Phosphorus nuclear magnetic resonance studies of phosphorus metabolites in frog muscle. 31P-nuclear magnetic resonance was applied to living muscles of bullfrogs, and the time courses of metabolic changes of ATP, creatine phosphate, inorganic phosphate, and sugar phosphates were studied under anaerobic and aerobic conditions. A decrease in creatine phosphate was observed in the resting muscle under anaerobic conditions with a concomitant decrease in the intracellular pH, while the ATP level remained constant. With the use of 2,4-dinitro-1-fluorobenzene and iodoacetic acid, ATP disappeared quickly. When the resting muscle was perfused with oxygen-saturated glucose-Ringer's solution, the amount of creatine phosphate increased gradually. These findings indicate that anaerobic glycolysis is insufficient for even the resting energy consumption whereas oxidative phosphorylation is sufficient. The effects of tetanic stimulation on living muscles were also studied. When glycolysis and oxidative phosphorylation were suppressed, the intracellular energy store was depleted by the tetanic contraction. Anaerobic glycolysis produced rapid recovery of the energy store level, although it was insufficient to reach the initial level. Aerobic oxidative phosphorylation produced sufficient energy to reach the initial level, and this level was never exceeded. This finding suggests the existence of a regulatory mechanism for the energy store level.


Purification and some properties of a specific nuclease which cleaves transfer RNA precursors from the posterior silk gland of Bombyx mori. A specific endonuclease involved in the processing of tRNA precursors was isolated and partially purified from the posterior silk gland of Bombyx mori, and designated as RNase P.Bmo. This enzyme was shown to catalyze the conversion of 4.5 S precursor RNA to 4.1 S RNA by trimming the 5'-additional segment from the precursor RNA. RNase P.Bmo required divalent cations, Mg2+ or Mn2+. In the presence of these divalent cations, K+ or NH4+ activated the RNase P.Bmo reaction. Optimum pH was observed around 8.0. Ribosomal RNA's and mature tRNA from the silk gland were not cleaved by RNase P.Bmo. A 4.5 S precursor RNA fraction containing formycin, an adenosine analog, was less susceptible to RNase P.Bmo than the normal one. These results indicate that RNase P.Bmo has a high substrate specificity. An additional nuclease(s) was isolated. This activity was assumed to remove the extra 3'-segment of the 4.5 S precursor RNA.


Purification and properties of acetate kinase from Bacillus stearothermophilus. 1. Acetate kinase [EC 2.7.2.1] from an thermophile, B. stearothermophilus, was purified and crystalized. 2. This enzyme was shown to be a tetramer of identical subunits which had a molecular weight of about 40,000. Amino acid analysis showed no SH group. By analyzing the CD spectrum it was deduced that this enzyme is composed of 36% beta-structure, 21% alpha-helix and 43% unordered structure. 3. This enzyme shared many common enzymatic properties with the counterpart from mesophiles, i.e. pH optimum, substrate specificity, requirement of metal ions and essential amino acid residues necessary for the catalytic activity. However, this enzyme was remarkably thermostable. 4. A plot of the reaction velocity against the concentration of acetate, ADP or acetyl phosphate gave a curve of the Michaelis-Menten type. However, such a plot against ATP gave a sigmoid curve, suggesting a homotropic allosteric nature of the enzyme. 5. From the results of chemical modification it was deduced that an amino group and an imidazole group, at least, are involved in the active site of the enzyme.


Studies of a calcium-activated neutral protease from chicken skeletal muscle. I. Purification and characterization. A calcium-activated neutral protease was purified 2,700-fold over the crude extract from chicken skeletal muscle. The purified protease migrated as a single band on polyacrylamide gel electrophoresis with or without SDS. Its molecular weight was 80,000 and pH optimum for activity was 7.7. The activity required strictly the presence of calcium (optimum concentration: 1.8 mM) or strontium (optimum concentration: 10 mM) ions. The protease was inhibited by leupeptin, which is known to be a strong inhibitor of papain, cathepsin B, trypsin, and plasmin.


Studies on cellulases of a phytopathogenic fungus, Pyricularia oryzae cavara. II. Purification and properties of a beta-glucosidase. Three components (GA, GB-1, and GB-2) of beta-glucosidase were detected in the culture filtrate of Pyricularia oryzae grown in a cellulose or cellulose derivative medium. Among them, GB-1 was induced most strongly. Purified GB-1 was homogeneous on polyacrylamide gel electrophoresis and showed an approximately 1,400-fold increase of specific activity over the starting material. The molecular weight was determined to be 240,000 by sodium dodecyl sulfate-gel electrophoresis. A similar value was also obtained by sucrose density gradient centrifugation. The enzyme contained a high proportion of acidic amino acids and mannose, and the isoelectric point of the enzyme was pH 4.15. The enzyme had a pH optimum of 5.5 and a temperature optimum at 55 degrees C. beta-Glucosidase activity was inhibited by Mn2+, Cu2+, Hg2+, p-chloromercuribenzoate, and glucono-delta-lactone. The enzyme split off glucose units one by one from the nonreducing ends of not only beta-glucooligosaccharides but also some beta-glucans, such as carboxymethylcellulose, laminaran, pustulan, and zeagallan. The affinity for cello- and laminari-oligosaccharides tended to increase in parallel with the chain length.


Pyridine nucleotide interaction with rat liver dihydropteridine reductase. The interactions of a homogeneous preparation of rat liver dihydropteridine reductase with NADH, NADPH, NAD+, NADP+, and the 1-N6-ethenoadenine derivative of NAD+ have been investigated by fluorescence titration, circular dichroism, equilibrium dialysis, Sephadex G-25 chromatography, and polyacrylamide gel electrophoresis. The procedures indicate that the dimeric enzyme has a definite preference for NADH, but binds only 1 mol of this nucleotide per mol of enzyme. The binary complex of enzyme with NADH is only partially stable to exhaustive dialysis and gel electrophoresis, where it shows greater mobility (0.26) than the free enzyme (0.21); however, the complex can be isolated by Sephadex G-25 chromatography, and characterized with respect to its absorbance spectrum. No ternary complexes are observed when samples of reductase, preincubated with excess NADH, and either the reaction product, 2-amino-4-hydroxy-6,7-dimethyl-5,6,7,8-tetrahydropteridine, or the inhibitor, methotrexate, are subjected to polyacrylamide gel electrophoresis.


Membrane bioenergetic parameters in uncoupler-resistant mutants of Bacillus megaterium. Mutants of Bacillus megaterium displaying malate-driven ATP synthesis resistant to uncouplers of oxidative posphorylation are further characterized. Both the pH gradient and electrical potential generated across the membrane by malate respiration are equally sensitive to uncouplers in the wild type and uncoupler-resistant mutants. The mutants possess 0 to 10% of the wild type ATPase activity which is not activated by pretreatment with heat or trypsin. Despite this inability to measure ATPase activity, the mutants demonstrate acid-pulse-driven ATPase synthesis which is sensitive to uncouplers as well as malate-driven ATP synthesis which becomes uncoupler sensitive at pH 5.5. N,N' -Dicyclohexylcarbodiimide and valinomycin plus potassium inhibition of ATP synthesis is reversed by uncouplers in the mutants but not in the wild type. The data support the existence of a specific site on the ATPase complex for uncoupler binding which, if altered by mutation, affects uncoupler binding to the complex. The retention of malate-driven ATP synthesis in the absence of a significant pH gradient or electrical potential suggests that an alternative intermediate is involved in coupling oxidation to phosphorylation.


Subunit interactions of Glycera dibranchiata hemoglobin. The coelomic cells of the polychaete annelid Glycera dibranchiata contain two hemoglobins. The monomer hemoglobin fraction is composed of one major component and two minor components as determined by starch gel electrophoresis and isoelectrofocusing, but is homogeneous as to subunit size as demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The polymer hemoglobin fraction has and initial molecular weight of Mn = 125,000 as determined by osmometry, but exhibits an increased state of aggregation upon storage. The quaternary structure of the polymer is constituted of monomeric subunits in a non-covalent state of aggregation as demonstrated by its subunit dissociation inthe presence of propyl urea. The oxygen affinity of the polymer is lower than the monomer but increases with deaggregation. The Bohr effect is present only in the polymer. Cooperativity is also characteristic of the polymer and is pH-dependent. Interestingly, cooperativity increases with intermediate states of polymer deaggregation. By far, the main organic phosphate component of the coelomic red cells is ATP accompanied by small amounts of ADP and GTP. No modulating effect of ATP on the oxygen equilibrium of either polymer or total hemolysate was found.


A spectral probe near the subunit catalytic site of glutamine synthetase from Escherichia coli. Reduced pyridoxal 5'-phosphate.enzyme complexes. In order to label phosphate binding sites, unadenylylated glutamine synthetase from Escherichia coli has been pyridoxylated by reacting the enzyme with pyridoxal 5'-phosphate followed by reduction of the Schiff base with NaBH4. A complete loss in Mg2+-supported activity is associated with the incorporation of 3 eq of pyridoxal-P/subunit of the dodecamer. At this extent of modification, however, the pyridoxylated enzyme exhibits substantial Mn2+-supported activity (with increased Km values for ATP and ADP). The sites of pyridoxylation appear to have equal affinities for pyridoxal-P and to be at the enzyme surface, freely accessible to solvent. At least one of the three covalently bound pyridoxamine 5'-phosphate groups is near the subunit catalytic site and acts as a spectral probe for the interactions of the manganese.enzyme with substrates. A spectral perturbation of covalently attached pyridoxamine-P groups is caused also by specific divalent cations (Mn2+, Mg2+ or Ca2+) binding at the subunit catalytic site (but not while binding to the subunit high affinity, activating Me2+ site). In addition, the feedback inhibitors, AMP, CTP, L-tryptophan, L-alanine, and carbamyl phosphate, perturb protein-bound pyridoxamine-P groups. The spectral perturbations produced by substrate and inhibitor binding are pH-dependent and different in magnitude and maximum wavelength. Adenylylation sites are not major sites of pyridoxylation.


The internal pH of isolated serotonin containing granules of pig platelets. The [14C]methylamine distribution method was utilized to measure the internal pH of isolated serotonin containing granules of pig blood platelets under varying conditions. The granules used were isolated by a new protocol which stressed platelet rupture under controlled conditions and preservation of isotonicity throughout the isolation procedure, In a well buffered external medium, pH 6.85, The deltapH was measured as 1.11 with the internal pH being found acidic (pH 5.74). Increasing the external pH produced a corresponding increase in the deltaH. The pH gradient could be collapsed by the addition of ionophores and uncouplers which are known to transport protons across biological membranes. In addition, the deltapH was constant for granules suspended in various ionic media, thus suggesting that the deltaH did not arise secondarily due to the establishment of a Donnan equilibrium. The existence of the acidic intragranular space is discussed with respect to previous ancillary findings. Also, an explication of the possible physiological significance of the deltaH is presented.


