Contribution of a net transmembrane HCO3- flux to intracellular acid-base regulation. Experiments were performed to determine the relative effects of a net extracellular-to-intracellular HCO3- flux and of elevated carbon dioxide tension (PCO2) on cellular acid-base regulation. Isolated rabbit hearts were perfused by recirculating a small volume of Ringer solution in which the PCO2 and the HCO3- concentration could be independently altered. Net HCO3- flux was assessed by the disappearance of HCO3- from perfusate. Between 40 and 100 Torr PCO2, a HCO3- flux into the cell occurs only when perfusate HCO3- concentration is increased. Therefore, by selective manipulation of perfusate HCO3- and PCO2 it is possible to induce hypercapnia with or without an accompanying HCO3- flux. When perfusate HCO3- concentration was increased from 20 to 36 mM, cellular HCO3- concentration increased from 22.5 +/- 0.8 to 26.1 +/- 1.0 mM at 40 Torr PCO2 and from 27.8 +/- 0.7 to 34.1 +/- 1.4 mM at 98 Torr PCO2. These increases can be accounted for by the amount of HCO3- that disappeared from the perfusate. The results suggest that most of the initial cell CO2 buffering is provided by the net HCO3- flux in addition to the passive physicochemical buffering.


Effect of pH on cardiorespiratory and metabolic responses to exercise. Five male subjects performed exercise at 33, 66, and 95% of their maximum power output on three occasions in random order. Each study was preceded by a 3-h period in which capsules were taken by mouth, containing either CaCO3 (control, NH4Cl (acidosis), or NaHCO3 (alkalosis) in a dose of 0.3 g/kg body wt; preexercise blood pH was 7.38 +/- 0.015, 7.21 +/- 0.033, and 7.43 +/- 0.029, respectively. Exercise was continuous and maintained for 20 min at the two lower power outputs and for as long as possible at the highest. Compared with control (270 +/- 13 s), endurance time at the highest power output was reduced in acidosis (160 +/- 22 s) and increased in alkalosis (438 +/- 120 s). No differences were observed for central cardiovascular changes in exercise (cardiac output, frequency, or stroke volume). The respiratory changes expected from changes in blood pH were observed, with a higher alveolar ventilation in acidosis. At all power outputs arterialized venous lactate was lowest in acidosis and highest in alkalosis. Plasma glycerol and free fatty acids were lowest in acidosis. Changes in blood [HCO3-] and pH were shown to have major effects on metabolism in exercise which presumably were responsible for impaired endurance.


Catecholamine-synthesizing enzymes in adrenals of seasonally acclimatized voles. Tyrosine hydroxylase (TH) and phenylethanolamine-N-methyltransferase (PNMT) activities were assayed in adrenal glands of the following groups of the Alaskan red-backed vole (Clethrionomys rutilus dawsoni): 1) laboratory reared at 20 degrees C and 2) exposed to 5 degrees C for 1, 3, 7, and 28 days; 3) wild, summer acclimatized; 4) wild, fall acclimatized; and 5) wild, winter acclimatized. TH activity in laboratory-acclimated voles exposed to 5 degrees C was increased by 2 times after 3 days and remained elevated after 28 days. PNMT activity in these same voles was increased after 7 days and also remained elevated after 28 days of cold exposure. In wild-acclimatized voles TH activity and PNMT activity in summer were equivalent to levels in 28-day cold-acclimated laboratory voles. In fall, TH activity was increased to 2.5 times the summer value. It decreased by midwinter, but remained elevated above the summer level. In contrast, PNMT activity appeared unchanged from summer through fall and winter. Pregnant summer voles had markedly increased TH activity. Adrenal norepinephrine and epinephrine did not change significantly with cold acclimation or seasonal acclimatization. Thus, acclimatization of wild voles to fall and winter conditions involved aquisition of a greater capacity to synthesize adrenal catecholamines than that produced by exposing laboratory-reared voles to an extended period of cold.


A32390A, a new biologically active metabolite. I. Discovery and fermentation studies. A32390A is an isonitrile-containing derivative of diacyl D-mannitol. The compound is produced in fermentation as the major component of a metabolic complex known as A32390. A32390A inhibits dopamine-beta-hydroxylase reduces heart and adrenal norepinephrine levels, lowers blood pressure in hypertensive rats, and possesses antibiotic activity vs. Gram-positive bacteria and fungi, including Candida albicans. A32390 is produced in submerged culture by a mold, a species of Pyrenochaeta, NRRL-5786. Glucose and sucrose are among the best carbon sources for the biosynthesis of A32390. Mannitol, although a substituent of the A32390A molecule, supports little or no biosynthesis of the compound when employed as the major carbon source for the fermentation. The addition of crotonic acid derivatives. ethanol, or L-histidine to the fermentation medium enhances the level of A32390 produced.


Regulation of gamma-aminobutyric acid degradation in Escherichia coli by nitrogen metabolism enzymes. The possible role of glutamate dehydrogenase, glutamate synthase, and glutamine synthetase in the regulation of enzyme formation in the gamma-aminobutyric acid (GABA) catabolic pathway of Escherichia coli K-12 was investigated. Evidence is presented indicating that glutamine synthetase acts as a positive regulator in the E. coli GABA control system. Mutations impairing glutamate synthase activity prevent the depression of the enzymes of the GABA pathway in ammonia-limited glucose media. However, mutations resulting in constitutive synthesis of glutamine synthetase (GlnC) restore the ability of the glutamate synthase-less mutants to grow in glucose-GABA media and result in depressed synthesis of the GABA enzymes. It is suggested that the loss of glutamate synthesis activity affects the GABA control system indirectly by lowering glutamine synthetase levels.


Mitochondrial adenosine triphosphatase of wild-type and poky Neurospora crassa. We have compared the adenosine triphosphatase (ATPase) activity of mitochondria prepared from wild-type Neurospora crassa and from poky, a maternally inherited mutant known to possess defective mitochondrial ribosomes and reduced amounts of cytochromes aa3 and b. poky contains two distinct forms of mitochondrial ATPase. The first is normal in its Km for ATP, specificity for nucleotides and divalent cations, pH optimum, cold stability, and sensitivity to inhibitors (oligomycin, N,N-dicyclohexyl carbodiimide, and adenylyl imidodiphosphate). The fact that membrane-bound, cold-stable, oligomycin-sensitive ATPase activity is present in poky (with an activity of 1.93 +/- 0.03 mumol/min-mg of protein compared with 1.33 +/- 0.07 mumol/min-mg of protein in the wild-type strain) and also in chloramphenicol-grown wild-type cells suggests that products of mitochondrial protein synthesis play only a limited role in the attachment of the mitochondrial ATPase to the membrane in Neurospora. poky also contains a second form of mitochondrial ATPase, which has an activity of 1.5 +/- 0.2 mumol/min-mg of protein, is oligomycin sensitive but cold labile, and presumably is attached less firmly to the mitochondrial membrane. The two forms, added together, represent a substantial overproduction of mitochondrial ATPase by poky.


Catabolic N2-acetylornithine 5-aminotransferase of Klebsiella aerogenes: control of synthesis by induction, catabolite repression, and activation by glutamine synthetase. Klebsiella aerogenes formed two N2-acetylornithine 5-aminotransferases (ACOAT) which were separable by diethylaminoethyl-cellulose chromatography. One ACOAT was repressed when the cells grew on arginine-containing medium, indicating its function in arginine biosynthesis. The second ACOAT was induced when arginine or ornithine was present in the medium as the sole source of carbon or nitrogen, suggesting its function in the catabolism of these compounds. The induced enzyme was purified almost to homogeneity. Its molecular weight is 59,000; it is a pyridoxal 5-phosphate-dependent enzyme and exhibits activity with N2-acetylornithine (Km = 1.1 mM) as well as with ornithine (Km = 5.4 mM). ACOAT did not catalyze the transamination of putrescine or 4-aminobutyrate. The best amino acceptor was 2-ketoglutarate (Km = 0.7 mM). ACOAT formation was subject to catabolite repression exerted by glucose when ammonia was present in excess. When the cells were deprived of nitrogen, ACOAT escaped from catabolite repression. This activation was mediated by glutamine synthetase as shown by the fact that mutants affected in the regulation or synthesis of glutamine synthetase were also affected in the control of ACOAT formation.


Effect of glucose starvation on the nicotinamide adenine dinucleotide phosphate-dependent glutamate dehydrogenase of yeast. Yeast cells growing on mineral medium plus ammonia and glucose contained high levels of nicotinamide adenine dinucleotide phosphate-dependent glutamate dehydrogenase activity, as measured in crude extracts. After suspension of cells in fresh medium lacking glucose, there was a loss of the glutamate dehydrogenase activity. Loss of activity was inhibited by 2,4-dinitrophenol, sodium azide, iodoacetic acid, and cycloheximide. The enzyme activity was restored when glucose was added back to the medium, and this recovery was fully prevented in the presence of cycloheximide.


Evidence for the degradation of nicotinamide adenine dinucleotide phosphate-dependent glutamate dehydrogenase of Candida utilis during rapid enzyme inactivation. The nicotinamide adenine dinucleotide phosphate-dependent glutamate dehydrogenase (NADP-GDH) from the food yeast Candida utilis was found to be rapidly inactivated when cultures were starved of a carbon source. The addition of glutamate or alanine to the starvation medium stimulated the rate of inactivation. Loss of enzyme activity was irreversible since the reappearance of enzyme activity, following the addition of glucose to carbon-starved cultures, was blocked by cycloheximide. A specific rabbit antibody was prepared against the NADP-GDH from C. utilis and used to quantitate the enzyme during inactivation promoted by carbon starvation. The amount of precipitable antigenic material paralleled the rapid decrease of enzyme activity observed after transition of cells from NH(4) (+)-glucose to glutamate medium. No additional small-molecular-weight protein was precipitated by the antibody as a result of the inactivation, suggesting that the enzyme is considerably altered during the primary steps of the inactivation process. Analysis by immunoprecipitation of the reappearance of enzyme activity after enzyme inactivation showed that increase of NADP-GDH activity was almost totally due to de novo synthesis, ruling out the possibility that enzyme activity modulation is achieved by reversible covalent modification. Enzyme degradation was also measured during steady-state growth and other changes in nitrogen and carbon status of the culture media. In all instances so far estimated, the enzyme was found to be very stable and not normally subject to high rates of degradation. Therefore, the possibility that inactivation was caused by a change in the ratio of synthesis to degradation can be excluded.


