ca2+ to monolayers and bilayers of phosphatidylserine has been investigated as a function of pH, ionic strength (NaCl concentration) and Ca2+ concentration using surface and colloid chemical techniques. The molar ratio of lipid to bound calcium decreases to 2 as the Ca2+ concentration is increased to about 0.1 mM. At [Ca2+] greater than 0.1 mM a 1:1 complex is formed. The apparent binding constant Ka ranges from about approximately 10(6) - 10(4) l/mol depending on the Ca2+ concentration. After allowing for electrostatic effects and neighbour group interactions, the intrinsic binding constant Ki of the phosphorylserine polar group at pH 7 (I = 0.01 M), where it carries a net negative charge of one, is approximately 10(4) l/mol; consistent values for Ki were obtained using several independent approaches. Ka for Ca2+ binding decreases with increasing NaCl concentration because the monovalent cations compete with Ca2+ for the same binding site. Na+ and K+ are equally effective in displacing 45Ca2+ adsorbed to monolayers of phosphatidylserine, both with respect to the kinetics and the equilibrium of the displacement. Ka for the reaction between phosphatidylserine and monovalent cations is about 10(3)-fold smaller than that of Ca2+. An investigation of the binding of Mn2+ to phosphatidylserine by both surface chemical and nuclear magnetic resonance methods shows that this cation has a similar binding constant to that of Ca2+. The Ca2+-binding capabilities of monolayers containing only carboxyl groups (i.e. arachidic acid) and phosphodiester groups (i.e. dicetyl phosphate) have also been determined; the apparent pK for the - COOH group in monolayers is larger than or equal to 9 and that for the phosphodiester group is less than 4. Since these groups do not retain the same pK values when they are in close proximity in the phosphorylserine group, the relative contributions of the two groups to the binding of Ca2+ to phosphatidylserine is not obvious. | [
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PMID:3419 | Action of H1 and H2 inhibitors on the response of histamine sensitive adenyly cyclase from guinea-pig mucosa. | In the guinea-pig, it has been shown that homogenates of mucosa from the fundus contain an adenylyl cyclase system that is activated by histamine as well as by prostaglandins PGE1 and PGA1. The effects of burimamide, an H2-inhibitor, and mepyramine and chlorpheniramide, both H1-inhibitors, were tested. Both H1 and H2 inhibitors behaved kinetically as competitive inhibitors of histamine, but the Km derived for burimamide (2.5 - 4.1 . 10(-5)) was significantly lower than that for either chlorpheniramine (0.9 - 1.9 . 10(-4)) or mepyramine (1.3 - 1.4 . 10(-4)). On the other hand none of the three inhibitors influenced the cyclase activation by PGE1 and PGA1. These results suggest that there are at least two types of receptors in the preparation studied, one responsive to histamine and the other to the prostaglandins, and that the specificity of H1- and H2-receptors is not absolute in the broken cell preparation. | Action of H1 and H2 inhibitors on the response of histamine sensitive adenyly cyclase from guinea-pig mucosa. In the guinea-pig, it has been shown that homogenates of mucosa from the fundus contain an adenylyl cyclase system that is activated by histamine as well as by prostaglandins PGE1 and PGA1. The effects of burimamide, an H2-inhibitor, and mepyramine and chlorpheniramide, both H1-inhibitors, were tested. Both H1 and H2 inhibitors behaved kinetically as competitive inhibitors of histamine, but the Km derived for burimamide (2.5 - 4.1 . 10(-5)) was significantly lower than that for either chlorpheniramine (0.9 - 1.9 . 10(-4)) or mepyramine (1.3 - 1.4 . 10(-4)). On the other hand none of the three inhibitors influenced the cyclase activation by PGE1 and PGA1. These results suggest that there are at least two types of receptors in the preparation studied, one responsive to histamine and the other to the prostaglandins, and that the specificity of H1- and H2-receptors is not absolute in the broken cell preparation. | [
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PMID:3420 | Anti-arrhythmic action of nadolol, a beta-adrenergic receptor blocking agent. | The anti-arrhythmic action of 2,3-cis-1,2,3,4-tetrahydro-5-[(2-hydroxy-3-tert-butylamino)propoxy]2,3-naphthalenediol (nadolol) was evaluated and compared with that of propranolol in several experimental models of cardiac arrhythmias. Both nadolol and propranolol antagonized isoproterenol-induced tachycardia and ouabain-induced arrhythmias in cats, antagonized coronary artery ligation-induced ventricular fibrillation and suppressed ventricular ectopic activity during vagal stimulation in dogs. In contrast to propranolol, nadolol was considerably weaker in suppressing existing digoxin-induced arrhythmias, lacked local anesthetic activity and did not depress the heart in dogs. Because of these findings, it is concluded that the anti-arrhythmic activity of nadolol is apparently related to blockade of beta-adrenergic receptors. | Anti-arrhythmic action of nadolol, a beta-adrenergic receptor blocking agent. The anti-arrhythmic action of 2,3-cis-1,2,3,4-tetrahydro-5-[(2-hydroxy-3-tert-butylamino)propoxy]2,3-naphthalenediol (nadolol) was evaluated and compared with that of propranolol in several experimental models of cardiac arrhythmias. Both nadolol and propranolol antagonized isoproterenol-induced tachycardia and ouabain-induced arrhythmias in cats, antagonized coronary artery ligation-induced ventricular fibrillation and suppressed ventricular ectopic activity during vagal stimulation in dogs. In contrast to propranolol, nadolol was considerably weaker in suppressing existing digoxin-induced arrhythmias, lacked local anesthetic activity and did not depress the heart in dogs. Because of these findings, it is concluded that the anti-arrhythmic activity of nadolol is apparently related to blockade of beta-adrenergic receptors. | [
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PMID:3421 | Effects of chemical stimulation of the mesolimbic dopamine system upon locomotor activity. | The effects of local injections of drugs into terminal areas of the mesolimbic dopamine system were investigated. Bilateral administration of dopamine, but not of noradrenaline and serotonin, into the nucleus accumbens of non-pretreated rats resulted in stimulation of locomotor activity. No clear or only minor effects were seen after injections of the dopamine metabolites 3-methoxytyramine, DOPAC and HVA and after injections of media with different pH and osmolality. d-Amphetamine proved more effective than dopamine in producing locomotor stimulation, whereas both stimulant and depressant effects were observed following injection of apomorphine into the nucleus accumbens. ET 495 and the noradrenaline agonists clonidine, phenylephrine and isoprenaline did not enhance locomotor activity, but theophylline was effective. Pretreatment with haloperidol, but not with clozapine, significantly reduced the effects of dopamine and theophylline. Locomotor stimulation was also found following bilateral administration of dopamine, d-amphetamine and apomorphine into the tuberculum olfactorium, whereas noradrenaline, serotonin and ET 495 produced no, or rather depressant effects. These results provide further evidence for an important role of the mesolimbic dopamine system with respect to locomotor activity. | Effects of chemical stimulation of the mesolimbic dopamine system upon locomotor activity. The effects of local injections of drugs into terminal areas of the mesolimbic dopamine system were investigated. Bilateral administration of dopamine, but not of noradrenaline and serotonin, into the nucleus accumbens of non-pretreated rats resulted in stimulation of locomotor activity. No clear or only minor effects were seen after injections of the dopamine metabolites 3-methoxytyramine, DOPAC and HVA and after injections of media with different pH and osmolality. d-Amphetamine proved more effective than dopamine in producing locomotor stimulation, whereas both stimulant and depressant effects were observed following injection of apomorphine into the nucleus accumbens. ET 495 and the noradrenaline agonists clonidine, phenylephrine and isoprenaline did not enhance locomotor activity, but theophylline was effective. Pretreatment with haloperidol, but not with clozapine, significantly reduced the effects of dopamine and theophylline. Locomotor stimulation was also found following bilateral administration of dopamine, d-amphetamine and apomorphine into the tuberculum olfactorium, whereas noradrenaline, serotonin and ET 495 produced no, or rather depressant effects. These results provide further evidence for an important role of the mesolimbic dopamine system with respect to locomotor activity. | [
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PMID:3426 | Effect of PGE1 on lipogenesis in perfused rat liver. | In perfused livers of 24 hour-fasted rats, PGE1 (prostaglandin E1) infused continuously into the perfusate, was found to cause a 45% increase in the incorporation of 1-14C acetate into liver fatty acids. PGE1 was found to have no effect, however, on the activity of the key lipogenic enzymes. | Effect of PGE1 on lipogenesis in perfused rat liver. In perfused livers of 24 hour-fasted rats, PGE1 (prostaglandin E1) infused continuously into the perfusate, was found to cause a 45% increase in the incorporation of 1-14C acetate into liver fatty acids. PGE1 was found to have no effect, however, on the activity of the key lipogenic enzymes. | [
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PMID:3428 | Influence of mono- and multivalent cations on the electrokinetic properties of normal human lymphoid and burkitt lymphoma cells. | Various mono- and multivalent cations, at constant ionic strength, affected the surface electrokinetic properties of normal human lymphoid and Burkitt lymphoma cells in a manner which reflected more the physicochemical properties and binding affinities of the cation than its valence. The effect was expressed in the changes of the magnitude of the net surface charge and in the shift of the isoelectric point of the surface. These changes were greater at the surface of Burkitt's lymphoma cells than at the surface of their normal cell-line counterparts. | Influence of mono- and multivalent cations on the electrokinetic properties of normal human lymphoid and burkitt lymphoma cells. Various mono- and multivalent cations, at constant ionic strength, affected the surface electrokinetic properties of normal human lymphoid and Burkitt lymphoma cells in a manner which reflected more the physicochemical properties and binding affinities of the cation than its valence. The effect was expressed in the changes of the magnitude of the net surface charge and in the shift of the isoelectric point of the surface. These changes were greater at the surface of Burkitt's lymphoma cells than at the surface of their normal cell-line counterparts. | [
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PMID:3435 | 1,8-naphthyridine derivatives: synthesis and pharmacological evaluation of beta-receptor blocking activity. | The synthesis of a number of 1-alkyl-7-(2-hydroxy-3-alkylaminopropoxy)-1,8-naphthyridin-2-ones is described. The compounds studied were prepared by reaction of 1-alkyl-7-hydroxy-1,8-naphthyridin-2-ones with epichlorohydrin. The substituted epoxy intermediates obtained were allowed to react with amines and gave the desired products. All the compunds prepared were devoid of beta-blocking activity. | 1,8-naphthyridine derivatives: synthesis and pharmacological evaluation of beta-receptor blocking activity. The synthesis of a number of 1-alkyl-7-(2-hydroxy-3-alkylaminopropoxy)-1,8-naphthyridin-2-ones is described. The compounds studied were prepared by reaction of 1-alkyl-7-hydroxy-1,8-naphthyridin-2-ones with epichlorohydrin. The substituted epoxy intermediates obtained were allowed to react with amines and gave the desired products. All the compunds prepared were devoid of beta-blocking activity. | [
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PMID:3433 | [Pharmacology of methvin--a new ganglionic blocking preparation of short-term effect]. | Methvin (vincanin chlormethylate) is an active ganglion blocking agent of a short-term action. As regards its gangliolytic action it is approximately 6 times superior to afronad. In test animals the drug produces a well-marked and short-term (easily controllable) hypotensive effect, without causing any histamine-like and direct vasodilation action. When used in relatively high doses methvin blocks the neuro-muscular conduction, potentiates the action of major muscle relaxants. A study of methvin in clinical conditions confirmed its high gangliolytic activity previously revealed in experiments. | [Pharmacology of methvin--a new ganglionic blocking preparation of short-term effect]. Methvin (vincanin chlormethylate) is an active ganglion blocking agent of a short-term action. As regards its gangliolytic action it is approximately 6 times superior to afronad. In test animals the drug produces a well-marked and short-term (easily controllable) hypotensive effect, without causing any histamine-like and direct vasodilation action. When used in relatively high doses methvin blocks the neuro-muscular conduction, potentiates the action of major muscle relaxants. A study of methvin in clinical conditions confirmed its high gangliolytic activity previously revealed in experiments. | [
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PMID:3434 | [Heightened anticholinesterase activity of chlorophos in its interaction with the TMB-4 reactivator in vitro]. | Experiments in vitro demonstrated that a thermostatic treatment of an aqueous chlorophos solution at 38 degrees is attended by its increased cholinesterase activity. The speed of the process is much higher in an alkaline medium. A joint incubation of chlorophos with the TMB-4 reactivator of pH of 7.5 and 38 degrees not only fails to result in decomposition of the poison, but on the contrary, tends to speed up the progressive accretion of the inhibitory properties of this organophosphorus compound. The author considers the data obtained as one of the proofs pointing to the formation during direct interaction of TMB-4 with chlorophos or the product of its transformation of a stable complex appearing to be a powerful anticholinesterase agent. | [Heightened anticholinesterase activity of chlorophos in its interaction with the TMB-4 reactivator in vitro]. Experiments in vitro demonstrated that a thermostatic treatment of an aqueous chlorophos solution at 38 degrees is attended by its increased cholinesterase activity. The speed of the process is much higher in an alkaline medium. A joint incubation of chlorophos with the TMB-4 reactivator of pH of 7.5 and 38 degrees not only fails to result in decomposition of the poison, but on the contrary, tends to speed up the progressive accretion of the inhibitory properties of this organophosphorus compound. The author considers the data obtained as one of the proofs pointing to the formation during direct interaction of TMB-4 with chlorophos or the product of its transformation of a stable complex appearing to be a powerful anticholinesterase agent. | [
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PMID:3445 | The temperature, pH, and partial pressure of oxygen in the cervix and uterus of women and uterus of rats during the cycle. | Changes in the temperature, pH, and partial pressure of oxygen (PO2) in the cervical canal and cavum uteri were measured in women with or without an intrauterine device and in the uteri of rats throughout the cycle. Only the PO2 exhibited significant variations, rising during the ovulatory phase in both cervices of women and uteri in rats. It is speculated that the rise in PO2 is related to the function of these organs as reservoirs for spermatozoa. | The temperature, pH, and partial pressure of oxygen in the cervix and uterus of women and uterus of rats during the cycle. Changes in the temperature, pH, and partial pressure of oxygen (PO2) in the cervical canal and cavum uteri were measured in women with or without an intrauterine device and in the uteri of rats throughout the cycle. Only the PO2 exhibited significant variations, rising during the ovulatory phase in both cervices of women and uteri in rats. It is speculated that the rise in PO2 is related to the function of these organs as reservoirs for spermatozoa. | [
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PMID:3446 | [Responses of small intestine tissue chemoreceptors to change in the pCO2, pH and (HCO3-) in perfusion solutions]. | Perfusion of the small intestine of anesthetized cats with a solution having excessive CO2 and H+ concentration (pCO2 60 mm Hg; pH; 7.2; [HCO3-] 25 MM) produced a threshold reflex increase in the blood pressure. The subsequent increase of pCO2 to 380 mm Hg and decrease of pH to 6.4 evoked a gradual raise of the blood pressure (8.0+/-0.6 mm Hg) followed by the sharp increase of pressor reflexes amplitude within the range of pH 6.4--6.1. Tissue receptors were found to be essentially sensitive to solutions imitating metabolic acidosis (decrease of [HCO3-] within the physiological range of pH changes (pH 7.1--6.8). Solutions with pH 6.4--6.1 imitating respiratory acidosis (increase of pCO2) were more effective than those imitating metabolic acidosis. The possible role of interstitial pH changes in responses of the tissue chemoreceptors to CO2, is discussed. | [Responses of small intestine tissue chemoreceptors to change in the pCO2, pH and (HCO3-) in perfusion solutions]. Perfusion of the small intestine of anesthetized cats with a solution having excessive CO2 and H+ concentration (pCO2 60 mm Hg; pH; 7.2; [HCO3-] 25 MM) produced a threshold reflex increase in the blood pressure. The subsequent increase of pCO2 to 380 mm Hg and decrease of pH to 6.4 evoked a gradual raise of the blood pressure (8.0+/-0.6 mm Hg) followed by the sharp increase of pressor reflexes amplitude within the range of pH 6.4--6.1. Tissue receptors were found to be essentially sensitive to solutions imitating metabolic acidosis (decrease of [HCO3-] within the physiological range of pH changes (pH 7.1--6.8). Solutions with pH 6.4--6.1 imitating respiratory acidosis (increase of pCO2) were more effective than those imitating metabolic acidosis. The possible role of interstitial pH changes in responses of the tissue chemoreceptors to CO2, is discussed. | [
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PMID:3448 | Identification of dimethylnitrosoamine metabolites in vitro. | The incubation of dimethylnitrosoamine (DMNA) in the presence of rat liver microsomes leads to production of formaldehyde, formic acid, methylamine, and N-methylhydrazine. When pH 5-enzymes are added to the medium there is also the formation of N-methylhydroxylamide and N,N-dimethylhydrazine. The last compound is the only metabolite produced, to a lesser extent, by the pH 5-enzymes. Thus, the denitrosated or non-denitrosated metabolites are produced either by an oxidative dealkylation and by a reduction of DMNA, catalysed by microsomal and cellular soluble enzymes. | Identification of dimethylnitrosoamine metabolites in vitro. The incubation of dimethylnitrosoamine (DMNA) in the presence of rat liver microsomes leads to production of formaldehyde, formic acid, methylamine, and N-methylhydrazine. When pH 5-enzymes are added to the medium there is also the formation of N-methylhydroxylamide and N,N-dimethylhydrazine. The last compound is the only metabolite produced, to a lesser extent, by the pH 5-enzymes. Thus, the denitrosated or non-denitrosated metabolites are produced either by an oxidative dealkylation and by a reduction of DMNA, catalysed by microsomal and cellular soluble enzymes. | [