Nonrandom distribution of sialic acid over the cell surface of bristle-coated endocytic vesicles of the sinusoidal endothelium cells. Previous studies with protein tracers have shown that the luminal surface of the vascular endothelium of the bone marrow is endocytic. The endocytosis occurs through the formation of large bristle-coated vesicles (LCV). The anionic charge distribution in this process was examined at the luminal surface of the endothelial cell, At pH 1.8, colloidal iron (CI), native ferritin, and polycationic ferritin (PCF) are bound by the luminal surface of the endothelial cell, but not at the sites of LCV formation. PCF used over a pH range of 1.8--7.2 (CI is unstable at higher pH levels) revealed LCV binding of this agent in increasing manner from pH 3.5 upwards. PCF binding at low pH (1.8) at the endothelial cell surface was markedly reduced by neuraminidase. Neuraminidase did not reduce PCF binding by the endothelial cell surface nor by the LCV at higher pH levels. It is concluded that the luminal surface of the endothelial cell has exposed sialic acid groups which are absent or significantly diminished at endocytic sites. The free surface of the endothelial cells as well as the sites of endocytosis have, in addition, anionic material with a pKa higher than that of sialic acid (pKa 2.6). These anionic materials may be different at the sites of endocytosis as compared to those present at the free cell surface.


Hemagglutination with simian papovavirus SA12. Simian papovavirus SA12 agglutinated human, guinea pig, and chicken erythrocytes. SA12 hemagglutinin was most effectively released from debris of infected tissue culture cells at an alkaline pH.


Relationship between the rate of H+ transport and pathways of glucose metabolism by turtle urinary bladder. The urinary bladder of the fresh-water turtle acidifies its contents by actively transporting H(+) ions across the luminal membrane. It is known that the H(+) transport system is dependent upon oxidative metabolism and the substrate glucose; however, the specific biochemical events resulting in H(+) translocation have not been identified. This study examines the relationship between active H(+) transport and a specific oxidative pathway of glucose metabolism, the pentose phosphate shunt. To investigate this relationship the metabolic and transport rates were simultaneously measured under several well-defined conditions. When H(+) transport was inhibited by either the application of an opposing pH gradient or by acetazolamide, glucose metabolism by the pentose phosphate shunt declined. Conversely, stimulation of H(+) transport by either imposing a more favorable pH gradient or by CO(2) addition resulted in an increase in pentose phosphate shunt metabolism. Glycolytic activity, in contrast, was invariant with the maneuvers which altered the rate of H(+) transport. Additional experiments localized pentose phosphate shunt enzyme activity to the mucosal fraction of the bladder which is the cell layer responsible for acid secretion. The finding that the rate of glucose metabolism by the pentose phosphate shunt is related to the rate of H(+) transport suggests but does not prove that the pentose phosphate shunt may be an important metabolic pathway for H(+) transport by the turtle urinary bladder.


Tissue guanosine-3',5'-cyclic monophosphate levels and soluble guanylate cyclase activity: a positive correlation during unilateral cryptorchidism in the rat testis. The relationship between the subcellular distribution of guanylate cyclase and tissue guanosine-3',5'-cyclic monophosphate (cGMP) levels was investigated in rat testes after surgically induced unilateral cryptorchidism. Placement of one of a testis pair in the abdominal cavity results in loss of testicular weight and function in the abdominal testis whereas the remaining scrotal testis appears to be functionally normal. Within 5 days after surgery, tissue cGMP levels were increased by twofold in the abdominal testis. A fourfold elevation was noted from 10 to 30 days after surgery. Whereas the homogenate guanylate cyclase activity was only slightly elevated 10 and 20 days postoperatively, a 200% increase in the soluble guanylate cyclase activity was seen at 5 days. Between 10 and 30 days, the rise in activity was >250% (P < 0.01). An increase in soluble guanylate cyclase activity was noted when the data were expressed as per milligram protein, per milligram DNA or per whole testis. Conversely, particulate guanylate cyclase activity was reduced by 40% in the cryptorchid testis. Kinetic analysis of the soluble enzyme prepared from abdominal and scrotal testes yielded linear Line-weaver-Burke plots for both enzyme preparations with an identical K(m) for guanosine triphosphate, but a three-fold higher maximal velocity for the abdominal enzyme. When the soluble guanylate cyclases from both testes were mixed and assayed together, the activities were additive rather than exhibiting synergism or inhibition. These experiments indicate that the altered V(max) is not due to a transferable activator or inhibitor.An immunocytochemical technique was used to assess the cell type in which the alterations in cGMP metabolism occurred. Comparison of the scrotal and abdominal testes revealed that the abdominal testis exhibited enhanced cGMP immunofluorescence within the cells lining the inner aspect of the seminiferous tubule as well as tubular elements and interstitial cells. Thus, it is inferred that the correlated changes in soluble guanylate cyclase activity and cGMP levels occur in several of the cell types that remain viable within the cryptorchid testis.


Human llamas: adaptation to altitude in subjects with high hemoglobin oxygen affinity. To assess the adaptive value of the right-shift of the oxyhemoglobin dissociation curve (decreased affinity for oxygen) observed in humans upon altitude exposure, the short-term physiologic responses to altitude-induced hypoxia were evaluated in two subjects with a high oxygen affinity hemoglobin (Hb Andrew-Minneapolis) and in two of their normal siblings. In striking contrast to normal subjects, at moderately high altitude (3,100 m) the high affinity subjects manifested: (a) lesser increments in resting heart rate; (b) minimal increases in plasma and urinary  erythropoietin; (c) no decrement in maximal oxygen consumption; and (d) no thrombocytopenia. There was no difference between subject pairs in 2,3-diphosphoglycerate response to altitude exposure. These results tend to contradict the belief that a decrease in hemoglobin oxygen affinity is of adaptive value to humans at moderate altitudes. Rather, they support the hypothesis that, despite disadvantages at low altitude, a left-shifted oxyhemoglobin dissociation curve may confer a degree of preadaptation to altitude.


Direct evidence of participation of rat lung carbonic anhydrase in CO2 reactions. Isolated rat lungs were ventilated with air and perfused with a blood-free Krebs-Ringer bicarbonate solution under conditions of net CO2 elimination in the lung. Some of the effluent perfusate was drawn through a stop-flow pH electrode apparatus, arriving at the electrode within 4 s after passing through the pulmonary capillaries. pH and temperature of the fluid in the electrode chamber were continuously monitored both before and after withdrawal was suddenly stopped. Little or no change was observed in the pH of the perfusate after flow was stopped, despite the fact that CO2 was eliminated in the lung, suggesting that the conversion of H2CO3 to CO2 in the blood-free perfusion fluid was markedly accelerated and the rise in pH was complete by the time the perfusate reached the electrode. Because the effluent perfusate was shown to be free of carbonic anhydrase activity, the catalysis must have occurred during transit through the isolated lung. When acetazolamide was added to the perfusate, a rise in the pH of the perfusate after stopping flow was consistently seen. These results suggest that the carbonic anhydrase of isolated lungs accelerates the conversion of H2CO3 to CO2 and enhances COW elimination as perfusate passes through the pulmonary capillaries, and that the enzyme may be present on the capillary endothelial surface.


Hereditary male pseudohermaphroditism associated with an unstable form of 5 alpha-reductase. The properties of 5alpha-reductase have been compared in genital skin fibroblasts cultured from five patients from three families (Los Angeles, Dallas, and Dominican Republic) in which hereditary male pseudohermaphroditism has been established to result from deficient conversion of testosterone to dihydrotestosterone. Despite the fact that 5alpha-reductase was immeasurable in a homogenate of epididymis removed from one of the Los Angeles patients, 5alpha-reductase activity was normal in intact fibroblasts and fibroblast extracts from both patients from the Los Angeles family. Although the apparent K(m) for testosterone was also near normal, the apparent K(m) for NADPH in these mutants is elevated some 40-fold above normal. Furthermore, the enzyme is not protected against denaturation at 45 degrees C by concentrations of NADPH that stabilize normal 5alpha-reductase, and in intact fibroblasts from these patients (but not from controls), enzyme activity decreases promptly when protein synthesis is inhibited. We conclude that the mutation in this family results in an unstable enzyme. In contrast 5alpha-reductase activity in fibroblast extracts from a patient from the Dominican Republic family is similar to that previously described in two members of the Dallas family, namely total enzyme activity is low at the optimal pH for the normal reaction, and the apparent K(m) for testosterone is some 20-fold higher than that of the controls. We conclude that the mutations in the Dallas and Dominican Republic families are similar and result in low activity of the enzyme as the result of a decreased affinity for testosterone.Thus, two distinct types of mutations can produce male pseudohermaphroditism due to deficient dihydrotestosterone formation.


Taped vs. "live" desensitization and level of autonomic arousal. Investigated whether variations in method of presentation of anxiety items (Es voice, E's taped voice, S's taped voice) will result in differences in autonomic arousal (as measured by self-report, galvanic skin responses, cardiac rate, blood pressure). Ss were 24 Naval Academy midshipmen. The procedure involved (1) training in deep muscle relaxation; and (2) measuring arousal after the presentation of anxiety-eliciting stimuli. Statistical analysis indicated that the influence of method of presentation was significant for galvanic skin response and self-reported anxiety. Comparison of cell means showed that verbal presentations were more effective in eliciting autonomic arousal than taped presentations. The results of this study provide partial evidence to support the conventional form of systematic desensitization as more effective than the newer automated or taped versions.


Neuroleptic-induced attenuation of brain stimulation reward in rats. In 30-min free-operant tests, the dopamine receptor blockers pimozide (.125, .25, and .50 mg/kg) and (+)-butaclamol (.1, .2, and .4 mg/kg) attenuated lever pressing for lateral hypothalamic brain stimulation. When discrete self-stimulation trials were offered in a straight alleyway, pimozide increased start box latencies, slowed running speeds, and reduced lever-pressing rates. However, performance early in both lever-pressing and runway sessions was normal; performance deteriorated as testing progressed, following patterns that paralleled those seen when animals were tested with reductions in the amplitude of stimulating current. Spontaneous recovery was obtained in both situations; experimenter-imposed 10-min time-outs caused renewed lever pressing and running. In contrast, alpha-noradrenergic receptor blockade by phenoxybenzamine (5, 10, and 20 mg/kg) failed to produce extinction-like response patterns. These data support the view that central dopaminergic systems are important components of the neural mechanisms mediating reward.


