Growth of Candida utilis on a mixture of monosaccharides, acetic acid and ethanol as a model of waste sulphite liquor. Candida utilis cultivated under batch conditions in a synthetic medium with a mixture of different carbon sources utilized first D-glucose and then D-galactose, D-mannose, D-xylose, L-arabinose, ethanol and acetic acid. The effect of acetic acid was primarily a function of pH and the physiological state of the inoculum. At pH 4.5, acetic acid at a concentration of 1 g/l increased the specific growth rate, reduced time of cultivation and increased yield of the yeast dry weight. The yield from acetic acid was 61%. In the presence of a higher content of acetic acid (3--6 g/l) the yield was only 18--26%. The yield calculated only from monosaccharides increased but the yield with respect to total carbon sources was lower. The specific growth rate decreased as well. The addition of ethanol also resulted in an increase of the production and yield of the yeast dry weight but the cultivation time was prolonged. The simultaneous utilization of carbon sources of the studied mixture modelling a sulphite fermentation medium with ethanol is advantageous. However, due to physiology of the yeast, it is most suitable to cultivate a strain adapted to utilizable carbon sources in a continuous way, in the presence of their limiting concentrations in the cultivation medium.


Growth of Aureobasidium pullulans on waste water hemicelluloses. Strain Aureobasidium pullulans capable of utilizing hemicelluloses and xylan was cultivated on processed waste dialysis liquor from the production of viscose fibres, containing about 1.5% hemocelluloses. Basic conditions of biomass production were tested on a laboratory scale. The dialysis waste liquor adjusted with mineral acids to pH 4--5 and supplemented with 0.05% yeast autolyzate and 0.2% ammonium sulphate affords protein yields of about 0.8 g/l, corresponding to 4.0--4.5 g dry biomass. Biomass is isolated together with residual water-insoluble hemicelluloses which are not utilized by the microorganism. The total utilization of hemicelluloses attains about 70%.


Factors affecting protein synthesis during biotin deficiency in Aspergillus nidulans. Biotin deficiency in Aspergillus nidulans resulted in a 70% increase of the protein content and increased levels of free and bound aspartate, glutamate, serine, leucine and methionine. Likewise, the activities of NADP+ glutamate dehydrogenase, NAD+ gluatmate dehydrogenase, aspartate aminotransferase and alanine aminotransferase were significantly increased. The total RNA content increased while the DNA content was unaffected. The rRNA/tRNA ratio remained higher in biotin-deficient cells. Supplementation of glutamate, aspartate, serine, leucine and methionine to the culture medium raised the rRNA/tRNA ratio, and the difference observed in the qualitative and the quantitative patterns of protein and dry cell mass between normal and biotin-deficient cultures was abolished.


Testicular biopsy in the study of male infertility: its current usefulness, histologic techniques, and prospects for the future. Testicular biopsy has been widely used for the diagnosis of male infertility for more than three decades. During that time, with advances in cytogenetics, radioimmunoassay, and endocrinology, the role of testicular biopsy has changed. Testicular biopsy is still useful in the diagnosis of azoospermic and oligospermic males without stigmata of gonadotropic insufficiency or Klinefelter's syndrome. A classification and description is presented of pathologic changes in the testis as seen on testicular biopsy by light microscopy. The present rationale for testicular biopsy in infertility, the processing and staining of histologic material, and the role of testicular biopsy in infertility and infertility related situations, including cryptorchidism, malignant disease, and chemotherapy related changes, are discussed. Further understanding of testicular function and disease will depend upon the correlation of histologic and ultramicroscopic changes, immunohistologic localization of hormones, and epidemiologic and endocrinologic data.


Mannose-sensitive stimulation of human leukocyte chemiluminescence by Escherichia coli. Escherichia coli organisms with mannose-sensitive adherence factors (adhesins) are known to associate with human peripheral leukocytes (WBCs) in vitro in the absence of serum. To determine whether the WBC respiratory burst is activated during the interaction with E. coli, WBC chemiluminescence was measured. E. coli with mannose-sensitive adhesins stimulated a sharp burst of chemiluminescence which peaked 15 to 30 min after the bacteria and WBCs were mixed. Stimulation of chemiluminescence could be abrogated by including 10 mM alpha-methyl-D-mannoside in the test suspension. The addition of alpha-methyl-D-mannoside up to 20 min after the E. coli and WBCs were combined caused a rapid decrease in chemiluminescence. E. coli stimulation of chemiluminescence could not be inhibited by pretreating the WBCs with purified type 1 pili (fimbriae). E. coli lacking mannose-sensitive adhesins failed to stimulate chemiluminescence. The results emphasize the importance of mannose-sensitive adhesins in the association of E. coli with WBCs and suggest that the E. coli-WBC interaction system may be a useful tool for studying the mechanisms involved in the activation of the respiratory burst during phagocytosis.


Effect of L-histidine pretreatment on methamphetamine induced sterotyped behaviour in rats. Pretreatment with L-histidine precursor of histamine was found to delay the onset, shorten the duration and decrease the intensity of stereotyped behaviour induced by methamphetamine while chlorcylizine a H1 receptor blocker was found not only to potentiate methamphetamine induced sterotyped behaviour but also to block the inhibitory effect of L-histidine on methamphetamine stereotypy. Our results indicate that the central histaminergic system by exerting a modulatory influence on the striatal dopaminergic system may influence the functioning of the extrapyramidal motor system.


Bacterial aggregating activity in human saliva: simultaneous determination of free and bound cells. Two new assays for saliva-mediated aggregation of oral bacteria have been developed, based on the use of [3H]thymidine-labeled cells. One assay separates free cells from aggregated cells by centrifugation through sucrose, whereas the other utilizes membrane filters (8 micrometers, Nuclepore) to effect the separation. Comparison of these assays with the turbidity method reveals that they are faster (X20 to 40) and require 10 times less saliva and bacteria. The aggregation of Streptococcus sanguis M5, as determined with these assays, is complete in 5 min and is dose dependent on added cells and saliva. The reaction exhibits a temperature optimum of 42 degrees C with no reaction at 0 degrees C. If the pH is reduced to below 5, saliva-dependent aggregation is inhibited. The salivary factor(s) are heat labile, losing 100% of their activity after 100 degrees C, 10 min or 70 degrees C, 30 min.


Central histaminoceptor-adrenoceptor interrelations in the release of adrenocorticotrophic hormone. Intracerebroventricular administration of adrenaline, noradrenaline phenylephrine, clonidine and histamine produced a significant rise in plasma cortisol concentration whereas isoprenaline had no effect. alpha-Adrenoceptor blockers (yohimbine or piperoxon) per se did not alter the plasma cortisol level. Central pretreatment with yohimbine or piperoxin, blocked the rise in plasma cortisol level induced by icv noradrenaline, phenylephrine and clonidine. In another set of experiments, both H1 and H2 receptor antagonists (mepyramine, and metiamide) per se had not significant effect on plasma cortisol concentration. Central histamine induced rise in plasma cortisol concentration was significantly blocked by icv pretreatment with both H1 and H2 receptor blockers. Furthermore, yohimbine also significantly prevented the rise of plasma cortisol level induced by icv histamine.


Phenotypic stability of the cell wall of Streptococcus mutans Ingbritt grown under various conditions. Quantitative analyses of cell walls from Streptococcus mutans Ingbritt grown under carbohydrate limitation in the chemostat showed that growth conditions had no statistically significant effect on the composition of polysaccharide, peptidoglycan, or the proportion of polysaccharide in the cell wall. Lysis of cell wall preparations with a muramidase supported this conclusion and further indicated that there was little difference in their overall structure. In contrast, there was a consistent difference between the rates of lysis by this enzyme of organisms grown in 0.2% glucose and 0.5% glucose. Extremes of pH or dilution rate essentially did not influence the immunogenicity of type c antigen in whole organisms irrespective of whether the carbohydrate source was glucose or sucrose. However, differences were found in the immunogenicity of lipoteichoic acid under similar circumstances. The results indicated there was an inherent phenotypic stability in the cell walls of S. mutans Ingbritt despite changes in pH, generation time, and carbohydrate source, and that any changes that did occur were probably due to associated cell-surface components.


Effect of carbohydrate source and growth conditions on the production of lipoteichoic acid by Streptococcus mutans Ingbritt. Streptococcus mutans Ingbritt was grown in a chemostat at defined dilution rates and pH values and under carbohydrate limitation. At a constant dilution rate of D = 0.1 h-1 and with either 0.5% glucose or 0.5% sucrose, the amounts of both cellular and extracellular lipoteichoic acid increased as the culture pH increased from 5.0 to 7.5. At a constant pH of 6.0, the amount of cellular lipoteichoic acid formed by cultures growing in 0.2% or 0.5% glucose was relatively constant over a range of dilution rates, although the amount of extracellular lipoteichoic acid formed in 0.2% glucose at intermediate dilution rates was less than that formed in 0.5% glucose. Organisms grown in 0.5% sucrose at pH 6.0 contained increasing amounts of cellular lipoteichoic acid as the dilution rate was increased. A comparison of the amounts of cellular lipoteichoic acid formed by organisms growing at D = 0.5 h-1 and pH 6.0 in glucose, sucrose, fructose, or mixtures of glucose and fructose in limiting amounts suggested that the enhanced production of lipoteichoic acic by sucrose-grown organisms was due to the fructose component. The culture fluids from both glucose- and sucrose-grown organisms contained detectable amounts of serotype c antigen, whereas glucose-grown cultures also contained significant amounts of an extracellular hexose-containing polymer.


Opsonic activity of normal human cerebrospinal fluid for selected bacterial species. The opsonic activity of normal human cerebrospinal fluid (CSF) has not been well defined. In this study, the opsonic activity of normal CSF for laboratory and blood culture isolates of Staphylococcus aureus, Streptococcus pneumoniae, Escherichia coli, Hemophilus influenzae type b, and Neisseria meningitidis was measured by a quantitative assay employing radiolabeled bacteria and polymorphonuclear leukocytes. All isolates of S. aureus, except the Wood 46 strain, were opsonized in undiluted CSF (>50% uptake by polymorphonuclear leukocytes.) There was heat-stable and heat-labile opsonic activity in CSF for S. aureus. Only one blood culture isolate of E. coli was moderately well opsonized in undiluted CSF (26% uptake). None of the remaining laboratory or clinical isolates were opsonized in undiluted CSF. The S. aureus isolates were more readily opsonized in dilute normal serum than were the other bacterial species, and complement appeared to be the heat-labile opsonin in serum. However, complement may not be the heat-labile opsonin in normal CSF for S. aureus. In contrast to serum, complement C3 was not visualized on the staphylococcal cell surface by immunofluorescence microscopy and chelation of CSF did not diminish opsonic activity. This study demonstrates that normal CSF is opsonic for S. aureus but not for bacterial species that more commonly cause meningitis. These species differences in opsonic requirements may be important in the pathogenesis of meningitis.