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PMID:3449 | Degradation of dimethylnitrosoamine catalysed by physical and chemical agents. | Decomposition of dimethylnitrosoamine (DMNA) by chemical and physical agents was further investigated. Both photoirradiation with sunlight or ultraviolet ray and reductive reactions under acid conditions (likely occurring in the stomach) led to the formation of formaldehyde, formic acid, and N-methydrazine, in addition to denitrosated compounds such as methylamine, dimethylamine, and N-methylhydroxylamine. N-Methylhydrazine was the only compound which was not detected by photoirradiation under neutral conditions. The agreement between physiochemical and metabolic degradation products and the possible biological meaning are discussed together with the problem of environmental contamination by the nitroso compound. | Degradation of dimethylnitrosoamine catalysed by physical and chemical agents. Decomposition of dimethylnitrosoamine (DMNA) by chemical and physical agents was further investigated. Both photoirradiation with sunlight or ultraviolet ray and reductive reactions under acid conditions (likely occurring in the stomach) led to the formation of formaldehyde, formic acid, and N-methydrazine, in addition to denitrosated compounds such as methylamine, dimethylamine, and N-methylhydroxylamine. N-Methylhydrazine was the only compound which was not detected by photoirradiation under neutral conditions. The agreement between physiochemical and metabolic degradation products and the possible biological meaning are discussed together with the problem of environmental contamination by the nitroso compound. | [
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PMID:3450 | Properties of aryl hydrocarbon hydroxylase in microsomes of Morris hepatoma 5123D and the host liver. | Properties of aryl hydrocarbon hydroxylase in the microsomes were compared between Morris hepatoma 5123D and the host liver from rats bearing this tumor. Requirement of NADPH for the assay of the enzyme activity was observed, compared to that of NADH, and also the additive effect of NADH on the requirement of NADPH was found in the tumor and liver. Curve of pH optimum of the enzyme activity in tumor and liver differed between the rats treated with corn oil and those with 3-methylcholanthrene, indicating a slight shift of the peak value to alkaline pH in the latter. The same values of the apparent Km for NADPH and NADH were shown for the enzyme from the liver and tumor even 24 hr after the treatment with 3-methylcholanthrene, but a difference in the apparent Km for benzo[a]pyrene was demonstrated between the tumor and the host liver, showing 3.6 approximately 6.6 muM in the former and 9.1 approximately 20 muM in the latter. By the addition of 7,8- or 5,6-benzoflavone to the assay medium for the tumor, the induced enzyme was inhibited noncompetitively, and the constitutive enzyme was enhanced, as demonstrated in the host liver. As observed in the induced enzyme in both tissues, cyclohexene oxide and 1,1,1-trichloropropane oxide slightly increased the activity of the constitutive enzyme in the tumor, in contrast to its inhibition in the host liver. | Properties of aryl hydrocarbon hydroxylase in microsomes of Morris hepatoma 5123D and the host liver. Properties of aryl hydrocarbon hydroxylase in the microsomes were compared between Morris hepatoma 5123D and the host liver from rats bearing this tumor. Requirement of NADPH for the assay of the enzyme activity was observed, compared to that of NADH, and also the additive effect of NADH on the requirement of NADPH was found in the tumor and liver. Curve of pH optimum of the enzyme activity in tumor and liver differed between the rats treated with corn oil and those with 3-methylcholanthrene, indicating a slight shift of the peak value to alkaline pH in the latter. The same values of the apparent Km for NADPH and NADH were shown for the enzyme from the liver and tumor even 24 hr after the treatment with 3-methylcholanthrene, but a difference in the apparent Km for benzo[a]pyrene was demonstrated between the tumor and the host liver, showing 3.6 approximately 6.6 muM in the former and 9.1 approximately 20 muM in the latter. By the addition of 7,8- or 5,6-benzoflavone to the assay medium for the tumor, the induced enzyme was inhibited noncompetitively, and the constitutive enzyme was enhanced, as demonstrated in the host liver. As observed in the induced enzyme in both tissues, cyclohexene oxide and 1,1,1-trichloropropane oxide slightly increased the activity of the constitutive enzyme in the tumor, in contrast to its inhibition in the host liver. | [
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PMID:3451 | Electronic interaction of carcinogenic and noncarcinogenic benz(c)acridines and DNA. | Electronic interaction of DNA with nine benzacridine derivatives was studied. The interaction system of these benzacridines and DNA was found to show a marked hypochromism in the ultraviolet region. The orderly double helical structure of DNA was found to play an essential role in this spectroscopic change. A high concentration of the interactant, low environmental ionic strength, low pH, and low temperature were beneficial in this interaction. The degree of hypochromism expressed by the integrated and pKa of the benzacridines were found to be in parallel. The hypochromism of the interaction system, in which the carcinogenic benz[c]acridine derivatives took part, was larger than that of the system in which the noncarcinogenic derivatives were involved. | Electronic interaction of carcinogenic and noncarcinogenic benz(c)acridines and DNA. Electronic interaction of DNA with nine benzacridine derivatives was studied. The interaction system of these benzacridines and DNA was found to show a marked hypochromism in the ultraviolet region. The orderly double helical structure of DNA was found to play an essential role in this spectroscopic change. A high concentration of the interactant, low environmental ionic strength, low pH, and low temperature were beneficial in this interaction. The degree of hypochromism expressed by the integrated and pKa of the benzacridines were found to be in parallel. The hypochromism of the interaction system, in which the carcinogenic benz[c]acridine derivatives took part, was larger than that of the system in which the noncarcinogenic derivatives were involved. | [
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PMID:3452 | Purification of an immunosuppressive principle from Ehrlich carcinoma ascites. | An immunosuppressive factor was purified from Ehrlich carcinoma ascites by the combination of ultrafiltration and Sephadex chromatography. The resulting product showed 50% reduction in the number of splenic plaque-forming cells in mice immunized with sheep red blood cells when as low as 50 mug dose was given twice intraperitoneally before erythrocyte injection. The molecular size of the product was between 30,000 and 100,000, and it was relatively heat unstable. | Purification of an immunosuppressive principle from Ehrlich carcinoma ascites. An immunosuppressive factor was purified from Ehrlich carcinoma ascites by the combination of ultrafiltration and Sephadex chromatography. The resulting product showed 50% reduction in the number of splenic plaque-forming cells in mice immunized with sheep red blood cells when as low as 50 mug dose was given twice intraperitoneally before erythrocyte injection. The molecular size of the product was between 30,000 and 100,000, and it was relatively heat unstable. | [
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PMID:3453 | [Inhalation analgesia with methoxyfluran (penthrane) in obstetrics (author's transl)]. | In 105 normal spontaneous deliveries the effect of inhalation analgesia with Methoxyfluran (Penthrane) was studied. Penthrane was administered intermittently with the Penthrane oxilator (Rod inhalator of Abbott). The initial concentration was 0.25 volume%. The maximal concentration was 0.35 volume% of Penthrane. Fetal monitoring records were obtained in all deliveries. In 49 cases (46.7%) intra- and post-partum microblood studies were obtained. In 85 women (81.0%) the result of the analgesia was good. Nausea occurred in 21 women (20%). There were no abnormal fetal monitoring patterns. The Apgar scores of the newborns were between 8 and 10. The actual pH's were between 7.24 and 7.41. The third stage of labour was normal in all cases. | [Inhalation analgesia with methoxyfluran (penthrane) in obstetrics (author's transl)]. In 105 normal spontaneous deliveries the effect of inhalation analgesia with Methoxyfluran (Penthrane) was studied. Penthrane was administered intermittently with the Penthrane oxilator (Rod inhalator of Abbott). The initial concentration was 0.25 volume%. The maximal concentration was 0.35 volume% of Penthrane. Fetal monitoring records were obtained in all deliveries. In 49 cases (46.7%) intra- and post-partum microblood studies were obtained. In 85 women (81.0%) the result of the analgesia was good. Nausea occurred in 21 women (20%). There were no abnormal fetal monitoring patterns. The Apgar scores of the newborns were between 8 and 10. The actual pH's were between 7.24 and 7.41. The third stage of labour was normal in all cases. | [
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PMID:3456 | Post-prandial changes in PH and electrolyte concentration, in the upper jejunum after truncal vagotomy and drainage in man. | The changes in pH and concentration of electrolytes in the jejunal lumen after a hypertonic fluid meal have been studied after truncal vagotomy and drainage, with and without diarrhoea. The results show that, in these respects, there are no specific changes in the jejunal content associated with post-vagotomy diarrhoea, but that these measurements are markedly affected by the completeness of vagotomy, as judged by the insulin test. | Post-prandial changes in PH and electrolyte concentration, in the upper jejunum after truncal vagotomy and drainage in man. The changes in pH and concentration of electrolytes in the jejunal lumen after a hypertonic fluid meal have been studied after truncal vagotomy and drainage, with and without diarrhoea. The results show that, in these respects, there are no specific changes in the jejunal content associated with post-vagotomy diarrhoea, but that these measurements are markedly affected by the completeness of vagotomy, as judged by the insulin test. | [
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PMID:3457 | Vitamin B12 absorption--a study of intraluminal events in control subjects and patients with tropical sprue. | The intraluminal fate of orally administered radioactive vitamin B12 has been studied in control subjects with normal vitamin B12 absorption and those with vitamin B12 malabsorption due to tropical sprue. In control subjects 1 to 21% of the dose was bound to sedimentable material and 37 to 75% was bound to immunoreactive intrinsic factor. In subjects with vitamin B12 malabsorption due to tropical sprue, the results were identical with the control subjects. Bacteriological studies showed a statistically significant correlation between both the number of flora in the jejunum and the number of bacteroides in both the jejunum and ileum and vitamin B12 malabsorption. In patients with tropical sprue who have normal intrinsic factor secretion, the vitamin B12 absorptive defect is not due to binding of the vitamin to bacteria or to alteration to the intrinsic factor vitamin B12 complex in the intestinal lumen. The lesion appears to be one of the mucosal cell receptors or of the cells themselves, possibly caused by bacterial toxins. | Vitamin B12 absorption--a study of intraluminal events in control subjects and patients with tropical sprue. The intraluminal fate of orally administered radioactive vitamin B12 has been studied in control subjects with normal vitamin B12 absorption and those with vitamin B12 malabsorption due to tropical sprue. In control subjects 1 to 21% of the dose was bound to sedimentable material and 37 to 75% was bound to immunoreactive intrinsic factor. In subjects with vitamin B12 malabsorption due to tropical sprue, the results were identical with the control subjects. Bacteriological studies showed a statistically significant correlation between both the number of flora in the jejunum and the number of bacteroides in both the jejunum and ileum and vitamin B12 malabsorption. In patients with tropical sprue who have normal intrinsic factor secretion, the vitamin B12 absorptive defect is not due to binding of the vitamin to bacteria or to alteration to the intrinsic factor vitamin B12 complex in the intestinal lumen. The lesion appears to be one of the mucosal cell receptors or of the cells themselves, possibly caused by bacterial toxins. | [
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PMID:3461 | [Experimental testing of a new gastrotherapeutic agent]. | A new gastrotherapeutical preparation, AcT 72, has been evaluated. This preparation contains the anticholinergic drug propantheline bromide, the psychotherapeutical agent perazine, and antacids. In two different series of tests it was demonstrated that after a peroral dose of ACT72, gastric motility was significantly reduced when compared with the control situation. Dosages used were in accordance with normal therapeutic doses, and, because of the influence on gastric motility, the duration of action in the stomach was prolonged, thus reducing daily dose requirements. Reduction of stomach motility per se, has therapeutic value in patients with gastric ulcers. | [Experimental testing of a new gastrotherapeutic agent]. A new gastrotherapeutical preparation, AcT 72, has been evaluated. This preparation contains the anticholinergic drug propantheline bromide, the psychotherapeutical agent perazine, and antacids. In two different series of tests it was demonstrated that after a peroral dose of ACT72, gastric motility was significantly reduced when compared with the control situation. Dosages used were in accordance with normal therapeutic doses, and, because of the influence on gastric motility, the duration of action in the stomach was prolonged, thus reducing daily dose requirements. Reduction of stomach motility per se, has therapeutic value in patients with gastric ulcers. | [
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PMID:3462 | [The disulfide bridges of the trypsin-kallikrein inhibitor K from snails (Helix pomatia). Thermal inactivation and proteolysis by thermolysin (author's transl)]. | Isoinhibitor K is the main component of the complex mixture of isoinhibitors of broad specificity secreted into the mucus by the Roman snail (Helix pomatia). The disulfide pairing was determined after the amino acid sequence had been elucidated. Two cystine-containing peptides with the disulfide bridges Cys32-Cys53 and Cys32-Cys53 plus Cys7-Cys57 were obtained after thermolytic hydrolysis of the native inhibitor at 80 degrees C and chromatographic separation of the peptides using SE-Sephadex. The Cys16-Cys40 disulfide bridge could be reduced selectively by sodium borohydride with no loss in biological activity. This property and the covalent structure correspond to that of the intracellular inhibitor from bovine organs, which is largely homologous in its amino acid sequence to the secretory inhibitor from the snail. The complete covalent structure of isoinhibitor K will be presented. The snail inhibitor is less stable against proteolytic inactivation by thermolysin and against thermal denaturation at pH 8.0 than the inhibitor from bovine organs (Kunitz inhibitor). | [The disulfide bridges of the trypsin-kallikrein inhibitor K from snails (Helix pomatia). Thermal inactivation and proteolysis by thermolysin (author's transl)]. Isoinhibitor K is the main component of the complex mixture of isoinhibitors of broad specificity secreted into the mucus by the Roman snail (Helix pomatia). The disulfide pairing was determined after the amino acid sequence had been elucidated. Two cystine-containing peptides with the disulfide bridges Cys32-Cys53 and Cys32-Cys53 plus Cys7-Cys57 were obtained after thermolytic hydrolysis of the native inhibitor at 80 degrees C and chromatographic separation of the peptides using SE-Sephadex. The Cys16-Cys40 disulfide bridge could be reduced selectively by sodium borohydride with no loss in biological activity. This property and the covalent structure correspond to that of the intracellular inhibitor from bovine organs, which is largely homologous in its amino acid sequence to the secretory inhibitor from the snail. The complete covalent structure of isoinhibitor K will be presented. The snail inhibitor is less stable against proteolytic inactivation by thermolysin and against thermal denaturation at pH 8.0 than the inhibitor from bovine organs (Kunitz inhibitor). | [
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PMID:3463 | [The inter-alpha-trypsin inhibitor as precursor of the acid-stable proteinase inhibitors in human serum and urine]. | A small amount of antitryptic activity is detectable in the supernatant of deproteinized human serum. Preincubation of serum with trypsin causes an increase in acid-stable antitryptic activity. This rise in activity depends on the inter alpha-trypsin inhibitor concentration. The native inhibitor present in normal sera, and in higher concentrations in sera of patients with nephropathies, and the trypsin-liberated inhibitor show immunological cross reaction with antibodies to the serum inter-alpha-trypsin inhibitor. The two inhibitors differ in molecular weight and electrophoretic mobility. The physiological inhibitor (I-34), with a molecular weight of 34 000 and a high carbohydrate content, can be transformed by trypsin into an inhibitor (I-17) with a molecular weight of 17 000. This inhibitor is identical with the inhibitors liberated by trypsin from serum or from purified inter-alpha-trypsin inhibitor. The acid-stable inhibitor from urine is identical with the physiological serum inhibitor. Analogously, this inhibitor is transformed by trypsin into the inhibitor with a molecular weight of 17 000. We conclude that the inter-alpha-trypsin inhibitor is the precursor of both the physiological and the trypsin-liberated inhibitor. By a mechanism as yet unknown, but most likely a limited proteolysis, the secreted inhibitor is liberated from the high molecular weight precursor. In contrast to the monospecific trypsin-inhibiting precursor, the physiological and artificially liberated inhibitors are trypsin/chymotrypsin/plasmin inhibitors. | [The inter-alpha-trypsin inhibitor as precursor of the acid-stable proteinase inhibitors in human serum and urine]. A small amount of antitryptic activity is detectable in the supernatant of deproteinized human serum. Preincubation of serum with trypsin causes an increase in acid-stable antitryptic activity. This rise in activity depends on the inter alpha-trypsin inhibitor concentration. The native inhibitor present in normal sera, and in higher concentrations in sera of patients with nephropathies, and the trypsin-liberated inhibitor show immunological cross reaction with antibodies to the serum inter-alpha-trypsin inhibitor. The two inhibitors differ in molecular weight and electrophoretic mobility. The physiological inhibitor (I-34), with a molecular weight of 34 000 and a high carbohydrate content, can be transformed by trypsin into an inhibitor (I-17) with a molecular weight of 17 000. This inhibitor is identical with the inhibitors liberated by trypsin from serum or from purified inter-alpha-trypsin inhibitor. The acid-stable inhibitor from urine is identical with the physiological serum inhibitor. Analogously, this inhibitor is transformed by trypsin into the inhibitor with a molecular weight of 17 000. We conclude that the inter-alpha-trypsin inhibitor is the precursor of both the physiological and the trypsin-liberated inhibitor. By a mechanism as yet unknown, but most likely a limited proteolysis, the secreted inhibitor is liberated from the high molecular weight precursor. In contrast to the monospecific trypsin-inhibiting precursor, the physiological and artificially liberated inhibitors are trypsin/chymotrypsin/plasmin inhibitors. | [