Effect of abdominal radiation therapy on drug absorption in humans. The absorption of oral digoxin and of desmethyldiazepam, from its precursor clorazepate, was studied in seven patients who had received abdominal and/or pelvic radiation therapy for neoplastic disease. All patients were in remission and had normal renal function and no evidence of malabsorption. Single 0.5-mg doses of digoxin tablets and 15-mg doses of clorazepate were administered in the fasting state. Concentrations of digoxin (by radioimmunoassay) and desmethyldiazepam (by gas chromatography) were determined in multiple plasma samples and all urine collected during 24 hours after dosage. The mean (+/- S.E.) weight-normalized area under the 24-hour plasma digoxin concentration curve (WtN-AUC-24) in the patients (722 +/- 40 ng/ml-hr-kg) was similar to that in five normal controls (713 +/- 57 ng/ml-hr-kg), but 24-hour urinary excretion of digoxin in patients (54.5 +/- 4.4 microgram) was significantly less (P less than 0.025) than in controls (83.4 +/- 11.4 microgram). Neither age, sex, nor renal function explained the difference. In the clorazepate study, WtN-AUC-24 for desmethyldiazepam in the patients (187 +/- 19 microgram/ml-hr-kg) was significantly less (P less than 0.01) than in 15 normal control subjects (230 +/- 5 microgram/ml-hr-kg). Age and sex did not explain the difference. Thus, radiation therapy, or the underlying disease, is associated with malabsorption of these two drugs, possibly because of damage to gastric acid-secreting cells.


Selective sensitivity to hydrocortisone of regulatory functions that determine the magnitude of the antibody response to type III pneumococcal polysaccharide. Previous studies on the basis for the immunosuppressive potential of adrenal corticosteroids have stressed that the effects of these agents on immune functions depend on the animal species being considered, as well as the subpopulations of lymphocytes involved in the expression of immune functions examined. In the present work, we have evaluated the effect of a single dose of hydrocortisone on three different immunoregulatory functions that can influence the magnitude of an antibody response to Type III pneumococcal polysaccharide (SSS-III) in mice; these functions include suppressor, amplifier, and helper activity that are dependent upon the presence of distinct subpopulations of thymus-derived (T) cells. The results obtained show that a single injection of a relatively large dose of hydrocortisone, when given at the time of priming with carrier, eliminated all evidence of carrier-specific helper T cell activity; hydrocortisone was also found to eliminate a significant amount of helper T cell activity when given after such activity had been generated. But, under the same experimental conditions, suppressor and amplifier T cell activities were unaffected, even in this steroid-sensitive species. Such selective sensitivity may account for some of the immunosuppressive potency of steroids.


Hepatic lipid metabolism. Age-related changes in triglyceride metabolism. Age-related changes in hepatic triglyceride formation have been described in developing rats. Triglyceride formation was measured in vitro in the presence of [14C]glycerol-3-phosphate, palmitate, ATP, CoA, and Mg2+ by using liver homogenates and microsomal fractions derived from various age groups of animals. Triglyceride formation was most active in one-day-old rats and then decrease with age. The increase in triglyceride formation following birth was prevented by the administration of puromycin or by denying suckling. In addition, changes in plasma and hepatic triglyceride concentrations, were also determined as functions of age. These studies suggest that the age of the animal significantly influences triglyceride metabolism.


R-factor mediated dihydrofolate reductases which confer trimethoprim resistance. Six different R-factors conferring trimethoprim resistance had been isolated from a variety of sources. The trimethoprim-resistant dihydrofolate reductases (EC 1.5.1.3) from strains containing these R-factors were purified by ammonium sulphate precipitation and DEAE-cellulose ion-exchange chromatography. The enzymes showed no significant differences in molecular weight, pH profile, substrate profile, heat sensitivity, inhibition profile and Michaelis-Menten kinetics. There was, however, considerable variation in the specific activity of these enzymes in the same bacterial host. When two Escherichia coli trimethoprimsensitive dihydrofolate reductases were examined as controls, considerable differences between their properties and those of the enzymes mediated by R-factors were detected. The data suggest that one trimethoprim resistance gene could be spreading through the bacterial population, possibly situated on a transposon.


An RNA polymerase activity in purified rabies virions. An RNA polymerase activity has been demonstrated in purified rabies virions. Efficiency of the reaction is low since the rate of incorporation was equal to 3 to 5 pmol of uridine per hour, per mg of protein. As with other mammalian rhabdoviruses the optimal temperature was 31 degrees C. Unlike vesicular stomatitis virus, manganese could be substituted for magnesium as a divalent cation, at an optimum concentration of 10 to 20 mM.


Increase by calcium in production of interferon by L929 cells induced with polyriboinosinate-polyribocytidylate complex. Calcium chloride (5 to 20 mM) potentiated interferon production induced by rIn:rCn in L929 mouse fibroblasts up to a thousand-fold. Higher concentrations of calcium (20 to 65 mM) mixed with rIn:rCn were associated with increased cytotoxicity and a more acidic medium, but were effective in enhancing interferon production if preparations were adjusted to a uniform pH. Although calcium increased cellular binding of 3H-rCn:rIn, only a partial correlation between binding and interferon production was observed.


Poly (A) polymerase activity in L cells following encephalomyocarditis virus infection. Poly (A) polymerase activity has been measured in crude cytoplasmic extracts of mouse L cells infected with encephalomyocarditis (EMC) virus. After infection there is first a decrease in enzyme activity followed by an increase which itself precedes detectable virus RNA and protein synthesis. The activity of the enzyme then declines before the release of mature virions and cell death take place. The early inhibition of poly (A) polymerase activity is correlated with the virus-induced shut-off of cellular protein synthesis but it is not due to inhibition of the synthesis of cellular enzyme and occurs in the absence of virus replication. The poly (A) polymerase is not synthesized after infection and modification of its activity can be reversed late in the virus cycle. These results indicate that host poly (A) polymerase activity can be regulated by the virus and further show that there is a correlation between the modification of poly (A) polymerase activity and the biosynthesis of poly (A).


Cerebral transmitter precursors and metabolites in advanced renal disease. Patients with chronic renal disease had low plasma total tryptophan but an abnormally high proportion of this was in the free state. The subjects with encephalopathy had raised plasma free tryptophan, CSF tryptophan, and CSF 5-hydroxyindoleacetic acid. CSF tryptophan correlated better with plasma free than with plasma total tryptophan. Plasma and CSF tyrosine concentrations were normal but CSF homovanillic acid was raised especially in subjects with encephalopathy. The possible significance of these changes in advanced renal disease is discussed.


Guanylate cyclase activity in normal and diseased human muscle. Guanylate cyclase activity has been studied in muscle of normal subjects and of patients suffering from muscular and neuromuscular diseases. In normal tissue a guanylate cyclase activity was found in both soluble and particulate fractions of homogenate. We found also that the kinetic analysis of the enzyme of soluble differed from that of particulate fraction. A decrease of guanylate cyclase activity in crude homogenate was observed in muscular dystrophies, in neuromuscular atrophies, and in inflammatory forms of muscle disease.


Neurosurgery: considerations for strength and quality. The 1978 AANS presidential address. The President of the American Association of Neurological Surgeons reviews the organization of the Association designed to represent neurosurgery and neurosurgeons. He summarizes the pertinent problems that the specialty has faced and with which it continues to deal, in persistent pursuit of its objective: to assure the highest quality of neurosurgical care for all.


Selective defect of precursor T cells associated with apparently normal B lymphocytes in severe combined immunodeficiency disease. Two patients, one with an autosomal and the other a sex-linked form of severe combined immunodeficiency, had more than 95% B cells in their peripheral blood. Despite an increased absolute number of B lymphocytes, the patients were unable to produce serum antibodies. In each patient, geno- or pheno-identical bone marrow transplantation was followed by the visualization of a thymus shadow and the appearance of both cellular and humoral functions. Chromosome of allotype studies showed that the T cell originated from the donor whereas serum immunoglobulins were synthesized by host B cells. In these patients the pathogenesis appears to be a selective defect of bone marrow precursor T cells without concomitant intrinsic B cell defect. The successful outcome of the graft in these two patients, who are now, respectively, 5 years and 11 months of age and free of infections, indicates that the preferred form of therapy in such patients is transplantation of bone marrow stem cells, which populate the thymus and mature slowly into T cells that cooperate fully with host B cells in synthesis of antibody.


The effect of microcapsule size on the oxidative decomposition of core material. The factors that affect the size of microcapsules and the oxidation of their benzaldehyde core have been examined. The pH of the preparation changed the overall size of the microcapsules which reached a maximum diameter at pH 4.1. The size of the core droplets also varied with the preparative pH and their oxidation rate depended on the bulk droplet size rather than their surface area. A rapid oxidation of benzaldehyde associated with the microcapsule wall resulted in a preliminary peak in the oxidation curve. Explanations for these phenomena are discussed.


The wetting of powders of acetylsalicylic acid, salicylic acid, phenacetin and paracetamol. The wetting of powders of acetylsalicylic acid, salicylic acid, phenacetin and paracetamol has been assessed using methanol--water mixtures to give a range of surface tensions. The results have been interpreted in terms of the critical surface tension, adhesion tension and spreading coefficients. The critical surface tension values are surprisingly low which may be due to adsorption of the methanol at the solid surface, exposing its CH3 group to the liquid. The adhesion tension and spreading coefficient values could be useful guides in formulation.


Structure activity relation for some 1,4-benzodiazepinones: correlation between rate constants for reduction by sodium borohydride and antileptazol ED50. Significant correlation in 11 different 1,4-benzodiazepinones has been established between log k2 (the second order rate constant for the reduction of the "azepinones" by sodium borohydride) and their ED50 against leptazol-induced seizures in mice. The results suggest a possible involvement of the carobnyl group at the receptor site.


The use of 13C-nmr spectroscopy for the detection and identification of metabolites of carbon-13 labelled amitriptyline. The antidepressant drug amitriptyline and two of its metabolites, nortriptyline and desmethylnortriptyline, each containing two 13C atoms, have been used to determine the sensitivity and selectivity of 13C-nmr spectroscopy for the detection of unchanged amitriptyline and N-desmethyl metabolites in the urine of animals dosed orally with the labelled drug. The resonance signals from the 13C atoms detected in the 13C-nmr spectrum of entire extract from a control 12 h rat urine sample to which 1 mg of each labelled compound had been added were easily detected, using an instrument accumulation time of 1 h. The 13C-nmr spectrum of an extract of hydrolysed urine from a dog that had received an oral dose of  [13C2]amitriptyline (30mg) exhibited signals that could be assigned to metabolites resulting from N-dealkylation and N-oxidation, as well as those bearing the intact amitriptyline side-chain. These assignments were confirmed by analysis of the same extract by g.c.--ms and h.p.l.c.