Effect of external environmental factors on the morphology of Spiroplasma citri. Spiroplasma citri was examined by electron microscopy for morphological changes when maintained under a variety of conditions. PPLO serum fraction maintained spiral and helical morphology of S. citri at pH values of 8.0, 7.5, and 7.0, but only partially at pH 6.0 and 5.0. The absence of PPLO serum fraction resulted in round, deteriorated cells at all pH values tested. Bovine serum albumin (BSA), Phytone, soluble starch, potato starch, spermine, lipid-extracted PPLO serum fraction, and lipid-extracted BSA could substitute for PPLO serum fraction in maintaining spiral and helical morphology at pH 7.5. At pH 5.0, only BSA, lipid-extracted BSA, and lipid-extracted PPLO serum fraction were effective. Only BSA supported growth of S. citri for more than two transfers, whereas all other substitutes could not support growth longer than two transfers.


The ATPase reaction in the steady state and in the initial burst catalyzed by chicken gizzard myosin in 0.6 M KCl1. On studying the steady-state activity in 0.6 M KCl, it was found that Mg-ATPase of chicken gizzard myosin was identical with that of rabbit skeletal myosin in the pH-activity profile, Michaelis-Menten constant, and maximum velocity. As regards the "initial burst" of ATP splitting in the presence of Mg (0.6 M KCl), it was found that gizzard and skeletal myosins were identical both in the size of the initial burst and in the velocity-ATP concentration relationship. The only difference we observed was that the Ca- and EDTA-ATPase activities of gizzard myosin were, as reported by other investigators, approximately one-half to one-third of those of skeletal myosin, although the pH-activity profiles for the ATPase of gizzard myosin was essentially the same as that of skeletal myosin.


Modification of rabbit muscle phosphorylase b by a water-soluble carbodiimide. Glycogen phosphorylase b from rabbit muscle was rapidly inactivated by incubation with 1-cyclohexyl-3-(2-morpholinyl-(4)-ethyl)carbodiimide metho-p-toluenesulfonate (CMC) at pH 5.1. The inactivation was pH-dependent and was not restored by treatment with hydroxylamine. The addition of glycine ethyl ester or N-(2,4-dinitrophenyl)-ethylenediamine (DNP-EDA) markedly increased the rate of inactivation. Of the various amino analogs of glucose tested, only glucosyl amine accelerated the inactivation, although they are all bound to the glucose 1-phosphate site of the enzyme. In the absence of amines, incorporation of about 3 mol of [metho-14C]CMC per protein monomer was observed on complete inactivation. In the presence of DNP-EDA, however, only 2 mol of [metho-14C]CMC and 1 mol of DNP-EDA were incorporated before the activity was completely lost. The treatment of phosphorylase b with CMC did not change the Km values of the enzyme for glucose 1-phosphate and AMP, in spite of the 56% inactivation. It is suggested that, in the phosphorylase-catalyzed reaction, an essential carboxyl group of the enzyme plays a role in the protonation of the glucosidic oxygen of glucose 1-phosphate.


Kinetic studies on redox reactions of hemoproteins. II. Reduction of thermoresistant cytochrome c-552 and horse heart cytochrome c by ascorbic acid. The reductions of thermoresistant cytochrome c-552 and horse heart cytochrome c by ascorbic acid were studied by the stopped-flow method between pH 4 and 10. The results were as follows (1) The reduction of horse heart cytochrome c showed two relaxation decays above pH 8.5, one of which was pseudo-first order, as was the case below pH 8, while the other was nearly concentration-independent. These results were consistent with those reported by Greenwood and Palmer (J. Biol. Chem. (1965) 240, 3660-3663). (2) For the reduction of cytochrome c-552, only a single relaxational decay that obeyed pseudo-first order kinetics was observed. (3) It seems most reasonable to assume that the concentration-independent relaxation process can be attributed to the isomerization reaction accompanying ligand exchange, since it is known that only horse heart cytochrome c exhibits ligand exchange, involving a residue with pK 9.3.


Genetic expression of microsomal electron transport in mice. Different patterns of dependence of constitutive cytochrome P-450 reductase activity of pyridine nucleotide concentrations. Cytochrome P-450 reductase and aryl hydrocarbon hydroxylase activities were investigated in hepatic microsomes from untreated C57BL/6J, DBA/2J, B6D2F1, and (B6D2) D2 mice. The dependence of the rate of P-450 reduction on the concentration of added pyridine nucleotide (NADPH or NADH) was biphasic in DBA/2J microsomes but monophasic in C57BL/6J microsomes. Analogous strain-specific patterns were observed when the dependence of the rate of benzpyrene hydroxylation on NADPH concentration was examined. In crosses between the two inbred strains and between B6D2F1 mice and DBA/2J mice, the biphasic pattern for both the reductase and the hydroxylase activities was found to co-segregate with the recessive allele for aromatic hydrocarbon responsiveness. These results might reflect an architectural difference between the microsomal electron transport systems of responsive and nonresponsive mice.


Oxygen equilibria of hybrid-heme hemoglobins containing proto- and mesoheme groups. On the nonequivalence of alpha and beta chains. Hybrid-heme hemoglobins, alpha(meso)2beta(proto)2 and alpha(proto)2beta(meso)2, were prepared, and the O2 equilibria of their alpha and beta chains were measured separately at the isosbestic points of the partner chains at different pH values and in the presence and absence of inositol hexaphosphate. The Adair equation was extended to distinguish between the O2 saturations of the alpha and beta chains, and the seven equilibrium parameters were obtained by curve fitting to those equations. The results showed that the beta chains have an affinity slightly higher than the alpha chains in the binding of the first O2 molecule. For the second O2 molecule, the molecular species that has been oxygenated on the alpha chain has a higher affinity than that carrying O2 on the beta chain. The slopes of the Hill plots were higher for the alpha chain. The O2 saturation curves for the alpha and beta chains were calculated from the parameters averaged for the hybrids alpha(meso)2beta(proto)2 and alpha(proto)2beta(meso)2 in order to cancel the effects of the heme replacement. The curves showed that the difference in O2 saturation between the two kinds of chains depends on the conditions and on the degree of O2 saturation. It was concluded that the functional difference between the chains is small enough so that it is not required to modify the models already accepted for the cooperativity of hemoglobin.


Dibutyryl cyclic AMP increases the amount of functional messenger RNA coding for tyrosine aminotransferase in rat liver. The administration of N6,O2-dibutyryl cyclic AMP and theophylline to adrenalectomized rats results in an increase in the amount of functional mRNA coding for tyrosine aminotransferase that can be isolated from liver. The induction of this specific mRNA, as quantitated in a mRNA-dependent reticulocyte lysate system, and using poly(A)+ mRNA extracted from total tissue and polysomes, is very rapid. Within an hour after the intraperitoneal injection of the cyclic AMP derivative there is a 5- to 7-fold elevation of functional mRNA coding for tyrosine aminotransferase (mRNATAT), and by 3 h this has returned to basal levels. In contrast, the 4- to 5-fold induction of tyrosine aminotransferase catalytic activity is maximal at 2 h and is still significantly greater than the basal level at 5 h. In the basal state, tyrosine aminotransferase mRNA codes for 0.019 +/- 0.003% of the protein synthesized in the in vitro system, whereas after cyclic nucleotide treatment this value 0.115 +/- 0.015%, hence the increase in mRNATAT activity is relatively specific. Cordycepin, at a concentration which prevents the accumulation in cytoplasm of poly(A)+ mRNA, completely blocks the increase in both the catalytic and mRNA activity of this enzyme. The marked increase in functional mRNA, the requirement for continued synthesis of poly(A)+ RNA, and the rapid induction and deinduction suggest that the cyclic nucleotide is enhancing specific mRNA synthesis and/or, processing, however an effect on mRNA degradation cannot be excluded.


Purification and subunit structure of rat mammary gland acetyl coenzyme A carboxylase. 1. Acetyl coenzyme A carboxylase from lactating rat mammary gland has been purified to apparent homogeneity. 2. The purified enzyme has the following characteristics: (a) its specific activity approaches 15 units/mg of protein, (b) the sedimentation constants of the protomeric and polymeric forms of the enzyme are 12 to 13 S and greater than or equal to 40 S, respectively, (c) the polymeric form of the enzyme shows filamentous structures in the electron microscope, and (d) the polypeptide(s) arising from its dissociation reveals a single major component of Mr = 240,000 to 260,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 3. The enzyme contains 1 mol of biotin and approximately 6 mol of phosphate/240,000 g of protein.


Prune-bell syndrome: a report of twelve cases and review of the literature. Prune-belly syndrome is a rare congenital disease involving a deficiency of abdominal muscles, genitourinary abnormalities, and cryptorchidism. About half of the patients with the disease have associated musculoskeletal abnormalities, including club foot and dislocation of the hip. We saw twelve patients with this disease, four of whom required extensive orthopaedic treatment.


Survival of Streptococcus pneumoniae in sputum from patients with pneumonia. The isolation rate of Streptococcus pneumoniae in sputum cultures from patients with pneumococcal pneumonia is low. An investigation was made to determine whether this low yield might be due to loss of pneumocci and/or overgrowth by pharyngeal flora before the specimen is plated. Pneumococcal survival times and pharyngeal overgrowth at 4 degrees C and at room temperature were determined in sputum obtained from 42 patients with pneumococcal pneumonia. It was found that pneumococci survived for long periods in sputum--2.2 +/- 1.4 days at room temperature and 9.5 +/- 3.6 days at 4 degrees C. Overgrowth by pharyngeal flora occurred in only 6 of 42 specimens kept at 4 degrees C and 31 of 42 specimens kept at room temperature. The low yield of S. pneumoniae in sputum from patients with pneumococcal pneumonia is not explained by decreased viability of the organism.


Quantitative evaluation of three commercial blood culture media for growth of anaerobic organisms. The ability of three different commercial blood culture media--brain heart infusion broth (Pfizer), thiol broth (Difco), and PRS-peptone broth (Becton, Dickinson & Co.)--to support the growth of five different anaerobes is described. Inocula of 100 and 1,000 colony-forming units per ml were used to evaluate potential differences in survival, lag time, growth rate, and doubling times of each anaerobe in each medium. In addition, each medium was evaluated for its ability to neutralize the antibacterial effects of whole blood. The results of this study indicate that the PRS-peptone broth is superior to brain heart infusion and thiol broths. Shorter lag times and accelerated generation times and growth rates were noted for more different anaerobes in the PRS-peptone broth. Neither the size of inoculum nor the addition of normal whole blood appeared to alter the survival or growth characteristics of the anaerobes in any medium. However, the addition of normal whole blood did extend the lag time of each anaerobe by approximately 1 to 2 h in each medium.


A double-blind controlled study of pipothiazine palmitate in the maintenance treatment of schizophrenic outpatients. In a double-blind controlled study lasting 9 months, pipothiazine palmitate, a long-acting neuroleptic that can be administered intramuscularly once a month, was compared with fluphenazine enanthate in the maintenance treatment of 32 schizophrenic outpatients. Before the trial commenced, all patients were being treated with fluphenazine enanthate. The results indicate that pipothiazine palmitate was not as potent a neuroleptic as fluphenazine enanthate. Pipothiazine appears to resemble fluphenazine enanthate in its capacity to induce parkinsonism and tardive dyskinesia.