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PMID:3464 | Plant microbody proteins, I. Purification and characterization of catalase from leaves of lens culinaris. | 1) Catalase from green leaves of Lens culinaris (lentils) was investigated with respect to isoenzyme patterns. In contrast to other plants, which have been reported to contain multiple forms of catalase, only one form of this enzyme was revealed when crude extracts were subjected to starch gel electrophoresis or to polyacrylamide disc-gel electrophoresis. Furthermore, catalases from leaves, stems and cotyledons were electrophoretically identical. 2) The leaf enzyme has been purified by conventional methods to apparent homogeneity. It has a molecular weight of 225 000 (ultracentrifuge) and is composed of four identical subunits of molecular weight 54 000 (sodium dodecylsulphate gel electrophoresis). The ratio A280/A405 of the pure enzyme was found to be 1.5. The isoelectric point is at pH 5.5. The enzyme, very labile at pH-values below 7.0, is stable in Tris chloride and potassium phosphate buffers between pH 7.5 and 9.5. It is slowly inactivated by 1mM dithiothreitol and is rapidly inactivated by 1mM mercaptoethanol. 3) The catalase was shown to be the major protein component of the peroxisomal matrix. It could not be detected at the membranes of the leaf peroxisomes. | Plant microbody proteins, I. Purification and characterization of catalase from leaves of lens culinaris. 1) Catalase from green leaves of Lens culinaris (lentils) was investigated with respect to isoenzyme patterns. In contrast to other plants, which have been reported to contain multiple forms of catalase, only one form of this enzyme was revealed when crude extracts were subjected to starch gel electrophoresis or to polyacrylamide disc-gel electrophoresis. Furthermore, catalases from leaves, stems and cotyledons were electrophoretically identical. 2) The leaf enzyme has been purified by conventional methods to apparent homogeneity. It has a molecular weight of 225 000 (ultracentrifuge) and is composed of four identical subunits of molecular weight 54 000 (sodium dodecylsulphate gel electrophoresis). The ratio A280/A405 of the pure enzyme was found to be 1.5. The isoelectric point is at pH 5.5. The enzyme, very labile at pH-values below 7.0, is stable in Tris chloride and potassium phosphate buffers between pH 7.5 and 9.5. It is slowly inactivated by 1mM dithiothreitol and is rapidly inactivated by 1mM mercaptoethanol. 3) The catalase was shown to be the major protein component of the peroxisomal matrix. It could not be detected at the membranes of the leaf peroxisomes. | [
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PMID:3465 | The activator of human cerebroside sulphatase. Activating effect on the acidic forms of the sulphatases from invertebrates. | 1) Acidic forms of the sulphatase were partially purified from the following invertebrate species: Tethya aurantium (Porifera), Patella vulgata (mollusca), Maja squinado (Arthropoda), Marthasterias glacialis (Echinodermata) and Microcosmus sulcatus (Tunicata). Enzyme preparations thus obtained cleaved cerebroside sulphates (sulphatides) only in the presence of either specific detergents (e.g. taurodeoxycholate) or an activator protein isolated from human liver. This corresponds to the findings on purified sulphatase A of human origin. 2) At low concentrations, the activating effect was proportional to the amount of activator protein applied; at higher concentrations, proportionality was obtained only in some cases. On a molar basis, less of the activator protein was required to achieve the same activation as taurodeoxycholate. At optimum concentrations of the detergent however, the activation was much higher. 3) The enzyme specificity of the activator and some evolutionary implications are discussed. | The activator of human cerebroside sulphatase. Activating effect on the acidic forms of the sulphatases from invertebrates. 1) Acidic forms of the sulphatase were partially purified from the following invertebrate species: Tethya aurantium (Porifera), Patella vulgata (mollusca), Maja squinado (Arthropoda), Marthasterias glacialis (Echinodermata) and Microcosmus sulcatus (Tunicata). Enzyme preparations thus obtained cleaved cerebroside sulphates (sulphatides) only in the presence of either specific detergents (e.g. taurodeoxycholate) or an activator protein isolated from human liver. This corresponds to the findings on purified sulphatase A of human origin. 2) At low concentrations, the activating effect was proportional to the amount of activator protein applied; at higher concentrations, proportionality was obtained only in some cases. On a molar basis, less of the activator protein was required to achieve the same activation as taurodeoxycholate. At optimum concentrations of the detergent however, the activation was much higher. 3) The enzyme specificity of the activator and some evolutionary implications are discussed. | [
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PMID:3466 | Multiple forms of boar acrosin and their relationship to proenzyme activation. | Further evidence is presented that the acrosomal proteinase acrosin exists as a zymogen precursor in freshly ejaculated boar spermatozoa. Autoactivation of proacrosin to acrosin takes place optimally at slightly alkaline pH and in the presence of calcium ions. Activation is considerably accelerated by catalytic amounts of trypsin or highly purified acrosin. A significant acceleration of the activation is also achieved by porcine pancreatic and urinary kallikrein, whereas chymotrypsin, plasmin, thrombin or urokinase showed no effect. Activation can be inhibited by p-amino-benzamidine and p-nitrophenyl p'-guanidino-benzoate. Electrophoretic analysis at different stages of activation revealed that during this process various molecular forms of acrosin are produced, apparently by limited proteolysis. | Multiple forms of boar acrosin and their relationship to proenzyme activation. Further evidence is presented that the acrosomal proteinase acrosin exists as a zymogen precursor in freshly ejaculated boar spermatozoa. Autoactivation of proacrosin to acrosin takes place optimally at slightly alkaline pH and in the presence of calcium ions. Activation is considerably accelerated by catalytic amounts of trypsin or highly purified acrosin. A significant acceleration of the activation is also achieved by porcine pancreatic and urinary kallikrein, whereas chymotrypsin, plasmin, thrombin or urokinase showed no effect. Activation can be inhibited by p-amino-benzamidine and p-nitrophenyl p'-guanidino-benzoate. Electrophoretic analysis at different stages of activation revealed that during this process various molecular forms of acrosin are produced, apparently by limited proteolysis. | [
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PMID:3471 | [Conditions for the efficacy of nutritional therapy]. | This is to discuss regulatory mechanisms which enable the organism to respond optimally to special nutritive conditions. Those regulatory mechanisms are often in disorder with patients who need nutritive therapy. However, normal functions of the body are required for nutrition. Disarrangements in microcirculation, oxygen supply, water and electrolyte metabolism and acid-base metabolism are described and the way they will influence nutrition therapy. Furthermore, we try to describe special conditions of metabolism in stress and its influence to efficiency of nutritive therapy. | [Conditions for the efficacy of nutritional therapy]. This is to discuss regulatory mechanisms which enable the organism to respond optimally to special nutritive conditions. Those regulatory mechanisms are often in disorder with patients who need nutritive therapy. However, normal functions of the body are required for nutrition. Disarrangements in microcirculation, oxygen supply, water and electrolyte metabolism and acid-base metabolism are described and the way they will influence nutrition therapy. Furthermore, we try to describe special conditions of metabolism in stress and its influence to efficiency of nutritive therapy. | [
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PMID:3472 | [Clinical testing of bronchospasmolytics in combined alternating test schedules]. | NAB 365, a new beta2-adrenergic bronchodilator was tested in oral doses of 0.01, 0.02, 0.03 and 0.04 mg. An acute trial was carried out in outpatients suffering from chronic obstructive airways disease. Airway resistance was measured by bodyplethysmography. For doses from 0.02 to 0.04 mg NAB 365 a quick, dose related and statistically significant bronchodliation without remarkable side effects could be found. The application of Fenoterol and Sch 1000 (ipratropiumbromide) by a metered dose inhaler was followed by a further and finally equal bronchodilation. To avoid adrenergic side effects the combination of NAB 365 with Sch 1000 should be preferred to the combination of NAB 365 with Fenoterol. | [Clinical testing of bronchospasmolytics in combined alternating test schedules]. NAB 365, a new beta2-adrenergic bronchodilator was tested in oral doses of 0.01, 0.02, 0.03 and 0.04 mg. An acute trial was carried out in outpatients suffering from chronic obstructive airways disease. Airway resistance was measured by bodyplethysmography. For doses from 0.02 to 0.04 mg NAB 365 a quick, dose related and statistically significant bronchodliation without remarkable side effects could be found. The application of Fenoterol and Sch 1000 (ipratropiumbromide) by a metered dose inhaler was followed by a further and finally equal bronchodilation. To avoid adrenergic side effects the combination of NAB 365 with Sch 1000 should be preferred to the combination of NAB 365 with Fenoterol. | [
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PMID:3473 | The first transplantation of a Fallopian tube of frozen material in woman. | There is an ever-present possibility that the children or husband of a sterilized woman may die. In the latter case, and after remarriage, she and her new husband may want children of their own, which presents difficulties, as the low success rate of reconstructive surgery in the sterilised woman is well known. A case history is presented of the first sterilized woman to undergo Fallopian tube transplantation. | The first transplantation of a Fallopian tube of frozen material in woman. There is an ever-present possibility that the children or husband of a sterilized woman may die. In the latter case, and after remarriage, she and her new husband may want children of their own, which presents difficulties, as the low success rate of reconstructive surgery in the sterilised woman is well known. A case history is presented of the first sterilized woman to undergo Fallopian tube transplantation. | [
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PMID:3474 | An objective method for evaluating Angus and Hereford sperm motility. | Angus and Hereford sperm motility was evaluated by an objective spectrophotometric procedure and by a conventional subjective ranking system. Semen was collected by electroejaculation and divided so that one group contained 6 Angus samples and one group contained 12 Hereford samples. Objective procedures indicated that Angus sperm was twice motile as Hereford sperm in 2.9% sodium citrate. This objective procedure depends on the orientation of sperm in a flowing liquid and then spectrophotometrically measuring the sperm's ability to return to randomness when the flow is stopped. During preparation for freezing with liquid nitrogen, microscopic subjective ranking of sperm motility was carried out. Three conditions were studied subjectively: (I) arrival at the laboratory, (II) pre-freeze, and (III) thawed samples. No significant differences in subjective evaluation of sperm motility was found between conditions or breeds. Preliminary results indicate that this objective procedure can distinguish between the sperm motility in 2.9% sodium citrate from two breeds of cattle. Use of this objective procedure for studies relating to fertilization and artificial insemination is evident and there is no theoretical reason why the procedure can not be used with other species. | An objective method for evaluating Angus and Hereford sperm motility. Angus and Hereford sperm motility was evaluated by an objective spectrophotometric procedure and by a conventional subjective ranking system. Semen was collected by electroejaculation and divided so that one group contained 6 Angus samples and one group contained 12 Hereford samples. Objective procedures indicated that Angus sperm was twice motile as Hereford sperm in 2.9% sodium citrate. This objective procedure depends on the orientation of sperm in a flowing liquid and then spectrophotometrically measuring the sperm's ability to return to randomness when the flow is stopped. During preparation for freezing with liquid nitrogen, microscopic subjective ranking of sperm motility was carried out. Three conditions were studied subjectively: (I) arrival at the laboratory, (II) pre-freeze, and (III) thawed samples. No significant differences in subjective evaluation of sperm motility was found between conditions or breeds. Preliminary results indicate that this objective procedure can distinguish between the sperm motility in 2.9% sodium citrate from two breeds of cattle. Use of this objective procedure for studies relating to fertilization and artificial insemination is evident and there is no theoretical reason why the procedure can not be used with other species. | [
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PMID:3475 | Sperm antibodies in men from infertile couples. Analysis of sperm agglutinins and immunofluorescent antibodies in 657 men. | Sera from 657 men from infertile couples were tested for sperm agglutinins and spermatozoal antibodies detectable by the indirect immunofluorescense technique (IFT), and the results were correlated to the clinical examinations of the couples. Sperm agglutinins were found in 6.7%. Spontaneous agglutination of the ejaculated spermatozoa was observed only among these men, most commonly among those with high serum titres. IF-antibodies against the four spermatozoal antigens located beneath the cell membrane occurred in 15.2% of the patients. Antibodies against the front part of the acrosome and the postnuclear cap were mainly IgM. Antibodies against the equatorial segment of the acrosome were predominantly IgG and in a few cases IgA, whereas straining of the main tail piece was caused by IgG antibodies. Considering the clinical fertility status of the couples, sperm agglutinins in high titres (greater than or equal to 10) against the equatorial segment and the main tail piece of the spermatozoa were found significantly more often among men from couples with unexplained infertility than among clinically normal men from couples where the findings in the women could be assumed to cause infertility. These results support the view that sperm agglutinins can cause infertility, whereas the significance of the IF-antibodies is still unclarified as, in some cases, these can be found even in high titres in men with proven fertility. The possible mechanism of autosensitization were evaluated by means of an anamnestic study. | Sperm antibodies in men from infertile couples. Analysis of sperm agglutinins and immunofluorescent antibodies in 657 men. Sera from 657 men from infertile couples were tested for sperm agglutinins and spermatozoal antibodies detectable by the indirect immunofluorescense technique (IFT), and the results were correlated to the clinical examinations of the couples. Sperm agglutinins were found in 6.7%. Spontaneous agglutination of the ejaculated spermatozoa was observed only among these men, most commonly among those with high serum titres. IF-antibodies against the four spermatozoal antigens located beneath the cell membrane occurred in 15.2% of the patients. Antibodies against the front part of the acrosome and the postnuclear cap were mainly IgM. Antibodies against the equatorial segment of the acrosome were predominantly IgG and in a few cases IgA, whereas straining of the main tail piece was caused by IgG antibodies. Considering the clinical fertility status of the couples, sperm agglutinins in high titres (greater than or equal to 10) against the equatorial segment and the main tail piece of the spermatozoa were found significantly more often among men from couples with unexplained infertility than among clinically normal men from couples where the findings in the women could be assumed to cause infertility. These results support the view that sperm agglutinins can cause infertility, whereas the significance of the IF-antibodies is still unclarified as, in some cases, these can be found even in high titres in men with proven fertility. The possible mechanism of autosensitization were evaluated by means of an anamnestic study. | [
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PMID:3476 | The value of the basal temperature chart in the management of infertility. | The basal temperature chart (BTC) has been used as a means of screening the ovarian function of infertile patients and for monitoring their response to ovarian stimulation. During three years in a busy infertility clinic, specifically designated for cases of functional infertility, 491 patients were seen and 96 per cent of them kept valid basal temperature charts. The use of the BTC enabled therapy to be instituted early on in the course of investigations in those cases of infertility in which there was an endocrine factor. There were 420 patients in this category; of those who received treatment a high percentage became pregnant in contrast to those who were being observed. It is recommended that the BTC should be used early and more frequently in the management of cases of infertility, since it has been shown to serve as an inexpensive and convenient parameter of ovarian function. | The value of the basal temperature chart in the management of infertility. The basal temperature chart (BTC) has been used as a means of screening the ovarian function of infertile patients and for monitoring their response to ovarian stimulation. During three years in a busy infertility clinic, specifically designated for cases of functional infertility, 491 patients were seen and 96 per cent of them kept valid basal temperature charts. The use of the BTC enabled therapy to be instituted early on in the course of investigations in those cases of infertility in which there was an endocrine factor. There were 420 patients in this category; of those who received treatment a high percentage became pregnant in contrast to those who were being observed. It is recommended that the BTC should be used early and more frequently in the management of cases of infertility, since it has been shown to serve as an inexpensive and convenient parameter of ovarian function. | [