Comparative immunogenicity of group 6 pneumococcal type 6A(6) and type 6B(26) capsular polysaccharides. The comparative immunogenicity of the two cross-reacting group 6 pneumococcal capsular polysaccharides, type 6A(6) and type 6B(26), was studied with hyperimmune rabbit typing antisera and with sera from adult volunteers injected with polyvalent pneumococcal vaccines containing either 50 mug of type 6A (U.S. designation, type 6) or 50 mug each of type 6A and type 6B (U.S. designation, type 26) polysaccharides. Both group 6 polysaccharides were linear copolymers composed of 1 mol each of d-galactose, d-glucose, l-rhamnose, and d-ribitol phosphate. They differed only in that type 6A had a rhammopyranosyl-(1 --> 3)-d-ribitol bond and the type 6B had a rhamnopyranosyl-(1 --> 4)-d-ribitol bond. Quantitative precipitation and absorption analyses with rabbit hyperimmune antisera induced by simultaneous injection with type 6A and type 6B organisms revealed extensive cross-reactions between the two group 6 polysaccharides. There was less, although still quite extensive, cross-reactivity between the two group 6 polysaccharides examined with antisera from rabbits injected with only one of the group 6 pneumococci. In a radioimmunoassay, using (14)C internally labeled type 6A or type 6B polysaccharide antigens, there was no difference in the serum antibody level to either type of volunteer injected with polyvalent pneumococcal vaccines containing type 6A or both type 6A and type 6B polysaccharides. These studies indicate that the structural similarity of the pneumococcal group 6 polysaccharides confers extensive cross-reactivity with hyperimmune typing antisera prepared with whole organisms or after injection of purified polysaccharides in adult volunteers. With our current polysaccharides, it appears that a polyvalent pneumococcal vaccine formulation that contains only type 6A will serve to induce the maximum amount of serum antibodies to both group 6 organisms.


Effect of growth rate and glucose concentration on the biochemical properties of Streptococcus mutans Ingbritt in continuous culture. A comparison was made of the properties of Streptococcus mutans Ingbritt grown in continuous culture under conditions of excess glucose (nitrogen limitation) and limiting glucose at mean generation times of 1.7 to 14 h. Only low levels of glucoamylase-specific glycogen were formed in cells from either culture, and the total carbohydrate content of the cells under excess glucose was only at most 1.6-fold higher than in the glucose-limited culture. A negligible amount of cell-free polysaccharide was formed in either culture, although a significant level of glucosyltransferase activity was observed in both, with the highest activity at D = 0.2 and 0.4 h(-1) with a glucose limitation. Other differences were observed. (i) Lactate was the main end product of the glucose-excess culture, whereas acetate, formate, and ethanol were the main products of the glucose-limited culture except at a mean generation time of 1.5, when lactate represented 30% of the products. (ii) The yield (in grams per mole of glucose) of the latter culture was 2.6- to 4.0- fold higher than the yield of the glucose-excess culture. (iii) Washed cells from the glucose-limited culture were much more acidogenic (1.7- to 6.2-fold) than the glucose-excess cells when incubated with glucose, sucrose, and fructose. Endogenous glycolytic activity by the latter cells was significant, being 31 to 92% of the exogenous glucose rate at the four dilution rates. (iv) Cells from the glucose-excess culture were more insensitive to fluoride than cells from the glucose-limited culture. The NaF 50% inhibition dose values for the effect of fluoride on the metabolism of glucose, sucrose, and fructose were calculated for the four dilution rates at four pH values. This analysis indicated that rapidly metabolizing cells were more sensitive to fluoride than cells that metabolized the sugars more slowly.


Effects of heat-stable enterotoxin of Yersinia enterocolitica on ion transport and cyclic guanosine 3',5'-monophosphate metabolism in rabbit ileum. Strains of Yersinia enterocolitica produce a heat-stable enterotoxin which is positive in the suckling mouse bioassay. Partial purification by a procedure previously worked out for heat-stable Escherichia coli enterotoxin yielded a substance which increases particulate guanylate cyclase activity and short-circuit current and inhibits active Cl-absorption in rabbit ileal mucosa. These effects of Y. enterocolitica enterotoxin are similar to those of heat-stable E. coli enterotoxin, suggesting a common mechanism of action.


Effects of the administration of artroglobina suppositories in healthy volunteers. Forty-nine healthy volunteers were treated daily with Artroglobina suppositories (anticartilage antiparathyroid immunoglobulins) for twelve days, in order to prove the absence of side effects. Volunteers were examined daily. Blood and urine samples were taken before the treatment, twelve days, twenty-four days and six months after the beginning of the treatment. Slight changes were noted in some parameters though remaining in the normal range and disappearing six months later. No symptoms of intolerance, toxicity or side effects were registered.


Congenital defect in intracellular cobalamin metabolism resulting in homocystinuria and methylmalonic aciduria. II. Biochemical investigations. Biochemical investigations are reported in an infant with methylmalonic aciduria and homocystinuria who died at 4 months of age. Postmortem analysis of liver obtained 2 weeks after the child was treated with vitamin B12 revealed deficient activity of both cobalamin dependent enzymes: N5-methyltetrahydrofolate: homocysteine methyltransferase (requiring Me-Cbl), and methylmalonyl CoA mutase (requiring Ado-Cbl). MMA-CoA mutase activity could be restored to normal in vitro by added Ado-Cbl, but MeTHF-HCy transferase activity was not significantly enhanced by addition of Me-Cbl. Though the serum total cobalamin was normal, total cobalamin in liver and kidney was abnormally low. In the kidney Me-Cbl and Ado-Cbl were disproportionally decreased whereas in the liver only Ado-Cbl was significantly reduced. This suggests that at least some of the CN-Cbl administered was converted to the coenzymes in liver which would explain the reduction of MMA- and HCy-excretion during therapy. The results show 1. that this infant suffered from a congenital defect in one of the steps of intracellular cobalamin metabolism or transport common to the synthesis of both coenzymes, 2. that life-long treatment with vitamin B12 (OH-Cbl) may be of value in similar cases, particularly if given early in the course of the disease.


Immunocytochemical localization of tyrosine hydroxylase, dopamine-beta-hydroxylase and phenylethanolamine-N-methyltransferase in the adrenal glands of the frog and rat by a peroxidase-antiperoxidase method. The subcellular localizations of tryrosine hydroxylase (TH), dopamine-beta-hydroxylase (DBH) and phenylethanolamine-N-methyltransferase (PNMT) in the adrenal glands of the frog and rat have been examined by a peroxidase-antiperoxidase (PAP) method. TH was localized in the ground substance of the adrenaline-containing cells and noradrenaline-containing cells, but not in the nucleus or in the mitochondria. TH was also located on the outside of the membrane of the chromaffin granules. DBH was observed only inside the granules. PNMT was found not only in the ground substance but also on the membrane of some adrenaline-containing granules. Cortical lipid cells of the frog adrenals did not show TH-, DBH-, and PNMT-reactions. The negative reactions to TH-, DBH-, and PNMT-antiserum exhibited by the summer cells of the frog adrenals prove that they belong to the cortical cells.


A cohort study of canine testicular neoplasia. A prospective epidemiologic study of canine testicular neoplasia was undertaken in the Philadelphia area in 1971, with the cooperation of private veterinary practitioners. By the end of 1975, 938 dogs had been monitored for an average of 2 years. The cohort consisted of 609 cryptorchid and 329 age- and breed-matched controls. The incidence of testicular neoplasia in the cryptorchid subcohort was 12.7/1,000 dog-years at risk. Testicular neoplasms did not develop in controls. A large proportion of the dogs were below the average age at onset for this neoplasm. Among dogs over 6 years of age, the incidence was 68.1/1,000 dog-years at risk. The incidence of Sertoli cell tumors and seminoma was approximately twice as high in dogs with unilaterally retained inguinal testicles as in abdominal cryptorchids. Sertoli cell tumors developed in 10 dogs and seminoma developed in 6. One half of the testicular neoplasms that developed did so within the first year of observation. This study demonstrated the feasibility of conducting prospective epidemiologic studies of canine diseases with the assistance of practicing veterinarians.


Acidic degredation of cephaloglycin and high performance liquid chromatographic determination of deacetylcephaloglycin in human urine. In order to provide fundamental knowledge about the determination of deacetylcephaloglycin excreted in human urine as an active metabolite of cephaloglycin, the degradation of cephaloglycin in acidic media has been investigated with varying reaction temperature between 30 degrees and 50 degrees C and pH between 1.2 and 2.8. The degradation pathway observed under these conditions was the elimination of the 3-acetyl group yielding deacetylcephaloglycin followed by formation of deacetylcephaloglycin lactone. Estimation of first order rate constants revealed that deacetylation is the rate-determining step for the degradation of cephaloglycin to the lactone. It is found from the kinetic results that reproducible assays of deacetylcephaloglycin excreted in urine can be achieved by a quantitative conversion to deacetylcephaloglycin lactone in a medium of pH 1.4 at 37 degrees C for 2 hours, followed by a reversed-phase ion-pair high-performance liquid chromatography. The utility of the present method is demonstrated by determining the time course of urinary excretion of deacetylcephaloglycin after oral administration of cephaloglycin capsule.


Biosynthesis of leupeptin. II Purification and properties of leupeptin acid synthetase. An enzyme which condenses acetyl-L-leucyl-L-leucine and L-arginine into acetyl-L-leucyl-L-leucyl-L-leucyl-L-arginine (leupeptin acid) was partially purified from a cell extract of Streptomyces roseus MA839-A1. With respect to this catalytic activity, the enzyme showed the following characteristics: ATP is essential; optimum pH is 9.5; the activity is inhibited either by EDTA or pyrophosphate or N-ethylmaleimide. The molecular weight of the enzyme is about 260,000 daltons. It also catalyzes some other extension reactions, such as, acetyl-L-leucine+L-leucine+L-arginine leads to leupeptin acid, and acetyl-L-leucine+L-leucine leads to acetyl-L-leucyl-L-leucine, but neither L-leucine+L-arginine leads to (L-leucyl)1--2-L-argining, nor acetyl-L-leucine+L-arginine leads to acetyl-L-leucyl-L-arginine. ATP-PPi exchange, catalyzed by this enzyme, proceeds with either acetyl-L-leucine, or acetyl-L-leucyl-L-leucine or L-leucine, but not with acetate or arginine.