Effects of antianxiety drug and personality on stress-inducing psychomotor performance test. The present study was carried out to clarify the effects of an antianxiety drug and of personality characteristics on a psychomotor performance test. Forty-eight healthy women college students were chosen from 64 volunteers as having either high or low levels of trait anxiety, neuroticism, or extroversion. Subjects with high trait anxiety and/or neuroticism tended to show a decrease in both speed and accuracy of the mirror drawing test (MDT) in the initial nondrug trials. Bromazepam, 5 mg, a benzodiazepine derivative, decreased this decrement in highly anxious subjects but worsened the speed in less anxious subjects. The personality traits of subjects, as well as the degree to which a performance test will induce stress, must be considered when evaluating the effects of antianxiety drugs on the performance of normal volunteers. The clinical anxiety-reducing efficacy of drugs may be predicted by using the MDT in subjects with high levels of anxiety and/or neuroticism.


The effect of Skylab on the chemical composition of saliva. The levels of specific proteins and electrolytes in stimulated whole saliva were monitored in Skylab crew members before and after each mission. With few exceptions, mission-associated compositional changes in saliva were relatively minimal. There were no changes in(formula see text), Cl-, albumin, or IgG concentrations. There were slight decreases in total protein coinciding with moderate saliva flow rate increases immediately before and after each flight. Other changes included diminutions in Na+ and lysozyme, and elevations in Mg++ and IgA. The IgA increase was the most pronounced mission-associated change observed.


Tetramethyl benzidine for horseradish peroxidase neurohistochemistry: a non-carcinogenic blue reaction product with superior sensitivity for visualizing neural afferents and efferents. Tetramethyl benzidine (TMB) is a presumptively non-carcinogenic chromogen which yields a blue reaction-product at sites of horseradish peroxidase activity. Sixty-six distinct procedures were performed in rats and monkeys in order to determine the optimal incubation parameters for TMB. As a result, a procedure is recommended whose sensitivity greatly surpasses that of a previously described benzidine dihydrochloride method. Indeed, the sensitivity of this new method in demonstrating retrograde transport is markedly superior to that of the previously described benzidine dihydrochloride method. Furthermore, as a consequence of this enhanced sensitivity, many efferent connections of the injection site are also visualized. The injection site demonstrated by this TMB procedure is significantly larger than the one demonstrated when benzidine dihydrochloride or diaminobenzidine is used as a chromogen. Finally, this TMB procedure has been compared to two other TMB procedures and found to provide superior morphology and sensitivity.


The pH of the hamster sperm acrosome.

The pH of the hamster sperm acrosome. The pH of the hamster sperm acrosome was estimated by a method based on the distribution of monoamines between membrane enclosed volumes maintaining pH gradients. A fluorescent amine, 9-aminoacridine, was used to permit both microscopic and fluorometric measurements of amine distribution. Cauda epididymal hamster sperm incubated with 9-aminoacridine accumulated the amine in the acrosomal volume. In the presence of NH4Cl or the ionophore Nigericin (compounds which discharge pH gradients) 9-aminoacridine fluorescence disappeared from the acrosome. Amine distribution between the acrosome and external volume was estimated by fluorometric measurement of sperm filtrates in the presence and absence of NH4Cl and Nigericin. These values, together with an estimated acrosomal volume of 0.4mu3 were used to calculate an acrosomal pH of less than 5. In addition, an acrosomal pH of 5 or less was obtained with 14C-methylamine. We suggest that such an acidic acrosomal pH of 5 or less could serve to inhibit the activation or autoactivation of the acrosomal zymogen proacrosin to acrosin, a trypsin-like enzyme involved in fertilization.


Effects of calcium on the absorption and retention of lead. An inverse relationship between lead retention and dietary calcium content has been known to exist for many years, but the reasons for this association remained unknown. In rats, the manipulation of dietary calcium had no significant effect upon the absorption of lead, but calcium-deprived animals had decreased excretion and thus increased body retention of lead. Intraluminal calcium decreased the absorption of test doses of lead from the small intestine in a dose-related manner. We postulated that this occurred because the two metals competed for similar binding sites on intestinal mucosal proteins which were important in the absorptive process. In vivo, lead bound to two heat-stable intestinal mucosal fractions which have been shown to bind calcium. Although more lead bound to the higher molecular weight fraction and more calcium bound to the lower molecular weight vitamin D-induced CaBP, substantial amounts of lead and calcium were found in both fractions. Further, the addition of calcium to test doses of lead markedly diminished the amount of lead bound by both fractions. Shared binding sites on absorptive proteins would explain why dietary calcium decreases lead absorption.


[Enzyme-immunological tests: determination of the activity of peroxidase with the aid of the "Trinder reagent" (author's transl)]. Most of the currently used enzyme-immunological tests employ horse radish peroxidase as a marker enzyme. A new method is described for the determination of extremely small quantities of peroxidase. This largely prevents the inactivation of the peroxidase by H2O2 and thereby permits a much longer incubation time. In the presence of 25 mmol/l phenol, 2 mmol/l 4-amino-antipyrin and 0.8 mmol/l H2O2, peroxidase catalyses the formation of a red quinonimine, whose increase in adsorption is directly proportional to the enzyme concentration.


Thin layer chromatographic screening for methaqualone, phenothiazines, opiates and benzodiazepines. A method is described which permits the simultaneous detection of methaqualone, phenothiazines, opiates and benzodiazepines in urine. Its diagnostic application is discussed. After cleavage of conjugates with hydrochloric acid, the substances are extracted and identified by thin-layer chromatography. In most cases analysis can be carried out using 2 solvent systems, phenothiazines, methaqualone and opiates being visualised using a three stage spray sequence. Since phenothiazines can interfere with the detection of methaqualone, a specific eluant is used to ensure reliable detection of the latter. Methaqualone can be positively identified by its characteristic metabolite pattern, whereas phenothiazines can only be detected as a group.


Glutamate dehydrogenase from coelenterates is NADP specific. Glutamate dehydrogenases detected in tissue extracts of a broad sample of coelenterate species all require NADP(H) as a co-substrate, rather than being capable of using either NAD(H) or NADP(H). In this respect, the coelenterate phyla appear to be unique in the animal kingdom.


Purification and properties of beta-hydroxybutyrate dehydrogenase from Mycobacterium phlei ATCC354. beta-Hydroxybutyrate dehydrogenase (EC 1.1.1.30) was purified 145-fold from Mycobacterium phlei ATCC354 by ammonium sulphate fractionation and DEAE-cellulose chromatography. The pH optima for oxidation and reduction reactions were 8.4 and 6.8 respectively. The purified enzyme was specific for NAD, NADH, acetoacetate and D(-)-beta-hydroxybutyrate. Km values for DL-beta-hydroxybutyrate and NAD were 7.4 mM and 0.66 mM respectively. The enzyme was inactivated by mercurial thiol inhibitors and by heat, but could be protected by NADH, Ca2+ and partially by Mn2+. The enzyme did not require metal ions and was insensitive to EDTA, glutathione, dithiothreitol, beta-mercaptoethanol and cysteine.


Virulence and resistance to superoxide, low pH and hydrogen peroxide among strains of Mycobacterium tuberculosis. Six strains of Mycobacterium tuberculosis of different virulence in guinea-pigs were compared with regard to their resistance to low pH, to hydrogen peroxide (H2O2) at different pH values and to superoxide (O2-). Low virulence was associated with susceptibility to H2O2 in native and isoniazid-resistant strains but not in laboratory-attenuated strain H37Ra. H2O2 resistance was only partly related to catalase content. Low virulence was not associated with susceptibility to an acid environment but the tuberculocidal effect of H2O2 was significantly increased at low pH. The strains were uniformly resistant to O2- and contained similar amounts of superoxide dismutase. The implications of these observations are discussed in the context of mechanisms of host defence in tuberculosis.


Exopolysaccharide production by Pseudomonas NCIB11264 grown in continuous culture. Exopolysaccharide formation by Pseudomonas NCIB11264 in a single-stage continuous culture was maximal under nitrogen limitation with excess carbohydrate substrate at 30 +/- 1 degrees C and pH 7.0 +/- 0.1. Polysaccharide production was not enhanced by phosphate limitation but was dependent on the dilution rate. Steady states were maintained for up to 500 h without deterioration of the culture or the development of mutant strains. The efficiency of conversion of the glucose substrate utilized into exopolysaccharide by the chemostat cultures was as high as 73%.


Detection of a complex intermediate in the oxidation of ascorbic acid by the copper (II) ion. A complex intermediate of the copper (II) ion with the ascorbate anion was detected by using a rapid scanning spectrophotometer with a stopped-flow apparatus. The intermediate was confirmed to be a copper (II) complex ion by electron paramagnetic resonance measurements. The dependence of the absorbance of the complex on the concentrations of reactants and on the pH of the reaction solutions indicates that the complex is Cu(II)Ha+ where Ha is the ascorbate anion ligand. The absorption maximum, the molar absorption coefficient, and the formation constant of the complex are 410 nm, 33 +/- 1 M-1 cm-1 at 420nm, and 42 +/- 6 M-1, respectively. The values are comparable to those in the other metal complexes of the ascorbate anion.


Salicylic acid fails to inhibit generation of thromboxane A2 activity in platelets after in vivo administration to the rat. Arachidonic acid-induced aggregation of rat platelets and the accompanying generation of thromboxane A2 activity were inhibited by aspirin, whereas 20 times higher doses of salicylic, gentisic and salicyluric acids were inactive. Salicylic acid administered to the rats before aspirin prevented the inhibition of the cyclo-oxygenase-mediated effects of arachidonic acid. These results do not support the hypothesis that the anti-inflammatory activity of salicylic acid is due to inhibition of prostaglandin systhetase (cyclo-oxygenase) by an unknown metabolite and indicate that salicylic acid displays an anti-inflammatory activity independent inhibition of prostaglandin biosynthesis.


Studies on the role of ACTH and of 5-HT in anxiety, using an animal model. The effects of ACTH (5 & 7.5 microgram/100 g) were studied using a new animal model of anxiety. ACTH had an anxiogenic effect that was maximal during a 10 min test period starting 3 min after injection. The behavioural effects of ACTH were counteracted by chronic administration of chlordiazepoxide (5 mg kg-1 for 5 days) and by acute administration of ethanol (0.4 g kl-1). These anxiolytic drugs decreased the turnover of 5-HT in the midbrain, hypothalamus and cerebral cortex, whereas ACTH increased 5-HT turnover in the midbrain and hypothalamus. Is it therefore proposed that anxiety results from the action of ACTH, possibly on 5-HT pathways in the midbrain and hypothalamus.