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PMID:3477 | Absorption activity of the blood group substances in various fractions of split ejaculates from man. | Antigens of the ABO system have been determined in the various fractions of split ejaculates from three A and two O (H) men who were secretors. The activity of the antigen was determined by a hemagglutination inhibition test. Within a split ejaculate the various fractions contained almost the same antigen activity, indicating that the antigens were secreted from all the glands contributing to the seminal plasma. | Absorption activity of the blood group substances in various fractions of split ejaculates from man. Antigens of the ABO system have been determined in the various fractions of split ejaculates from three A and two O (H) men who were secretors. The activity of the antigen was determined by a hemagglutination inhibition test. Within a split ejaculate the various fractions contained almost the same antigen activity, indicating that the antigens were secreted from all the glands contributing to the seminal plasma. | [
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PMID:3478 | The polycystic ovary. I. Estradiol/testosterone ratio and the use of antiestrogens. | Several treatments have been used to induce ovulation in the polycystic ovary. In the following study, estradiol and testosterone plasma concentration were measured and evaluated in 18 women with polycystic ovaries, as one of the factors involved in the antiestrogenic response to Clomiphene Citrate. An estrogen plasma concentration baseline of 50 pg/ml is proposed in order to obtain the antiestrogenic effects. Nevertheless, induction of ovulation was achieved when the estradiol/testosterone ratio was 0.09 or higher (P less than 0.001). | The polycystic ovary. I. Estradiol/testosterone ratio and the use of antiestrogens. Several treatments have been used to induce ovulation in the polycystic ovary. In the following study, estradiol and testosterone plasma concentration were measured and evaluated in 18 women with polycystic ovaries, as one of the factors involved in the antiestrogenic response to Clomiphene Citrate. An estrogen plasma concentration baseline of 50 pg/ml is proposed in order to obtain the antiestrogenic effects. Nevertheless, induction of ovulation was achieved when the estradiol/testosterone ratio was 0.09 or higher (P less than 0.001). | [
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PMID:3479 | The polycystic ovary. II. Progesterone plasma levels after the use of antiestrogens. | In the present study, the changes observed in the length of the cycle, basal body temperatures, endometrial histology and plasma progesterone in a group of 18 patients with polycystic ovaries, treated with Clomiphene Citrate, is reported. Two distinctive responses were observed depending on the estradiol/testosterone mean concentration found. Regardless of the induction of ovulation, a clear change in the length of the cycle was observed. Progesterone concentration of 2.1-2.9 ng/ml gave a definite rise in BBT, but no endometrial secretory changes were found in parallel with that concentration. Progesterone concentration above 6.1 ng/ml did induce secretory changes in the endometrium. | The polycystic ovary. II. Progesterone plasma levels after the use of antiestrogens. In the present study, the changes observed in the length of the cycle, basal body temperatures, endometrial histology and plasma progesterone in a group of 18 patients with polycystic ovaries, treated with Clomiphene Citrate, is reported. Two distinctive responses were observed depending on the estradiol/testosterone mean concentration found. Regardless of the induction of ovulation, a clear change in the length of the cycle was observed. Progesterone concentration of 2.1-2.9 ng/ml gave a definite rise in BBT, but no endometrial secretory changes were found in parallel with that concentration. Progesterone concentration above 6.1 ng/ml did induce secretory changes in the endometrium. | [
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PMID:3480 | Offspring of subfertile parents. A preliminary survey. | This preliminary survey reports findings among offspring of subfertile parents. Combining neonatal death, low birth weight and major anomalies, 11 of 60 livebirths (18%) were affected, indicating that subfertility adversely influences neonatal morbidity and mortality. Information was available for 49 of 56 children and there were none with developmental delay nor was there an increase in malformations or minor behavioral or learning disorders. Although numbers are small, these observations suggest that children of subfertile parents who survive the neonatal period do not differ from the normal population. | Offspring of subfertile parents. A preliminary survey. This preliminary survey reports findings among offspring of subfertile parents. Combining neonatal death, low birth weight and major anomalies, 11 of 60 livebirths (18%) were affected, indicating that subfertility adversely influences neonatal morbidity and mortality. Information was available for 49 of 56 children and there were none with developmental delay nor was there an increase in malformations or minor behavioral or learning disorders. Although numbers are small, these observations suggest that children of subfertile parents who survive the neonatal period do not differ from the normal population. | [
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PMID:3481 | Increased fibrinolytic activity in the endometrium of patients using copper-iud (gravigard). | The use of an IUD often causes an increase in the menstrual blood loss, and this is the most common reason for removing the device. The fibrinolytic activity of the endometrium was studied histochemically in 15 women before and after the use of copper-IUD for 2-5 as well as for 8-12 months. The fibrinolytic activity in the endometrium was found to be increased at both examinations. This increase is probably a contributory cause of the menorrhagia in patients with IUD. No significant difference in fibrinolytic activity was found between the two examinations after insertion of the IUD, which thus suggests that the increase in endometrial fibrinolytic activity was steady. | Increased fibrinolytic activity in the endometrium of patients using copper-iud (gravigard). The use of an IUD often causes an increase in the menstrual blood loss, and this is the most common reason for removing the device. The fibrinolytic activity of the endometrium was studied histochemically in 15 women before and after the use of copper-IUD for 2-5 as well as for 8-12 months. The fibrinolytic activity in the endometrium was found to be increased at both examinations. This increase is probably a contributory cause of the menorrhagia in patients with IUD. No significant difference in fibrinolytic activity was found between the two examinations after insertion of the IUD, which thus suggests that the increase in endometrial fibrinolytic activity was steady. | [
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PMID:3482 | Effects of aminophylline, imidazole and indomethacin on spontaneous and prostaglandin induced ovarian contractions in vitro. | Imidazole, acetylcholine, phenylephrine prostaglandin F2alpha, and methyl prostaglandin F2alpha increased the contractility of guinea pig and human ovaries in vitro. This effect was suppressed by aminophylline. Indomethacin inhibited ovarian contractions. The inhibitory effect of indomethacin was reversed by prostaglandin F2alpha or by its methyl derivative. Prostaglandin E2 decreased the amplitude and frequency of the spontaneous and prostaglandin F2alpha induced contractions of guinea pig ovaries in vitro. The study shows that compounds which interfere with cyclic AMP and prostaglandin metabolism affect ovarian contractility in vitro. | Effects of aminophylline, imidazole and indomethacin on spontaneous and prostaglandin induced ovarian contractions in vitro. Imidazole, acetylcholine, phenylephrine prostaglandin F2alpha, and methyl prostaglandin F2alpha increased the contractility of guinea pig and human ovaries in vitro. This effect was suppressed by aminophylline. Indomethacin inhibited ovarian contractions. The inhibitory effect of indomethacin was reversed by prostaglandin F2alpha or by its methyl derivative. Prostaglandin E2 decreased the amplitude and frequency of the spontaneous and prostaglandin F2alpha induced contractions of guinea pig ovaries in vitro. The study shows that compounds which interfere with cyclic AMP and prostaglandin metabolism affect ovarian contractility in vitro. | [
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PMID:3483 | Effect of low doses of alpha-chlorohydrin on fertility and semen characteristics and binding of the drug of spermatozoa in swine. | Daily feeding of 1 mg of alpha-chlorohydrin per kg body weight to boars prevented fertility completely when the ejaculate was used for insemination. The semen charactreated than in untreated boars, but the sperm morphology was otherwise normal. In vitro addition of 5 mg/epididymal contents from the treated boars revealed normal Na+, K+ and glycerylphosphorylcholine concentrations. The movement of sperm cytoplasmic droplets was completed on all spermatozoa more distally in treated than in untreated boars, but the sperm morphology was otherwise normal. In vitro addition of 5 mg/100 ml of alpha-chlorohydrin to ejaculate boar semen completely inhibited and 2.5 mg/100 ml decreased fertility. Removal of the alpha-chlorohydrin prior to insemination partially restored fertility. 14C-alpha-chlorohydrin was shown to be more firmly bound to boar spermatozoa than 14C-carboxyinulin and could not be removed from the spermatozoa with 3 washings. The contraceptive mechanism of the drug is suggested to be alkylation of the sperm membrane by free alpha-chlorohydrin in the epididymis. | Effect of low doses of alpha-chlorohydrin on fertility and semen characteristics and binding of the drug of spermatozoa in swine. Daily feeding of 1 mg of alpha-chlorohydrin per kg body weight to boars prevented fertility completely when the ejaculate was used for insemination. The semen charactreated than in untreated boars, but the sperm morphology was otherwise normal. In vitro addition of 5 mg/epididymal contents from the treated boars revealed normal Na+, K+ and glycerylphosphorylcholine concentrations. The movement of sperm cytoplasmic droplets was completed on all spermatozoa more distally in treated than in untreated boars, but the sperm morphology was otherwise normal. In vitro addition of 5 mg/100 ml of alpha-chlorohydrin to ejaculate boar semen completely inhibited and 2.5 mg/100 ml decreased fertility. Removal of the alpha-chlorohydrin prior to insemination partially restored fertility. 14C-alpha-chlorohydrin was shown to be more firmly bound to boar spermatozoa than 14C-carboxyinulin and could not be removed from the spermatozoa with 3 washings. The contraceptive mechanism of the drug is suggested to be alkylation of the sperm membrane by free alpha-chlorohydrin in the epididymis. | [
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PMID:3484 | Active transport system in human spermatozoa. | The presence of potassium influx into human spermatozoa was investigated through the use of radioactive K42 and Rb86. A gradient of potassium and sodium has been found between the spermatozoal midpiece-tail region and seminal plasma. The influx of potassium seemed to correlate with spermatozoal motility. | Active transport system in human spermatozoa. The presence of potassium influx into human spermatozoa was investigated through the use of radioactive K42 and Rb86. A gradient of potassium and sodium has been found between the spermatozoal midpiece-tail region and seminal plasma. The influx of potassium seemed to correlate with spermatozoal motility. | [
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PMID:3485 | Sperm antibodies in serum and seminal plasma. | The relation between sperm antibodies (agglutinins and IF-antibodies) in serum and seminal plasma was studied in three selected groups of male partners in infertile couples with respect to specificity, concentrations and immunoglobulin classes. Agglutinins were found in seminal plasma only when they were also present in serum, but there was no strict correlation between the titres in seminal plasma and serum although serum titres were always the higher ones. However, in cases with tail-to-tail agglutinins a difference of more than 2 fourfold steps between the titres was never found; spontaneous agglutination in the ejaculate generally occurred when the seminal plasma titre was 64 or more, and in 6 patients with high seminal plasma titres IgA was detected in the midpiece region of ejaculated spermatozoa. In contrast, head-to-head agglutinins were found in seminal plasma in only one of 3 patients with these agglutinins in serum, and only in a low titre. Absorption of IgG of IgG with protein-A-producing Staphylococcus aureus showed that tail-to-tail agglutinins in serum were IgG antibodies, whereas they could be characterized as "non-IgA-in seminal plasma, suggesting a local production of these antibodies. IF-antibodies were rarely found in seminal plasma and seemed to be of little significance in relation to infertility in men. | Sperm antibodies in serum and seminal plasma. The relation between sperm antibodies (agglutinins and IF-antibodies) in serum and seminal plasma was studied in three selected groups of male partners in infertile couples with respect to specificity, concentrations and immunoglobulin classes. Agglutinins were found in seminal plasma only when they were also present in serum, but there was no strict correlation between the titres in seminal plasma and serum although serum titres were always the higher ones. However, in cases with tail-to-tail agglutinins a difference of more than 2 fourfold steps between the titres was never found; spontaneous agglutination in the ejaculate generally occurred when the seminal plasma titre was 64 or more, and in 6 patients with high seminal plasma titres IgA was detected in the midpiece region of ejaculated spermatozoa. In contrast, head-to-head agglutinins were found in seminal plasma in only one of 3 patients with these agglutinins in serum, and only in a low titre. Absorption of IgG of IgG with protein-A-producing Staphylococcus aureus showed that tail-to-tail agglutinins in serum were IgG antibodies, whereas they could be characterized as "non-IgA-in seminal plasma, suggesting a local production of these antibodies. IF-antibodies were rarely found in seminal plasma and seemed to be of little significance in relation to infertility in men. | [
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PMID:3489 | [Clinical and experimental observations on idiopathic urticaria due to the contact with heat]. | This study describes the probably eleventh case, mentioned in literature, of acquired heat contact uticaria in an otherwise healthy young woman. With regard to true contact induction heat contact urticaria clearly differs from the more common cholinergic uticaria. On the other hand, heat contact urticaria is completely analogous to cold uticaria because of the exposure area, reversible blocking by unphysiological prolonged heating of the skin, sensitivity to antihistamines and resistance to corticosteroids. In this case, whealing of the skin occurred on exposure to heating of 39 degrees C for 5 min. With a temperature of 44-46 degrees C, The shortest time for wheal induction was 3-5 sec. At 70 degrees C, the shortest time for maximal reaction was only a split second. An "optimal temperature" for wheal induction could not be determined. Local anaesthesia with 2% Xylocain caused a considerable blocking of wheals. Histamine and cholinergic drugs showed normal skin reactions after intradermal injection. Antihistamines administered parenterally or perorally were highly effective. Corticosteroids, however, given systemically in high doses proved to be ineffective. During our observations, a spontaneous remission appeared with a clinical symptom-free state; on unphysiological high temperature stimulus, however, contact uticaria could still be demonstrated. The pathogenetic uniformity of sporadic heat contact urticaria and problems of therapeutical controls are discussed. | [Clinical and experimental observations on idiopathic urticaria due to the contact with heat]. This study describes the probably eleventh case, mentioned in literature, of acquired heat contact uticaria in an otherwise healthy young woman. With regard to true contact induction heat contact urticaria clearly differs from the more common cholinergic uticaria. On the other hand, heat contact urticaria is completely analogous to cold uticaria because of the exposure area, reversible blocking by unphysiological prolonged heating of the skin, sensitivity to antihistamines and resistance to corticosteroids. In this case, whealing of the skin occurred on exposure to heating of 39 degrees C for 5 min. With a temperature of 44-46 degrees C, The shortest time for wheal induction was 3-5 sec. At 70 degrees C, the shortest time for maximal reaction was only a split second. An "optimal temperature" for wheal induction could not be determined. Local anaesthesia with 2% Xylocain caused a considerable blocking of wheals. Histamine and cholinergic drugs showed normal skin reactions after intradermal injection. Antihistamines administered parenterally or perorally were highly effective. Corticosteroids, however, given systemically in high doses proved to be ineffective. During our observations, a spontaneous remission appeared with a clinical symptom-free state; on unphysiological high temperature stimulus, however, contact uticaria could still be demonstrated. The pathogenetic uniformity of sporadic heat contact urticaria and problems of therapeutical controls are discussed. | [
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PMID:3487 | Fluorescence polarization studies on the binding between glutamate dehydrogenase and cytoplasmic aspartate aminotransferase. | Polarization of fluorescence measurements on aspartate aminotransferase (from pig heart cytosol) labeled with fluorescein isothiocyanate have been used to detect the formation of a soluble complex of this protein wich glutamate dehydrogenase from bovine liver. The binding of the labeled transaminase to dehydrogenase is detectable at catalytic concentrations of the enzymes. | Fluorescence polarization studies on the binding between glutamate dehydrogenase and cytoplasmic aspartate aminotransferase. Polarization of fluorescence measurements on aspartate aminotransferase (from pig heart cytosol) labeled with fluorescein isothiocyanate have been used to detect the formation of a soluble complex of this protein wich glutamate dehydrogenase from bovine liver. The binding of the labeled transaminase to dehydrogenase is detectable at catalytic concentrations of the enzymes. | [