An analysis of the buffer system in the rumen of dairy cattle. A method is presented for the analysis of buffer systems in the rumen using the first derivation of titration curves. Bicarbonate and volatile fatty acids (VFA) are the main components of the buffering system in the rumen fluid of dairy cattle under widely different feeding conditions. Phosphate from saliva is of little importance as a buffer, but neutralizes acids produced in the rumen. After studying five cows during the peripartal period a spontaneous and transient increase in the concentrations of VFA and a soluble marker (PEG) as well as a drop in pH and in the bicarbonate concentrations not related to feeding was observed in two animals that were sampled several hours before parturition. The potential risk of provoking rumen disturbances upon feeding animals close to the time of parturition, when buffering capacity may be minimal, is stressed.


Effect of dietary lactic acid on rumen lactate metabolism and blood acid-base status of lambs switched from low to high concentrate diets. Two experiments were conducted with ruminally fistulated wether lambs to determine the effect of lactic acid addition to a hay diet on rumen lactate metabolism, blood acid-base status and subsequent adaptation to a high concentrate diet. In Exp. 1, lambs were fed mature brome hay (H), H plus 5% (w/w) D,L lactic acid (H5L) or H plus 10% lactic acid (H10L) (three lambs per treatment) for 14 days (phase I) then switched to a 90% concentrate diet for 2 days (phase II). In Exp. 2, lambs were fed alfalfa-brome hay (H) (six lambs), H plus 2.5% lactic acid (H2.5L) (six lambs) or H plus 5% lactic acid (H5L) (four lambs) during phase I, then switched to a 70% concentrate diet (3 days) followed by a 90% concentrate diet (10 days) (phase II). During both experiments rumen fluid samples were taken periodically for pH and lactate analyses and in vitro L- or D-lactate disappearance (IVLD) studies. Blood samples were taken to measure acid-base status, serum lactate, and serum calcium, magnesium and phosphorus. Dietary lactic acid enhanced IVLD during phase I of both experiments. L and D isomer IVLD rates were similar and followed zero-order kinetics. In Exp. 2, IVLD increased rapidly during phase II in response to increased concentrate level in the diet; the enhanced rates of H2.5L and H5L lambs were sustained for the first 3 days of phase II. Blood data from both experiments indicated a deleterious effect of dietary lactic acid on blood acid-base balance; however, this treatment effect was not manifested in any symptoms of acute acidosis. There was a decrease (P less than .05) in serum calcium during phase II of both experiments. In Experiment 1, serum calcium increased linearly (P less than .05) in response to dietary lactic acid level. In Exp. 1, rumen fluid total lactate and L-lactate were lower (P less than .05) for H5L vs H lambs during phase II. However, all lambs in Exp. 1 experienced acute acidosis; four of the nine lambs subsequently died. There was evidence of acidosis in Exp. 2, but there were no clear treatment effects during phase II on rumen fluid pH or lactate, or feed intake. All lambs adapted to the high concentrate diets as evidenced by rumen lactate levels and feed intakes. In both experiments, the proportion of L-lactate in rumen fluid decreased from almost 100 to about 50% of total lactate by the end of phase II.


Quantitative and dynamic aspects of nitrogen metabolism in the rumen: a modeling analysis. A modeling approach to analyses of ruminant digestive function is described. The approach provides for evaluations of current concepts and data for adequacy as explanations of behavior of the real system. It also provides for evaluation of hypotheses for probable adequacy as explanations of real behavior where proven concepts fail, and for identification of critical experiments. Analyses of several aspects of microbial maintenance and growth and dietary protein degradation are emphasized.


Colorimetric and spectrophotometric determination of total and non-phenolic alkaloids in ipeca and its formulations. A simple method, requiring no chromatographic separation, is presented for the determination of the total and non-phenolic alkaloids in ipeca and its preparations. The complex formed between the alkaloid and methyl orange at pH 5.0 is extracted with chloroform and treated with 0.1N NaOH. The liberated dye, determined at 460 nm, is a measure of the total alkaloids. The chloroform phase remaining is treated with 0.1N H2SO4, and the acid extract is measured at 283 nm for the non-phenolic alkaloids, calculated as emetine. The proposed method was successfully applied to samples of ipeca powder, ipeca tincture, and 3 British Pharmaceutical Codex mixtures containing ipeca tincture, namely, ipecacuanha mixture, pediatric; ipecacuanha and ammonia mixture, pediatric; and belladonna and ipecacuanha mixture, pediatric. The proposed method compares favorably with the Egyptian Pharmacopoeia, British Pharmacopoeia, and USP methods and has a relative standard deviation of 1.54%. The present procedure is less time-consuming and requires about 45 and 90 min for the assay of ipeca tincture and powder, respectively. Only a small sample (0.2 mL tincture of 1.0 g powder) is required.


Ion-exchange chromatographic separation and fluorometric detection of guanidino compounds in physiologic fluids. A high-performance liquid chromatographic procedure has been developed for the separation and fluorometric detection of guanidino compounds in physiologic fluids. All guanidino compounds were separated on a 17 X 0.46 cm cation-exchange column using a stepwise pH gradient. The chromatographic system was designed to enable the use of the specific reagent 9,10-phenanthrenequinone as a means of monitoring the guanidino compounds of physiologic fluids. This new analytical method is so sensitive that it enables the analysis at the picomole level. Our automatic guanidino-compound analyzer was successfully applied to the quantitative determination of all guanidino compounds in physiologic fluids from normal controls and uremic patients.


Artifacts produced by using dichloromethane in the extraction and storage of some antihistaminic drugs. The use of dichloromethane for the extraction of amines may lead to the formation of artifacts. Dichloromethane reacts rapidly (37.5 degrees) with brompheniramine, diphenylpyraline, cyclizine, cyproheptadine but not with antazoline and lignocaine. This difference in reactivity as well as the thin-layer chromatographic, gas-liquid chromatographic, nuclear magnetic resonance, and mass spectrometric characteristics of cyclizine and diphenylpyraline chloromethochlorides are investigated.


Simple preparation of a bonded cation-exchange packing material and its application to the separation of phenothiazines by high-performance liquid chromatography. A method for introducing sulphonic acid groups on to a microparticulate silica is described. This involves formation of a mercaptopropyl bonded phase followed by oxidation to the corresponding sulphonic acid. The packing material so prepared displays cation-exchange properties and has been used to separate phenothiazines.


Determination of butaperazine in biological fluids by gas chromatography using nitrogen specific detection system. A simple and sensitive gas chromatographic method for the quantitative determination of butaperazine in biological fluids is described. The use of a nitrogen specific detector reduces the number of interfering peaks, thereby increasing the number of samples that can be analyzed. When butaperazine is extracted from 2 ml of plasma, the coefficient of variation is 7.4% over the concentration range of 5-180 ng/ml. The method was used to measure the levels in plasma and red blood cells in schizophrenic patients treated with butaperazine. It was also extended to measure butaperazine levels in rat red blood cells, plasma, liver, and brain after intraperitoneal injection (15 mg/kg).


Enzyme-linked immunosorbent assay for measurement of antibodies against pneumococcal polysaccharide antigens: comparison with radioimmunoassay. An enzyme-linked immunosorbent assay (ELISA) for measuring antibodies against each of 14 polysaccharides in contemporary pneumococcal vaccine is described, and the findings of tests of paired sera from vaccinated human subjects are compared with those obtained by radioimmunoassay. The findings were in very poor agreement, and this appears to be due to the lesser ability of the ELISA procedure to measure antibody of low avidity. The ELISA procedure described here is not considered to be a satisfactory substitute for radioimmunoassay for measuring antibody responses to pneumococcal vaccine.


Effects of blood on blood culture medium. The morphological and biochemical changes that occur after inoculation of sterile blood into a blood culture medium (tryptic soy broth) with sodium polyanetholesulfonate and CO(2) were investigated. Cellular changes, pH, PCO(2), and PO(2) were monitored and evaluated. Erythrocytes became crenated and developed precipitated hemoglobin inclusions within 4 h. The lymphocytes appeared morphologically intact at 24 h, and by 48 h a few cells had undergone transformation. Many neutrophils were vacuolated at 24 h. Neutrophils capable of phagocytizing Staphylococcus aureus were observed after 18 h of incubation. Identifiable eosinophiles were present on day 6 of the study. A decrease in PO(2) in the unvented bottles from 44.4 to 8 mm of Hg occurred by 24 h. PO(2) remained low for 6 days, after which a slight increase occurred. An increase in PO(2) in the vented bottle from 51 to 58 mm of Hg occurred by 24 h of incubation. In both the vented and unvented bottles the PCO(2) increased. This increase was markedly more rapid in the unvented bottle. From a pH of 7.06 a decrease occurred for the first 24 h after inoculation, with the pH stabilizing at 6.8 in the vented bottles and at 6.6 in the unvented bottles. The biochemical changes that occurred in the vented culture bottles stabilized more rapidly than those of the unvented bottles. Changes caused by the addition of sterile blood to a blood culture medium resulted in conditions which departed considerably from accepted optima for the isolation of clinically important microorganisms. The phagocytosis of organisms that occurred may also have reduced the yield.


Suppression of histamine-induced pruritus by hydroxyzine and various neuroleptics. This study evaluates the ability of hydroxyzine and various neuroleptics to suppress histamine-induced pruritus in ten volunteer subjects with the use of a double-blind crossover protocol. The itch threshold was determined in each volunteer by intradermal injection of gradually increasing concentrations of histamine. Volunteers were then given the study drugs and placebo at the same interval of time, under near identical conditions, and the itch threshold was determined. Thiothixene, hydroxyzine hydrochloride, chlorpromazine, thioridazine, and a lactose placebo were evaluated. Compared to other drugs, hydroxyzine alone was more effective in the suppression of histamine-induced itch. Consequently, hydrozyzine may be more effective in histamine-induced pruritus. The neuroleptic drugs used in this study do not significantly suppress histamine-induced pruritus, but they may be beneficial in nonhistamine-induced pruritus or psychogenic pruritus.


Clostridium perfringens type A: certain characters of epidemiologic significance. Survival of spores of Cl. perfringens type A was significantly greater than that of vegetative cells in acid pH (pH 1.2). Survival of spores in soil varied from strain to strain. Time required for 5 million spores to be reduced to 500 per gram of soil varied from 2 to 8 months with an average of 4.5+/-2.3 months. Quantitative and qualitative heat resistance studies revealed that a majority of the Indian and all the American strains tested were heat-sensitive. These characters of Cl. perfringens have an important bearing on the epidemiology of food poisoning due to this agent.