Acidic in vivo metabolites of cannabinol isolated from rat faeces. Six acidic metabolites were isolated from rat faeces and identified by gas chromatographymass spectrometry and proton magnetic resonance. Cannabinol-7-oic acid was the most abundant acidic metabolite isolated. Others present in decreasing order of prominence were 1"-hydroxy-, 4"-hydroxy-, 3"-hydroxycannabinol-7-oic acid, cannabinol-3"-one-7-oic acid and 2"-hydroxycannabinol-7-oic acid.


Comparison between spontaneously beating atria from control and streptozocin-diabetic rats. Isolated spontaneously beating atria from streptozocin diabetic rats were compared with those from controls. Diabetic atria were found to have reduced rates, increased forces of contraction and reduced sensitivity to the inotropic effects of noradrenaline, isoprenaline, tyramine and calcium. Positive chronotropic responses to tyramine were also reduced but those to noradrenaline and isoprenaline were increased suggesting that tyramine releasable stores of noradrenaline were reduced. Elevation of glucose concentration in the medium from 5.6 to 27 mM resulted in decrease of inotropic sensitivity to the agents used in both control and diabetic rat atria. Resting contractile force of control rat atria was reduced by the inclusion of either 22 mM 2-deoxyglucose, 10(-3) i.u. insulin ml-1 or 5 mM acetate in the medium. The rate was also reduced by medium containing 2-deoxyglucose but increased by insulin. 2-Deoxyglucose also reduced inotropic but increased chronotropic sensitivity to isoprenaline. Possible mechanisms responsible for the changes observed are discussed.


The antitussive actions of the drug Ru 20201 given as an aerosol to cats. The effect of the drug Ru 20201 (1,2,3,4,4a,9b-hexahydro,-8,9b-dimethyl-4-[3-(4-methylpiperazin-1-yl)propionamido]dibenzofuran-3-one upon mechanically-evoked cough from the laryngopharyngeal and tracheobronchial areas in nine unanaesthetized cats has been examined. Inhalation of 2 ml of an aerosol of a 10% solution in water suppressed coughing for 30 min. The effect was greatest on the number of cough efforts. The expiratory component of cough was suppressed more than was the inspiratory one. The effect was greater on cough from the laryngopharyngeal than from the tracheobronchial area.


Characterization of pharmacologically important prototropic species derived from a pyridinemethanol antimalarial by electronic absorption and fluorescence spectroscopy. Variations of the absorption and fluorescence spectra of the experimental antimalarial drug, alpha-dibutylaminomethyl-2,6-bis(p-trifluoromethylphenyl)-4-pyridinemethanol, were investigated throughout the pH region in concentrated sulfuric acid media and in n-hexane. The predominant prototropic species at physiological pH is the singly charged cation. In the pH 6--12 region, the structured fluorescence of the monocation is quenched with the concomitant appearance of a diffuse, long wavelength emission while the corresponding absorption spectra shift only slightly to longer wavelengths. Furthermore, the dibutylamino group exhibits an unusually low basicity. This behavior is explained as due to the formation of an intramolecular hydrogen bond in the neutral molecule in the ground and lowest excited singlet states. A similar intramolecular hydrogen bond in the monocation is not spectroscopically visible.


pH-solubility profiles or organic carboxylic acids and their salts. The solubilities of naproxen, 7-methylsulfinyl-2-xanthonecarboxylic acid, 7-methylthio-2-xanthonecarboxylic acid, and their sodium, potassium, calcium, and magnesium salts were determined as a function of pH. The results on the solubility of naproxen and its salts were in excellent agreement with the theoretical profiles describing the relationship between pH values of the solutions and the dissociation constant of the acid. The solubilities of the two xanthonecarboxylic acids were higher at higher pH values than the values calculated when complete dissociation in solution was assumed. The influence of the salt species on the solubility of organic carboxylic acids, at and above pH values of complete ionization, cannot be predicted even qualitatively from equations used for alkali and alkaline earth metal salts.


Synthesis and beta-adrenergic blocking action of a new thiazolylthiopropanolamine derivative. The synthesis of (+/-)-2-(3'-tert-butylamino-2'-hydroxypropylthio)-4-(5'-carbamoyl-2'-thienyl)thiazole hydrochloride is described. The new compound antagonized the cardiovascular effects, such as positive chronotropic, positive inotropic, or depressor arterial blood pressure responses, elicited by intravenous isoproterenol; it was 9--14 times as potent as propranolol in anesthetized open chest dogs. The oral administration of the compound reduced isoproterenol tachycardia in conscious dogs. It was about five times as potent as propranolol in this test, with maximal action after 1 hr, and its duration was significantly longer than that of propranolol.


GLC determination of atenolol and beta-blocking agents in biological fluids. A rapid, sensitive, and specific method of analysis for atenolol is described. Metoprolol is used as the internal standard. Atenolol and metoprolol are extracted into 1-butanol--benzene. Interfering components present in palsma and urine, but not discolored saliva, are removed during an acid wash and reextraction into ether. Drug and internal standard are converted to the pentafluoropropionate derivatives, which are quantitated by GLC with electron-capture detection and characterized by chemical-ionization mass spectrometry. The method should be applicable to measurement of other beta-adrenergic blocking agents with similar structures.


An ATPase dependent, radiosensitive acidic microclimate essential for intestinal folate absorption. 1. 5-Methyltetrahydrofolic acid transport was studied across everted sacs of rat jejunal segments from control and whole body X-irradiated (700 rad) rats at 10(-5)M concentrations (at which optimum transport occurs) at various pHs.2. The folate transport from mucosal to serosal compartment was inhibited by about 55% in irradiated rat at the pH of the intestinal chyme (6.5). Extraneous ATP in the incubation system could restore the defective transport of the irradiated intestine.3. The maximum folate transport which occurred at pH 4.0 was not adversely affected by whole body irradiation. An acidic, pH dependent, passive uptake of 5-methyltetrahydrofolic acid was observed.4. The normal absorption barrier of the small bowel was not disrupted by the acidification process as practically no uptake was observed with irradiated segments pretreated at pH 4.0 except in the presence of ATP.5. Leucine and serine transport at a zero concentration gradient indicated active transport mechanisms which were not affected by acidification. Their uptake was additively increased in the presence of glucose and ATP, further indicating that the normal physiology of the intestines was not affected by the acidification process.6. An intestinal mucosal cell surface ATPase was observed which was Mg(2+) dependent. It could hydrolyse solution phase ATP and thus generate the protons necessary for the acidification of a microenvironment where passive uptake of the neutral folate species could occur.7. The ATPase activity was inhibited about 90% by 50 mM-Na azide at pH 6.5. Below this concentration folate transport was also inhibited.8. Na azide did not inhibit folate transport at pH 4.0, suggesting that its inhibition of folate uptake at pH 6.5 is related to its inhibitory effect on ATPase, rather than on folate transport per se. ATPase activity was therefore essential for folate transport at the pH of the intestinal chyme.


beta-Adrenergic blocking agents. 18. 1-(Aryloxy)-3-(arylthioalkylamino)propan-2-ols and 1-substituted alkylthioamino-3-(aryloxy)propan-2-ols. The synthesis is described of a seris of derivaties of 1-(aryloxy)-3-(arylthioalkylamiho)propan-2-ols and 1-(alkylthioamino)- and 1-(aralkylamino)-3-(aryloxy)propan-2-ols. These compounds were investigated for their beta-adrenoreceptor blocking properties and their selectivity of action for the cardiac beta1 receptor. The structure-activity relationships are discussed with particular reference to the effects of the sulfur, sulfoxide, and sulfone groups on beta-adrenoreceptor blocking potency and selectivity.


A series of hexahydro[1,4]oxazino[3,4-a]isoquinolines as potential neuroleptics. The synthesis and stereochemistry of trans-N,N-diethyl-9,10-dimethoxy-1,3,4,6,7,11b-hexahydro[1,4]oxazino[3,4-a]isoquinoline-3-carboxamide hydrochloride (16) and a series of analogues are described. 16 and its (+) isomer had neuroleptic properties in the Sidman avoidance test in gerbils. A few closely related amides of the trans series were active but cis amides were inactive as neuroleptics.


Facile syntheses of potent dopaminergic argonists and their effect on neurotransmitter release. The facile syntheses of important intermediates used in the preparation of the two potent dopaminergic argonists, 2-amino-6,7-dihydroxytetrahydronaphthalene (11) (referred to by some authors as ADTN) and its 5,6-dihydroxyl isomer 12, are described. Thus 6,7-dimethyoxy-2-tetralone has been prepared in two steps and 5,6-dimethoxy-2-tetralone in three steps both from commercially available materials. The effects of 11, 12, and the noncatechol analogue, 2-aminotetrahydronaphthalene (ATN), on radioactive neurotransmitter release have been studied in vitro using rat brain slices. It has been shown that both 11 and 12, at a concentraiton of 2 micron, cause a release of [3H]-DA and NA, 11 being more potent than 12 in releasing [3H]-DA. ATN (2 micron) was found to be inactive in these experiments which shows the importance of the catechol function in this uptake--release process.


Synthesis and biologic distribution of radioiodinated beta-adrenergic antagonists. Iodinated analogues 2, 7, and 8 were prepared from propranolol, practolol, and acebutolol in 30--50% yields. Radioisotopic exchange between carrier-free Na125I and the molten iodinated beta-adrenergic antagonist yielded the corresponding 125I-labeled product. The biodistribution in rats, determined at 15 and 60 min postinjection, indicated that the radioiodinated analogues of the cardioselective drugs practolol and acebutolol localized to a greater degree in the liver and heart than the analogue of propranolol. Conversely, [125I]iodopropranolol (2) was concentrated to a greater extent in the lungs than [125I]iodopractolol (7) or [125I]iodoacebutolol (8). Therefore, 123I- or 131I-labeled cardioselective beta-adrenergic antagonists, such as 7 or 8, may prove useful as radiodiagnostic agents for the external imaging of the myocardium.