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PMID:3490 | Lubrication and cartilage. | Mechanisms of lubrication of human synovial joints have been analysed in terms of the operating conditions of the joint, the synovial fluid and articular cartilage. In the hip and knee during a walking cycle the load may rise up to four times body weight. In the knee on dropping one metre the load may go up to 25 time body weight. The elastic modulus of cartilage is similar to that of the synthetic rubber of a car tyre. The cartilage surface is rough and in elderly specimens the centre line average is 2-75 mum. The friction force generated in reciprocating tests shows that both cartilage and synovial fluid are important in lubrication. The viscosity-shear rate relationships of normal synovial fluid show that it is non-Newtonian. Osteoarthrosic fluid is less so and rheumatoid fluid is more nearly Newtonian. Experiments with hip joints in a pendulum machine show that fluid film lubrication obtains at some phases of joint action. Boundary lubrication prevails under certain conditions and has been examined with a reciprocating friction machine. Digestion of hyaluronate does not alter the boundary lubrication, but trypsin digestion does. Surface active substances (lauryl sulphate and cetyl 3-ammonium bromide) give a lubricating ability similar to that of synovial fluid. The effectiveness of the two substances varies with pH. | Lubrication and cartilage. Mechanisms of lubrication of human synovial joints have been analysed in terms of the operating conditions of the joint, the synovial fluid and articular cartilage. In the hip and knee during a walking cycle the load may rise up to four times body weight. In the knee on dropping one metre the load may go up to 25 time body weight. The elastic modulus of cartilage is similar to that of the synthetic rubber of a car tyre. The cartilage surface is rough and in elderly specimens the centre line average is 2-75 mum. The friction force generated in reciprocating tests shows that both cartilage and synovial fluid are important in lubrication. The viscosity-shear rate relationships of normal synovial fluid show that it is non-Newtonian. Osteoarthrosic fluid is less so and rheumatoid fluid is more nearly Newtonian. Experiments with hip joints in a pendulum machine show that fluid film lubrication obtains at some phases of joint action. Boundary lubrication prevails under certain conditions and has been examined with a reciprocating friction machine. Digestion of hyaluronate does not alter the boundary lubrication, but trypsin digestion does. Surface active substances (lauryl sulphate and cetyl 3-ammonium bromide) give a lubricating ability similar to that of synovial fluid. The effectiveness of the two substances varies with pH. | [
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PMID:3491 | Urea transport-defective strains of Saccharomyces cerevisiae. | Experiments characterizing the urea active transport system in Saccharomyces cerevisiae indicate that (i) formamide and acetamide are strong competitive inhibitors of urea accumulation, (ii) uptake is maximal at pH 3.3 and is 80% inhibited at pH 6.0, and (iii) adenosine 5'-triphosphate generated by glycolysis in conjunction with formation of an ion gradient is likely the driving force behind urea transport. Mutant strains were isolated that are unable to accumulate urea at external concentrations of 0.25 mM. These strains also exhibit a depressed growth rate on 10 mM urea, indicating existence of a relationship between the active transport and facilitated diffusion modes of urea uptake. | Urea transport-defective strains of Saccharomyces cerevisiae. Experiments characterizing the urea active transport system in Saccharomyces cerevisiae indicate that (i) formamide and acetamide are strong competitive inhibitors of urea accumulation, (ii) uptake is maximal at pH 3.3 and is 80% inhibited at pH 6.0, and (iii) adenosine 5'-triphosphate generated by glycolysis in conjunction with formation of an ion gradient is likely the driving force behind urea transport. Mutant strains were isolated that are unable to accumulate urea at external concentrations of 0.25 mM. These strains also exhibit a depressed growth rate on 10 mM urea, indicating existence of a relationship between the active transport and facilitated diffusion modes of urea uptake. | [
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PMID:3492 | Physiological factors affecting transformation of Azotobacter vinelandii. | Cells of Azotobacter vinelandii (ATCC 12837) can be transformed by exogenous deoxyribonucleic acid towards the end of exponential growth. Transformation occurs at very low frequencies when the deoxyribonucleic acid is purified or when the transformation is carried out in liquid medium. Optimal transformation occurs on plates of Burk nitrogen-free glucose medium containing either high phosphate (10 mM) or low calcium (0 to 0.29 mM) content. Higher levels of calcium are inhibitory, whereas magnesium ions are essential for transformation and growth. Extracellular polymer and capsule are increasingly inhibitory to transformation and are most abundant when the calcium content of the medium is high. Transformation is optimal at pH 7.0 to 7.1 and at 30 C, conditions which also coincide with minimal extracellular polymer production. Nonencapsulated strains are excellent transformation recipients. Glycine-induced pleomorphism reduces the transformation frequency and the degree of inhibition is dependent on the phosphate concentration of the medium. Rifampin resistance and shifts from adenine, hypoxanthine, uracil, and nitrogenase auxotrophy to prototrophy can be achieved. Although single marker transfer is always greater than double marker transfer, the data suggest that rifampin resistance is linked to hypoxanthine, adenine and uracil protorophy at intervals of increasing distance. Rifampin resistance did not appear to be linked to nitrogenase. | Physiological factors affecting transformation of Azotobacter vinelandii. Cells of Azotobacter vinelandii (ATCC 12837) can be transformed by exogenous deoxyribonucleic acid towards the end of exponential growth. Transformation occurs at very low frequencies when the deoxyribonucleic acid is purified or when the transformation is carried out in liquid medium. Optimal transformation occurs on plates of Burk nitrogen-free glucose medium containing either high phosphate (10 mM) or low calcium (0 to 0.29 mM) content. Higher levels of calcium are inhibitory, whereas magnesium ions are essential for transformation and growth. Extracellular polymer and capsule are increasingly inhibitory to transformation and are most abundant when the calcium content of the medium is high. Transformation is optimal at pH 7.0 to 7.1 and at 30 C, conditions which also coincide with minimal extracellular polymer production. Nonencapsulated strains are excellent transformation recipients. Glycine-induced pleomorphism reduces the transformation frequency and the degree of inhibition is dependent on the phosphate concentration of the medium. Rifampin resistance and shifts from adenine, hypoxanthine, uracil, and nitrogenase auxotrophy to prototrophy can be achieved. Although single marker transfer is always greater than double marker transfer, the data suggest that rifampin resistance is linked to hypoxanthine, adenine and uracil protorophy at intervals of increasing distance. Rifampin resistance did not appear to be linked to nitrogenase. | [
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PMID:3493 | Role of D-tryptophan oxidase in D-tryptophan utilization by Escherichia coli. | Mutants of Escherichia coli K-12 that require L-tryptophan (trp) are normally unable to utilize D-tryptophan to fulfill their requirement. However, secondary mutations (dadR) that confer this ability can be isolated. In such strains two distinct enzymes are found to be produced at high levels: D-amino acid oxidase (EC 1.4.3.3) and D-tryptophan oxidase. A convenient assay procedure for D-tryptophan oxidase is described. The two enzymes could be distinguished on the basis of their sensitivity to inhibition by L-phenylalanine and L-tyrosine. Strains that were trp dadR could not grow with D-tryptophan in the presence of L-phenylalanine, but further mutations, Fyo, could be isolated that allowed growth under these conditions. Some of them were characterized by further increases in the level of D-tryptophan oxidase activity and a sharp decrease in D-amino acid oxidase. These kinds of Fyo mutations lay in or near the dadR gene. The substrate specificity of the two enzymes toward a large number of compounds was examined. The transamination of aromatic keto acids was investigated. In the wild-type strain only a single enzyme, transaminase A (EC 2.6.1.5), was found, and it was irreversibly activated when subjected to elevated temperatures. The present state of our knowledge on D-amino acid utilization in E. coli is summarized. | Role of D-tryptophan oxidase in D-tryptophan utilization by Escherichia coli. Mutants of Escherichia coli K-12 that require L-tryptophan (trp) are normally unable to utilize D-tryptophan to fulfill their requirement. However, secondary mutations (dadR) that confer this ability can be isolated. In such strains two distinct enzymes are found to be produced at high levels: D-amino acid oxidase (EC 1.4.3.3) and D-tryptophan oxidase. A convenient assay procedure for D-tryptophan oxidase is described. The two enzymes could be distinguished on the basis of their sensitivity to inhibition by L-phenylalanine and L-tyrosine. Strains that were trp dadR could not grow with D-tryptophan in the presence of L-phenylalanine, but further mutations, Fyo, could be isolated that allowed growth under these conditions. Some of them were characterized by further increases in the level of D-tryptophan oxidase activity and a sharp decrease in D-amino acid oxidase. These kinds of Fyo mutations lay in or near the dadR gene. The substrate specificity of the two enzymes toward a large number of compounds was examined. The transamination of aromatic keto acids was investigated. In the wild-type strain only a single enzyme, transaminase A (EC 2.6.1.5), was found, and it was irreversibly activated when subjected to elevated temperatures. The present state of our knowledge on D-amino acid utilization in E. coli is summarized. | [
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PMID:3494 | Transduction of chromosomal genes between enteric bacteria by bacteriophage P1. | We have used P1 transduction to create intergeneric hybrid strains of enteric bacteria by moving the genA and hut genes between Klebsiella aerogenes, Escherichia coli and Salmonella typhimurium. The use of E. coli as the recipient in such transductions permits the construction of episomes and specialized transducing phage containing non-E. coli material. The effect of host restriction modification and deoxyribonucleic acid homology on the frequency of intergeneric transduction of these loci has been examined. | Transduction of chromosomal genes between enteric bacteria by bacteriophage P1. We have used P1 transduction to create intergeneric hybrid strains of enteric bacteria by moving the genA and hut genes between Klebsiella aerogenes, Escherichia coli and Salmonella typhimurium. The use of E. coli as the recipient in such transductions permits the construction of episomes and specialized transducing phage containing non-E. coli material. The effect of host restriction modification and deoxyribonucleic acid homology on the frequency of intergeneric transduction of these loci has been examined. | [
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PMID:3495 | Galactoside accumulation by Escherichia coli, driven by a pH gradient. | Acidification of the external medium results in thiomethylgalactoside accumulation in an energy-depleted adenosine triphosphatase-negative mutant of Escherichia coli. | Galactoside accumulation by Escherichia coli, driven by a pH gradient. Acidification of the external medium results in thiomethylgalactoside accumulation in an energy-depleted adenosine triphosphatase-negative mutant of Escherichia coli. | [
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PMID:3496 | Adenosine 5'-monophosphate-stimulated cyanide-insensitive respiration in mitochondria of Moniliella tomentosa. | Mitochondria of the yeastlike fungus Moniliella tomentosa oxidize reduced nicotinamide adenine dinucleotide, reduced nicotinamide adenine dinucleotide phosphate, succinate, isocitrate, and lactate. These oxidations are completely inhibited by cyanide or antimycin A in mitochondria isolated from cells grown in the standard medium. On the other hand, the oxidation of all substrates, except lactate, is almost completely insensitive to cyanide or antimycin A in mitochondria from cells grown in the presence of ethidium bromide. In this instance, the oxidation is mainly mediated by an alternate oxidase which can be blocked by salicyl hydroxamic acid. The alternate oxidase can be specifically stimulated by adenosine 5'-monophosphate and this provides a new method for the characterization of the alternate oxidase in mitochondria of M. tomentosa. | Adenosine 5'-monophosphate-stimulated cyanide-insensitive respiration in mitochondria of Moniliella tomentosa. Mitochondria of the yeastlike fungus Moniliella tomentosa oxidize reduced nicotinamide adenine dinucleotide, reduced nicotinamide adenine dinucleotide phosphate, succinate, isocitrate, and lactate. These oxidations are completely inhibited by cyanide or antimycin A in mitochondria isolated from cells grown in the standard medium. On the other hand, the oxidation of all substrates, except lactate, is almost completely insensitive to cyanide or antimycin A in mitochondria from cells grown in the presence of ethidium bromide. In this instance, the oxidation is mainly mediated by an alternate oxidase which can be blocked by salicyl hydroxamic acid. The alternate oxidase can be specifically stimulated by adenosine 5'-monophosphate and this provides a new method for the characterization of the alternate oxidase in mitochondria of M. tomentosa. | [
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PMID:3497 | Sulfate-reducing pathway in Escherichia coli involving bound intermediates. | Although a sulfate-reducing pathway in Escherichia coli involving free sulfite and sulfide has been suggested, it is shown that, as in Chlorella, a pathway involving bound intermediates is also present. E. coli extracts contained a sulfotransferase that transferred the sulfonyl group from a nucleosidephosphosulfate to an acceptor to form an organic thiosulfate. This enzyme was specific for adenosine 3'-phosphate 5'-phosphosulfate, did not utilize adenine 5'-phosphosulfate, and transferred to a carrier molecule that was identical with thioredoxin in molecular weight and amino acid composition. In the absence of thioredoxin, only very low levels of the transfer of the sulfo group to thiols was observed. As in Chlorella, thiosulfonate reductase activity that reduced glutathione-S-SO3- to bound sulfide could be detected. In E. coli, this enzyme used reduced nicotinamide adenine dinucleotide phosphate and Mg2+, but did not require the addition of ferredoxin or ferredoxin nicotinamide adenine dinucleotide phosphate reductase. Although in Chlorella the thiosulfonate reductase appears to be a different enzyme from the sulfite reductase, the E. coli thiosulfonate reductase and sulfite reductase may be activities of the same enzyme. | Sulfate-reducing pathway in Escherichia coli involving bound intermediates. Although a sulfate-reducing pathway in Escherichia coli involving free sulfite and sulfide has been suggested, it is shown that, as in Chlorella, a pathway involving bound intermediates is also present. E. coli extracts contained a sulfotransferase that transferred the sulfonyl group from a nucleosidephosphosulfate to an acceptor to form an organic thiosulfate. This enzyme was specific for adenosine 3'-phosphate 5'-phosphosulfate, did not utilize adenine 5'-phosphosulfate, and transferred to a carrier molecule that was identical with thioredoxin in molecular weight and amino acid composition. In the absence of thioredoxin, only very low levels of the transfer of the sulfo group to thiols was observed. As in Chlorella, thiosulfonate reductase activity that reduced glutathione-S-SO3- to bound sulfide could be detected. In E. coli, this enzyme used reduced nicotinamide adenine dinucleotide phosphate and Mg2+, but did not require the addition of ferredoxin or ferredoxin nicotinamide adenine dinucleotide phosphate reductase. Although in Chlorella the thiosulfonate reductase appears to be a different enzyme from the sulfite reductase, the E. coli thiosulfonate reductase and sulfite reductase may be activities of the same enzyme. | [
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PMID:3498 | An endonuclease from Escherichia coli that introduces single polynucleotide chain scissions in ultraviolet-irradiated DNA. | An endonuclease that makes single polynucleotide chain scissions in ultraviolet-irradiated DNA has been purified from Escherichia coli. The activity has the following properties: (a) unirradiated DNA is attacked very little if at all; (b) single strand DNA is not attacked, whether irradiated or not; (c) there is no requirement for divalent cations and the activity is not affected by the addition of EDTA; (d) the pH optimum is approximately 7; (e) the activity is inhibited by 1 M NaCl, single strand DNA, transfer RNA and double strand DNA; (f) the sedimentation coefficient, S20,w, is approximately 2.6; (g) it is a basic protein. The enzyme is tentatively named E. coli endonuclease III. The physiological function of the endonuclease has not yet been established. | An endonuclease from Escherichia coli that introduces single polynucleotide chain scissions in ultraviolet-irradiated DNA. An endonuclease that makes single polynucleotide chain scissions in ultraviolet-irradiated DNA has been purified from Escherichia coli. The activity has the following properties: (a) unirradiated DNA is attacked very little if at all; (b) single strand DNA is not attacked, whether irradiated or not; (c) there is no requirement for divalent cations and the activity is not affected by the addition of EDTA; (d) the pH optimum is approximately 7; (e) the activity is inhibited by 1 M NaCl, single strand DNA, transfer RNA and double strand DNA; (f) the sedimentation coefficient, S20,w, is approximately 2.6; (g) it is a basic protein. The enzyme is tentatively named E. coli endonuclease III. The physiological function of the endonuclease has not yet been established. | [