Experimental otitis media in chinchillas following nasal colonization with type 7F Streptococcus pneumoniae: prevention after vaccination with pneumococcal capsular polysaccharide. Chinchillas were colonized intranasally with type 7F Streptococcus pneumoniae, and pneumococcal otitis media developed in greater than 50% of the animals during the first week after negative middle ear pressure (-25 mm Hg) was briefly applied. Twenty-three chinchillas were vaccinated subcutaneously with the capsular polysaccharde of type 7F S. pneumoniae to determine whether vaccination could prevent the development of experimental otitis media. Following vaccination, 14 animals seroconverted with at least a twofold rise in serum antibody concentration; nine animals that were vaccinated did not seroconvert. All of 23 vaccinated animals and 42 of 42 unvaccinated control animals became colonized after intranasal inoculation with pneumococci. Only one (7%) of the vaccinated seroconverting animals developed pneumococcal otitis media, whereas 26 (62%) of the control animals developed middle ear infection with type 7F pneumococci. Four (44%) of nine vaccinated nonseroconverting animals developed pneumococcal otitis media. Protection was associated with high levels of serum antibody prior to intranasal inoculation. Higher antibody levels were found in sterile middle ear effusions than in S. pneumoniae-infected effusions. Vaccination with type 7F pneumococcal capsular polysaccharide significantly reduced the incidence of pneumococcal otitis media following intranasal inoculation of type 7F S. pneumoniae in chinchillas.


Triploids and male determination in the mosquito, Anopheles culicifacies. One family of Anopheles culicifacies was isolated in which all individuals examined (4 out of 13) were triploid. The one male examined was XXY, suggesting that unlike Drosophila the Y chromosome in this species may be important in male determination.


Failure of pyridoxine to effect neuroleptic-induced hyperprolactinemia in psychotic patients. The effects of acute and repeated administration of pyridoxine on serum prolactin levels were studied in 18 chronic schizophrenics, 10 women and 8 men, in whom hyperprolactinemia had been induced by long-term treatment with phenothiazines, haloperidol, sulpiride or clopentixol. The patients were divided into 5 groups: group 1 received 300 mg of the vitamin per os in a single dose; group 2 received 300 mg of the vitamin per os for 7 days; group 3 received 300 mg of the vitamin iv as a single bolus; group 4 received 600 mg of the vitamin iv as a single bolus; group 5 received 1200 mg of the vitamin per os in a single dose. Prolactin levels were examined before and 15, 30, 60, 90, 120 and 150 min after the single administration of pyridoxine, either iv or per os, and on days 3,5, 7 of the chronic administration. There was no decrease in prolactin levels either after oral or iv administration of the drug, given in single or repeated doses. Therefore, this treatment is not useful for the suppression of hyperprolactinemia induced by neuroleptics.


Comparative gastrointestinal and biliary tract effects of morphine and butorphanol (Stadol). Butorphanol, levo-N-cyclobutylmethyl-3,14 beta-dihydroxy-morphinan, is a new agonist-antagonist type analgetic agent which is 4 to 8 times more potent than morphine in experimental animals and man. Butorphanol and morphine were evaluated in mice and dogs for their gastrointestinal and biliary tract smooth muscle effects. Morphine decreased the propulsion of a charcoal meal through the gastrointestinal tract of the mouse in a dose related manner with the maximal inhibition obtainable being 90%. Butorphanol produced a maximal inhibition of motility of only 40% with very high doses producing less of an inhibitory effect than lower doses. In dogs, morphine caused a dose-dependent increase in duodenal smooth muscle activity and a dose-dependent decrease in bile duct flow. A clinically effective i.v. dose of morphine (0.1 mg/kg) produced a statistically significant spasmogenic effect on dog biliary tract and gastrointestinal smooth muscle, while a clinically effective equianalgetic i.v. dose of butorphanol (0.025 mg/kg i.v.) had little or not effect on the biliary or gastrointestinal systems. These findings indicate that at equianalgetic doses, butorphanol should produce less constipation and less biliary tract and gastrointestinal smooth muscle spasm than morphine.


Glutamine metabolism in nitrogen-starved conidia of Neurospora crassa. During nitrogen deprivation, de novo synthesis of glutamine synthetase was induced in non-growing conidia of Neurospora crassa. When ammonia or glutamine was added to conidia which had been deprived of nitrogen, glutamine and arginine accumulated at a higher rate than in condia not deprived of nitrogen. The degradation of exogenous glutamine to glutamate is apparently a necessary step in the accumulation of glutamine and arginine within the conidia. In non-growing conidia, a cycle probably operates in which glutamine is degraded and resynthesized. The advantages of such a cycle would be that the carbon and nitrogen could be used to synthesize amino acids in general, as well as for the synthesis and accumulation of arginine and/or glutamine in particular.


Adenosine triphosphatase activity of Tritrichomonas foetus. Homogenates of Tritrichomonas foetus exhibited a Mg2+-dependent adenosine triphosphatase (ATPase) activity, with a pH optimum in Tris buffers of 8.2 to 8.3. The activity was not sensitive to oxygen. At high concentrations, quercetin and 4-chloro-7-nitrobenzofurazan inhibited ATPase activity in the cytoplasmic extract by 20 and 70%, respectively, whereas oligomycin, venturicidin, triethyltin, leucinostatin, dibutylchloromethyltin chloride, spegazzinine, efrapeptin, citreoviridin and sodium azide had no effect and N,N'-dicyclohexylcarbodi-imide stimulated the activity somewhat. The activity was localized in a population of small cytoplasmic particles which also contained an acid phosphatase. There was no indication of an association of ATPase with hydrogenosomes. The ATPase activity (or activities) in this aerotolerant anaerobe is different from the ATPases characteristic of mitochondria or of anaerobic bacteria.


Purification of pili from Bacteroides nodosus and an examination of their chemical, physical and serological properties. Pili from Bacteroides nodosus were purified to greater than 99% homogeneity by precipitation at pH 4.0 and in MgCl2 followed by chromatography on BioGel A150. The pili were composed entirely of one type of polypeptide subunit, pilin. No carbohydrates, nucleic acid, lipid, lipopolysaccharide or phosphate could be detected in purified pili preparations. The molecular weight of pilin from B. nodosus strains 91B and 198 was 18,400 and from strain 80 was 19,300. The isoelectric points of pili from B. nodosus strains 91B and 80 were both 4.5. The buoyant densities of pili from strains 91B, 80 and 198 were 1.287, 1.284 and 1.286 g ml-1, respectively. The three strains of B. nodosus did not cross-react in K-agglutination tests and produced pili which did not cross-react in immunodiffusion tests. Antiserum to highly purified pili caused a characteristic K-type agglutination reaction. It was concluded that pili are the K-agglutinogen.


Taxonomic and serologic studies on Micropolyspora faeni and Micropolyspora strains from soil bearing the specific epithet rectivirgula. The results of serological studies on six strains of Micropolyspora faeni from hay, sputum and plant debris, and five strains of Mip. rectivirgula from soil indicated no significant differences between the two species. Antisera raised in rabbits against purified antigens of the type strains were used to compare the 11 strains by immunoelectrophoresis. The detailed antigenic composition of the type strains was also determined by two-dimensional immunoelectrophoresis against specific rabbit antisera and pooled serum samples from patients suffering from farmer's lung. Cross-reacting antigens were identified by intermediate gel immunoelectrophoresis. The close similarity of the two species was confirmed by the results of 60 morphological physiological and biochemical tests applied to the 11 strains. We consider that the strains belong to a single species and propose that the specific epithet faeni be conserved for the taxon.


Purification and partial characterization of hyaluronate lyase (EC 4.2.2.1) from Propionibacterium acnes. Hyaluronidase from Propionibacterium acnes has been purified 13,000-fold from the culture supernatant to homogeneity (as determined by polyacrylamide disc gel electrophoresis). The molecular weight of the purified enzyme was 85,110 as determined by gel filtration. The purified enzyme had a pH optimum at 6.4, was stable between pH 5 and 5.8 and was completely inactivated after 15 min at 50 degrees C. Preliminary studies suggested that the enzyme is active against chondroitin 4- and 6-sulphates, but not against dermatan sulphate. Analysis by paper chromatography of the reaction products from the degradation of hyaluronic acid by bacterial, testicular and P. acnes enzymes suggested that the P. acnes enzyme is similar in its mode of action to other bacterial hyaluronate lyases. The enzyme from P. acnes may thus be tentatively classified as a hyaluronate lyase.


Properties of a gonococcal inhibitor produced by Escherichia coli. Strains of Escherichia coli can inhibit the in vitro growth of Neisseria gonorrhoeae. One E. coli strain released a potent agar-diffusible gonococcal growth inhibitor which was extracted and assayed in an agar well assay system. The culture conditions necessary to produce the inhibitor were determined. The inhibitor was bacteriostatic, in most cases, for N. gonorrhoeae. Based on ultrafiltration and column chromatography, the inhibitor appeared to have a molecular weight in the range of 1200 to 2000. Evidence that the molecule contained charged sites was obtained by membrane binding and column chromatography. The inhibitor was stable to extremes of heat, cold and pH. It was not volatile or susceptible to proteolytic enzymes, lysozyme, lipase, DNAase, RNAase or certain chelating agents. Its activity was completely blocked by ferric ammonium citrate. This inhibitor is dissimilar to previously reported gonococcal inhibitors of bacterial origin.


Physico-chemical characterization and partial purification of mouse immune interferon. Mouse immune (type T) interferon was produced from suspensions of spleen cells (1 X 10(7) cells/ml) treated with 3 micrograms/ml of phytohaemagglutinin. The crude interferon was chromatographed on four sorbents with varying affinities, namely concanavalin A-Sepharose, Affi-Gel 202, Blue Sepharose CL-6B and Phenyl-Sepharose CL-4B. With each of these the interferon activity was observed to have considerable heterogeneity. By means of affinity chromatography, mouse immune interferon was purified 100 to 200 times with concomitant complete recovery of activity.


Biophysical properties of a non-cultivable 29-nm enteric virus. A 29 nm non-cultivable virus (NCV) was detected in faecal extracts from children hospitalized for gastroenteritis. The NCV had a density of 1.35 g/ml in glycerol-potassium tartrate density gradients and was resistant to degradation by proteolytic enzymes, non-ionic detergents and pH extremes. The surface of these virus particles had knob-like projections which appeared to have a symmetrical arrangement. When heated to 56 degrees C, the virus was completely degraded to soluble components which could not be seen by electron microscopy.


Direct and reflex effects of nitroglycerin on the blood volume distribution, evaluated by regional weighing in the cat. The effects of nitroglycerin on the blood volume distribution were studied with a method of regional weighing in the anaesthetized cat. An i.v. bolus injection of nitroglycerin produced a dose-dependent decrease in arterial pressure accompanying a decrease in the thoracic blood volume. The latter change was associated with blood volume increases principally in the abdomen, and slightly in the hindquarters. Elimination of the cardiovascular reflex effects by carotid sinus denervation and cervical vagotomy signficantly enhanced and prolonged the following changes: the hypotension, the decrease in thoracic blood volume and the volume pooling in the abdomen. The magnitude of increase in hindquarters blood volume was not significantly affected by the denervation procedures, but the duration was much prolonged. The results indicate that the major site of nitroglycerin-induced venous pooling is in the splanchnic circulation. The peripheral venous pooling is produced at the expense of a decrease in the central or pulmonary blood volume. The secondary reflex adjustments tend to minimize the direct effects of nitroglycerin on the blood pressure and blood volume distribution.