The formation of peptides from the 2'(3')-glycyl ester of a nucleotide. We have synthesized 2'(3')-O-(glycyl)-adenosine-5'-(O-methylphosphate), an analogue of the 3'-terminus of aminoacylated tRNA. A 0.4M solution of this compound maintained at pH 8.2, yields 5.5% of diglycine and 11.5% of diketopiperazine, in addition to the hydrolysis products glycine and adenosine-5'-(O-methylphosphate). Under the same conditions, glycine ethyl ester reacts much more slowly, but ultimately gives similar yields of diglycine and diketopiperazine. The aminolysis of 2'(3')-O-(glycyl)-adenosine-5'-(O-methylphosphate) by free glycine is relatively inefficient, but serine reacts 20 times more rapidly and yields up to 50% of N-glycylserine. The prebiotic significance of these reactions is discussed.


Muscarinic cholinergic activation of mouse spleen cells cytotoxic to tumor cells in vitro. Carbamylcholine, acting via a pharmacologically specific receptor, had the ability to activate effector populations of spleen cells from female BALB/cfC3H and BALB/c mice; those cell populations were then significantly reactive in vitro against syngeneic tumor target cells but were only minimally reactive to normal syngeneic target tissues. The induced reactivity was inhibited by the muscarinic cholinergic antagonists atropine, scopolamine, and isopropamide, but not by the nicotinic antagonist d-tubocurare, and it appeared to involve both T-cell and non-T-cell effectors.


Marke's disease herpesviruses. III. Purification and characterization of Marek's disease herpesvirus B antigen. Sera from chickens naturally infected with Marek's disease herpesvirus (MDHV) form preciptin lines with at least two immunologically distinct soluble antigens designated MDHV-A and MDHV-B. Partial purification and characterization of the glycoprotein MDHV-A antigen was previously reported. MDHV-B was found predominantly in the sonically treated extracts of infected cells, in contrast to the predominantly extracellular MDHV-A. Analysis of these extracts from [14C]glucosamine-labeled cells by immunodiffusion with chicken anti MDHV-B serum negative for MDHV-A followed by autoradiography confirmed that MDHV-B was a common antigen between MDHV and herpesvirus of turkeys and revealed that it was also a glycoprotein. Because of their glycoprotein nature, both MDHV-A and MDHV-B bound to concanavalin A affinity chromatography columns and could then be eluted by alpha-methyl-D-mannoside and recovered for further analysis. Concanavalin A affinity chromatography was an excellent technique for initial purification of MDHV-A and MDHV-B, since approximately 5- and 15- fold purification, respectively, was achieved in a single simple step. MDHV-B was resistant to trypsin under conditions where MDHV-A was sensitive, but was similar to MDHV-A in resistance to pH 2.0 and to 1.0 or 2.0 M urea and 0.05% Brij 35. Partially purified MDHV-B was analyzed by sucrose gradient sedimentation, isoelectric focusing, and gel filtration on Sephadex G-200 in the presence of 1.0 or 2.0 M urea and 0.05% Brij 35 to purify the antigen and to determine its physical and chemical properties in comparison with those already reported for MDHV-A. MDHV-B had a much lower isoelectric point in pH 4,54, a higher sedimentation coefficient of 4.4S, and a greater molecular weight of 58,250. These data indicate that MDHV-B is physically distinct from MDHV-A antigen, although the size difference is not sufficient to allow for effective separation. In contrast, the isoelectric point difference of greater than 2 pH units makes isoelectric focusing an effective means of purifying the antigens free of one another. The four-step purification procedure achieved greater than 200-fold purification of MDHV-B. Immunization of rabbits with this highly purified antigen results in the preparation of antisera that appeared monospecific for MDHV-B in immunodiffusion.


Effects of phosphorylation and pH on the association of NS protein with vesicular stomatitis virus cores. The proteins of vesicular stomatitis virus (VSV) were analyzed on the basis of charge as well as size in polyacrylamide gels containing urea and acetic acid. The phosphorprotein NS was resolved into two major species. The less phosphorylated NS1 species contained about 10% fewer phosphate residues than the second species, NS2. These two phosphorylated forms were compartmentalized both in the virus and in the infected cell cytoplasm. Cores from virions and the core-containing fraction of the infected cell cytoplasm contained only the NS1 form. All of the more highly phosphorylated NS2 form and some of the NS1 form were found to be free of cores, whether they were derived from virions or from the infected cell. Therefore, the degree of phosphorylation appeared to determine whether or not the NS protein became bound to VSV cores. Moreover, the amount of bound NS1 protein relative to nucleocapsids increased as the pH of the culture medium was raised from 6.6 to 7.4. Because an increased in pH increases VSV replication (Fiszman et al., J. Virol. 13:801-808, 1974; Palma and Huang, in W.S. Robinson and C.F. Fox, ed., Mechanisms of Virus Disease, ICN-UCLA Symposia, p. 87-100, 1974), the NS1 protein may either regulate overall VSV RNA synthesis or regulate the switch between transcription and replication.


Methyprylon-induced bone marrow suppression in siblings. An inherited defect? Transient bone marrow suppression in two sisters followed ingestion of the sedative-hypnotic drug methyprylon. No other pharmacologic or environmental inciting factor common to both patients was identified. The mechanisms responsible could not be defined; abnormal suppression fo granulocyte progenitors by the drug in vitro was not demonstrable. It is postulated that a pharmacogenetic interaction may have been responsible for methyprylon-related bone marrow suppression in these two siblings.


Clinical evaluation of a new anticoagulant therapy in prosthetic valve replacement. Thromboembolic complication was correlated with the methods of anticoagulant therapy used in the patients with Starr-Edwards non cloth-covered or cloth-covered ball valve. A new anticoagulant therapy combining Bucolome with Warfarin has been used for the patients who were unable to be maintained in a suitable range of thrombotest Owren with Warfarin alone. The results obtained in this new therapy showed that the incidence of thromboembolism was reduced to 0.02 per patient year or less in the patients with Starr-Edwards ball valve as well as Björk-Shiley tilting disc valve.


Antisecretory effect of imidazole and its derivatives in an isolated  gastric mucosa preparation and an anesthetized young chicken preparation; comparison with a histamine H2-receptor antagonist. We invesigated the influences of imidazole on the basal and the secretagogue-stimulated gastric acid secretion in isolated bullfrog gastric mucosa preparations and in anesthetized young chickens. Imidazoles (1 x 10(-4) g/ml) readily depressed the basal acid secretion in gastric mucosa in vitro. The inhibitory effect of imidazole was diminished considerably after washing out of the drug. The maximum acid secretion elicited by tetragastrin or bethanechol was completely antagonized by imidazole (1 x 10(-4) g/ml). The stimulatory action of histamine or dibutyryl cyclic AMP was also remarkably depressed in the presence of imidazole (3 x 10(-4) g/ml). after dibenamine pretreatment (5 x 10(-5) g/ml) for 60 min, the isolated gastric mucosa preparation became refractory to tetragastrin, bethanechol and histamine, but responded to dibutyryl cyclic AMP. Imidazole protected the histamine sensitivity against dibenamine blockade in the concentration of 5 x 10(-4) g/ml. In anesthetized young chickens, imidazole (200 mg/kg, s.c.) depressed tetragastrin- and histamine-stimulated gastric acid secretion. The effects of the imidazole derivatives and several antagonists (metiamide, atropine, diphenhydramine, acetazolamide and 2,4-dinitrophenol) on acid production were compared with that of imidazole. From these results, it is concluded that imidazole has a potent antisecretory effect on the basal and the secretagogue-stimulated acid secretion.


Manufacture and utilization of antimony pH electrodes. A new technique for manufacturing single-barreled and double-barreled antimony pH microelectrodes is described. The results of investigations into the accuracy of antimony as a pH sensor disclosed that the pH-voltage response is: 1) within the physiologic range, principally the result of the hydrogen ion activity of the solution in which the voltage is being developed, 2) in part, qualitatively anion-dependent, 3) modified by the presence of significant amounts of at least carbon dioxide, oxygen, and nitrogen gases, and 4) markedly offset by fluctuations in temperature. Our results further indicate that the accuracy of antimony as a pH sensor is determined by the quality of the calibration procedure. We conclude that if the antimony electrode is to accurately determine the pH of a biological fluid, the pH calibration solutions must closely resemble the unknown biological fluid with respect to temperature, PO2, PN2, and types of buffering anions. A calibration procedure is described which can minimize errors with antimony pH estimations when measuring the pH of proximal tubular fluid of the mammalian kidney.


Primary malignant melanoma of the small intestine and the APUD cell concept. A primary melanoma of the small intestine is reported. A possible explanation of its origin according to the APUD cell concept is suggested. This concept may lessen the doubts and confusion whenever an intestinal melanoma is discovered and a primary tumor at one of the more common sites can reasonably be excluded.


Age-associated changes in glutamine synthetase activity in WI-38 cells. The human fibroblast cell line WI-38 shows a gradual decline in glutamine synthetase specific activity with increasing age in culture. However, the level of functional enzyme per cell does not change with age. Heat inactivation profiles indicate that glutamine synthetase from younger cell cultures is more heat labile than that from older cultures. A possible explanation for these observations is that alterations in the glutamine synthetase molecule occur with increasing age of WI-38 cells in culture.


Developmental restrictions on hormone modulated gene transcription. II. Hormone induced interactions of RNA polymerase with chromatin. Chromatin-bound and soluble RNA polymerase subspecies have been isolated and fractionated by isoelectric focusing at various times (0, 6, 12 and 18 h) following auxin treatment of 4 day (responsive) and 8 day (unresponsive) soybean hypocotyls. Young 4 day seedlings displayed two well defined phases of auxin induced gene transcription. Phase I (6 h) evidenced the selective dissociation of many RNA polymerase subspecies from the chromatin complex which was accompanied by the retention of three class II enzymes. Phase II occurred after 12 h of treatment when the dissociated enzymes including some species which were soluble in the 0 h controls became re-associated with chromatin. These induced RNA polymerases may be responsible for the synthesis of auxin induced RNAs. In contrast, the unresponsive 8 day hypocotyl did not display two phases of auxin induction. Phase one, the dissociation of the chromatin bound enzymes, occurred at 12 h (compared to 6 h for the 4 day seedling) and was not followed by the later translocation of any soluble enzymes towards the chromatin complex. The results support earlier findings suggesting that the developmental "phasing out" of RNA polymerase subspecies limits the hormone induced growth response of this tissue and thus is regarded as an off switch for the transcription of such hormone controlled gene sequences.