Problems associated with analysis and interpretation of small molecule/macromolecule binding data. In the analysis of binding data, arbitrary transformations such as the Scatchard plot, may give misleading estimates of the binding parameters. The statistically correct approach is to determine values of K and n by non-linear regression of the actual dependent variable against the actual independent variable. In the case of the spectrophotometric titration method the dependent variable is the absorbance and the independent variable is the composition of the drug/macromolecule mixture. The method relies on an accurate estimate of the extinction coefficient of the bound drug and this is best treated as a parameter to be estimated in the regression analysis. In testing models by data fits alone it is emphasized that whilst a model may be rejected if it does not fit the data, a good fit does not ensure uniquieness and confirmatory, independent evidence must be sought.


The effect of drug concentration on the thermal (dark) degradation of promethazin hydrochloride in aqueous solution. The thermal (dark) degradation of promethazine hydrochloride in aqueous solution presents a complex kinetic picture. The process is oxygen dependent and is modified by EDTA. In citrate buffer, pH 4.0, ionic strength 0.5M, containing 0.1% EDTA, the thermal degradation at 90 degrees can be fitted to first order rate plots at drug concentrations up to 1.56 x 10.27 (0.5%) and to zero order rate plots at drug concentrations greater than 9.35 x 10.2M (3.0%). At intermediate concentrations no simple equation can describe the data. These effects have been correlated with the formation of drug micelles and the rate date have been interpreted on the basis of a first order monomer process and a half order micellar process occurring simultaneously.


The determination of salicylic acid and benzoic acid in pharmaceutical formulations by spectrofluorimetry. Methods of extraction from pharmaceutical formulations and subsequent determination of benzoic acid and salicylic acid by spectrofluorimetry are described. The recovery of benzoic acid, in the presence of salicylic acid, was 99.3%, with a coefficient of variation of 1.04%, while the recovery of salicylic acid, in the presence of benzoic acid, was 97.7%, with a coefficient of variation of 0-68%.


Gastric erosions induced by analgesic drug mixtures in the rat. Gastric erosions after oral administration of analgesics separately and in admixture have been examined in adult rats. After administration of acetylsalicylic acid (aspirin), phenacetin, paracetamol and caffeine as single drugs, gastric erosions were only observed with aspirin. The combination of aspirin with phenacetin did not change, that of aspirin with caffeine significantly increased, and aspirin with paracetamol significantly decreased the incidence of gastric lesions compared with aspirin alone. The results for aspirin with paracetamol did not differ from those for the vehicle. Addition of caffeine to the combination of aspirin and phenacetin caused a significant increase in erosions, but when given with aspirin and paracetamol no erosions occurred. The mechanisms underlying the effects of these drugs on aspirin-induced erosions are discussed.


Effect of tolerance to morphine, ethanol and barbiturate on amphetamine circling in rats with a striatal dopamine lesion. The (+)-amphetamine circling rate of rats with unilateral 6-OHDA lesions in the striatum was recorded. Morphine tablets were implanted subcutaneously for chronic treatment. In the morphine-dependent animal the circling rate to amphetamine given 4 days after morphine was first implanted was depressed but after withdrawal with naloxone a day later the rate increased, returning to normal after 21 days. Barbiturate physical dependence was induced by adding increasing amounts of barbitone to the drinking water of lesioned rats over four weeks after which the amphetamine circling response was depressed and remained so after the barbituate was withdrawn. Ethanol tolerance was induced by adding ethanol to the drinking water of lesioned rats for four weeks. Neither the induction of tolerance over this period nor ethanol withdrawal had any effect on the circling response to amphetamine. The change in the response of striatal dopamine neurons to amphetamine that occurs after chronic morphine treatment, cannot be produced by chronic treatment with either barbitone or ethanol. The neurochemical bases of barbiturate and ethanol tolerance are different from morphine tolerance.


Effects of bromocriptine on adenylate cyclase and phosphodiesterase activities of rat striatum. The activity of 2-bromo-alpha-ergokryptine (bromocriptine) (5 mg kg-1, i.p.) on adenylate cyclase and on phosphodiesterase (PDE-PDE II) of rat striatum, has been examined both in vitro and in vivo. In vitro and in vivo bromocriptine stimulated adenylate cyclase activity, but reduced the stimulating effect of dopamine on adenylate cyclase activity. Bromocriptine showed a dose-dependent biphasic action on phosphodiesterases in vitro while in vivo it stimulated them. The results obtained proved bromocriptine to have an agonist-antagonist action at striatal dopamine receptor level, with a relevant effect on the cAMP system.


A comparison of the binding constant (KD) of 125I-labelled 3-(4-iodophenoxy)-1-isopropylamino-propan-2-ol obtained on beta-adrenoceptors in guinea-pig myocardial membranes, with its dissociation constants (KB) obtained on guinea-pig isolated atria and trachea. The dissociation constant of binding (KD) of 125I-labelled 3-(4-iodophenoxy)-1-isoproplyaminopropan-2-ol (IIP) to guinea-pig myocardial membrane preparations was 2.2 x 10-8M. In pharmacological experiments with the non-labelled material and 60 min contact time, IIP produced a parallel shift in the orciprenaline concentration-response line on guinea-pig isolated tracheal and atrial preparations. The dissociation constant (KB) of IIP was 2.9 x 10-8M on atria (pA2 7.54) and 3.3 x 10-8M on trachea (pA2 7-48). These values indicate that IIP is not a selective beta-adrenoceptor blocking drug. In addition, agreement was found between the affinity constant of this antagonist for beta-adrenoceptors as determined by a direct binding study and an indirect pharmacological study.


Inhibition of hydrochloric acid and pepsin secretion from gastric pouches by antral pouch acidification in sheep. 1. Secretion of acid and pepsin from separated pouches of the body of the abomasum was studied in sheep during perfusion of antral pouches with acid solutions. 2. Resting secretion of acid and pepsin was reduced by acidification of antral pouches to pH 2.7 or less. 3. Increases in the secretion of HCl and pepsin normally obtained on feeding were reduced or abolished by perfusion of antral pouches with solutions at pH 1.3--2.0. 4. Perfusion of antral pouches with acetylcholine at pH 2.5 failed to stimulate acid secretion as it did at a higher pH. 5. Pentagastrin stimulated acid and pepsin secretion during the inhibition of secretion produced by antral pouch acidification. 6. Increases in reticular motility occurred on antral pouch acidification with solutions of pH 1.1--1.3. 7. The contribution of antral pH in regulating abomasal acid secretion is discussed.


Osmotic behaviour of human red blood cells: an interpretation in terms of negative intracellular fluid pressure. 1. The observation that human red blood cells do not shrink in hypertonic media as much as expected for ideal osmometers has previously been explained in terms of either a marked increase in the osmotic coefficient of the cell contents or an increase in the chloride content of the cells.2. Changes in suspension pH and haematocrit have been observed when the concentration of the unbuffered NaCl medium was doubled. The small increases in external pH, and the size of the volume decreases, are inconsistent with variations in the Cl content as a significant factor in the non-ideal osmotic responses.3. Membrane potentials of red cells in buffered media were followed using the fluorescent dye, diS-C(3)-(5). On shrinking at pH 7.4, the cells hyperpolarized ca. 5 mV as predicted if changes in the osmotic coefficient rather than in Cl content explained the osmotic behaviour.4. Regarding haemoglobin in concentrated solution as a solute with high osmotic coefficient is formally correct but is little help in understanding the properties of the solution. We have found it useful to consider separately haemoglobin and the rest of the contents of the cell. The haemoglobin then supports part of the total hydrostatic pressure on the cell leaving the crystalloid solution to experience a reduced fluid pressure. In greatly shrunken cells the contents act like a gel with the matrix of haemoglobin under compression and the fluid which fills the spaces within the matrix under tension.


Effects of adrenoceptor stimulating and blocking agents on carotid body chemosensory inhibition. 1. The effects of alpha- and beta-adrenoceptor agonists and antagonists on chemosensory discharges originating from carotid bodies in situ were studied in anaesthetized cats.2. Noradrenaline (NA) injections commonly resulted in increased frequency of carotid nerve chemosensory discharge, an effect ascribed to reduced blood flow through the glomus, and reduced or eliminated by alpha-adrenergic block.3. NA injections occasionally produced an initial reduction of chemosensory discharge frequency, which was however less intense and of shorter duration than that caused by dopamine. This effect of NA is not mediated by alpha-adrenoceptors, since it is not blocked by dibenamine, but probably by low affinity for dopamine receptors.4. Dopamine and apomorphine-elicited chemosensory inhibition were not affected by low doses of phenoxybenzamine, which blocked NA-evoked hypertensive reactions.5. Higher doses of phenoxybenzamine and dibenamine produced a displacement to the right of dose-response curves for dopamine- and apormorphine-elicited chemosensory inhibition. However, this interference by alpha-adrenergic blockers was attributed to the resultant hypotension, since it was reversed upon restoration of blood pressure.6. Isoprenaline, a beta-adrenergic agonist, did not induce chemosensory inhibition, whilst beta-adrenergic blockers (propranolol and dichloroisoproterenol) did not modify dopamine- and apomorphine-elicited chemosensory inhibition.7. These results provide further support for the hypothesis that chemosensory inhibition could be mediated by specific dopamine receptors, distinct from alpha- and beta-adrenoceptors.


Acid secretion and intracellular pH in isolated oxyntic cells. This study demonstrates that isolated oxyntic cells are capable of secreting hydrochloric acid. Transitory peaks of medium acidification were observed when isolated oxyntic cells were stimulated with diffferent secretagogues. The duration of these peaks was of about 3 min and had a magnitude of about 0.5 muEquiv/hr mg dry weight. These peaks were abolished by selective inhibitors of acid secretion and/or secretagogue action. Cell pH, as measured by the DMO14C technique increases 0.13 pH units upon stimulation with db-cAMP. This change was abolished by pretreatment with SCN-. From acidification and cell pH experiments an increase of 5-10mM in base concentration inside the cell upon stimulation is calculated. These results are interpreted on the basis of the existence of a neutral anion exchange mechanism at the serosal side that serves to control the cell acid-base balance.


Transport of heterocyclic acids across rat small intestine in vitro. A study has been made of the steady-state fluxes of barbituric acid, six of its substituted derivatives, and 5,5-dimethyloxazolidinedione (DMO) across the wall of rat jejunum in vitro. For each of the compounds tested the mucosal (M) to serosal (S) flux was significantly larger than the S to M flux. Both M to S and S to M fluxes increased linearly with concentration, and the transport of one acid was not influenced by the presence of a tenfold greater concentration of a second heterocyclic acid. The fluxes decreased as the pH of the incubation saline was increased, but neither the M to S, nor the S to M fluxes could be described in terms of simple nonionic diffusion. It was found that the relation between the flux ratios of the transported acids and their pKalpha values could be described by an equation derived from consideration of the transport of a weak acid in a series three compartment system, and it has been concluded that the three compartment system provides a good working hypothesis for the mechanism of heterocyclic acid transport across rat jejunum. It was found that the best fit of the theoretical curve to the experimental data was obtained when the ratio of permeabilities to the ionized and nonionized forms of a weak acid at one of the barriers was assigned the value 5 X 10(-1). It is suggested that this value may be characteristic of a noncellular restriction to diffusion, such as a layer of connective tissue, and substantiates previous suggestions that the intermediate compartment of the intestinal three compartment system is a component of the sub-epithelial extracellular space.