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PMID:3499 | Quaternary conformational changes in human hemoglobin studied by laser photolysis of carboxyhemoglobin. | These experiments indicate that absorbance changes observed at the 425 nm isosbestic point of the Hb and HbCO following laser photolysis of HbCO provide a direct measure of the rates of quaternary conformational changes between rapidly reacting Hb (the immediate product of full photolysis) and slowly reacting normal deoxyhemoglobin. Hb, first observed by Gibson (Gibson, Q.H. (1959) Biochem. J. 71, 293-303), Has been interpreted as deoxyhemoglobin remaining in the liganded quaternary conformation following rapid removal of ligand by a light pulse. In borate buffers between pH 8.4 and 9.6 particularly simple pH-independent results were obtained which allowed the use of a Monod. Wyman, and Changeux model (Monod, J., Wyman, J., and Changeux, J (1965) J. Mol. Biol. 12, 88-118) to fit the data. In this case Hb is taken to be R state deoxyhemoglobin. Partial photolysis experiments at 425 nm show that the rate of the R - T conformational change at 20 degrees decreases by about a factor of 2 for each additional bound ligand. The rate of the ligand-free conformational change is found to be 920 +/- 60s(-1), 6400 +/- 600s(-1), and 15,700 +/- 700(-1) respectively at 3 degrees, 20 degrees, and 30 degrees. The previously uninterpreted effects of flash length and partial photolysis on the CO recombination kinetics can be explained in terms of the present model. Kinetic results obtained below pH 8 are found to be inconsistent with a two-state model. It appears that binding of inositol hexaphosphate produces a new rapidly reacting quaternary conformation of HbCO. | Quaternary conformational changes in human hemoglobin studied by laser photolysis of carboxyhemoglobin. These experiments indicate that absorbance changes observed at the 425 nm isosbestic point of the Hb and HbCO following laser photolysis of HbCO provide a direct measure of the rates of quaternary conformational changes between rapidly reacting Hb (the immediate product of full photolysis) and slowly reacting normal deoxyhemoglobin. Hb, first observed by Gibson (Gibson, Q.H. (1959) Biochem. J. 71, 293-303), Has been interpreted as deoxyhemoglobin remaining in the liganded quaternary conformation following rapid removal of ligand by a light pulse. In borate buffers between pH 8.4 and 9.6 particularly simple pH-independent results were obtained which allowed the use of a Monod. Wyman, and Changeux model (Monod, J., Wyman, J., and Changeux, J (1965) J. Mol. Biol. 12, 88-118) to fit the data. In this case Hb is taken to be R state deoxyhemoglobin. Partial photolysis experiments at 425 nm show that the rate of the R - T conformational change at 20 degrees decreases by about a factor of 2 for each additional bound ligand. The rate of the ligand-free conformational change is found to be 920 +/- 60s(-1), 6400 +/- 600s(-1), and 15,700 +/- 700(-1) respectively at 3 degrees, 20 degrees, and 30 degrees. The previously uninterpreted effects of flash length and partial photolysis on the CO recombination kinetics can be explained in terms of the present model. Kinetic results obtained below pH 8 are found to be inconsistent with a two-state model. It appears that binding of inositol hexaphosphate produces a new rapidly reacting quaternary conformation of HbCO. | [
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PMID:3500 | Muscarinic acetylcholine receptor from rat brain. Partial purification and characterization. | A protein capable of binding atropine and (3H)propylbenzilylcholine mustard was solubilized and purified (200-fold) from rat brain. Pronase and trypsin, but not phospholipases, diminished the binding capacity of the solubilized receptor. The molecular weight of the salt-solubilized receptor as determined by gel filtration in the absence of detergents is 30,000. The purified protein showed specificity of binding toward muscarinic ligands. the high and low affinity dissociation constants of the receptor.atropine complex are 0.3 nM and 0.15 muM. Binding of atropine is pH-dependent with an optimum at 7.1. Ca2+ influences the binding of atropine and maximal binding occurs at 0.5 mM Ca2+. The subcellular distribution of the receptor was also examined. | Muscarinic acetylcholine receptor from rat brain. Partial purification and characterization. A protein capable of binding atropine and (3H)propylbenzilylcholine mustard was solubilized and purified (200-fold) from rat brain. Pronase and trypsin, but not phospholipases, diminished the binding capacity of the solubilized receptor. The molecular weight of the salt-solubilized receptor as determined by gel filtration in the absence of detergents is 30,000. The purified protein showed specificity of binding toward muscarinic ligands. the high and low affinity dissociation constants of the receptor.atropine complex are 0.3 nM and 0.15 muM. Binding of atropine is pH-dependent with an optimum at 7.1. Ca2+ influences the binding of atropine and maximal binding occurs at 0.5 mM Ca2+. The subcellular distribution of the receptor was also examined. | [
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PMID:3501 | Quantitative relationships between phosphorylation, electron flow, and internal hydrogen ion concentrations in spinach chloroplasts. | 1. Further evidence that the uptake of [14C]hexylamine, determined by centrifugal filtration of spinach chloroplast thylakoids through silicone fluid layers, gives precise estimations of light-induced H+ concentration gradients (deltapH) is presented. DeltapH was independent of the amount of thylakoids used or of the concentration of hexylamine. Moreover, hexylamine uptake was sensitive to the osmolarity of the suspending medium. 2. Internal H+ concentration ([H+]in) is proportional to the rate of electron flow when light intensity was used to vary these parameters. Proportionality was still observed in the presence of 0.1 and 1.0 muM gramicidin D. When, however, [H+]in and electron flow were varied by increasing the concentration of gramicidin D, at constant light intensity the rate of electron flow was approximately proportional to 1/[H]in. 3. The phosphorylation efficiency (P/e2 ratio) falls with decreasing light intensity or increasing concentrations of the phosphorylation inhibitor, 4'-deoxyphlorizin. The proportionality between the rate of electron flow and [H+]in allows the calculation of the rate of nonphosphorylating (basal) electron flow if [H+]in under phosphorylating conditions is known. The contribution of basal electron flow, a consequence of passive efflux of H+ from the thylakoids, to the overall rate of electron flow increases as the rate of phosphorylation decreases. P/e2 ratios calculated using rates of electron flow from which the basal component has been subtracted are constant. A calculated P/e2 ratio of about 1.3 is obtained. 4. It is shown that the reciprocal of the phosphorylation efficiency should be proportional to 1/[H+]in2 when these parameters are varied using light intensity. This relationship was verified and provided an estimate of the P/e2 at infinite [H+]in. This value was 1.3. These results provide further evidence that a H+ electrochemical gradient serves to couple photophosphorylation to electron flow and that the rate of phosphorylation is proportional to [H+]in3. That is, three H+ are translocated out of thylakoids for each adenosine triphosphate formed. | Quantitative relationships between phosphorylation, electron flow, and internal hydrogen ion concentrations in spinach chloroplasts. 1. Further evidence that the uptake of [14C]hexylamine, determined by centrifugal filtration of spinach chloroplast thylakoids through silicone fluid layers, gives precise estimations of light-induced H+ concentration gradients (deltapH) is presented. DeltapH was independent of the amount of thylakoids used or of the concentration of hexylamine. Moreover, hexylamine uptake was sensitive to the osmolarity of the suspending medium. 2. Internal H+ concentration ([H+]in) is proportional to the rate of electron flow when light intensity was used to vary these parameters. Proportionality was still observed in the presence of 0.1 and 1.0 muM gramicidin D. When, however, [H+]in and electron flow were varied by increasing the concentration of gramicidin D, at constant light intensity the rate of electron flow was approximately proportional to 1/[H]in. 3. The phosphorylation efficiency (P/e2 ratio) falls with decreasing light intensity or increasing concentrations of the phosphorylation inhibitor, 4'-deoxyphlorizin. The proportionality between the rate of electron flow and [H+]in allows the calculation of the rate of nonphosphorylating (basal) electron flow if [H+]in under phosphorylating conditions is known. The contribution of basal electron flow, a consequence of passive efflux of H+ from the thylakoids, to the overall rate of electron flow increases as the rate of phosphorylation decreases. P/e2 ratios calculated using rates of electron flow from which the basal component has been subtracted are constant. A calculated P/e2 ratio of about 1.3 is obtained. 4. It is shown that the reciprocal of the phosphorylation efficiency should be proportional to 1/[H+]in2 when these parameters are varied using light intensity. This relationship was verified and provided an estimate of the P/e2 at infinite [H+]in. This value was 1.3. These results provide further evidence that a H+ electrochemical gradient serves to couple photophosphorylation to electron flow and that the rate of phosphorylation is proportional to [H+]in3. That is, three H+ are translocated out of thylakoids for each adenosine triphosphate formed. | [
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PMID:3502 | Kinetic properties of human placental aromatase. Application of an assay measuring 3H2O release from 1beta,2beta-3H-androgens. | The rapid and sensitive assay of 1beta,2beta-3H-androgen aromatization by measurement of 3H2O release (Thompson, E.A., Jr., and Siiteri, P.K. (1974) J. Biol. Chem. 249, 5364-5372) has been analyzed to determine its applicability to initial rate studies. It was found that aromatization is the sole reaction catalyzed by lyophilized placental microsomes that causes a loss of tritium from position 1 or 2 of androstenedione and testosterone. Tritium is, however, removed from position 2 of the estrogen products, presumably in 2-hydroxylation, but this does not invalidate use of the assay for initial rate measurements; it was therefore used to characterize the catalytic properties of aromatase. Aromatization by the freeze-dried preparation was stimulated by K+, EDTA, and dithiothreitol, and was maximally active at pH 7.5 TO 8.0. With incubation conditions optimized for these factors, the apparent Km for NADPH is approximately 1 muM. The maximum velocity of androstenedione aromatization exceeds that of testosterone, and the affinity of the substrate binding site is higher for the former substrate, the apparent Km values being 0.1 muM and 0.4 muM, respectively. Mutual competition experiments with the androgen substrates showed that each gives simple competitive inhibition of the other's aromatization; furthermore, the apparent Ki values for each are in close agreement with their respective Km values. Androst-1,4,6-triene-3,17-dione competitively inhibits the aromatization of both androstenedione and testosterone, the apparent Ki, in both cases being 0.2 muM. It is concluded that the two androgen substrates are aromatized at a single, identical site. | Kinetic properties of human placental aromatase. Application of an assay measuring 3H2O release from 1beta,2beta-3H-androgens. The rapid and sensitive assay of 1beta,2beta-3H-androgen aromatization by measurement of 3H2O release (Thompson, E.A., Jr., and Siiteri, P.K. (1974) J. Biol. Chem. 249, 5364-5372) has been analyzed to determine its applicability to initial rate studies. It was found that aromatization is the sole reaction catalyzed by lyophilized placental microsomes that causes a loss of tritium from position 1 or 2 of androstenedione and testosterone. Tritium is, however, removed from position 2 of the estrogen products, presumably in 2-hydroxylation, but this does not invalidate use of the assay for initial rate measurements; it was therefore used to characterize the catalytic properties of aromatase. Aromatization by the freeze-dried preparation was stimulated by K+, EDTA, and dithiothreitol, and was maximally active at pH 7.5 TO 8.0. With incubation conditions optimized for these factors, the apparent Km for NADPH is approximately 1 muM. The maximum velocity of androstenedione aromatization exceeds that of testosterone, and the affinity of the substrate binding site is higher for the former substrate, the apparent Km values being 0.1 muM and 0.4 muM, respectively. Mutual competition experiments with the androgen substrates showed that each gives simple competitive inhibition of the other's aromatization; furthermore, the apparent Ki values for each are in close agreement with their respective Km values. Androst-1,4,6-triene-3,17-dione competitively inhibits the aromatization of both androstenedione and testosterone, the apparent Ki, in both cases being 0.2 muM. It is concluded that the two androgen substrates are aromatized at a single, identical site. | [
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PMID:3503 | Apparent oxidation-reduction potential of Clostridium acidi-urici ferredoxin. Effect of pH, ionic strength, and amino acid replacements. | The effects of pH and ionic strength on the midpoint reduction potential (Emp) of Clostridium acidi-urici ferredoxin were determined using hydrogen gas and hydrogenase. The Emp of native ferredoxin at 24-25 degrees in 0.1 M Tris-chloride buffer, pH 7.0, is--0.434 V. In the pH range examined, the Emp becomes approximately 13 mv more negative per each pH unit increase. A plot of the log of ionic strength versus the apparent Emp of ferredoxin in 0.1 M Tris-chloride buffer, pH 7.5, Was linear over the range of 1.0 to 0.01 ionic strength with Emp values of--0.414 and--0.475 V, respectively, at these extremes. This effect is the same with sodium chloride, sodium bromide, or ammonium sulfate. Potassium phosphate buffer caused a similar change, but the absolute values of Emp differed from those obtained in the presence of the other salts. This effect of pH and ionic strength on Emp may be general for clostridial-type (Fe4S4)2-ferredoxins, since the apparent Emp of Clostridium pasteurianum ferredoxin is affected in a similar manner by these two variables. The Emp of this ferredoxin in 0.1 M Tris-chloride buffer pH 7.0, is--0.405 V. Since the NH2-terminal amino acid residue, Ala1, and Tyr2 of C. acidi urici ferredoxin are near an (Fe4S4)2-cluster in the protein, the apparent Emp of derivatives that contained amino acid replacements in these two positions were determined. Under similar conditions, the Emp of most of the 13 derivatives examined, including those of [Leu2]- and[3-NH2-Tyr30]ferredoxin, is approximately the same as that of native ferredoxin. However, the Emp of [His2]ferredoxin is approximately 15 mv more positive, whereas that of [Trp2]ferredoxin is 22 mv more negative than that of native C. acidi-urici ferredoxin. Variations in sodium chloride concentration and pH also affected the apparent Emp of the derivatives. It is suggested that the changes observed in the Emp of C. acidi-urici ferredoxin are caused by protein conformational changes. | Apparent oxidation-reduction potential of Clostridium acidi-urici ferredoxin. Effect of pH, ionic strength, and amino acid replacements. The effects of pH and ionic strength on the midpoint reduction potential (Emp) of Clostridium acidi-urici ferredoxin were determined using hydrogen gas and hydrogenase. The Emp of native ferredoxin at 24-25 degrees in 0.1 M Tris-chloride buffer, pH 7.0, is--0.434 V. In the pH range examined, the Emp becomes approximately 13 mv more negative per each pH unit increase. A plot of the log of ionic strength versus the apparent Emp of ferredoxin in 0.1 M Tris-chloride buffer, pH 7.5, Was linear over the range of 1.0 to 0.01 ionic strength with Emp values of--0.414 and--0.475 V, respectively, at these extremes. This effect is the same with sodium chloride, sodium bromide, or ammonium sulfate. Potassium phosphate buffer caused a similar change, but the absolute values of Emp differed from those obtained in the presence of the other salts. This effect of pH and ionic strength on Emp may be general for clostridial-type (Fe4S4)2-ferredoxins, since the apparent Emp of Clostridium pasteurianum ferredoxin is affected in a similar manner by these two variables. The Emp of this ferredoxin in 0.1 M Tris-chloride buffer pH 7.0, is--0.405 V. Since the NH2-terminal amino acid residue, Ala1, and Tyr2 of C. acidi urici ferredoxin are near an (Fe4S4)2-cluster in the protein, the apparent Emp of derivatives that contained amino acid replacements in these two positions were determined. Under similar conditions, the Emp of most of the 13 derivatives examined, including those of [Leu2]- and[3-NH2-Tyr30]ferredoxin, is approximately the same as that of native ferredoxin. However, the Emp of [His2]ferredoxin is approximately 15 mv more positive, whereas that of [Trp2]ferredoxin is 22 mv more negative than that of native C. acidi-urici ferredoxin. Variations in sodium chloride concentration and pH also affected the apparent Emp of the derivatives. It is suggested that the changes observed in the Emp of C. acidi-urici ferredoxin are caused by protein conformational changes. | [
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PMID:3504 | A deoxyribonucleic acid kinase from nuclei of rat liver. Purification and properties. | A DNA kinase has been partially purified from rat liver nuclei by a procedure which also yields DNA ligase. The kinase uses ATP to phosphorylate specifically the 5'-hydroxyl termini of oligodeoxynucleotides and of single- or double-stranded DNA, yielding 5'-phosphate termini and ADP. The kinase is inactive on RNA, or on oligodeoxynucleotides of chain length less than approximately 10 to 12 residues. The kinase requires a divalent cation (Mg2+, Mn2+, Co2+, Zn2+, Ni2+, or Ca2+) for activity and has an acidic pH optimum. It is inhibited by a variety of nucleotides as well as by very low levels of inorganic and organic sulfate compounds and sulfate analogues. The molecular weight of the kinase is estimated to be 8 times 10(4) from gel filtration. | A deoxyribonucleic acid kinase from nuclei of rat liver. Purification and properties. A DNA kinase has been partially purified from rat liver nuclei by a procedure which also yields DNA ligase. The kinase uses ATP to phosphorylate specifically the 5'-hydroxyl termini of oligodeoxynucleotides and of single- or double-stranded DNA, yielding 5'-phosphate termini and ADP. The kinase is inactive on RNA, or on oligodeoxynucleotides of chain length less than approximately 10 to 12 residues. The kinase requires a divalent cation (Mg2+, Mn2+, Co2+, Zn2+, Ni2+, or Ca2+) for activity and has an acidic pH optimum. It is inhibited by a variety of nucleotides as well as by very low levels of inorganic and organic sulfate compounds and sulfate analogues. The molecular weight of the kinase is estimated to be 8 times 10(4) from gel filtration. | [
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PMID:3505 | Hydrogen exchange at the amide group of reduced pyridine nucleotides and the inhibition of that reaction by dehydrogenases. | Stopped flow ultraviolet spectroscopy has been used to measure the rate of hydrogen exchange with solvent at the amide group of reduced nicotinamide nucleotide coenzymes. Several mechanisms for the exchange reaction are considered in the light of the kinetic data. Complex formation between the coenzyme and any of four dehydrogenases markedly slows the rate of hydrogen exchange. Hydrogen bond formation and/or hydrophobic interactions within these complexes are thought to be the reasons for the decreased rate of exchange. | Hydrogen exchange at the amide group of reduced pyridine nucleotides and the inhibition of that reaction by dehydrogenases. Stopped flow ultraviolet spectroscopy has been used to measure the rate of hydrogen exchange with solvent at the amide group of reduced nicotinamide nucleotide coenzymes. Several mechanisms for the exchange reaction are considered in the light of the kinetic data. Complex formation between the coenzyme and any of four dehydrogenases markedly slows the rate of hydrogen exchange. Hydrogen bond formation and/or hydrophobic interactions within these complexes are thought to be the reasons for the decreased rate of exchange. | [