Reduction in blood pressure in normal and spontaneously hypertensive rats by lergotrile mesylate. The acute administration of lergotrile mesylate to normal or spontaneously-hypertensive rats (SHR) causes a prompt reduction in blood pressure. In SHR, doses as small as 0.05 mg kg-1 are effective; maximal reductions in blood pressure are obtained at a dose of 0.25 mg kg-1. Lergotrile may be administered intraperitoneally or orally. Its efficacy as an antihypertensive agent in rats does not diminish significantly when administered twice daily for 7 days. Although lergotrile has been reported to be a dopamine agonist, the present data do not establish the mechanism by which the drug lowers blood pressure.


Release of [3H]noradrenaline induced by 5-hydroxytryptamine from cat pial arteries. Pial arteries of cats were used to analyse the effects of 5-hydroxytryptamine (5-HT) on the release of [3H]noradrenaline. To achieve this the vessels were preincubated with [3H]noradrenaline and the effect of different concentrations of 5-HT (10(-6), 10(-5), 10(-4) M) on the release of tritium was studied. 5-HT elicited release of radioactivity in a dose-dependent manner. Removal of both superior cervical sympathetic ganglia 15 days before the experiment of pretreatment of the animals with reserpine (3 mg kg-1, total dose) produced a significant decrease in the outflow of tritium induced by 5-HT. In these arteries, the amount of radioactivity retained at the end of the experiment was much diminished. Cocaine (10(-6) M) caused a significant decrease in the tritium efflux induced by 5-HT (1"0(-5) M). These results show that 5-HT has an indirect adrenergic effect in the pial arteries of the cat only at high doses of 5-HT, and confirm that sympathetic innervation of these vessels mainly comes from the superior cervical ganglia.


Methysergide induces selective potentiation in cholinergic contractions of the guinea-pig vas deferens by facilitating acetylcholine release. Methysergide (3 x 10(-6) M) enhanced the contractile responses of the isolated stripped vas deferens of guinea-pig to acetylchline(ACh) and arecoline, but not those to noradrenaline, tyramine and bradykinin. Methysergide (3 x 10(-5) M) suppressed the contraction elicited by noradrenaline or histamine. The methysergide-induced potentiation of the response to ACh was prevented by pre-addition of hemicholinium but not by tetrodotoxin or morphine. The augmentation of the response to ACh by physostigmine was unaffected by hemicholinium. The phasic contraction of the tissue elicited by 30 mM KCl was also enhanced by methysergide, and this enhancement was prevented by the pre-addition of atropine (1.4 x 10(-7) M). In the depolarized vas deferens after exposure to 30 mM KCl, methysergide occasionally induced a sustained tonic contraction which was inhibited by atropine. These findings suggest that methysergide facilitates a release of ACh by acting on the cholinergic nerve terminals and selectively potentiates the cholinergic response.


The effects of bromocriptine on pre-synaptic and post-synaptic alpha-adrenoceptors in the mouse vas deferens. Noradrenaline (NA) and dopamine (DA) contracted the mouse vas deferens and reduced the responses to low frequency nerve stimulation (0.1 Hz). The relative potencies of antagonists suggested that these effects were due to stimulation of post-synaptic and pre-synaptic alpha-adrenoceptors respectively. Bromocriptine produced a non-competitive antagonism of contractile responses to NA (pD2' = 7.6) and DA (pD2' = 8.0) but had no effect on responses to carbachol. Bromocriptine also reduced single twitch responses of the vas to low frequency field stimulation (0.1 Hz), but did not affect stimulation at higher frequencies (1--20 Hz). Yohimbine selectively and rapidly reversed the inhibiting effects of bromocriptine on single twitches, although they could not easily be reversed by washing. Bromocriptine produced a yohimbine-reversible reduction in the stimulated overflow of tritium from vasa previously loaded with 3H--NA. Thus the mouse vas deferens does not appear to contain specific DA receptors and the results suggest that bromocriptine acts as a pre-synaptic alpha-adrenoceptor agonist and post-synaptic alpha-adrenoceptor antagonist in this tissue.


The neuromuscular and autonomic blocking activities of pancuronium, Org NC 45, and other pancuronium analogues, in the cat. Twenty-six mono- or bis-quaternary salts of 3,17-dioxy-2 beta, 16 beta-dipiperidino-5 alpha-androstanes (including pancuronium) and one 17-desoxy congener were tested for neuromuscular blocking and autonomic blocking activities in the chloralose-anaesthetized cat. The 17 beta-acetoxy series, all the members of which contain an acetylcholine-like fragment in the steroidal D-ring, was most selective for effecting neuromuscular blockade. The salient member of this series is 3 alpha, 17 beta-diacetoxy-2 beta, 16 beta-dipiperidino-5 alpha-androstane 16 beta-N-monomethobromide (Org NC 45) which is highly selective in blocking neuromuscular transmission in that a dose approximately sixty times greater than the neuromuscular blocking dose was required to block responses to vagal stimulation. In contrast, in doses sufficient to produce neuromuscular block, pancuronium simultaneously blocked responses to vagal stimulation. Moreover, pancuronium and Org NC 45 exhibited the same order of neuromuscular blocking activity and therefore the latter potentially represents a useful addition to the armamentarium of neuromuscular blocking agents currently in clinical use.


Stereoselectivity of some oxotremorine antagonists containing two chiral centres. The stereoisomers of some analogues of oxotremorine containing two chiral centres, one in the 1-position of the butynyl chas are oxotremorine antagonists. They show a marked stereoselectivity, which depends mainly on the configuration of the chiral centre in the butynyl chain and to a lesser extent on the configuration of that in the pyrrolidine ring.


Apparent dose-dependent absorption of chlorothiazide in dogs. The purpose of this study was to determine the effect of oral dose on the absorption of chlorothiazide in the dog. Chlorothiazide was quantitatively excreted in the urine after administration of 50-mg and 250-mg intravenous doses. In contrast, the urinary recovery of chlorothiazide after oral administration showed appreciable interanimal variation and decreased from 70.4% to 26.7% on the average as the oral dose was increased from 125 mg to 750 mg. Oral administration of a single 15-mg dose of propantheline bromide (a potent inhibitor of gastric emptying and intestinal motility) at--1 hr increased the absorption of a 250 mg oral dose of chlorothiazide in three out of four dogs. These results suggest that chlorothiazide absorption is dose dependent and apparently site specific.


Plasma levels and beta-blocking effect of alpha-hydroxymetoprolol--metabolite of metoprolol--in the dog. The plasma levels and the beta-blocking effect of metoprolol and its active metabolite alpha-hydroxymetoprolol have been studied after i.v. bolus injections of the substances to dogs. For both substances the beta-blockade increased with the dose, and there was a linear relationship between percent reduction in exercise heart rate and the logarithm of plasma concentration. The dose of the metabolite, however, had to be 5 times higher than that of metoprolol to induce the same degree of beta-blockade. Because of differences in the volume of distribution, 2.0 liters/kg for alpha-OH-metoprolol and 3.5 liters/kg for metoprolol, the 5 times higher dose of alpha-OH-metoprolol resulted in 10 times higher plasma levels of the metabolite than of metoprolol. alpha-OH-Metoprolol was more slowly eliminated (t1/2 approximately 7.0 hr, total body clearance approximately 3.5 ml-kg-1-min-1) than metoprolol (t 1/2 approximately 2.0 hr, total body clearance approximately 20.0 ml-kg-1-min-1). Approximately 5% of an i.v. dose of metoprolol was metabolized to alpha-OH-metoprolol. The half-life of the endogenously formed metabolite was the same as after an i.v. dose of the compound.


Effect of interaction of aluminum hydroxycarbonate gel and magnesium hydroxide gel on acid neutralization. Acid neutralization by mixtures of aluminum hydroxycarbonate gel and magnesium hydroxide gel differs from the sum of the acid neutralization of each gel. Acid neutralization by magnesium hydroxide gel in the mixture is not observed until after a substantial portion of the aluminum hydroxycarbonate gel has reacted with acid, even though magnesium hydroxide gel is the faster reacting of the two gels. It is hypothesized that amorphous aluminum hydroxycarbonate forms a coating on the crystalline magnesium hydroxide particles due to electrostatic attraction. This coating prevents protons from reaching the highly reactive magnesium hydroxide until the coating is dissolved by the acid neutralization of aluminum hydroxycarbonate.


Use of an iodide-specific electrode to study lactoperoxidase-catalyzed iodination of l-tyrosine. An in vitro method employing an iodide-specific electrode for monitoring lactoperoxidase-catalyzed iodination is described. The method utilized lactoperoxidase, potassium iodide, and a glucose--glucose oxidase system for the generation of hydrogen peroxide and l-tyrosine. As iodination of l-tyrosine proceeded, the free iodide concentration in solution decreased and was monitored by an iodide-specific electrode. The iodide electrode was reliable when compared to a 131I-method for measuring free iodide changes in solution. Increasing concentrations of resorcinol, a well-known inhibitor of thyroid peroxidase-catalyzed iodination, in the reaction mixture resulted in graded inhibition of the initial rate of lactoperoxidase-catalyzed l-tyrosine iodination. This in vitro system can be used to assess inhibitory activity of various antithyroid substances.


Colorimetric determination of 1-(4'-nitrophenyl)-2-aminopropane-1,3-diol with 2,4,6-trinitrobenzenesulfonic acid in the presence of chloramphenicol. A colorimetric method based on the interaction between the chloramphenicol degradation product 1-(4'-nitrophenyl)-2-aminopropane-1,3-diol and the 2,4,6-trinitrobenzenesulfonic acid reagent was developed. Analytical solutions were reacted with the reagent at pH 9.1 for 20 min at room temperature, and the resulting color was measured at 340 nm. A linear relationship between absorbance and concentration occurred within the 5--25-micrograms/ml range under the conditions studied. Replicate analyses were in good agreement. An average recovery of 99.4 +/- 0.4% was obtained for the synthetic mixtures.