Deoxypyrimidine nucleoside metabolism in varicella-zoster virus-infected cells. Noninfected and varicella-zoster virus (VZV)-infected human foreskin fibroblasts were examined for thymidine kinase activity. The specific activity of VZV-infected cell extracts was approximately 7.5-fold greater than that of mock-infected cells and 3-fold greater than that of actively growing cells. The pH optimum of VZV-infected cell thymidine kinase activity was found to be 8.0, whereas thymidine kinase activity in noninfected cells exhibited a sharp pH optimum at 7.4. Electrophoretic analysis of cellular enzymes involved in pyrimidine nucleoside phosphorylation revealed at least three enzymes distinguishable by electrophoretic mobility and substrates used. These enzymes were presumed to be thymidine kinase, deoxycytidine kinase, and uridine kinase. The relative mobilities of these enzymes on 5% polyacrylamide gels were 0.18, 0.91, and 0.54, respectively. In VZV-infected cells, a single band of activity catalyzing the phosphorylation of thymidine, deoxyuridine, deoxycytidine, and cytidine was observed with a relative mobility of 0.48. Cellular pyrimidine-phosphorylating enzymes were not detected in VZV-infected cells. The molecular weight of the VZV-induced enzyme was determined to be 72,000 +/- 7%.


Protein kinase from avian myeloblastosis virus. A protein kinase associated with purified virions of avian myeloblastosis BAI strain A was partially purified by ion-exchange chromatography and gel filtration. The transfer of phosphate catalyzed by this enzyme required a divalent metal ion and ATP as phosphate donor. GTP could not be substituted for ATP, and the reaction was unaffected by either cyclic AMP or beef-heart protein-kinase inhibitor. Of the virus and nonvirus proteins tested as phosphate acceptors, only acidic proteins were phosphorylated. In particular, purified preparations of reverse transcriptase from avian myeloblastosis virus did not accept phosphate. The enzyme is a basic protein (pI = 9.3), and, on the basis of molecular sieving through Sephadex G-200 and velocity sedimentation on glycerol gradient, the protein kinase has a molecular weight of 45,000.


Increased lysyl oxidase activity in blood vessels of hypertensive rats and effect of beta-aminopropionitrile on arteriosclerosis. The activity of lysyl oxidase which catalyzes the initial step of cross-linking of collagen and elastin polypeptides was measured in blood vessels of the hypertensive rat. The enzyme activity was increased in the aorta and mesenteric artery when hypertension was induced in 8-week-old rats with administration of deoxycorticosterone acetate (DOCA) and 1% saline. Reserpine diminished this increase in vascular lysyl oxidase activity concomitant with reduction in blood pressure. When beta-aminopropionitrile, a specific inhibitor of lysyl oxidase, was administered before the onset of DOCA-salt hypertension, the aortic collagen content was reduced markedly. Concomitant with reduction in the aortic collagen content, the development of hypertension and arteriosclerotic changes in the kidney was partially prevented. These results would indicate that hypertension increases the amount and the degree of cross-linking of vascular collagen and that the deposition of excess collagen in the vascular wall contributes to the development of hypertension and arteriosclerosis.


Pathogenicity of a vaccine strain of vaccina virus in rabbits administered by intravenous route. The pathogenicity of the Lister (Elstree) strain, a vaccinia virus, in rabbits after intravenous injection was studied by histopathological and immunofluorescent methods. Inoculation of 1 X 10(7) PFU (pock-forming unit) virus into weanling and young adult rabbits caused severe emaciation and high mortality within 2 weeks. Pathological findings were characterized by vesicular lesions along muco-cutaneous junction areas of the eyes, nose and mouth and by inflammatory changes in the brain, mainly in the meninges and choroid plexus. Immunofluorescent staining of the tissues of animals sacrificed at intervals demonstrated the accumulation of vaccinia viral antigen (s) in the loci of pathological changes. The suitability of this model system for the study of pathogenesis of human postvaccinal meningoencephalitis is discussed.


Correction of hyperkalemia by bicarbonate despite constant blood pH. Patients having hyperkalemia often are given bicarbonate to raise blood pH and shift extracellular potassium into cells. Blood pH in many hyperkalemic patients, however, is compensated. To determine whether bicarbonate, independent of its pH action, affects plasma potassium, 14 hyperkalemic patients were treated with bicarbonate in 5% dextrose. In five patients (changed pH group), blood pH rose at least 0.08, while in nine (constant pH group), it changed less than 0.04. In the first group, pH rose 0.12, bicarbonate rose 5.9 mEq/liter, and plasma potassium fell 1.6 mEq/liter, and plasma potassium fell 1.4 mEq/liter. The correlation between changes in plasma potassium and bicarbonate was identical in the two groups and independent of urinary potassium excretion. Four additional patients, who were treated with 5% dextrose alone, did not significantly lower their plasma potassium, although subsequent treatment with bicarbonate in 5% dextrose lowered their plasma potassium. Thus, bicarbonate lowers plasma potassium, independent of its effect on blood pH, and despite a risk of volume overload, should be used to treat hyperkalemia in compensated acid-base disorders, even in the presence of renal failure, provided the plasma bicarbonate concentration is decreased.


Localization of some glycolipid glycosylating enzymes in the Golgi apparatus of rat kidney. Cell fractions from rat kidney were isolated and studied for their ability to synthesize several possible intermediates in the biosynthesis of sulfatides and gangliosides. The enzymes studied include UDP-Gal:ceramide galactosyltransferase, UDP-Gal:glucosylceramide galactosyltransferase, UDP-Gal:galactosylceramide galactosyltransferase, and CMP-NAN:lactosylceramide sialytransferase activities. The initial glycosylation of ceramide was found to be present in all of the kidney cell fractions studied. The remaining glycosylating enzymes were largely localized in the Golgi apparatus of kidney. Thus, in addition to modifying glycoproteins for secretion, the Golgi apparatus in kidney is involved in the modification of a number of glycolipids which are destined to form cell membrane components.


[Classical and modified intramedullary nailing in osteomyelitis pseudoarthroses]. 13 cases of osteomyelitic pseudarthroses following injury were treated by the authors, by means of intramedullary nailing. 5 of these patients already cured are introduced in detail in the article. Intramedullary nailing in the classic manner and its modification, using bolts as well, were performed alike. The authors have a favourable impression after these series. No septic complication occured. Intramedullary nailing is a good method in the management of this serious sequel to fracture treatment.


[Experience in the treatment of peripheral vascular injuries]. The authors report the results of the treatment of injuries to the peripheral blood vessels that were observed at the Semmelweis OTE in 16 years. As a result of primary operative treatment the peripheral pulse returned to the injured limb in 10 cases. A quick diagnosis and instant reconstructive surgery is absolutely necessary. Mistakes and complications in the treatment are listed in this paper, based on the own experiences of the authors. The authors stress that these complications should be recognized in time and be treated properly.


[Congenital knee luxation].

[Congenital knee luxation]. The authors discuss the frequency, etiology, symptoms and classification as well as treatment of congenital dislocation of the knee. 8 own cases are reported in the article. The authors suggest that congenital dislocation of the may be well influenced by non-operative methods. Etappe redressement by means of plaster does not interfere with the functional treatment of the hip dislocation usually treated with the Pavlik braces. Associated foot deformities may be easily corrected at the same time. The so called "earliest treatment" is most important in all the three deformities (knee-hip dislocation, club foot), the authors emphasise.


[Evaluation of operations on discoid meniscus]. A short survey of literature is given in the article. 1480 meniscectomies were performed at the Orthopedic Clinic of Semmelweis Medical University in the interval between 1952--75; 29 of these cases were carried out for discoid menisci. Symptoms are introduced and results of operative treatment are evaluated. The author makes a few suggestions based on his own experiences.


[Treatment of hand injuries on the principle of "delayed emergency"]. A certain number of drunk patients who suffered hand injury were not prone to operation on the day of admission, as usual, for different reasons (behaviour of the patient etc.). These patients were treated on the delayed urgency principle, that Iselin worked out. Results in the 266 cases were surprisingly good. There were much less complications with this method, compared to previous results achieved when primary treatment of wounds was practised. Yet more experience is necessary, to work out certain details and more exact indications, the authors state.


[Spontaneous rupture of the extensor pollicis longus tendon]. A rare case of "spontaneous" rupture of the extensor pollicis longus tendon with primary chronic polyarthritis in the background is reported in this paper. In this case a transposition of the extensor indicis proprius was carried out, with the modification by J. Böhler and Manninger. This method gives a very good functional result.


[2 cases of congenital humero-radial synostosis]. Two cases of a rare congenital deformity, humeroradial aplasia are reported in the article. In the first case the deformity was unilateral, in the second case a bilateral aplasia was associated to hip dislocation and aplasia of the os ischii.


[Double gunshot head injury with lesion of the upper cervical vertebrae in a case of attempted suicide]. The authors hold their case reported in this paper interesting for the following reasons: in several minutes two shots were fired by intention of suicide. Two entrance openings were found that had the characteristics of shots fired within close range, but no exit openings were to be seen. Considering the entrance openings, the bilateral fracture of the atlas and a comminuted unilateral fracture of the axis and spotting the jammed missiles inside the tunnel, one must deduce that the missiles went by vital structures without severing them.


[Surgical correction of a case of congenital proximal radio-ulnar synostosis]. The author reports a case of S.R.U.P.C. that was relieved by a Nigrisoli operation. There is a good function after two years. The author thought it worth publishing this cae because of its rarity: in the last twenty years only one article appeared on this problem in the Rumanian medical literature that reported 5 cases.


[A case of open carpo-metacarpal dislocation complicated by dislocation of the trapezoid bone]. A case of open carpo-metacarpal dislocation complicated by dislocation of the os trapesium is reported in this paper. The patient's wrist healed with good functional results, despite a partial mistake in diagnosis. The authors draw attention to the fact, that X-ray films taken at admission must be analysed carefully and intraoperative X-rays films must be taken. By demonstration their own mistake in diagnosis and treatment, the authors hope that a similar mistake will be avoided. Following early open or closed reduction, a temporary wire fixation of the metacarpal bones to the carpal bones is recommended in order to prevent redislocation.


[Bullet embolism of the median cerebral artery]. A shot injury to the middle cerebral artery caused occlusion to the vessel by thrombosis and emboly of the missile. The projectil had been removed, but a thrombus formed in the a. cerebri media as a postoperative complication or by progression of a thrombosis in the internal carotid artery and after 19 days the patient died. A focal emollition of the left cerebral hemisphere was found at autopsy.