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PMID:3508 | A Dacron wool packed-bed extracorporeal reactor: a kinetic study of immobilized Escherichia coli II L-asparaginase. | An extracorporeal reactor containing a packed bed of Dacron fibers has been developed. Escherichia coli II L-asparaginase was coupled to the Dacron using gamma-aminopropyltriethoxysilane and glutaraldehyde. The preparation had an activity of 37 IU per gram of Dacron (37 degrees C). The apparent Km was studied as a function of the flow rate. The data indicated that the apparent Km approached the Km of the native enzyme at flow rates of about 300 mg/min. In vivo use of L-asparaginase immobilized on the Dacron indicated effective lowering of plasmatic L-asparagine levels. | A Dacron wool packed-bed extracorporeal reactor: a kinetic study of immobilized Escherichia coli II L-asparaginase. An extracorporeal reactor containing a packed bed of Dacron fibers has been developed. Escherichia coli II L-asparaginase was coupled to the Dacron using gamma-aminopropyltriethoxysilane and glutaraldehyde. The preparation had an activity of 37 IU per gram of Dacron (37 degrees C). The apparent Km was studied as a function of the flow rate. The data indicated that the apparent Km approached the Km of the native enzyme at flow rates of about 300 mg/min. In vivo use of L-asparaginase immobilized on the Dacron indicated effective lowering of plasmatic L-asparagine levels. | [
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PMID:3509 | Isolation of intact megakaryocytes from guinea pig femoral marrow. Successful harvest made possible with inhibitions of platelet aggregation; enrichment achieved with a two-step separation technique. | Methods have been devised to harvest megakaryocytes from guinea pig femoral marrow and to isolate them in high yield. When marrow tissue was disaggregated the megakaryocytes underwent degenerative changes characterized by the loss of cytoplasmic granules and alterations in membrane topography, similar to the changes seen in aggregating platelets. These morphologic changes were interpreted to mean that megakaryocytes possessed functional attributes of platelets. The use of agents which inhibit platelt aggregation (0.38% sodium citrate. 10(-3) M adenosine, and 2 x 10(-3) M theophylline) in a medium free of bivalent cations prevented these changes. This solution resulted in both an excellent morphologic preservation and a significantly increased recovery of megakaryocytes from marrow tissue. A two-step purification of the intact megakaryocytes was carried out on the basis of their low density and large size, with equilibrium density gradient centrifugation followed by velocity sedimentation. This sequence gave approximately a 100-fold enrichment of megakaryocytes, significantly better than that achieved with either method alone. These techniques for harvesting and concentrating megakaryocytes make it possible for the first time to study megakaryocytes in vitro. | Isolation of intact megakaryocytes from guinea pig femoral marrow. Successful harvest made possible with inhibitions of platelet aggregation; enrichment achieved with a two-step separation technique. Methods have been devised to harvest megakaryocytes from guinea pig femoral marrow and to isolate them in high yield. When marrow tissue was disaggregated the megakaryocytes underwent degenerative changes characterized by the loss of cytoplasmic granules and alterations in membrane topography, similar to the changes seen in aggregating platelets. These morphologic changes were interpreted to mean that megakaryocytes possessed functional attributes of platelets. The use of agents which inhibit platelt aggregation (0.38% sodium citrate. 10(-3) M adenosine, and 2 x 10(-3) M theophylline) in a medium free of bivalent cations prevented these changes. This solution resulted in both an excellent morphologic preservation and a significantly increased recovery of megakaryocytes from marrow tissue. A two-step purification of the intact megakaryocytes was carried out on the basis of their low density and large size, with equilibrium density gradient centrifugation followed by velocity sedimentation. This sequence gave approximately a 100-fold enrichment of megakaryocytes, significantly better than that achieved with either method alone. These techniques for harvesting and concentrating megakaryocytes make it possible for the first time to study megakaryocytes in vitro. | [
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PMID:3510 | The polymerization of actin. III. Aggregates of nonfilamentous actin and its associated proteins: a storage form of actin. | When echinoderm sperm are treated with the detergent Triton X-100 at pH 6.4 in 10 mM phosphate buffer, the membranes are solubilized, but the actin which is located in the periacrosomal region remains as a phase-dense cup. These cups can be isolated free from the flagella and chromatin and can be solubilized by increasing the pH to 8.0 and by changing the ionic strength and type of buffer used. Since the actin does not exist in the "F" state in unreacted sperm, and since the actin remains as a unit that does not diffuse away, it must be present in the mature sperm in a bound or storage state. The actin is, in fact, associated with a pair of proteins whose mol wt are 250,000 and 230,000. When the isolated cups are digested with trypsin, these high molecular weight proteins are digested, thereby liberating the actin. The actin will polymerize if heavy meromyosin or subfragment 1 is added to a preparation of isolated cups. Evidence is presented that this pair of high molecular weight proteins is similar in molecular weight and properties to erythrocyte spectrin. Attempts at transforming the storage form of actin in the cup into filaments were only moderately successful. The best conditions for filament formation involve incubating the cup in ATP and divalent salts. Careful examination of these cups reveals that the actin polymerized preferentially on either end of oriented filaments that already exist in the cup, indicating that self-nucleation is inefficacious. I conclude that the actin can exist in the storage form by its association with spectrin-like molecules and that the actin in this state polymerizes preferentially onto existing filaments. | The polymerization of actin. III. Aggregates of nonfilamentous actin and its associated proteins: a storage form of actin. When echinoderm sperm are treated with the detergent Triton X-100 at pH 6.4 in 10 mM phosphate buffer, the membranes are solubilized, but the actin which is located in the periacrosomal region remains as a phase-dense cup. These cups can be isolated free from the flagella and chromatin and can be solubilized by increasing the pH to 8.0 and by changing the ionic strength and type of buffer used. Since the actin does not exist in the "F" state in unreacted sperm, and since the actin remains as a unit that does not diffuse away, it must be present in the mature sperm in a bound or storage state. The actin is, in fact, associated with a pair of proteins whose mol wt are 250,000 and 230,000. When the isolated cups are digested with trypsin, these high molecular weight proteins are digested, thereby liberating the actin. The actin will polymerize if heavy meromyosin or subfragment 1 is added to a preparation of isolated cups. Evidence is presented that this pair of high molecular weight proteins is similar in molecular weight and properties to erythrocyte spectrin. Attempts at transforming the storage form of actin in the cup into filaments were only moderately successful. The best conditions for filament formation involve incubating the cup in ATP and divalent salts. Careful examination of these cups reveals that the actin polymerized preferentially on either end of oriented filaments that already exist in the cup, indicating that self-nucleation is inefficacious. I conclude that the actin can exist in the storage form by its association with spectrin-like molecules and that the actin in this state polymerizes preferentially onto existing filaments. | [
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PMID:3511 | Degradation of abnormal proteins in HeLa cells. | The experiments show that abnoramal proteins are degraded faster than normal ones in HeLa cells. Among the fragmentary proteins made in the presence of puromycin, those with low molecular weight are least stable. Proteins made after incubation with 5-fluorouracil or in the presence of some amino acid analogues are also unstable. Breakdown of proteins made in the presence or absence of puromycin is nearly unaffected by cycloheximide and is independent of pH between 7 and 8. | Degradation of abnormal proteins in HeLa cells. The experiments show that abnoramal proteins are degraded faster than normal ones in HeLa cells. Among the fragmentary proteins made in the presence of puromycin, those with low molecular weight are least stable. Proteins made after incubation with 5-fluorouracil or in the presence of some amino acid analogues are also unstable. Breakdown of proteins made in the presence or absence of puromycin is nearly unaffected by cycloheximide and is independent of pH between 7 and 8. | [
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PMID:3512 | The effect of pH on incorporation of galactose by a normal human cell line and cell lines from patients with defective galactose metabolism. | Incorporation of radioactive galactose into TCA-insoluble material of galactosemic fibroblasts is more sensitive to low pH than is the incorporation by normal human fibroblasts. This study was undertaken to determine (1) whether there was any pH which could correct or counteract the galactosemic defect relative to galactose incorporation, and (2) whether the low pH effect was specific for galactose metabolism or whether general cellular metabolism in galactosemic cells was more sensitive to low pH than that in normal cells. The pH dependencies of incorporation of radioactive galactose and glucose into cellular macromolecules were investigated in galactosemic and normal cells. Normal cells have a biphasic curve with respect to galactose incorporation with peaks at pH 7.0 and 8.5. Galactosemic cells have only the high pH peak. The maximum incorporation by galactosemic cells was never more than about 30% that seen by normal cells under the conditions of these experiments. Thus manipulation of the pH alone cannot correct the galactosemic defect. The rate of incorporation of radioactive galactose was studied in normal, galactosemic and galactokinase deficient cells, at pH 7.2 and at pH 6.3. At pH 7.2, galactosemic cells incorporate galactose at a linear rate which is 30 to 40% that of normal cells while incorporation by kinase-deficient cells is between 5 and 10% of normal. At pH 6.3, the incorporation is also linear. However, galactosemic cells now exhibit the same rate as kinase-deficient cells in which the low level of incorporation is unaffected by pH. These results suggest that incorporation of galactose by galactosemic cells at low pH is not due to metabolic death of the cells, but may be due to the inhibition of some specific step or steps along a metabolic route of galactose metabolism other than the Leloir pathway. | The effect of pH on incorporation of galactose by a normal human cell line and cell lines from patients with defective galactose metabolism. Incorporation of radioactive galactose into TCA-insoluble material of galactosemic fibroblasts is more sensitive to low pH than is the incorporation by normal human fibroblasts. This study was undertaken to determine (1) whether there was any pH which could correct or counteract the galactosemic defect relative to galactose incorporation, and (2) whether the low pH effect was specific for galactose metabolism or whether general cellular metabolism in galactosemic cells was more sensitive to low pH than that in normal cells. The pH dependencies of incorporation of radioactive galactose and glucose into cellular macromolecules were investigated in galactosemic and normal cells. Normal cells have a biphasic curve with respect to galactose incorporation with peaks at pH 7.0 and 8.5. Galactosemic cells have only the high pH peak. The maximum incorporation by galactosemic cells was never more than about 30% that seen by normal cells under the conditions of these experiments. Thus manipulation of the pH alone cannot correct the galactosemic defect. The rate of incorporation of radioactive galactose was studied in normal, galactosemic and galactokinase deficient cells, at pH 7.2 and at pH 6.3. At pH 7.2, galactosemic cells incorporate galactose at a linear rate which is 30 to 40% that of normal cells while incorporation by kinase-deficient cells is between 5 and 10% of normal. At pH 6.3, the incorporation is also linear. However, galactosemic cells now exhibit the same rate as kinase-deficient cells in which the low level of incorporation is unaffected by pH. These results suggest that incorporation of galactose by galactosemic cells at low pH is not due to metabolic death of the cells, but may be due to the inhibition of some specific step or steps along a metabolic route of galactose metabolism other than the Leloir pathway. | [
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PMID:3514 | Chromatography of hemoglobins on CM-cellulose with bis-tris and sodium chloride developers. | CM-Cellulose as an ion-exchange medium with Bis-tris as buffer and a gradient of sodium chloride provides a versatile system for the chromatography of hemoglobins. Changes in pH, Bis-tris concentration, and slope of the sodium chloride grandient provide means for markedly altering chromatographic behavior for special separations. Examples are given of the application of the method to normal samples and to those with hemoglobinopathies. | Chromatography of hemoglobins on CM-cellulose with bis-tris and sodium chloride developers. CM-Cellulose as an ion-exchange medium with Bis-tris as buffer and a gradient of sodium chloride provides a versatile system for the chromatography of hemoglobins. Changes in pH, Bis-tris concentration, and slope of the sodium chloride grandient provide means for markedly altering chromatographic behavior for special separations. Examples are given of the application of the method to normal samples and to those with hemoglobinopathies. | [
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] |
PMID:3515 | The determination of phanquone in biological material by gas-liquid chromatography. | A gas-liquid chromatographic method for the quantitative determination of phanquone is described, based on the formation of a dimethoxine prior to its extraction from biological material. The sensitivity of the procedure is about 15 ng/ml in biological fluid. | The determination of phanquone in biological material by gas-liquid chromatography. A gas-liquid chromatographic method for the quantitative determination of phanquone is described, based on the formation of a dimethoxine prior to its extraction from biological material. The sensitivity of the procedure is about 15 ng/ml in biological fluid. | [
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PMID:3516 | Stability of fluorescent antibody conjugates stored under various conditions. | Two experiments were carried out to determine the stability of fluorescent antibody conjugates. In experiment 1, Francisella tularemia conjugates in the lyophilized state retained their original staining titer for 1,294 days when stored at 25, 4 to 5, and -20 C; at 37 C the conjugates were stable for at least 65 days. In the liquid state at pH 7.4 and 8.0 these conjugates were stable for 1,294 days at 4 to 5 and -20 C, whereas those stored at 25 C remained stable through days 473 and 160 of storage, respectively, after which the staining titer gradually dropped. In experiment 2 five previously lyophilized conjugates were rehydrated with three different diluents and stored at 4 to 5 C for up to 600 days at their working dilutions. All of these conjugates retained their original staining titer during the test period except an anti-human globulin conjugate rehydrated with phosphate-buffered saline. Recommendations are made for the long-term storage of fluorescent antibody conjugates. | Stability of fluorescent antibody conjugates stored under various conditions. Two experiments were carried out to determine the stability of fluorescent antibody conjugates. In experiment 1, Francisella tularemia conjugates in the lyophilized state retained their original staining titer for 1,294 days when stored at 25, 4 to 5, and -20 C; at 37 C the conjugates were stable for at least 65 days. In the liquid state at pH 7.4 and 8.0 these conjugates were stable for 1,294 days at 4 to 5 and -20 C, whereas those stored at 25 C remained stable through days 473 and 160 of storage, respectively, after which the staining titer gradually dropped. In experiment 2 five previously lyophilized conjugates were rehydrated with three different diluents and stored at 4 to 5 C for up to 600 days at their working dilutions. All of these conjugates retained their original staining titer during the test period except an anti-human globulin conjugate rehydrated with phosphate-buffered saline. Recommendations are made for the long-term storage of fluorescent antibody conjugates. | [
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PMID:3517 | Inactivation of viruses in serum with binary ethyleneimine. | The inactivation os six strains from three different groups of viruses with 0.001 M binary ethyleneimine at 37 C proceeded at the same rate in either bovine serum or cell culture medium. The inactivant did not impair the growth-promoting capacity of bovine serum used in cell culture, nor did it affect the antibody activity of guinea pig hyperimmune serum. | Inactivation of viruses in serum with binary ethyleneimine. The inactivation os six strains from three different groups of viruses with 0.001 M binary ethyleneimine at 37 C proceeded at the same rate in either bovine serum or cell culture medium. The inactivant did not impair the growth-promoting capacity of bovine serum used in cell culture, nor did it affect the antibody activity of guinea pig hyperimmune serum. | [