Mode of stimulation by injection of cyclic AMP and external acidification of the sodium efflux in barnacle muscle fibres. 1. A study has been made in single barnacle muscle fibres of the effect of micro-injected pure protein kinase inhibitor (PKI) on the response of the Na efflux to injection of cyclic AMP and external acidification. 2. (i) Injection into fibres of 1.6 x 10(-4) M-pure PKI is without effect on the resting Na efflux. (ii) Injection of 1.6 x 10(4) M-pure PKI before 0.03 M-cyclic AMP causes a marked reduction in the magnitude of the response of the Na efflux to the nucleotide. The same is true when 10(-4) M-cyclic AMP is injected after PKI. (iii) Injection of partially pure catalytic subunits causes a sustained stimulation of the ouabain-insensitive Na efflux, which is almost completely reversed by injecting PKI. (iv) Injection of 100 mM-EGTA before PKI fails to alter the lowered response of the ouabain-insensitive Na efflux to injection of 10(-4) M-cyclic AMP. (v) Ouabain (10(-4) M) when applied following the injection of 10(-4) M-cyclic AMP causes a drastic fall in the stimulated Na efflux. 3. (i) Injection of 1.6 x 10(-4) M-pure PKI before or after external acidification fails to abolish or reduce the stimulatory response to acidification. (ii) Injection of 1.6 x 10(-4) M-pure PKI before acidification practically abolishes the response of the ouabain-insensitive Na efflux to 0.03 M-cyclic AMP in the presence of acidification. (iii) Radioimmunoassay of total cyclic AMP and cyclic GMP content in single fibres before and after acidification shows no appreciable alteration in nucleotide content following acidificiation. (iv) Injection of 100 mM-EGTA before acidification enhances the stimulatory response to acidification. (v) External application of Dantrolene (10(-5) M) fails to alter the size of the stimulatory response to acidification. 4. (i) Prior external application of 5 x 10(-4) M-benzolamide results in a marked reduction in the magnitude of the response of the ouabain-insensitive Na efflux to the injection of 3 x 10(-4) M-cyclic AMP. (ii) Benzolamide totally abolishes the response of the ouabain-insensitive Na efflux to the injection of catalytic subunits. 5. The evidence brought forward is compatible with the view that (a) The mechanism by which cyclic AMP stimulates the Na efflux involves activation by cyclic AMP of the cyclic AMP-dependent protein kinase system, and hence release of the catalytic subunit, and (b) the mechanism by which external acidification leads to stimulation of the Na efflux involves activation of a benzolamide-sensitive system, possibly carbonic anhydrase, rather than the adenyl cyclase system. The actions of cyclic AMP and catalytic subunits on the Na efflux are closely linked to activation of the benzolamide sensitive system.


Behavioral treatment of transsexualism: a case report. Patients who believe themselves to be transsexual seek only confirmation of their diagnosis so that they may proceed with their pre-chosen course of management: hormones and surgery. Their syntonic emotional set generates resistance to any other therapeutic direction. Despite this attitude, it is the therapist's responsibility to assess each case individually and to decide, with the patient, on realistic goals even if they be different from the original one. This case emphasizes this need since a probing history revealed underlying conflict and anxiety related to severe homophobia. With revelation of the homophobia, various behavioral techniques could be used therapeutically. These resulted in acceptance of lesbianism as a life style. Careful assessment of patients with self-diagnoses of transsexualism can sometimes uncover a different etiology to which appropriate therapy can be applied.


Basic Health Services in Nigeria: models for primary care in America. The basic problems in delivery of health care in the United States and in Nigeria are similar; the major differences are in magnitude. Nigeria's Basic Health Services Scheme, now being implemented, is a bold effort to make quality health care accessible to the entire population. American health planners should look to such developing countries for concepts adaptable to our own health care delivery system. In developing primary care programs in particular, they should consider three basic components of the Nigerian scheme: (1) delegation of appropriate responsibilities to non-physician health providers in order to augment physician manpower in underserved areas; (2) location of training centers in environments similar to those where the trainees will serve; and (3) use of home-based care records to increase patient participation in health care.


Measurement of the aquatic toxicity of volatile nitrosamines. The acute toxicity of N-nitrosodimethylamine (DMN) and N-nitrosodiethylamine (DEN) was determined for three groups of aquatic organisms: algae, invertebrates, and fish. Toxicity of DMN and DEN to algae was assessed as a repression in the growth rate of either Selenastrum capricornutum or Anabaena flos-aquae in static bioassay tests. DMN and DEN concentrations of 1-10 ppm depressed algal growth in all cases. Invertebrate toxicity was determined in 96-h static bioassay tests with Dugesia dorotocephala and Gammarus limnaeus. The data indicated that these organisms are not highly susceptible to nitrosamine toxicity. The 96-h LC50s for D. dorotocephala were 1365 and 1490 ppm for DMN and DEN, respectively. Similar studies with G. limnaeus indicated LC50s of 330 and 500 ppm for DMN and DEN, respectively. Fish toxicity was also determined in 96-h statis bioassays with the fathead minnow (Pimephales promelas). Acute toxicities were calculated as LC50s of 940 and 775 ppm for DMN and DEN, respectively. Algae were calculated as LC50s of 940 and 775 ppm for DMN and DEN, respectively. Algae were quite sensitive to relative low levels of volatile nitrosamines, but higher organisms (invertebrates and fish) were relatively insensitive.


Haemolysis of various mammalian erythrocytes in sodium chloride, glucose and phosphate-buffer solutions. Mouse, rat, rabbit, hamster, cow, pig, sheep, guinea-pig, dog and human erythrocytes were studied. A 0.9% or stronger solution of sodium chloride completely prevented haemolysis; sheep and pig erythrocytes appeared the more fragile, while human and dog erythrocytes were not haemolized in concentrations of 0.4% or more. Haemolysis of human, rabbit, cow, hamster, guineapig, pig and sheep erythrocytes was not observed in solutions of 0.4% or more of glucose. Except for sheep, human and dog erythrocytes, haemolysis was depressed in rate but not completely prevented by phosphate-buffer solution of pH 7.0.


[Results, problems and ways of future progress in the Hungarian trauma surgery (author's transl)]. Author after having given account of the importance of traumatology is dealing with the present situation of trauma care in Budapest and in the country side. The need of increasing the number of beds for trauma care is pointed out. Establishing regional centers would mean the up to date advancement in the management of injured. Out-patient treatment, instrumentation and postgraduated teaching is dealt with. Finally special profiles of trauma surgery are detailed the furthering of which seems especially important: intensive care, neurotraumatology, hand injuries and burns. As a final question the septic complications are mentioned meaning a problem not only for the traumatologist.


[Past and present of the Hungarian traumatology (author's transl)]. Author presents a consise picture about the history of the Hungarian traumatology. Remembering great Hungarian surgeons who recognised the significance of accident surgery and with their works contributed to the progress in traumatology, he delineates the development of the Hungarian trauma management. The second part of this paper is dealing with the present situation of the Hungarian accident surgery and outlines tasks to be realized for further development of the well advancing Hungarian traumatology.


[Why in the traumatologic research clinical results are not assessed in a satisfactory way? (author's transl)]. Author draws attention to the fact that in scientific works late results are not always taken in account in a satisfactory way. This is caused by the aversion of surgeons from the numbers by their ignoration of statistical laws and by the subjective judgment of some investigators. The importance of clinical investigations are emphasized in proving hypotheses and new methods. By exact evaluation of the result errors can be avoided.


[Primary treatment of complete fracture dislocation of the spine (author's transl)]. Author demonstrates from the point of view of primary treatment of three most severe cases of complete fracture dislocation of spine since the existence of the National Institute of Traumatology. All of them have been young persons which fact greatly contributed to their survival after polytraumatisation. The alteration of spine consisted in all three cases of complete disruption and slide indicating severe shearing force. In the literature available this type of injury and its treatment is hardly commented upon on reason that those injured used to die almost immediately after the trauma. The paper is dealing with primary treatment referring also to the great work of the Department of Paraplegics in the Central Institute of Rehabilitation where the rehabilitation of those three patients was carried out. One of the patients was over 16 years under the author's care, who treated him for late complications (pathological fractures etc.) as well.


[Microscopic surgery (author's transl)]. Author having reviewed the past of Hungarian hand surgery is dealing with the development and present state of microscopic surgery. The preconditions from point of view of the National Institute of Traumatology for realizing microscopic surgery in this country are summarized.


[Comparative analysis of the situation of intensive care (author's transl)]. Intensive care in Hungary in most of the medium size and major hospitals is provided on a proper level thanks for a great effort. The areal distribution of intensive care units is satisfactory. In case that a special intervention cannot be realized in one of the departments lack of instruments or expert personal, usually there is a possibility within 50--70 km distance in another unit to carry out this intervention. As a result of controlled development at the end of the sixth five year plan the units will come up quantitatively as well as qualitatively with all the needs for intensive care. In the present--let us hope transitory--situation the lack of expert personal, out of date hospital constructions and the great variety in types of instruments are meaning serious problems. The difficulties are exaggerrated by the fact that the work on an intensive care unit puts on an increased physical and psychological burden. As a result, in spite of all their interest and beauty the intensive care and its counter pair the anaesthesiology are among the less inviting professions with high incidence of migration and fluctuation of the personal.


[Treatment of pseudoarthrosis of the scaphoid bone by bone transplantation (modified Matti-Russe operation)]. Author reviews possibilities in the treatment of nonunited scaphoid fractures. Different techniques with cancellous bone grafting are described and his own function-stable grafting osteosynthesis presented. Indications for his operation are well delineated and with regards to good results their accurate application is emphasized. 352 cases were treated using his grafting method in 94 per cent good results was obtained passing by 14 per cent the average results of 80 per cent with other grafting techniques.


[The development in the treatment of supracondylar femur fractures (author's transl)]. Author compared the material of supracondylar femur fractures of the National Institute of Traumatology during a period of 5 years starting from 1962 until 1966 with that of a period of 6 years starting in 1972 until 1977. From among their 108 cases osteosynthesis was performed in 38 cases. He gives account of methods with wire pinning, medullar splinting, and nailing with Rush's, Jewett's and Küntscher's technique (transfixation of the knee in the latter) as well as of solutions with angulated nails performed mainly in the second period. For nonunion in this region external fixation was also used.


[X-ray and differential diagnosis of closed diaphragma ruptures (author's transl)]. In cases of thoraco-abdominal injuries and x-ray signs suspicious for diaphragma rupture, x-ray control should be as long repeated an all possible investigations carried out as the fact of diaphragma rupture is in a reassuring way not proved or ruled out.


[Management of trauma patients in the County of Veszprém (author's transl)]. Expert management of the increasing number of trauma patients is causing a problem all over the world. In Hungary centrally controlled, organized network of traumatology was set up. Author reviews mile stones in the management of injured in this country. The organization of the National Institute warranted the professional development and made available up to date treatment all over the country. As a basis for the treatment of trauma patients in the County Veszprém back in the year 1964 54 beds had been organized in the county hospital. Because of increased injury rate this number of beds was augmented to 66. Starting from 1972 a department with 66 beds in Ajka was established and since that time the management of injured is carried out in two departments with own territorial backgrounds. For an up to date treatment more surgeons would be needed. The postgraduate teaching of traumatology for surgeons and by this the provision for expert care of trauma patients in the County is due to professor Szántó.