H2-receptor antagonists in perspective.

H2-receptor antagonists in perspective. Cimetidine, like its predecessors burinamide and metiamide, has been shown in vitro to be a specific competitive histamine H2-receptor antagonist. In vivo, it is a potent inhibitor of histamine-stimulated gastric acid secretion in animals and man after both intravenous and oral administration. Doses sufficient to inhibit gastric secretions are without measureable effects on other physiologic systems. The main indication for cimetidine is in the treatment of duodenal ulcer and of the Zollinger-Ellison syndrome. Useful indications are treatment of gastric ulcer and pnacreatic insufficiency. Possible indications are prevention of gastrointestinal bleeding and treatment of peptic esophagitis. The neutrophil toxicity seen with metiamide has so far not been demonstrated with cimetidine; side effects with cimetidine have generally been trivial. In the future, H2-receptor antagonists are likely to become key therapeutic agents in diseases in which gastric acid-pepsin secretion plays a pathogenetic role.


Neural and endocrine development after chronic tryptophan deficiency in rats: I. Brain monoamine and pituitary responses. Caloric restriction and tryptophan deficient diets have been shown to delay aging in the immature laboratory rat. Studies of monoamine levels in the brain of developing female rats fed on these diets show changes in serotonin but not norepinephrine or dopamine levels: in tryptophan-restricted rats serotonin levels were reduced in all brain areas studied, whereas in caloric-restricted animals serotonin levels were increased in the cerebral hemispheres only. Another group of animals, in which growth and maturation was delayed by feeding d,1-parachlorophenylalanine (PCPA) showed decreases in serotonin, norepinephrine and dopamine concentrations in all brain regions investigated. All treatments employed to arrest growth and maturation resulted in pituitary alterations manifested by gross, histological and ultrastructural changes. It is postulated that there maturation- and age-retarding treatments delay the development of the central nervous system resulting in postponed maturation of the neuroendocrine axis, with consequent hypoactivity of certain pituitary functions and a resultant delay in the onset of maturation and senescence.


Reactions between peroxidizing lipids and histidyl residue analogues: enhancement of lipid oxidation and browning by 4-methylimidazole. As a part of our study on the interactions between peroxidizing lipids and the histidyl imidazole side-chain in simple, low-moisture model systems, 4-methylimidazole (4MI) was reacted with methyl linoleate (ML). This analogue was chosen to avoid interference from other functional groups in histidine (free base) or in proteins. Changes in the concentrations of lipid hydroperoxides, carbonyls, 4MI, and brown pigments were followed over a period of 24 days. The results indicate that 4MI exhibits significant prooxidative activity by reducing the induction period as well as by enhancing the formation of brown pigments. These effects are more pronounced at high 4MI/MI molar ratios and under basic pH's. Upon interactions with peroxidizing ML, as much as 44% of initially present 4MI was destroyed by the sixth day of incubation.


Patterns of the use of benzodiazepines in Australia. In six suburban areas of Sydney, chosen to provide a socioeconomic cross-section of the city, complete records of dispensing of benzodiazepine were collected over a four-week period. These drugs constituted 3.7% of all dispensing, female patients outnumbered males by 2.3:1, and 98% of patients were over 20 years of age. The predominance of females, and higher age groups was found in all the areas studied, but no socioeconomic correlation was detected in the use of the drugs. Analysis of national dispensing figures confirmed the higher consumption in higher age groups, and revealed no heterogeneity in per capita prescribing rates amongst the States.


The prevention of prematurity.

The prevention of prematurity. The prevention of prematurity requires identification of the patients most likely to go into labour prematurely and early recognition of threatened premature labour. There are unfortunately few clinical signs to help identify these patients, but certain features in the history are helpful. As soon as premature labour is diagnosed, energetic and active treatment with corticosteroids to advance pulmonary maturity and uterine suppression should be instituted. It is important to bear in mind that these patients should be managed in units, where not only is such treatment feasible, but neonatal intensive care is available, if labour cannot be suppressed.


Kinetic studies on 3-hydroxykynureninase from rat liver. 3-Hydroxykynureninase was purified from rat liver. The Michaelis constants for L-kynurenine and L-3-hydroxykynurenine were determined to be 2.33 X 10(-4)M and 6.85 X 10(-5)M, respectively, at pH 8.41 and 37 degrees. With L-kynurenine as substrate, the enzyme was competitively inhibited by L-alanine, 3-hydroxyanthranilic acid, and several other compounds which contained structural features of either amino acid or aryl portions of the substrate. The effect of pH on the initial velocity, maximal velocity, and Michaelis constant, using L-kynurenine as substrate, was studied. Maximal velocity was strongly pH-dependent, with a maximum at pH 8.4. The Michaelis constant decreased from 11.4 X 10(-4)M at pH 7.1 to 1.30 X 10(-4)M at pH 9.0. Logarithmic plots of these data showed pKa's for functional groups ionizing in the enzyme-substrate complex and free enzyme active center of 7.6 and 8.5, respectively. Possible groups responsible for these ionizations were discussed.


Structural evidence for gene duplication in the evolution of the acid proteases. X-ray studies of acid proteases indicate a bilobal structure with a well defined active site cleft. An intramolecular twofold symmetry axis relates two topologically similar domains and the active site residues. A possible mechanism for evolution by gene duplication, divergence and gene fusion is presented.


Disequilibrium pH and bicarbonate reabsorption: relevance to the pathogenesis of distal renal tubular acidosis. An augmented renal capacity to reabsorb bicarbonate (RHCO3) has been noted in patients with distal renal tubular acidosis (dRTA), and construed as evidence that the basic defect in dRTA is abnormal distal tubular permeability. According to this interpretation, the absence of a disequilibrium pH due to a back-leak of H2C03 permits increased distal H+ secretion and results in an increased RHCO3. To test this assumption, we have evaluated the effect of acute elimination of the disequilibrium pH by carbonic anhydrase infusion. The results establish that this maneuver doses not cause a rise in RHCO3. Thus, the elevated value of RHC3 described in dRTA cannot be the consequence of increased back-diffusion of H2CO3 and is more likely due to coexisting extracellular volume depletion and/or postassium deficiency.


[Hemodynamic and electrocardiographic effects caused by intravenous infusion of a tricyclic antidepressive agent dibenzepin]. The cardiovascular (blood pressure, heart rate, central venous pressure) and ECG changes caused by dibenzepine-infusion (720 mg/day for 48 hours) have been assessed. Mean blood pressure and heart rate did not show significant changes. In some patients, however, distinct changes in both variables were observed. Central venous pressure was not affected. The infusion elicited changes in the repolarisation phase of the ECG: it did not, however, alter either the automaticity or the conductivity of the heart. These results were obtained in 40 depressive patients, whose general conditions and cardiovascular state were excellent and must not be extrapolated for cardiopathic patients. The risks inherent in this therapeutic approach are discussed.


[Metabolic alkalosis. III. Spontaneous and experimental compensation mechanisms in plasma and erythrocytes]. Plasma and red cell acid-base balance were studied in 13 patients with metabolic alkalosis and 14 healthy subjects following the infusion of bicarbonate. Differences between the plasma and red cell patterns suggested that application of the results of plasma determinations to the body as a whole could be the cause of serious inaccuracies. Since the changes noted occurred in different combinations according to whether alkalinisation was acute or not, it is felt that cell response to a fall in (H+) varies in relation to the time available for compensation.


The use of relaxation and desensitisation techniques in eliminating a sleep problem. This study describes the treatment of a 16-year-old youth unable to sleep, failing to respond to sleeping pills, who on a programme of relaxation and desensitisation in six weeks was able to control his sleeping patterns. On followup four months, six months and twelve months later this ability had been maintained.


Prazosin in hypertension. Part I. Clinical experience in 100 patients. This paper reports the findings of an open evaluation of 100 patients treated with prazosin. When prazosin was added to existing hypotensive regimens in 50 patients whose blood pressure was poorly controlled, 36 (72 percent) became normotensive. Treatment was initiated with prazosin in a further 50 patients. Satisfactory control was achieved with prazosin alone in 24 and 20 of these became normotensive. The remaining 26 patients received in addition a beta-adrenoreceptor blocking agent together with a thiazide diuretic in 14. While prazosin alone caused a mean fall of 26/14mmHg in this group, the enchanced efficacy of combined therapy achieved a normal blood pressure in 19 (73 percent) and a total mean fall in pressure of 42/28mmHg. The most frequent side effect was dizziness or faintness at the start of therapy or, less often, when the dose was increased. This is minimised by using a low initial dose of 0.5mg two or three times daily. Prazosin is an effective hypotensive agent, used alone or in combination, in most patients with hypertension of all degrees of severity.


Effects of lorazepam on hyperactivity in ratarded children. Seven mentally-retarded children, diagnosed as hyperactive, participated in a double-blind trial of the drug lorazepam (Ativan). Statistical analysis of the behavioural records showed that hyperactivity was typically greater during lorazepam than during placebo periods. The degree of hyperactivity was not systematically related to dosage level. The drug is therefore contraindicated for the control of hyperactivity in mentally retarded children.


Initiation of human parturition. IX. Progesterone metabolism by placentas of early and late human gestation. In in vitro studies it was found that, in the presence of added NADPH, the human placenta metabolizes progesterone to 20alpha-hydroxy-4-pregnen-3-one at increasing rates as pregnancy advances. A fivefold increase in 20alpha-hydroxysteroid oxidoreductase activity is observed in term placentas compared with the enzymic activity of placentas of 12 to 20 weeks' gestation (368 +/- 53 pmoles/mg protein/hr vs 71 +/- 8 pmoles/mg protein/hr). The 5alpha-reductase and 3beta-hydroxysteroid oxidoreductase activities were similar in placentas obtained from early and late human pregnancies.


[Development of hematopietic and lymphoid tissues in conjoint bone marrow and thymus transplants]. The model of heterotopic transplantation of the mixture of bone marrow and thymus fragments was used to study the interaction of hemopoietic and lymphoid tissues under their direct contact. The bone marrow and thymus fragments of adult mice F1 (CBAXXC57BL) were transplanted separately or in the mixture under the kidney capsule of mice of the same strain. During the whole period of observation (from 10 days up to 14 months), the development of bone marrow and thymus fragments in the joint transplants proceeded independently, no "mixed" stroma appeared, and the stroma of each organ ensured the differentiation characteristic of its organ. The development of joint transplants somewhat differs from that of isolated transplants: on the 10th day a greater amount of hemopoietic tissues was noted in the former; the bone marrow component increases continuously up to 6 months (vs. 1--2 months in the isolated transplants); the bone and hemopoietic tissues predominate in the joint transplants by 14 months, the amount of thymic tissue markedly decreases but it does not disappear completely.