Emergence of multiply resistant pneumococci. Multiple antimicrobial resistance in pneumococci was detected in Johannesburg in July, 1977, and prompted an investigation of the prevalence of resistant strains in two hospitals. Carriers of Types 6A and 19A penicillin-resistant pneumococci, resistant to antibiotic concentrations ranging between 0.12 and 4 microgram per milliliter were found in 29 per cent of 543 pediatric patients and 2 per cent of 434 hospital staff members. Multiply resistant Type 19A strains, resistant to beta-lactam antibiotics, erythromycin, clindamycin, tetracycline and chloramphenicol, were isolated from 128 carriers, and were responsible for bacteremia in four patients. Isolates from 40 other carriers were resistant to penicillin alone or to penicillin and chloramphenicol or to penicillin, chloramphenicol and tetracycline. Pneumococci can be screened for penicillin resistance with a modified Kirby--Bauer technic; the strains with zones of less than 35 mm around 6-microgram penicillin disks or less than 25 mm around 5-microgram methicillin disks should be tested for sensitivity to penicillin by measurements of minimum inhibitory concentration.


Polygalacturonase, biomass and ascospore production by Byssochlamys fulva. II. Effects of sugars found in fruits. Polygalacturonase, biomass, and ascospore production by four strains of Byssochlamys fulva cultured in laboratory media supplemented with glucose, sucrose, or fructose was studied over a 20-day incubation period at 30 degrees C. The production of polygalacturonase was variable, but most activity was detected between 4 and 8 days in 1% sugar media at an initial pH of 4 or 5. The rate of biomass production was retarded early in the incubation period in media initially at pH 3 or 4 as compared to pH 5, but the amount of growth was about the same in media containing the test sugars after 20 days. Large numbers of ascospores were produced between 8 and 10 days in media containing 5% sugar initially at pH 5 and 4. Production of ascospores was retarded at pH 3 in media containing 5% sugar as compared to media initially at pH 5 and 4.


The distribution of geophilic dermatophytes in Kenyan soils. Two hundred and eighty one soil samples from different provinces of Kenya were examined for dermatophytes by the hair baiting technique. Dermatophytes were recovered from 84 samples. Microsporum gypseum constituted 75.8% of the total isolates while Keratinomyces ajelloi and M. cookei formed 21% and 3.2%, respectively. The distribution of the dermatophytes was influences by soil pH, being more prevalent in acidic than in alkaline soils.


Mycological profile of crew during 56-day simulated orbital flight. Mycological analyses were made on specimens obtained from the skin, oral rinse, urine, and feces of three subjects participating in a 56 day Skylab simulation test. Samples were taken over a 175 day period divided into three phases consisting of a prechamber, intrachamber, and postchamber period. A total of 69 species of filamentous fungi and 22 species of yeast and yeast-like fungi were recovered and identified. There was a marked decrease in the isolation incidence of both the filamentous and yeast and yeast-like fungi during the intrachamber period. This, taken with the fact that 71% of the filamentous species and 55% of the yeast and yeast-like species were isolated no more than twice, reflects the transitory nature of the relationship between the human body and most fungi. However, there was a relative increase in the potential pathogens Candida albicans and Candida parapsilosis during the intrachamber period. Over 50% of all fungi recovered were isolated from the nasal and oral cavities. There was no evidence of intercrew transfer of a particular species during the intrachamber period nor was there any evidence of microbial shock during the postchamber period.


Experiments to increase the selectivity of tumor chemotherapy by means of in vivo activation of transport forms of cancerostatics by exogenous enzymes. A significant tumor damaging effect (growth inhibition) on transplanted syngeneic sarcoma in mouse was obtained by means of pH-dependent activation of a transport form of a cancerostatic drug by an enzyme foreign to the organism. This effect was achieved by combined administration of 8-0-(alpha-L-arabinofuranosyl)beta-peltatin-A as a transport form of beta-peltatin-A and the exogenous enzyme alpha-L-arabinofuranosidase from Aspergillus niger and additional increase of the acidity of the tumor by injection of glucose. The combined application of the transport form plus enzyme showed a more favorable effect on selectivity than free peltatin when a quantitative comparison was made between the tumor growth inhibition and the damage to the blood picture.


[Neuroleptics with prolonged action].

[Neuroleptics with prolonged action]. In the last twenty years, the study of long-acting neuroleptic drugs has been active and promising progress in maintenance of psychotic patients. At present, there are several long-acting derivatives of phenothiazines, e.g. thioridazine, or drugs that allow the esterification of phenothiazine, e.g. perphenazine, fluphenazine and pipothiazine. Other esterifiable product is flupenthizol, which is a thioxanthene, and other long-acting drug is chlofluperol, which is a butyrophenone derivative. There is also a new series of neuroleptics, diphenilbultilpiperidine derivatives, such as fluspirilene, pimocide and penfluridol. Our clinical studies with these drugs allows us to state that there is a new revolution in pharmacological psychiatry, with important perspectives in the ways of helping patients.


[Treatment of epileptic crisis with chlorazepate dipotassium. Clinical study of 48 patients with the determination of serum levels of nordiazepam]. 48 patients with several types of seizures (following the international classification of epileptic seizures), were studied. Dipotassium chlorazepate was administered as a secondary antiepileptic drug. The cases were selected due to the severity of their seizures. The therapeutic results were evaluated with a daily record of seizures and attempt was made to correlate the serum levels of nordiazepam with the clinical results.


[Study of the antispastic activity of prifinium bromide in motor dysfunction of the esophagus]. Motor disturbances of the oesophagus are attributable to hypermotility or hypomotility, or to peristaltic uncoordination of the upper or lower sphincter, or of the corpus. Endoesophageal manometry has enabled considerable progress to be made in the study of oesophageal dysfunction by allowing the spasm to be quantified. Treatment is poorly effective and its results are uncertain, save in the very few cases where surgery leads to resolution. For this reason, the efficacy of a new drug with marked antispastic properties, prifinium bromide, was investigated in 12 cases of megaoesophagus, of hiatal hernia, 6 of diverticulum, and 4 of dyskinesia in the light of the manometric values observed before and after its administration. The drug relieved spasm of the sphincters and corpus.


[Effect of mouse age on the ability of hematopoietic stem cells to interact with thymus cells]. The effect of the thymus cells of the C57BL/6 mice on the colony forming ability of the stem hemopoietic cells of the embryonic liver and bone marrow of young (3 months) and old (2 years) mice was studied their joint transplantation into the mice (CBAXXC57BL/6) F1. The stimulating effect of the thymus cells on the colony forming ability of the stem hemopoietic cells of different age depends both on the dose of the stem hemopoietic cells of embryonic liver and the dose of T-lymphocytes. A suggestion is put forward that the stimulating effect of the thymus cells on the colony formation is due to their interaction with the stem cells in the G2 phase of the mitotic cycle.


[Relationship between hematopoietic stem differentiation and the functional state of the thymus]. The direction of differentiation of the stem cells with respect to the physiological activity of thymus determined by the age of an animal was studied by means of histological analysis of hemopoietic colonies in the spleen of lethally irradiated mice. The immaturity of thymus of its involution are characterized by the inhibition of differentiation of the stem cell along the granuloid path. An analysis of the data on differentiation of the stem cells in mice of different age, as well as in thymectomized mice allows to draw a conclusion that the process of differentiation of the hemopoietic stem cells is thymus-dependent.


Takayasu arteritis and renovascular hypertension in childhood. Takayasu arteritis with renal artery involvement was the cause of severe persistent hypertension in eight children under 12 years of age. Features of severe hypertension dominated the clinical picture. Unequal or absent pulses were found in three patients. An irregular contour of the descending thoracic aorta on the chest roentgenogram was an early clue to the diagnosis. On aortography both renal arteries were affected in five patients, although a discrepancy in kidney size on excretory urography had suggested a unilateral lesion. Total aortography is mandatory to outline the full extent of the vascular abnormalities. All the patients had strongly positive tuberculin skin tests, and, although mycobacteria were not isolated, all patients received antituberculous as well as antihypertensive therapy. Nephrectomy of the worse kidney is contraindicated if the renal arteries are asymmetrically involved, for fear of later extension of the arteritis. Takayasu arteritis is an important cause of severe persistent hypertension in nonwhite children.


The response of lymphatic smooth muscles to vasoactive substances. A study was made of the isotonic response of bovine mesenteric lymphatics to several physiological vasoactive substances. Contractions of lymphatic smooth muscles were induced by serotonin (5-HT), prostaglandin F2alpha (PGF2alpha), noradrenaline (NA), histamine, dopamine and acetylcholine (ACh). The smooth muscles were particularly sensitive to 5-HT. Excepting PGF2alpha no other substances could equal 5-HT in the magnitude of the maximum response. The majority of 5-HT receptors seemed to be the D receptors. The decreasing order of the contractile responses was as follows: 5-HT greater than PGF2alpha greater than NA greater than histamine greater than dopamine greater than ACh. The contractile response to ACh was observed only in specimens involving valvular region. It was very likely that, in the lymphatics, there were 2 kinds of receptors for catecholamines, i.e. alpha and beta receptors, and the stimulation of the former induced smooth muscle contraction and that of the latter relaxation. A difference was noticed between the responses of valvular and intervalvular segments to NA. Relaxations of lymphatic smooth muscles were induced not only by isoproterenol but also by adenosine and adenine nucleotides. The decreasing order of the relaxant responses was as follows: ISP greater than adenosine greater than ATP greater than ADP greater cyclic AMP greater than or equal to AMP. The relaxant responses to adenine nucleotides tended to reduce with decrease in the number of high energy phosphates.


High pyruvate kinase activity causes low concentration of 2,3-diphosphoglycerate in fetal rabbit red cells. 1. The high oxygen affinity of fetal blood in rabbits is due to a very low concentration of 2,3-diphosphoglycerate (2,3-DPG) in the red cells. In order to gather informations on the factors responsible for this characteristic we have studied synthesis and break-down of 2,3-DPG in fetal and adult rabbit red cells in vitro and examined possible regulative pathways which may lead to the low 2,3-DPG concentration in vivo. 2. Under conditions where 2,3-DPG and 3-phosphoglycerate (3-PGA) accumulate in adult erythrocytes, i.e. in a solution containing inosine, pyruvate and inorganic phosphate, the amount of 2,3-DPG synthetized in fetal red cells was only 40% of the adult value and 3-PGA was not measurable. Upon inhibition of enolase by NaF, however, both 2,3-DPG and 3-PGA increased to a similar extent in fetal and adult red cells. These findings point towards differences in the pyruvate kinase (PK) reaction which is one of the rate limiting steps of glycolysis. Direct measurements revealed an over tenfold higher PK activity in fetal compared to adult red cells. This higher activity of PK will lead to a decreased concentration of 3-PGA with a consecutive fall in 2,3-DPG concentration. 3. Other factors, like a decreased glucose utilization, a decreased activity of 2,3-DPG mutase or an increased 2,3-DPG phosphatase activity could be excluded as a cause for the low 2,3-DPG concentration in fetal red blood cells. The same holds for extraerythrocytic factors like glucose concentration or pH value in fetal blood. 4. During the postnatal development of rabbits the PK activity decreased. 50 days after birth, PK activity was 20% of the fetal value but still somewhat higher than in adult erythrocytes. This change is paralleled by an increase in 2,3-DPG concentration and half saturation oxygen pressure. With respect to the synthesis of 2,3-DPG and ATP, the fetal rabbit red cell is comparable to hereditary high PK activity in human erythrocytes.