[Operative treatment of acute acromioclavicular dislocation (author's transl)]. Author describes clinical appearance of the acromioclavicular dislocation. Discussing different surgical techniques, a new combination of surgical procedures is recommended. In case of acute acromioclavicular dislocation beside the osteosynthesis by means of traction wire loop, the reconstruction of the coracoclavicular ligament and the fixation of clavicula with tendon graft should be performed. Good results were obtained with the above method.


[Analysis of femoral shaft fractures (author's transl)]. Authors after having reviewed the literature analyse 157 healed cases from their own material of 198 femoral shaft fractures. Fractures between the subtrochanteric and supracondylar region has been considered. 2/3 part of the injured have been victims of traffic accidents, 1/6 part suffering severe polytraumatisation. From among the healed patients in 88 per cent an operation was carried out 76.8 per cent of which were medullary nailing following a broadened indication. In their patients treated surgically the fractures healed within 6 months in 75.4 per cent of cases. It is concluded that in the treatment of femoral shaft fractures the usual operative measures yield fast healing with good function. For multiple or comminuted fractures the use of other newer methods seems inevitable.


[Remarks to the operative treatment of arthritis of the knee (author's transl)]. Author deals with operative treatment in secondary arthritis following severe trauma of the knee. Review of operative techniques and their problemes are given. The results of and total prosthetic replacement are compared with those of arthrodesis. It is concluded that in order to obtain the best result in prevention of secondary arthritis exact reduction and stable fixation of the fracture should be performed.


[Infected pseudoarthroses of long tubular bones. (Principles of treatment and results in the material of the University Hospital) (author's transl)]. Based on clinical experiences author takes part for principles in the treatment of infected nonunions of long tubular bones. Main characteristics of his treatment are: Baside an approach saving the blood supply of bone, stable osteosynthesis, sequestrotomy and removal of necrotic tissues (focus elimination) filling in with autologous spongy bone chips the residual bone cavities, in acute stadium the application of PMMA balls as preparatory measure for a following cancellous bone, transplantation. Author comments at the same time on drainage, rinsing and sucking. Selective use of antibiotics may complete the therapy. Data of treatment in 42 cases of infected nonunions are given.


[Bolted medullary nailing (author's transl)]. Authors described the development of bolted medullary nailing and presents indications and technique. Points out the advantages of the procedure, but the difficulties in aiming, a basic part of the technique is also mentioned. With selfconstructed guide and other modifications the applicability of bolted medullary nailing was promoted. His experience of 112 operations is remarkable even in international relations. This material is analysed.


[Operative elimination of shoulder contracture maintained by subaxillary scarring (author's transl)]. In order to avoid irreversible changes the solving of shoulder contracture caused by subauxillary scarring is an urgent task. His method is the Z-plastic with many variables the choice and application of which are dependent from the extension and site of the subauxillary scar.


[Development in the treatment of congenital dislocation of hip (author's transl)]. Authors review the development in the treatment of congenital dislocation and dysplasy of hip and demonstrate the results with various methods. Results with treatment according to Lorenz (1040 cases) were unsatisfactory because of frequent occurance (66.3 percent) of osteochondritis. The treatment with Frejka cushion (360 cases) did not improve basically the results. Pavlik's strapping, especially when started under 1 month of age gave excellent results (3156 cases). In operative treated the results of 332 Colonna operations were unsatisfactory because of consecutive deformation of the hip and restriction of movements. Open reduction with varisation derotation osteotomy (474 cases) provided good acetabular development especially in cases where articular cartilage and upper limbus could be saved. Salter's operation is successful when proper indications are considered, but should be performed only after 1--2 years of observation of the acetabular development in non improving cases. The Chiari procedure can improve the weight bearing surfaces in cases of steep acetabular roof flat acetabulum and consecutive subluxation. In 557 cases of varisation derotation osteotomy subcapital coxa valga occured frequently. Basic principle of treatment remains prevention, early recognition and treatment after birth.


[The treatment of burns--an autonomous discipline (author's transl)]. In Europe Hungary was among the initiators to establish specialised surgical departments for the treatment of burnt patients. This new element in the structure of the hospital resulted in specialisation of surgeons for skin replacement and scar surgery. Thus the surgeon treating burnt patients is becoming the "surgeon of wounds" The treatment of thermal injury needs more detailed knowledge of biochemistry, pathophysiology and internal medicine compared with other field of surgery. The specialised thermist is in many respect similar to the intensivist.


[Choledochal obstruction due to Fasciola hepatica (author's transl)]. A 45-year-old woman was admitted in July, 1976 with an acute cholecystitis without jaundice. She had suffered from hepatic colic without fever, jaundice, diarrhea or allergic episodes for the past 8 years. The physical examination only revealed an elective pain on the cystic point. Laboratory data were unremarkable, except for a 12 percent eosinophils. The cholecystogram showed a cholelithiasis. The lithiasis was confirmed during the surgical operation and a fasciolasis was diagnosed after one and 10-12 parasites had been found into the cystic and common bile duct, respectively. A cholecistectomy and choledochoduodenostomy were performed. The patient was treated with 60 mg dehydroemetine during 10 days and 500 mg chloroquine during the other next 10 days. Eggs of Fasciola hepatica were found in the stool culture. The follow-up examinations 3 months and a year after surgery were completely normal. The national literature on this topic is reviewed and the clinical manifestations and therapy of this disease are commented on.


[Study of the acidification capacity of the urine in polyclonal hypergammaglobulinemia (author's transl)]. The renal capacity of acidification of urine was studied in 21 patients with hypergammaglobulinemia (polyclonal gammapathies) secondary to several conditions. The reabsorption threshold of bicarbonate was determined by Rodriguez-Soriano's technique; the acid and ammonium production was measured using Wrong and Davies' technique. Values for the bicarbonate threshold ranged from 18.5 to 29 mmol/liter of plasma bicarbonate with a mean value of 25 +/- 2.29; they were not significantly different from the control values. Urinary pH ranged from 4.6 to 5.8; ammonium excretion ranged from 18.4 to 114.5 microEq/min (mean value 63.01 +/- 25.36). Acidity values ranged from 13 to 65.05 microEq/min (mean value 33.61 +/- 19.36). Thhre was no statistical difference between these values and those obtained from the controls. We found no evidence that the acidification function of urine was limited in patients with hypergammaglobulinemia secondary to several conditions as compared with normal subjects. In the two cases in which this function was altered there was no reason why it should be attributed to hypergammaglobulinemia.


Phospholipid metabolism in Neisseria gonorrhoeae: phospholipid hydrolysis in nongrowing cells. Hydrolysis of cell envelope phospholipids was demonstrated in cells of both autolytic and nonautolytic strains of Neisseria gonorrhoeae that were labeled during growth in the presence of [3H] acetate. The label incorporated into the cellular phospholipids was located exclusively in the fatty acid acyl side chains. Labeled cells were incubated for 2 hr in N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid buffer, pH 8.5, containing various additions, and then examined for distribution of 3H in lipids. Ca++ selectively stimulated the deacylation of phosphatidylethanolamine (PE), whereas Mn++ stimulated the deacylation of phosphatidylglycerol (PG). Hydrolysis of phosphatidylethanolamine by phospholipase A was accompanied by the accumulation of lysophosphatidylethanolamine (LPE) and free fatty acids in the cells. Free fatty acids accumulated to a greater extent than lysophosphatidylethanolamine, suggesting that the latter was further hydrolyzed to glycerophosphorylethanolamine (GPE) and free fatty acids by a lysophospholipase. Methanol, ethanol, propanol, and isopropanol, added at concentrations which inhibited growth by 50%, stimulated phospholipase A, but not lysophospholipase activity. Differences in heat inactivation, metal ion requirements, and pH optima suggested that phospholipase A activities with phosphatidylethanolamine or phosphatidylglycerol as substrate and lysophospholipase may be separate enzymes.


Superhelical DNA in Streptococcus sanguis: role in recombination in vivo. Competent Streptococcus sanguis treated with non-lethal doses of coumermycin A1 immediately before or after uptake of radioactive transforming DNA were reduced in their capacity to yield transformants. This treatment did not alter bacterial ability to bind DNA in DNase I-resistant form, nor did it prevent the single-stranded donor DNA-recipient protein complexes formed upon uptake at the surface of the bacteria from translocating to chromosomal sites. Inhibition of transformation by heterospecific DNA was greater than that by homospecific DNA. The reduction in transformant yield was not accompanied by any loss of donor counts incorporated into the recipient chromosome, but rather by a loss of genetic activity of incorporated donor material indicating a failure of genetic integration and degradation of donor DNA as a consequence of coumermycin treatment. The inhibitory effect of coumermycin on transformation was associated with in vivo loss of chromosomal DNA superhelicity, The chromosomal DNA remained intact, however, indicative of inhibition of a gyrase-like enzyme responsible for the maintenance of negative supercoiling of the S. sanguis chromosome. Upon treatment with the drug, a coumermycin-resistant mutant strain showed neither loss of chromosomal superhelicity nor any inhibitory effect on genetic integration of donor DNA. The evidence supports the idea that chromosomal superhelicity promotes genetic recombination in vivo.


NADP-specific glutamate dehydrogenase is not involved in repression of yeast sporulation by ammonia. In Saccharomyces cerevisiae, the presence or absence of NADP-specific glutamate dehydrogenase does not affect inhibition of sporulation by ammonia, suggesting that the inhibition is not mediated by this enzyme.


Relationship between the antibacterial activity towards Escherichia coli NCTC 5933 and the physico-chemical properties of some esters of 3,4,5-trihydroxybenzoic acid (Gallic acid). Aqueous solubilities, oleyl alcohol: water and octanol: water partition coefficients, RM values, reduction in surface tension of water, relative antioxidant activities and pKa values, were determined for gallic acid and a series of its alkyl esters. Correlations were sought between these physico-chemical measurements and MIC, MBC and killing-rate determinations against Escherichia coli. Variations in antibacterial activity generally correlated well with partition parameters, but these correlations did not accurately predict the cut-off point in antibacterial activity.


[Effect of cultivation conditions on growth of Candida lipolytica yeasts and alpha-ketoacid biosynthesis in the presence of thiamine deficiency]. The effect of pH and aeration on the growth of Candida lipolytica and the biosynthesis of alpha-keto acids on acetate and glucose was studied in batch cultures at thiamine deficiency. If the initial thamine concentration was the same, then, irrespective of the carbon source, the yeast biomass in a medium saturated with oxygen by 5--10% was 1.5--2.0 times higher than in a medium with 60--90% [O2]. The rate of alpha-keto acid biosynthesis, on the contrary, decreased in the conditions of low aeration. The biomass increased and the rate of acid production decreased when pH was changed from 6.0 to 8.0 in a medium with acetate. However, at all studied values of pH and [O2] in a medium with either acetate or glucose, the growth at the deceleration phase was of a linear character, and was accompanied with the accumulation of alpha-keto acids in the cultural broth. The rate of acid production was maximal when the specific growth rate decreased to 0.04--0.06 hr-1. The presence of a linear phase in the conditions of thiamine deficiency suggests that here the growth of yeasts is determined by the constant activity of one of the thiamine dependent enzymes per unit volume of the cultural broth. However, the value of this activity seems to change depending on the cultivation conditions which, in turn, causes changes in the rates of biosynthetic processes.