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PMID:3518 | Inorganic phosphate homeostasis. Renal adaptation to the dietary intake in intact and thyroparathyroidectomized rats. | The possibility of renal tubular adaptation to variations in dietary inorganic phosphate (Pi) was investigated in intact and thyroparathyroidectomized (TPTX) rats pair-fed diets containing low, normal, and high amounts of Pi for periods up to 10 days. Clearances were measured before and during active i.v. infusions with Pi in conscious animals. Thus tubular reabsorption of phosphate (TRPi) could be assessed over a wide range of plasma phosphate concentrations ([Pi]P1). It was found that the renal tubule could adapt its capacity to transport Pi according to the dietary Pi: TRPi was always higher, for a given [Pi]P1, in the animals fed low than in those fed higher Pi diets. This diet-induced modification also occurred in the absence of thyroparathyroid glands, in the presence of the same calcemia and urinary pH, and during marked extracellular volume expansion. A time-course study in rats TPTX both before and during the administration of the experimental diets showed that a difference in the tubular handling of Pi was detectable as early as 3 days after switching the animals from a normal to low- or high-Pi diets. These results indicate that factors other than parathyroid hormone are implicated in the tubular response to variations in the dietary intake of inorganic phosphate. | Inorganic phosphate homeostasis. Renal adaptation to the dietary intake in intact and thyroparathyroidectomized rats. The possibility of renal tubular adaptation to variations in dietary inorganic phosphate (Pi) was investigated in intact and thyroparathyroidectomized (TPTX) rats pair-fed diets containing low, normal, and high amounts of Pi for periods up to 10 days. Clearances were measured before and during active i.v. infusions with Pi in conscious animals. Thus tubular reabsorption of phosphate (TRPi) could be assessed over a wide range of plasma phosphate concentrations ([Pi]P1). It was found that the renal tubule could adapt its capacity to transport Pi according to the dietary Pi: TRPi was always higher, for a given [Pi]P1, in the animals fed low than in those fed higher Pi diets. This diet-induced modification also occurred in the absence of thyroparathyroid glands, in the presence of the same calcemia and urinary pH, and during marked extracellular volume expansion. A time-course study in rats TPTX both before and during the administration of the experimental diets showed that a difference in the tubular handling of Pi was detectable as early as 3 days after switching the animals from a normal to low- or high-Pi diets. These results indicate that factors other than parathyroid hormone are implicated in the tubular response to variations in the dietary intake of inorganic phosphate. | [
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PMID:3519 | Effect of beta adrenergic blockade on renin response to renal nerve stimulation. | The ability of d,l-propranolol to block renin secretion in response to various extrarenal stimuli, such as hemorrhage and hypoglycemia, has been interpreted to indicate the presence of an intrarenal beta receptor regulating renin release. However, two problems complicate this interpretation: (a) the stimuli have effects outside the kidney, and (b) d,l-propranolol has a local anesthetic, as well as a beta adrenergic blocking, action. In the present study, the effects of a purely intrarenal stimulus, in the form of renal nerve stimulation (RNS), on renin secretion was examined. The effects of d,l-propranolol (anesthetic and beta-blocking activity), l-propranolol (beta-blocking activity only), and d-propranolol (local anesthetic activity only) on the renin response to RNS were examined. In a control group of animals, two sequential RNS increased mean renin secretion from 401 to 1,255 U/min (P less than 0.25) and from 220 to 2,179 U/min (P less than 0.01). In a second group the first RNS increased renin secretion from 201 to 1,181 U/min (P less than 0.01), but after d,l-propranolol was given RNS did not significantly alter renin secretion (33 to 55 U/min). In a third group the initial RNS increased renin secretion from 378 to 1,802 U/min (P less than 0.025), but after l-propranolol was given RNS had no significant effect on renin secretion (84 to 51 U/min). A fourth group of dogs showed a rise in renin secretion from 205 to 880 U/min (P less than 0.001) in response to the first RNS, while the second RNS, given after an infusion of d-propranolol, caused a rise in renin secretion from 80 to 482 (P less than 0.005). The nature of the electrical stimulus was consistent in all groups and caused no detectable changes in renal or systemic hemodynamics or in urinary electrolyte excretion. The results, therefore, indicate that renin secretion can be stimulated through intrarenal beta receptors independent of changes in systemic or renal hemodynamics or in tubular sodium reabsorption. Hence the effect of beta stimulation on renin secretion would appear to result from a direct action on the renin-secreting cells of the juxtaglomerular apparatus. | Effect of beta adrenergic blockade on renin response to renal nerve stimulation. The ability of d,l-propranolol to block renin secretion in response to various extrarenal stimuli, such as hemorrhage and hypoglycemia, has been interpreted to indicate the presence of an intrarenal beta receptor regulating renin release. However, two problems complicate this interpretation: (a) the stimuli have effects outside the kidney, and (b) d,l-propranolol has a local anesthetic, as well as a beta adrenergic blocking, action. In the present study, the effects of a purely intrarenal stimulus, in the form of renal nerve stimulation (RNS), on renin secretion was examined. The effects of d,l-propranolol (anesthetic and beta-blocking activity), l-propranolol (beta-blocking activity only), and d-propranolol (local anesthetic activity only) on the renin response to RNS were examined. In a control group of animals, two sequential RNS increased mean renin secretion from 401 to 1,255 U/min (P less than 0.25) and from 220 to 2,179 U/min (P less than 0.01). In a second group the first RNS increased renin secretion from 201 to 1,181 U/min (P less than 0.01), but after d,l-propranolol was given RNS did not significantly alter renin secretion (33 to 55 U/min). In a third group the initial RNS increased renin secretion from 378 to 1,802 U/min (P less than 0.025), but after l-propranolol was given RNS had no significant effect on renin secretion (84 to 51 U/min). A fourth group of dogs showed a rise in renin secretion from 205 to 880 U/min (P less than 0.001) in response to the first RNS, while the second RNS, given after an infusion of d-propranolol, caused a rise in renin secretion from 80 to 482 (P less than 0.005). The nature of the electrical stimulus was consistent in all groups and caused no detectable changes in renal or systemic hemodynamics or in urinary electrolyte excretion. The results, therefore, indicate that renin secretion can be stimulated through intrarenal beta receptors independent of changes in systemic or renal hemodynamics or in tubular sodium reabsorption. Hence the effect of beta stimulation on renin secretion would appear to result from a direct action on the renin-secreting cells of the juxtaglomerular apparatus. | [
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PMID:3520 | Identification and characterization of a bile acid receptor in isolated liver surface membranes. | It is generally assumed that hepatic transport of bile acids is a carrier-mediated process. However, the basic mechanisms by which these organic anions are translocated across the liver cell surface membrane are not well understood. Since carrier-mediated transport involved binding of the transported molecule to specific receptor sites, we have investigated the possibility that bile acid receptors are present in liver surface membranes. Isolated liver surface membranes were incubated at 4 degrees C with [14C]cholic acid and [14C]taurocholic acid, and membrane-boudn bile acid was separated from free by a rapid ultrafiltration technique through glass-fiber filters. Specific bile acid binding is rapid and reversible and represents approximately 80% of the total bile acid bound to liver surface membranes. Taurocholic acid binding is independent of the medium pH, while cholic acid binding demonstrates an optimum at pH 6.0. Analysis of equilibrium data for both cholic and taurocholic acid binding indicates that specific binding is saturable and consistent with Michaelis-Menten kinetics, while nonspecific binding is nonsaturable. Apparent maximal binding capacity and dissociation constant values indicate a large capacity system of receptors that have an affinity for bile acids comparable to that of the hepatic transport mechanism. Scatchard analysis of the saturation kinetics as well as inhibition studies suggest that bile acids bind to a single and noninteracting class of anion that competes with bile acids for hepatic uptake, also inhibits cholic acid binding. In contrast, no inhibition was demonstrated with indocyanine green and probenecid. Specific bile acid binding is enriched and primarily located in liver surface membranes and found only in tissues involved in bile acid transport. Specific bile acid binding is independnet of Na+, Ca2+, and Mg2+ and does not require metabolic energy. In addition, thiol groups and disulfide are not required for activity at the binding site. However, specific bile acid binding is markedly decreased by low concentrations of proteolytic enzymes and is also decreased by the action of neuraminidase and phospholipases A and C. These results are consistent with the existence of a homogeneous bile acid receptor protein in liver surface membranes. The primary surface membrane location of this receptor, its binding properties, and its ligand specificity suggest that bile acid binding to this receptor may represent the initial interaction in bile acid transport across liver surface membranes. | Identification and characterization of a bile acid receptor in isolated liver surface membranes. It is generally assumed that hepatic transport of bile acids is a carrier-mediated process. However, the basic mechanisms by which these organic anions are translocated across the liver cell surface membrane are not well understood. Since carrier-mediated transport involved binding of the transported molecule to specific receptor sites, we have investigated the possibility that bile acid receptors are present in liver surface membranes. Isolated liver surface membranes were incubated at 4 degrees C with [14C]cholic acid and [14C]taurocholic acid, and membrane-boudn bile acid was separated from free by a rapid ultrafiltration technique through glass-fiber filters. Specific bile acid binding is rapid and reversible and represents approximately 80% of the total bile acid bound to liver surface membranes. Taurocholic acid binding is independent of the medium pH, while cholic acid binding demonstrates an optimum at pH 6.0. Analysis of equilibrium data for both cholic and taurocholic acid binding indicates that specific binding is saturable and consistent with Michaelis-Menten kinetics, while nonspecific binding is nonsaturable. Apparent maximal binding capacity and dissociation constant values indicate a large capacity system of receptors that have an affinity for bile acids comparable to that of the hepatic transport mechanism. Scatchard analysis of the saturation kinetics as well as inhibition studies suggest that bile acids bind to a single and noninteracting class of anion that competes with bile acids for hepatic uptake, also inhibits cholic acid binding. In contrast, no inhibition was demonstrated with indocyanine green and probenecid. Specific bile acid binding is enriched and primarily located in liver surface membranes and found only in tissues involved in bile acid transport. Specific bile acid binding is independnet of Na+, Ca2+, and Mg2+ and does not require metabolic energy. In addition, thiol groups and disulfide are not required for activity at the binding site. However, specific bile acid binding is markedly decreased by low concentrations of proteolytic enzymes and is also decreased by the action of neuraminidase and phospholipases A and C. These results are consistent with the existence of a homogeneous bile acid receptor protein in liver surface membranes. The primary surface membrane location of this receptor, its binding properties, and its ligand specificity suggest that bile acid binding to this receptor may represent the initial interaction in bile acid transport across liver surface membranes. | [
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PMID:3521 | A brief anxiety rating scale in evaluating anxiolytics. | The Wang Anxiety Rating Scale (WARS) was designed to evaluate degrees of anxiety in patients receiving anxiolytic medication. WARS contains 12 pertinent symptoms of anxiety: nervousness, restlessness, excitability, irritability, worrying, disturbed concentration, palpitation, insomnia, hostility, tremors, smoking, and excessive perspiration. Frequently encountered side effects of anxiolytic medications are excluded. The validity of the WARS was determined by correlation with the Hamilton Anxiety Rating Scale (HARS) in a single-blind study in which 20 chronically anxious patients consecutively received placebo (three days), 15 mg clorazepate dipotassium (two weeks), and 22.5 mg clorazepate dipotassium (two weeks). Both anxiety scales, a side effect scale, and a global assessment were completed at regular intervals (periods 0-6). Results show (1) highly significant correlation (P less than 0.001) between WARS and HARS for periods 1-6; (2) greater correlation between HARS and side effect scale than between WARS and side effect scale; (3) greater correlation between WARS and global assessment than between HARS and global assessment; correlated changes in scores for WARS, HARS, and global assessment demonstrate efficacy of active medication. | A brief anxiety rating scale in evaluating anxiolytics. The Wang Anxiety Rating Scale (WARS) was designed to evaluate degrees of anxiety in patients receiving anxiolytic medication. WARS contains 12 pertinent symptoms of anxiety: nervousness, restlessness, excitability, irritability, worrying, disturbed concentration, palpitation, insomnia, hostility, tremors, smoking, and excessive perspiration. Frequently encountered side effects of anxiolytic medications are excluded. The validity of the WARS was determined by correlation with the Hamilton Anxiety Rating Scale (HARS) in a single-blind study in which 20 chronically anxious patients consecutively received placebo (three days), 15 mg clorazepate dipotassium (two weeks), and 22.5 mg clorazepate dipotassium (two weeks). Both anxiety scales, a side effect scale, and a global assessment were completed at regular intervals (periods 0-6). Results show (1) highly significant correlation (P less than 0.001) between WARS and HARS for periods 1-6; (2) greater correlation between HARS and side effect scale than between WARS and side effect scale; (3) greater correlation between WARS and global assessment than between HARS and global assessment; correlated changes in scores for WARS, HARS, and global assessment demonstrate efficacy of active medication. | [
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PMID:3545 | The changing pattern of bacterial sepsis since the introduction of antibiotic therapy. | During the six-year period, 1968-1973, sepsis developed in 1 of every 80 patients admitted to the Presbyterian Hospital, New York. In 1 of 133 patients the sepsis was due to Gram-positive organisms, and in 1 of 188 patients to Gram-negative organisms. The mortality rate for Gram-positive cases was 4.4 percent, for Gram-negative cases 19.1 percent, and for urologic cases 15.3 percent (versus 56.25 percent in 1959-1964). Data are presented on the relative incidences of involved pathogens in 1740 cases of Gram-positive sepsis /78 deaths), and in 1236 cases of Gram-negative sepsis (205 deaths). The lowering of the sepsis mortality rate has been the result of preventative measures, early diagnosis, and vigorous treatment. Treatment includes the correction of acidosis and anoxia, early administration of bactericidal antibiotics, and restoration of the microcirculation by administration of corticosteroids, beta-adrenergic drugs, and appropriate diuretics. | The changing pattern of bacterial sepsis since the introduction of antibiotic therapy. During the six-year period, 1968-1973, sepsis developed in 1 of every 80 patients admitted to the Presbyterian Hospital, New York. In 1 of 133 patients the sepsis was due to Gram-positive organisms, and in 1 of 188 patients to Gram-negative organisms. The mortality rate for Gram-positive cases was 4.4 percent, for Gram-negative cases 19.1 percent, and for urologic cases 15.3 percent (versus 56.25 percent in 1959-1964). Data are presented on the relative incidences of involved pathogens in 1740 cases of Gram-positive sepsis /78 deaths), and in 1236 cases of Gram-negative sepsis (205 deaths). The lowering of the sepsis mortality rate has been the result of preventative measures, early diagnosis, and vigorous treatment. Treatment includes the correction of acidosis and anoxia, early administration of bactericidal antibiotics, and restoration of the microcirculation by administration of corticosteroids, beta-adrenergic drugs, and appropriate diuretics. | [
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PMID:3546 | [Has the pH meter replaced the Apgar score?]. | The introduction of measuring the pH appears to place it in competition with Apgar scoring because of its precision. A study of this which has been carried out has illustrated that there are two different criteria for assessing the state of the infant at birth. The usual agreement between pH values and Apgar scoring can be broken when clinical fetal distress has become established before metabolic equilibrium of the infant has become modified. In these circumstances the Apgar score will be bad while the pH will be good. | [Has the pH meter replaced the Apgar score?]. The introduction of measuring the pH appears to place it in competition with Apgar scoring because of its precision. A study of this which has been carried out has illustrated that there are two different criteria for assessing the state of the infant at birth. The usual agreement between pH values and Apgar scoring can be broken when clinical fetal distress has become established before metabolic equilibrium of the infant has become modified. In these circumstances the Apgar score will be bad while the pH will be good. | [
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PMID:3604 | Kinetics of the antibody response to type III pneumococcal polysaccharide. I. Evidence that suppressor cells function by inhibiting the recruitment and proliferation of antibody-producing cells. | For the first 126 hr after immunization of mice with an optimally immunogenic dose (0.5 mug) of Type III pneumococcal polysaccharide (SSS-III), splenic antibody-forming PFC and serum antibody levels were measured at 2- and 8-hr intervals, respectively. PFC were detected at 28 hr after immunization and then increased through 86 hr after immunization; thereafter, the number of PFC remained nearly constant for the next 20 to 24 hr, and then began to decline. In contrast, serum antibody was first detected 60 hr after immunization. The accumulation of serum antibody continued to lag behind the increase in numbers of PFC by 16 to 20 hr until maximal serum antibody levels were attained; curves fitted to the values obtained for each parameter were nearly parallel. | Kinetics of the antibody response to type III pneumococcal polysaccharide. I. Evidence that suppressor cells function by inhibiting the recruitment and proliferation of antibody-producing cells. For the first 126 hr after immunization of mice with an optimally immunogenic dose (0.5 mug) of Type III pneumococcal polysaccharide (SSS-III), splenic antibody-forming PFC and serum antibody levels were measured at 2- and 8-hr intervals, respectively. PFC were detected at 28 hr after immunization and then increased through 86 hr after immunization; thereafter, the number of PFC remained nearly constant for the next 20 to 24 hr, and then began to decline. In contrast, serum antibody was first detected 60 hr after immunization. The accumulation of serum antibody continued to lag behind the increase in numbers of PFC by 16 to 20 hr until maximal serum antibody levels were attained; curves fitted to the values obtained for each parameter were nearly parallel. | [
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