Cooperation of peripheral and central chemosensitive mechanisms in the control of the extracellular pH in brain in non-respiratory acidosis. The mathematical model of the respiratory control system in man of Middendorf and Loeschcke (1976 a, b) opens the possibility to stimulate the constellation of parameters in non-respiratory acidosis. Several investigators agree that the pH in CSF or in the extracellular fluid of the brain stays remarkably constant in this situation and it can be shown that this is a result of a precise control rather than the consequence of a sluggishly reacting system. Application of the model assuming constant extracellular brain pH allowed to calculate the relative sensitivities to pH changes of the central and the peripheral sensory mechanisms generating respiratory drive. Assuming air breathing and a normal critical arterial O2-pressure and otherwise normal parameters of respiration, circulation and blood composition (except diminished buffer base) the central chemosensitivity to a pH change turned out to be 25 times the peripheral. This factor is critically dependent on the ratio of the bicarbonate change in extracellular brain fluid to that in arterial blood. The coinciding data of Fencl (1971) and of Kronenberg and Cain (1968) were used for the calculation.


Purification and properties of DNA-dependent DNA-polymerases from Neurospora crassa. Two DNA-dependent DNA-polymerases (E.C. 2.7.7.7) are partially purified from the high speed supernatant of mechanically disrupted hyphae of Neurospora crassa WT 74A. Some properties such as temperature and pH optimum and theoptimal concentrations for Mg2+, Zn2+, NH4+ and Na+ are very similar. On the other hand these enzymes show different properties on ion-exchange columns, are well distinguished by molecular weight (147 000 d and 110 000 d for A and B resp.) and the stimulation by K+ differs (K+ optimum for A: 5-70 mM and for B: 45 mM). Mn2+ and Zn2+ inhibit incorporation of deoxyribonucleoside monophosphates between 70 and 90%. Our best preparations so far have specific activities of 13 200 units/mg protein for A and 12 000 units/mg protein for B.


Observations on vascular neuropathies.

Observations on vascular neuropathies. A series of autopsy studies is presented of 13 cases of obliterative vascular disease of the vasa nervorum in a wide range of conditions, including polyarteritis nodosa, Wegener's granulomatosis, lymphomatoid granulomatosis, polymyositis and rheumatoid arthritis. In most cases, including 1 of rheumatoid neuropathy, the lesions were of the necrotising type. This supports the concept of polyarteritis nodosa as a pattern of reaction common to several diseases rather than that of a specific nosological entity. The remaining 4 cases of rheumatoid neuropathy, were associated with a bland endarteritis known as Bywater's arteritis. A comparison of these autopsy findings with clinical data and observations on sural nerve biopsies leads to the conclusion that all severe and most of the mild cases of rheumatoid neuropathy are due to occlusive vascular disease. Segmental demyelination independent of vascular lesions may be responsible for some of the mild cases and is frequently found in patients without clinical neurological manifestations.


Correlation of the biological activity and solution conformation of [Asp1,Ile5]- and [Phe4,Tyr8]angiotensin II. Angiotensin II is known to undergo a reversible conformational transition and a change in potency in rat uterus in vitro with pK approximately 6.5. We have shown by carbon-13 NMR that the conformational transition involves all-trans to partly cis isomerization of the His6-Pro7 peptide bond. Isomerization from all-trans at pH 6.8 to approximately 16% cis at pH 8.0 is therefore correlated with a 10-fold increase in biological activity for [Asp1,Ile5]-angiotensin II in rat uterus in vitro. Isomerization from all-trans at pH 6.8 to approximately 16% cis at pH 8.0 in the competitive inhibitor [Phe4,Tyr8]angiotensin II is correlated with exhibition of virtually no agonist activity at low pH to full agonist activity at high pH. An angiotensin II conformation with Pro7 in the cis form may therefore be the conformation with maximal binding or biological activity at the cellular receptor.


Role of a transmembrane pH gradient in epinephrine transport by chromaffin granule membrane vesicles. ATP-driven transport and accumulation of epinephrine in chromaffin granule membrane vesicles isolated from bovine adrenal medulla is inhibited by the proton ionophores carbonylcyanide p-trifluoromethoxyphenylhydrazone and nigericin, but not by valinomycin. Moreover, an artificially imposed pH gradient (interior acid) is able to drive this reserpine-sensitive transport system in the absence of ATP. Dicyclohexylcarbodiimide, an inactivator of the chromaffin granule membrane-bound ATPase, completely inhibits ATP-dependent epinephrine accumulation, but has much less effect when an imposed pH gradient is the driving force for epinephrine transport. The findings provide a strong indication that a pH gradient (interior acid) is the immediate driving force for epinephrine uptake in these storage granules and suggest that ATP-driven epinephrine transport is the result of two processes: (i) generation of a proton electrochemical gradient (interior acid and positive) by the membrane-bound, proton-translocating ATPase; and (ii) pH gradient-driven accumulation of the catecholamine.


Gaucher disease: isolation and comparison of normal and mutant glucocerebrosidase from human spleen tissue. Glucocerebrosidase was purified 26,000-fold from spleens from normal humans and from patients with Gaucher disease (Gaucher spleens). The specific activities of the purified normal and mutant enzymes with glucocerebroside as substrate were 8.5 X 10(5) and 5.4 X 10(4) nmol/mg of protein per hr, respectively. The ratio of enzymatic activities was constant throughout the isolation procedure. The two enzymes appeared to be similar by other parameters such as substrate affinity, heat lability, and pH optimum. Immunotitration with glucocerebrosidase antiserum showed equivalent quantities of crossreacting material in extracts of normal and Gaucher spleens. These data strongly suggest that the genetic basis of Gaucher disease is a strucutral mutation of glucocerebrosidase. The results of sodium dodecyl sulfate gel electrophporesis also indicate that there are differences between the normal and the Gaucher disease enzyme.


Potentiation by neuroleptic agents of the inhibitory action of intraperitoneally administered GABA on the locomotor activity of mice. The ability of several neuroleptics to potentiate the inhibitory action of IP administered GABA on the motor activity of mice has been investigated. Haloperidol, chlorpromazine, thioridazine, and clozapine, but not the apparently selective dopamine receptor-blocking agent spiperone, were found to possess such activity. Phenoxybenzamine also proved active in potentiating GABA. Thus blockade of dopamine receptors as well as alpha-adrenergic receptors may be responsible for neuroleptic-induced potentiation of GABA actions.


A new putative neuroregulator of the extrapyramidal system. A putative neurotransmitter, 2-phenylethylamine, which is most highly concentrated in the extrapyramidal system of human brain, is able to reverse reserpine-induced parkinsonism in animals and elicit stereotypy. This action is only partially antagonized after catecholamine depletion by pretreatment with a-methyl-para-tyrosine, and fully blocked by pretreatment with haloperidol, a dopamine receptor blocker. Therefore, via direct and/or indirect actions, 2-phenylethylamine may serve a neuroregulatory role in the extrapyramidal system.


The isolated nictitating membrane of the cat. A new model for the study of narcotic analgesics. Kinetic parameters of morphine and azidomorphine derivatives as well as methionine enkephalin methylester were determined on the electrically stimulated isolated medial smooth muscle of the cat nictitating membrane. The organ was found to be a promising in vitro model for testing the action of opiate agonists on opiate B receptors.


Chemical stability of prostacyclin (PGI2) in aqueous solutions. The rate constant for the hydrolysis of prostacyclin (PGI2) to 6-keto-PGF1alpha was measured by monitoring the UV spectral change, over a pH range 6 to 10 at 25 degrees C and the total ionic strength of 0.5 M. The first-order rate constant (kdegreesobs) extrapolated to zero buffer concentration follows an expression, kdegreesobs = kH+ (H+), where kH+ is a second-order rate constant for the specific acid catalyzed hydrolysis. The value of kH+ obtained (3.71 x 10(4) sec-1 M-1) Is estimated approximately 700-fold greater than a kH+ value expected from the hydrolysis of other vinyl ethers. Such an unusually high reactivity of PGI2 even for a vinyl ether is attributed to a possible ring strain release that would occur upon the rate controlling protonation of C5. A Brønsted slope (alpha) of 0.71 was obtained for the acid (including H3O+) catalytic constants, from which a pH independent first-order rate constant for the spontaneous hydrolysis (catalyzed by H2O as a general acid) was estimated to be 1.3 x 10(-6) sec-1. An apparent activation energy (Ea) of 11.85 Kcal/mole was obtained for the hydrolysis at pH 7.48, from which a half-life of PGI2 at 4 degrees C was estimated to be approximately 14.5 min. when the total phosphate concentration is 0.165 M (cf. 3.5 min. at 25 degrees C).


Retained inhibition of the beta receptor by propranolol after preincubation and washout. Mouse thymus cells possess a B2-adrenergic receptor and, when centrifuged through a solution of propanolol at 5 degrees and washed three times, will produce reduced amounts of cyclic AMP during a 5 minute challenge with isoproterenol at room temperature. Depending on the concentration and nature of the antagonist, the retained inhibition may be greater or less than that seen when it is added with the agonist. This retained inhibition is readily reversed at 37 degrees even in the absence of the agonist. These effects can also be demonstrated with rat erythrocytes and with their subsequently prepared ghosts making it most likely that they result from the specific and tight binding of the antagonist to the beta receptor.