[Fluorescent probe study of staphylococcal cell wall charges]. The effect of pH and MgCl2 concentration on the fluorescence of 1-anilinonaphthalene-8-sulfonate (ANS), an anion probe, was studied in suspensions of Staphylococcus aureus. The maximum of ANS fluorescence shifted towards the short wavelength region of the spectrum and its intensity increased when the cells were incubated in the medium with low pH and high ionic strength. The results are discussed in terms of electrostatic interactions between the anions of the probe and the negative space charge of the cell wall.


[Physiologo-biochemical features of the growth and development of Propionibacterium shermanii in egg albumin]. Propionibacterium shermanii was shown to be able of growing in such an unusual substrate as egg albumen. Incubation of the bacterium in egg albumen results in complete utilization of free carbohydrates in this substrate and accumulation in it of such bacterial metabolites as vitamins B2 and B12, free amino acids, bacterial protein, acetic and propionic acids which favour conservation of egg albumen. The bacterium does not possess proteolytic activity and does not utilize albumen; in the conditions of nitrogen deficiency, it can utilize free amino acids in the substrate. The growth of P. shermanii in egg albumen is stimulated by the addition of ammonium sulfate in combination with potassium phosphate or ammonium phosphate to the substrate. These substances change the direction of propionic fermentation toward the accumulation of propionic acid; at the same time, the content of glutamine, asparagine, alanine, methionine, cysteine and other free amino acids increases in the substrate.


Molecular aspects of cytochrome c oxidase: structure and dynamics. In the last few years much attention has been dedicated to the elucidation of some of the molecular aspects of cytochrome c oxidase. It has been shown conclusively that the enzyme from several sources (yeast, Neurospora, heart, liver) contains seven different subunits, which are asymmetrically inserted in the membrane. All of these are in contact with the lipid bilayer (except subunits V and VI) and to a greater or lesser extent with the water phase as well (except for subunit I). Subunit II of the enzyme appears to be involved in the formation of the binding site of cytochrome c. The location of the redox groups of the enzyme is still a matter of controversy. Their distance from the cytochrome c heme group is approximately 35 A such that electron tunneling appears to be the only possible mechanism for transporting electrons across such a distance. A proton pump appears to be associated with electron transport and approximately one proton is extruded per electron equivalent reducing oxygen via the enzyme. N,N', dicyclohexylcarbodiimide a well-established inhibitor of H+-translocating ATPases inhibits the proton pump and labels specifically subunit III of the enzyme.


Control of acetyl-CoA carboxylase by covalent modification. In this review, various experiments which establish the occurrence of covalent modification mechanisms, both in vivo and in vitro, in the control of acetyl-CoA carboxylase have been presented. It is interesting to note that phosphorylation of the carboxylase results in disaggregation of the active species. These studies indicate that aggregation and disaggregation of the enzyme are involved in the control of carboxylase activity. Our covalent modification mechanism and the allosteric control mechanism share a common ground in that both mechanisms affect the equilibrium between protomers and polymers of the enzyme. However, it is clear that the allosteric control mechanism cannot function alone under normal physiological conditions. Covalent modification of the carboxylase is prerequisite for efficient functioning of the allosteric mechanism. There are many aspects of the regulation of acetyl-CoA carboxylase which require further clarification. However, it is now established that short-term control of acetyl-CoA carboxylase involves the covalent modification mechanism.


Growth regulation by nerve growth factor. Although a great deal of descriptive information has been obtained about the actions of nerve growth factor on its target tissues, its structure, its receptors, and even its biosynthesis, there is no clear understanding, as yet, of the intracellular events mediating its transcriptional involvements. Work in this laboratory over the past five years has uncovered a number of nerve growth factor-initiated intracellular changes in sympathetic neurons and other nerve growth factor-sensitive systems, and has provided a framework into which they might fit. This article is written in an attempt to collect the data in a single communication and to suggest at least one mechanism by which the nerve growth factor may work.


Effects of ring substitution on the pre- and postjunctional alpha-adrenergic activity of aryliminoimidazolidines. The pre- and postjunctional alpha-adrenergic agonist potency of a series of aryliminoimidazolidines was determined in the isolated rabbit ear artery. This series included clonidine, an antihypertensive agent thought to act by stimulating brainstem alpha-receptors and known to be a preferentially prejunctional alpha-adrenergic agonist. Although all of the compounds acted preferentially on the prejunctional alpha-adrenoceptor, ring substitution had a dramatic effect on both potency and the degree of selectivity. 2-(3,4-Dihydroxyphenylimino) imidazolidine was both the most potent and most selective prejunctional alpha-agonist in this series.


Increase by alpha-adrenolytic drugs of acetylcholine release evoked by field stimulation of the guinea-pig ileum. The release of acetylcholine evoked by field stimulation of the guinea-pig ileum (3 Hz) is increased by yohimbine and tolazoline but not affected by phentolamine. It is proposed that yohimbine and tolazoline by blocking alpha-adrenoceptors of the cholinergic nerves abolish the inhibition caused by endogenous noradrenaline, and thus facilitate the output of acetylcholine.


Specific binding of 3H-adenosine to rat brain membranes. The binding of 3H-adenosine to rat brain membranes was studied by a microcentrifugation technique. Specific binding of 3H-adenosine was rapid, reversible, saturable and dependent on pH and temperature. Scatchard plots of equilibrium binding data were nonlinear suggesting the existence of two different binding sites for adenosine. The dissociation constants (Kd) were 1.7 muM and 13.6 muM and the maximal number of binding sites (Bmax) 31 and 165 pmol adenosine bound per mg of membrane protein. Ten adenosine derivatives were studied for their ability to compete with 3H-adenosine binding. The phosphorylated adenosine compounds 5'-AMP, cyclic AMP and ATP were most potent in displacing 3H-adenosine from its binding sites and the IC50-values ranged from 11--25 muM. N6-Phenylisopropyladenosine produced only partial inhibition (30%) of 3H-adenosine binding and no stereospecific difference between the (-)- and (+)isomer was observed. Several methylxanthines known as adenosine antagonists competed for the 3H-adenosine binding sites parallel with their pharmacological potency. The results offer a first approach for the study of adenosine binding sites in brain membranes.


Influence of dopamine synthesis on methamphetamine-induced changes in striatal and adrenal tyrosine hydroxylase activity. Methamphetamine in large doses decreases striatal tyrosine hydroxylase activity. This effect is prevented by neuroleptic agents such as chlorpromazine and haloperidol which would suggest that released dopamine may be involved in the response. To test this hypothesis, we have altered dopamine synthesis with alpha-methyl-p-tyrosine and L-Dopa and found that dopamine synthesis is necessary for the observed depression of striatal TH activity by methamphetamine. In the adrenal gland, however, the increase in TH activity by methamphetamine is not prevented by inhibition of catecholamine synthesis. It is possible that released dopamine may be inhibiting TH activity by activation of pre- or postsynaptic dopamine receptors in the neostriatum resulting in activation of the neuronal feedback pathway or released dopamine may act on dendrodendritic autoreceptors in the substantia nigra.


[Beta-blocking agents alone or combined with diuretics or alpha-adrenolytics in the treatment of essential arterial hypertension]. The Authors examine the antihypertensive effectiveness of the beta-adrenergic receptor blocking agents, according to their personal experience and a review of the bibliography. It is not cleared yet how these drugs reach their hypotensive effect. It is reasonable to assume however that several factors are involved: cardiac output, intravascular volume changes, plasma renin activity and peripheral resistance. Thirty patients suffering from essential hypertension not complicated by cardiac or renale failure were treated. Patients were allocated at random into one of three subsets of ten. In group A oxprenolol was given for 8 weeks and the dose was gradually increased up to 300 mgs daily. Oxprenolol was administered in combination with clortalidone in group B and with phentolamine in group C. A clinically satisfactory reduction in blood pressure was attained in no subset, despite the significant decrease of mean blood pressure. The blockade of beta-adrenergic receptors alone has proved to be less effective than the combined administration of oxprenolol and clortalidone or of oxprenolol and phentolamine. No differences were observed between the two combinations.


The influence of preganglionic nerves on the superior cervical ganglion of the rat. Sympathetic neurones grown in tissue culture with non-neuronal cells become cholinergic. Such a change from an adrenergic to cholinergic character does not occur in vivo and it has been suggested that this may be due to the determination of the adrenergic character by electrical activity. Electrical activity in the superior cervical ganglion by young rats were prevented by transection of the preganglionic nerve trunk. In no case did this operation result in an increase in the intrinsic choline acetyltransferase (CAT) of the ganglion. We conclude that electrical activity is not the factor responsible for the difference between in vivo and in vitro results.


Neuronal localisation of benzodiazepine receptors in cerebellum. The cellular localisation of benzodiazepine receptors was studied. Kainic acid induced neuronal lesions (2 x 0.25-2 x 2 micrograms; 2-26 days in rat cerebellum decreased specific binding of [3H]flunitrazepam down to 35% of controls. Specific binding of [3H]flunitrazepam was also decreased (to 80% of controls) in the cerebllum of mutant nervous mouse (nr/nr) where Purkinje cells are degenerated but in the mutant weaver mouse where granule cells are degenerated. These results show that benzodiazepine receptors are located mainly on neurons; both on Purkinje cells and other neurons, but not to a great extent on granule cells.


Benzodiazepine receptors: effect of tissue preincubation at 37 degrees C. [3H]Flunitrazepam binding to rat brain homogenates was assayed in two ways: by 37-0 degrees C incubations or 0 degrees C only incubations. The affinity of brain benzodiazepine receptors for [3H]flunitrazepam was significantly greater in 37-0 degrees C incubated samples than in 0 degrees C incubated samples, while there was no change in the number of benzodiazepine receptors. The possible mechanisms for this enhanced binding affinity as well as the methodological implications for this observation are discussed.


Anemone toxin discriminates between ionic channels for receptor potential and for action potential production in a sensory neuron. The effect of anemone toxin (ATX II), which slows sodium channel inactivation at electrically excitable membranes, was investigated in the slowly adapting stretch receptor organ of crayfish. The toxin affected the action potentials, but produced no changes in the stretch induced receptor potentials. This finding is further proof for the high selectivity of this toxin molecule for sodium channels which are gated by membrane depolarissations.