Stimulation of the activity of Schistosoma mansoni miracidia by snail-conditioned water. A dark-ground photographic technique was used to analyse the reactions of Schistosoma mansoni miracidia to homogeneous solutions of snail-conditioned water (SCW). The most significant effect of this water was to increase miracidial turning. This effect was maintained under both acid and alkaline conditions, after passage of the SCW through a mixed bed resin and after chelation of either calcium or both calcium and magnesium ions. The stimulant in the water was unaffected by trypsin but was protease-sensitive, suggesting its possible identity as a peptide. The importance of 'active spaces' rather than concentration gradients in miracidial host-location was emphasized.


Poisoning in children under age 5: identification and treatment. A rational approach to management of poisoning in young children requires assessment of what, when, and how much drug was ingested. Emesis, if indicated, is followed by general supportive measures, enhanced removal of absorbed poison, and antidote therapy.


The biochemistry of the basal ganglia and Parkinson's disease. The metabolic pathways for five transmitters in the basal ganglia are briefly described; the results of determinations of their concentrations, of their rate-limiting enzymes and of their degradation products are summarized. The changes found in Parkinson's disease are described. While dopamine synthesis in the basal ganglia is defective in this condition, abnormalities of other transmitters occur, and their possible significance is discussed.


Urticaria--current concepts.

Urticaria--current concepts. The mechanisms, clinical features and management of the various forms of urticaria are discussed. The importance of histamine receptors, complement and the kinin cascade are reviewed.


Evidence for acceleration of the rate of elongation of tyrosine aminotransferase nascent chains by dibutyryl cyclic AMP. Analogs of cyclic AMP elevate the synthesis of tyrosine aminotransferase (L-tyrosine:2-oxoglutarate aminotransferase; EC 2.6.1.5) in cultured hepatoma cells and rat liver at a post-transcriptional level but have no discernible effect on total soluble protein synthesis. In order to determine whether cyclic AMP exerts its effect on a step before or after initiation of the synthesis of this enzyme, we have analyzed the ribosomal transit times for both the aminotransferase and total soluble protein in hepatoma cells incubated in the presence or absence of N(6),O(2)'-dibutyryl cyclic AMP. The time required for one ribosome to translate one subunit of the "average" soluble protein (transit time) was about 2 min in cells incubated with or without the cyclic AMP analog. In contrast, the transit time for tyrosine aminotransferase was found to be reduced from 5-8 min under basal conditions to as low as 45 sec after exposure to dibutyryl cyclic AMP. Although the degree of effect varied from experiment to experiment, the relative rate of aminotransferase nascent chain elongation was found to be proportional to the stimulation of its activity. In contrast, dexamethasone did not alter the rate of aminotransferase elongation even though it elevated enzyme activity between 5- and 10-fold. These data are consistent with the hypothesis that induction of tyrosine aminotransferase with cyclic AMP analogs occurs by stimulation of the rate at which ribosomes translate pre-existing mRNA in contrast to adrenal steroids which act by increasing the level of translatable mRNA coding for this enzyme.


Agonist-induced increase in apparent beta-adrenergic receptor size. The properties of digitonin-solubilized beta-adrenergic receptors from frog erythrocyte membranes were studied by gel exclusion chromatography on AcA 34 Ultragel. beta-Adrenergic receptor binding activity in these membranes can be identified by both an agonist ligand, [(3)H]hydroxybenzylisoproterenol, and the antagonist ligands, [(3)H]dihydroalprenolol and (125)I-labeled hydroxybenzylpindolol. Occupancy of the beta-adrenergic receptors with the [(3)H]hydroxybenzylisoproterenol agonist prior to their solubilization from the membrane leads to an increase in apparent receptor size. Alterations in the molecular size of the receptor cannot be mimicked by occupancy of the binding site with the antagonist ligands. Exposure of frog erythrocyte membranes to [(3)H]hydroxybenzylisoproterenol agonist in the presence of 10 muM Gpp(NH)(p), a guanyl nucleotide analog that exerts multiple regulatory effects on the catecholamine-sensitive adenylate cyclase [ATP pyrophosphate-lyase (cyclizing); EC 4.6.1.1] system, results in the elution of the [(3)H]hydroxybenzylisoproterenol radioligand in both the region characteristic of the agonist-receptor complex and the region characteristic of the antagonist-receptor complex. The precise molecular interactions responsible for the agonist-induced increase in apparent beta-adrenergic receptor size are still unresolved. However, the low concentrations of agonist that are capable of altering apparent receptor size and the sensitivity of this effect to guanyl nucleotides suggest that these phenomena may be intimately involved in eliciting the physiological effects of beta-adrenergic catecholamines at the molecular level.


Myristic acid stimulation of bacterial bioluminescence in "aldehyde" mutants. The involvement of long chain aldehyde in bacterial luminescence was known both from its being required for light emission in the in vitro reaction with pure luciferase and from its ability to stimulate luminescence in vivo in a certain class of dark "aldehyde" mutants. We have found that the luminescence of some (but not all) of such aldehyde mutants is also stimulated by long chain aliphatic fatty acids, with a marked specificity for myristic (tetradecanoic) acid. This stimulation has been demonstrated in aldehyde mutants of two species of luminous bacteria, Beneckea harveyi and Photobacterium fischeri. The responses, both in intensity and yield, are proportional to the amount of added tetradecanoic acid over a 1000-fold range, down to 10 pmol ml-1. Unsaturated long chain fatty acids are potent inhibitors of the tetradecanoic acid stimulation, but they do not effect the in vivo luminescence of wild-type bacteria.


Self-association of hemoglobin betaSH chains is linked to oxygenation. Self-association of unliganded beta(SH) chains into tetramers (4 beta(1) [unk] beta(4)) is experimentally found to be energetically less favorable (DeltaG(0) = -19.05 +/- 0.30 kcal) than the corresponding oligomerization of fully oxygenated chains (4 beta(1)X [unk] beta(4)X(4); DeltaG(0) = -22.45 +/- 0.35 kcal). Hence the tetramers must bind oxygen with a higher affinity than that of dissociated chains. Calculations are presented showing why this affinity difference is not easily detected. The linkage is in a direction opposite to that exhibited by normal hemoglobin A, in which oligomerization of high-affinity unliganded dimers (2 alphabeta [unk] alpha(2)beta(2)) leads to tetramers with decreased oxygen affinity. In contrast, the oligomerization of high-affinity, unliganded beta(SH) chains leads to tetramers with even higher affinity. The results imply the existence of at least two conformational states for beta chains. Effects of inositol hexaphosphate on beta chain association were investigated. Inositol hexaphosphate was found to have no measurable effect at pH 7.4, in contrast to pH 7 where very pronounced effects have been observed. Some theoretical aspects of the linkages are presented and the relationship of the findings to concepts of structural transition and allosteric regulation is discussed. In contrast to the beta chains, self-association of alpha chains into dimers was found to occur with the same free energy in both unliganded and fully oxygenated states. Thus, the self-association of alpha chains is not linked to oxygenation.


Characterization of the residual adenosine deaminating activity in the spleen of a patient with combined immunodeficiency disease and adenosine deaminase deficiency. A number of infants with an autosomal recessive form of combined immunodeficiency disease also lack adenosine deaminase (adenosine aminohydrolase; EC 3.5.4.4) activity in their erythrocytes. Other tissues from these infants contain only a few percent of the adenosine-deaminating activity present in corresponding normal tissue. The residual adenosine-deaminating activity in extracts from the spleen of a combined immunodeficient, adenosine deaminase-deficient patient was compared with adenosine deaminase from normal spleen. Affinity and immunoadsorbant column chromatography revealed distinct differences between the adenosine-deaminating activity in the patient's spleen and adenosine deaminase from normal spleen. The point of maximum activity and general configuration of the pH optimum curves were also different. erythro-9-(2-Hydroxyl-3-nonyl)adenine, a potent inhibitor of adenosine deaminase from normal spleen, had relatively little effect on the activity from the patient's spleen. In contrast, adenine was a better inhibitor of the activity in the patient's spleen than it was of the enzyme from normal tissue. An adenosine-deaminating activity with the same characteristics and specific activity as that in the patient's spleen was also isolated from normal spleen. These results suggest that the adenosine-deaminating activity in the spleen of this patient is not due to a mutant form of adenosine deaminase.


Proliferative capacity of murine hematopoietic stem cells. The present study demonstrates a decrease in self-renewal capacity with serial transfer of murine hematopoietic stem cells. Production of differentiated cell progeny is maintained longer than stem cell self-renewal. In normal animals the capacity for self-renewal is not decreased with increasing donor age. The stem cell compartment in normal animals, both young and old, appears to be proliferative quiescent. After apparent recovery from the alkylating agent busulfan, the probability of stem cell self-renewal is decreased, there is a permanent defect in the capacity of the bone marrow for serial transplantation, and the stem cells are proliferatively active. These findings support a model of the hematopoietic stem cell compartment as a continuum of cells with decreasing capacities for self-renewal, increasing likelihood for differentiation, and increasing proliferative activity. Cell progress in the continuum in one direction and such progression is not reversible.


Genetic dissection of short-term and long-term facilitation at the Drosophila neuromuscular junction. Transmitter release at the Drosophila larval neuromuscular junction may be increased by previous activity of the nerve. This facilitation phenomenon involves at least two processes, one short-term and other long-term. These are shown to based on different mechanisms because (i) a mutant was found that had abnormal long-term facilitation but normal short-term  facilitation; and (ii) long-term facilitation was eliminated by tetrodotoxin or by removing external Na+ but short-term facilitation was not. In long-term facilitation, there was a prolonged release of transmitter due to a prolonged Ca2+ sensitivity of the presynaptic terminal after each nerve stimulus. The cause of this is probably accumulation of Na+ inside the nerve terminal.


Effect of antipsychotic and other classes of drugs on spontaneous locomotor activity and neurotoxicity in mice. Quantitative estimates were made of the effects of several classes of drugs on spontaneous activity and neurotoxicity in mice. Clinically effective antipsychotic agents had a more selective action on spontaneous activity than other classes of drugs with the exception of clonidine and a related compound.


Granulomatous ileocolitis (Crohn's disease). In a clinical survey, the symptoms, associated diseases, and local and metabolic complications of Crohn's disease are discussed. Medical treatment (with SASP, corticosteroids, and immunosuppressives), as well as treatment for malnutrition, is of special importance.


Prostaglandin synthesis in human placenta and fetal membranes. The conversion of exogenous arachidonic acid into prostaglandins was studied in human placenta and fetal membrane microsomes. Only one prostaglandin was formed, prostaglandin E2 (PGE2), in fetal membrane microsomes. In placental microsomes PGE2 was further transformed into 15 keto-PGE2. Cofactor requirements and some characteristics of the system were studied. 1 to 3% conversion of arachidonic acid into prostaglandins was observed in placental microsomes and 5 to 8% conversion in fetal membrane microsomes.


Tissue and blood gamma-glutamyl transferase distribution in the pig. Gamma-glutamyl transferase distribution was studied in 10 pigs with an average weight of 100 kg. It was shown that kidney had the highest concentration of the enzyme, followed by pancreas, spleen and liver. Blood activities measured in plasma and sera from 54 animals were very similar, ie, 35+/-21 iu/litre for plasma and 36+/-14 iu/litre for serum.


An investigation into the viability of broth cultures of the T1 strain of Mycoplasma mycoides sub-species mycoides. The T1 broth vaccine against contagious bovine pleuropneumonia can only be stored for four weeks at +4 degrees C because after that time the titre of Mycoplasma mycoides sub-species mycoides falls below the minimal vaccinating dose. An investigation into the death of these organisms during the stationary growth phase was made. The culture medium in which the vaccine was prepared was found to contain adequate nutrients. Control of the pH by the addition of KOH was shown to preserve the viability of the organism: there was no drop in titre during 336 h (14 days) at 37 degrees C. Use of this technique may prove valuable as a means of preserving the viability of T1 broth vaccine.


Activities of ketone body utilising enzymes in tissues of fed and fasted sheep. The activities of 3-hydroxybutyrate dehydrogenase, 3-ketoacid CoA-transferase and acetyl-CoA acetyltransferase have been measured in the kidney, heart and brain of fed and four-day fasted ewes. The results indicate tha there is a decrease in the capacity of these organs to catabolise ketone bodies in fasting ketosis.


Clinical and metabolic consequences to cattle of rumen overdosing with di-ureido iso-butane (DUIB). Di-ureido iso-butane (DUIB) was overdosed on three occasions to two year-old bullocks, under experimental conditions. Six g/kg live weight of DUIB was mixed manually into the rumen contents formed by a predominantly hay diet (10 kg) with added concentrates (2 kg). Marked clinical signs were observed for 72 h before recovery. Food and water intake were interrupted and a smell of ammonia saturated the accommodation. Blood concentrations of urea-nitrogen increased, while glucose fell; a metabolic alkalosis was present. Rumen VFA concentrations fell, but without altering the proportions of the individual acids present, pH increased, urea-nitrogen and ammonia-nitrogen concentrations fluctuated markedly. The pH of duodenal contents increased. Clinical recovery appeared complete after six days with biochemical homeostasis nearly restored.


A new method for determination of microsomal haem oxygenase (EC 1.14.99.3) based on quantitation of carbon monoxide formation. Microsomal haem oxygenase (MHO) catalyses the main pathway for haem degradation. Hitherto bilirubin formation rate (BFR) has been used for determination of MHO activity. In the present study a method is described where MHO activity is assessed from the rate of carbon monoxide formation (VCO) in tissue homogenates with methaemalbumin as substrate. Formed CO is bound to haemoglobin present in the tissue homogenate. CO is measured by gas chromatography after reduction to methane. CO amounts of 0.01 nmol could be measured. This corresponded to an MHO activity of 0.05 pkat when using standard incubation mixture and 12 min incubation time. The correlation with MHO activity determined with the BFR technique was good (r = 0.94; n = 41); MHOVCO = 0.94-MHOBFR-0.08). Advantages with the VCO method are a ten-fold increase in sensitivity compared to the BFR method and independence of biliverdin reductase.


Dissolution and uptake of cadmium from dental gold solder alloy implants. Pure metallic cadmium was irradiated by means of thermal neutrons. The irradiated cadmium (115Cd) was placed in bags of gold foil and the bags were implanted subcutaneously in the neck region of mice. Two and 3d respectively after implantation the mice were killed, the bags removed and the animals subjected to whole-body autoradiography. The autoradiograms revealed an uptake of 115Cd in liver and kidney. In another experiment specimens of a cadmium-containing dental gold solder alloy, a cadmium-free dental casting gold alloy and soldered assemblies made of these two alloys were implanted subcutaneously in the neck region of mice. The animals were killed after 6 months; cadmium analysis showed significant increases in the cadmium concentration in liver and kidney of those mice which had been given implants of gold solder alloy. The study clearly shows that due to electrochemical corrosion cadmium can be released from implants and accumulated in the kidneys and the liver.


Composition of the alkaline component of human gastric juice: effect of swallowed saliva and duodeno-gastric reflux. Five patients (four with vitiligo and one with pernicious anaemia) were subjected to the histamine infusion test; there were achlorhydric while the remaining two secreted quite small amounts of acid. [Na+],[Cl-)and[alkali]were determined in the alkaline gastric juice samples (pH greater than 7.0). In order to assess the contribution of swallowed saliva the histamine test was done twice in each patient: (A) with precautions to prevent swallowing of saliva; and (B) with the patient allowed to swallow saliva freely. In each sample reflux of duodenal juice was estimated so that its contribution to the alkaline gastric aspirate could be assessed. Such reflex was absent in one patient, negligible in another, while in the remaining three patients the mean pyloric reflux amounted to no more than 8% of the observed volume. Swallowed saliva had diluting effect on [Na+] and [Cl-] but raised K+ concentration in the alkaline gastric aspirate. The comparison of alkaline gastric juice, free to an appreciable extent of salivary contamination, was shown to be relatively constant. The results are consistent with the two-component hypothesis of gastric secretion.


A sensitive method for specific quantitation of secretory IgA. A method to quantitate specifically secretory IgA (SIgA) has been developed using the enzyme-linked immunosorbent assay. The IgA in the test sample was adsorbed to anti-alpha antibodies attached to plastic tubes via a cost of IgA myeloma protein. The reacted SIgA was determined using anti-secretory component antiserum conjugated with alkaline phosphatase. The technique permitted quantitation of secretory IgA in biological fluids like milk, urine, and saliva with a reproducibility of +/-7%, down to 0.03 mg/l. In contrast to earlier techniques, the presence of up to 157% of serum IgA without secretory component (SC) and free SC did not disturb the measurements of SIgA. Furthermore, variations in pH and osmolarity, within biological ranges in secretions, did not influence the estimations.


Detection of hepatitis B antigen in circulating immune complexes in acute and chronic hepatitis. By using polyethylene glycol precipitation at low concentration (PEG test) and the radiolabeled C1q binding test, immune complexes were detected sera from acute (23/28) and chronic (28/32) hepatitis patients, hemodialyzed patients with chronic hepatitis B surface (HBs) antigenemia (7/19), and asymptomatic HBs antigen carriers (2/11). After treatment of PEG precipitates with acidic pH, heating, or proteolytic enzyme (protease), electroimmunodiffusion or radioimmunoassay revealed the presence of HBs antigen or antibody in dissociated immune complexes in sera from several acute and chronic hepatitis patients. Electron microscopy showed immune complexes of HB virus in 9 of 12 PEG precipitates obtained from PEG-test-positive sera; these 9 precipitates were from patients with acute or chronic hepatitis and the other three from chronic HBs Ag carriers. Free HB virus particles were observed after protease digestion of PEG precipitates. Neither immune complexes nor virus particles were seen in precipitates from PEG-test-negative but HBs-Ag-positive sera from chronic carriers.


[Pipoxizin, a new bronchodilator].

[Pipoxizin, a new bronchodilator]. Pipoxizin is a new bronchodilator with the chemical name chlorhydrate-4-diphenyl-methylene-1-(2-/2-(2-hydroxy-ethoxy)-ethoxy/-ethyl/-ethyl)-piperidine. This substance exhibits powerful antihistaminic and antiserotonin properties but no parasympathetic or beta-adrenergic properties. The effects of Pipoxizin have been compared with those of the wellknown product hexoprenalin (Ipradol). Twelve male patients, each with bronchial obstruction, took part in the trials. Plethysmography before and after Alupent inhalation was carried out to determine the reversibility or irreversibility of the bronchial obstruction. On the following day, the plethysmorgraphic tests were repeated before and after intravenous injection of Pipoxizin, and on the third day, the tests were repeated again after intravenous injection of hexoprenalin. Pulse and blood pressure were monitored. Following the use of Pipoxizin, there was a significant decrease in the following parameters: RT, RE, RV, ITGV, pulse rate, and systolic blood pressure. A significant increase was found in FEV1. After hexoprenalin injection, there was a significant decrease only in RV and an increase in FEV1. A comparison of Pipoxizin and hexoprenalin shows a significant difference between the two products, Pipoxizin having the more favourable effect.


Nature and nurture in development of the autonomic neuron. Arguments are presented for the hypothesis that during an early stage of development the cells which become principal neurons of the autonomic nervous system possess information regarding the positions they will occupy within the body. A second stage of development, during which a decision is made regarding which neurotransmitter to employ, is delayed until each neuron has assumed its permanent position in the body and has sampled, presumably via its growing axons, the peripheral field which it will innervate. The development of cholinergic mechanisms takes precedence; adrenergic neurons may develop only when cholinergic sites have been occupied. An extended period during which the differentiation of transmitter mechanisms may be modulated permits the neuron to adequately sample the periphery prior to commitment to a specific transmitter economy.


Experimental Brugia timori and Wuchereria bancrofti infections in certain species of mosquitoes. Laboratory reared Aedes aegypti (black eye and Jakarta strains), Aedes togoi, (Taiwan), Aedes albopictus, (Jakarta), wild caught Anopheles barbirostris, (Java) and Mansonia uniformis, (Jakarta) were fed on a carrier with mixed infection of Brugia timori and Wuchereria bancrofti. B. timori and W. bancrofti were able to develop in A. aegypti (black eye) and A. togoi, with development proceeding more rapidly for of B. timori than W. bancrofti. Both species of parasites were readily distinguishable in each of their developmental stages. A. barbirostris from Java was able to support development of B. timori as well as A. barbirostris from Flores. B. timori and W. bancrofti did not develop in M. uniformis, A. aegypti (Jakarta strain) and A. albopictus.


Genetic analysis of the nitrogen fixation system in Klebsiella pneumoniae. Fine structure mapping of nif mutations of Klebsiella pneumoniae was accomplished by means of Pl-transductional crosses and the plasmid R144 drd mediated conjugations. The physical distance between nif mutations based on the percentage of co-transduction with hisD of the nif mutations was estimated. The maximal distance between two mutations was calculated about 3 Kb, and the average distance between different nif mutations was about 1 to 2 Kb. So no "silent region" was shown within the nif cluster nearby the histidine operon. Several hisD-unlinked nif mutants were isolated and investigated genetically and biochemically. They all differed from the glutamineless mutants, one of these mutants was tentatively assigned as a sort of N-assimilation mutant with little activity of glutamate synthetase. It differed from the known N-assimilation mutants in its absence of nitrogenase activity. Since the wild type hisD-linked nif genes carried by the plasmid RP4 failed to complement the defects of the hisD-unlinked nif genes in the recipient cells but they were effective to facilitate E. coli in acquiring the ability to fix nitrogen, which indicates that the hisD-unlinked nif genes necessary for the functioning of the hisD-linked nif genes are present in E. coli.