Effect of benzodiazepine derivatives on human blood cholinesterase in vitro. The in vitro effect of ten benzodiazepine derivatives on the cholinesterases of human plasma and red cells was determined. Flurazepam, temazepam, lorazepam, flunitrazepam and diazepam had an inhibitory effect on plasma cholinesterase of 60--90 per cent and, with the exception of lorazepam, an inhibitory effect of 40--50 per cent on red cell cholinesterase. Clonazepam, oxazepam, chlordiazepoxide, nitrazepam and bromazepam had comparatively minor effects on both enzymes


[Physiological trials with MIF-I in Parkinson's disease (author's transl)]. The writers present some physiological (and not therapeutic) acute and sub-acute trials with tripeptide: L-propyl-L-leucyl-glycine amide (M.I.F.-I) in Parkinson's Disease. This work confirms the earlier observation that M.I.F.-I employed alone or in combination with Levodopa is active against Parkinson's disease. The writers evoke the hypothesis that this action takes place at the postsynaptic receptors, whose configuration may be modified in the sense of a hypersensitivity. These studies justify undertaking a series of controlled therapeutic trials when the cost of the product permits and as well instituting development work on analogues active orally.


[Neuroleptics and sexual dysfunction in man. Neuroendocrine aspects]. Sexual behaviour and fertility of schizophrenic patients are discussed. The various sexual dysfunctions induced by neuroleptics, such as decrease in libido, troubles of ejaculation and impotence, are described and related to their various mechanisms of action. The central antidopaminergic effect of neuroleptics would be responsible for an overall non-specific decrease in libido. Their effect upon the autonomous nervous system would explain ejaculation disturbances. Their endocrine action (increase in prolactine) would produce impotence. The possibility of treatment of these endocrinological troubles by bromocriptine is discussed.


Lymphocyte changes after long-term therapy of some neurological diseases. Hematologic investigations were carried out in 50 patients with various forms of epilepsy, aged 20 to 50 years, who had been treated for different periods of time with antiepileptic and neuroleptic drugs such as primidon, luminal and prasine. All the cytomorphological and cytochemical studies concerned the white blood cells, especially lymphocytes, and their transformation into plasma cells and histiomonocytes. The cytochemical investigations referred particularly to nucleolar-RNA, glycogen and lipid cellular content. The results obtained showed that the lymphocyte count and mostly that of the large and medium lymphocytes, may reach twice the normal or more. The transition forms towards histiomonocytes and plasma cells were quite frequent. Cytochemically, a variable state of reactivity of the nucleolar apparatus was demonstrated, as well as certain changes of the carbohydrate and lipid metabolism. The influence of such changes on the cellular and humoral immunity of the respective patients is discussed. Emphasis is also laid on the utility of such investigations for the detection of the untoward effects of antiepileptic and neuroleptic drugs on hematopoiesis.


[Histamine H-2 receptor antagonists and medical treatment of Zollinger-Ellison syndrome]. Cimetidine was studied in 8 cases of Zollinger-Ellison syndrome. The average dose was 1,2 g a day for 14 to 195 days. The treatment was remarkably effective in 2 cases, but only partial or temporary improvement was noted in 3 other cases. The last 3 patients failed to respond to treatment. The possibility of a rebound phenomenon requires strict clinical and biological supervision.


Small cell carcinoma of the lung: cellular origin and relationship to other neoplasms. Many small cell carcinomas share morphological and physiological characteristics with normal and neoplastic cells of Pearse's APUD series, including pulmonary APUD cells and pulmonary carcinoid tumors. There is very likely more than one type of APUD cell in the lung, and conclusions that small cell carcinomas and carcinoids reflect neoplastic transformation of the same cell type are probably premature. The embryoogic lineage of pulmonary APUD cells is at present uncertain. The hypothesis that all APUD cells are derived from the neural crest is no longer tenable, and although some evidence does suggest a neural contribution to the pulmonary epithelium, additional embryologic studies are required. Some tumors that currently are classified as small cell carcinomas probably do not have APUD cell characteristics, and still others appear to have both APUD and non-APUD features. A  subclassification of small cell carcinomas based on a combination of physiological and morphological features might prove to be of prognostic and therapeutic value, but current knowledge probably would not provide a sufficient foundation for a reliable or practical subclassification. Multidisciplinary studies of the differentiation and function of normal and neoplastic APUD cells in the lungs and elsewhere are needed.


Maternal blood gas status of four racial groups in late pregnancy A preliminary study. The blood gas/acid-base status of normal healthy Black, Indian, Coloured and White pregnant women was measured. All groups exhibited evidence of hyperventilation (decreased) PaCO2 relative to the non-pregnant state, with compensatory metabolic adjustment. Arterial oxygen tensions (PaO2) were significantly lower in the Black and Indian groups compared with their White counterparts (P less than 0,01). Differences between the former two series and the Coloured group, however, just failed to reach statistically significant levels (0,1 less than P less than 0,05). The possible causes of a lowered PaO2 during pregnancy and their relevance to obstetric anaesthesia are discussed. A further study using a more sophisticated protocol is planned in an attempt to delineate the causes of these important differences.


Antacid properties of a sodium citrate preparation. A sodium citrate preparation (Citro-Soda; Abbott) given in single doses of 4 or 12 g, satisfactorily increased the pH level of urine without affecting the systemic acid-base balance. When the sodium citrate preparation was administered in the 12-g single dose the gastric pH level was raised above 3,0 in a significant number of subjects. Similar results were obtained with both single doses (4 and 12 g) after 3 - 6 days of continuous therapy at the appropriate dose given every 6 hours. It is concluded that the urine-alkalinizing sodium citrate preparation reduced intragastric acidity significantly.


Assessment of a diagnostic dip-stick for assaying plasma or serum pseudocholinesterase activity. A diagnostic dip-stick for measuring pseudocholinesterase activity has been assessed by 4 participants and compared with a standard spectrophotometric method. Duplicate dip-stick estimations (measured in kU/l) corresponded closely, and concurrence between participants was good. The dip-stick estimate of cholinesterase was consistently lower than the spectrophotometric value; the magnitude of this deviation depended on the absolute enzyme activity. With enzyme activities of up to 1 kU/l the dip-stick value was 1 unit lower. Between 1 and 4 kU/l the dip-stick estimate compared well with the spectrophotometric value. From 5 to 7 kU/l and beyond 7 kU/l the negative deviation recorded by the dip-stick increased to 2 units or more. Normal enzyme activity is 3 - 6 kU/l. Over the 1 - 4 kU/l range the method is most accurate, thus negative deviation below 1 kU/l or above 4 kU/l is generally diagnostically irrelevant. On the basis of cost, convenience, speed and accuracy this method will suffice in the absence of sophisticated assay facilities.


Clinical usefulness of the beta-adrenergic antagonists. The pharmacology of the beta-adrenergic antagonists, particularly their role as competitive antagonists at the receptor site, is discussed. The clinical use of these agents is listed and a detailed outline of their use in angina pectoris, hypertension and conditions associated with increased sympathetic activity is provided. The relative contraindications to the use of beta-adrenergic antagonists are summarized and the pharmacology of those drug interactions involving these agents is discussed. A thorough understanding of the pharmacology and therapeutics of these agents, which interfere with physiological function, is mandatory prior to their clinical use.


Teratogenic action of bis(dichloroacetyl)diamine on rats: patterns of malformations produced in high incidence at time-limited periods of development. Win 18,446, a bis(dichloroacetyl)diamine, is a drug that was previously shown to suppress spermatogenesis and to be an effective oral abortifacient in rats. The present study shows that the drug is capable of producing characteristic congenital malformations in high incidence, by a single treatment, and with high survival of fetuses to day 21. Gestation day 11 is the most sensitive time. The teratologies obtained after various schedules of treatment include malformations of the snout (100%), septal heart defects (100%) diaphragmatic hernias (100%), cryptorchism (100%), cervical pockets (100%) and absent or small irregular thymus (92%). Some of these data, namely of the  heart, face and thymus, cluster in patterns that indicate that the action of the drug is upon a time-resistricted developmental process. These periods of sensitivity are subsiding or have ended before primordia of these structures appear, but they coincide with the proliferation and migration of those mesenchyme cells that will eventually form or contribute to the structures affected. It is postulated that the drug acts on these mesenchyme cells, or on the extracellular matrix that provides the necessary framework for their dispersal.


Stabilization of interferons.

Stabilization of interferons. It is now obvious that there are a number of factors which can influence the stability of interferons. Since human fibroblast interferon is especially unstable both to heat and mechanical stress, the use of stabilizing conditions such as low pH or thioctic acid should facilitate purification and concentration of this interferon. Leukocyte interferon, on the other hand, seems to be more stable even when highly purified; however, losses in activity can occur in very dilute solutions containing low concentrations of protein. Inactivation during storage of interferons may be avoided by using freeze-dried preparations. In order to generate large quantities of purified, stable interferons for clinical use, there is a need to establish an armamentarium of different nontoxic techniques and approaches that will enable investigators to stabilize human interferons at each step in preparation, processing, concentration, purification, shipment and storage.


Study on dissolution and disintegration of calcium bilirubinate stone. When a slice of calcium bilirubinate stone was incubated in a solution of tetrasodium salt of ethylendiamine-tetraacetic acid (EDTA.4Na), a potent chelating agent, the solution exhibited a yellow brown tint, which was spectroscopically characteristic of bilirubin. Microscopic examination of the slice revealed dissolution of granules of calcium bilirubinate, leaving behind a reticular matrix of PAS-positive substance. The effect of EDTA.4Na was influenced by pH, being fully effective only at pH 10 or more, and by temperature and concentration as well. Simultaneous application of bile salt enhanced the activity of EDTA.4Na, hydrophilizing the gallstone surface to facilitate chelating reaction and also dissolving minor fatty components of the stone. Heparin at proper concentrations also promoted disintegration of the stone, changing surface potential of its constituent particles to the dispersion-prone charge. The effect of composite EDTA.4Na-bile salt-heparin was thus significantly greater than that of single EDTA.4Na. This mixture is promising for clinical application as a means of direct dissolution of residual gallstones.


[Concentration of acid and basic proteins in the nuclei of hippocampal neurons and neuroglia of rat brain following elaboration of a passive avoidance conditioned reflex]. By means of two-wave-length visible cytophotometry, the content of acidic and basic proteins per cell nucleus was shown to increase in CA3 hippocampal neurons only 3 hours after the learning session of conditioned reflex of passive avoidance. The protein content in the nuclei of perineuronal neuroglia cells of CA3 hippocampus increased only 6 hours after the learning session. Biochemical mechanisms dealing with the process of learning are discussed.


[Enzymologic study of the structural organization of the matrix or rat liver peroxisomes]. The effect of ionic strength and pH on the release of some enzymes of the matrix of peroxisomes in rat's liver was studied. Catalase, L ALpha-hydroxy acid oxidase, isocitrate dehydrogenase, glycerophosphate dehydrogenase and lactate dehydrogenase were easily released from the particles during their lysis and treatment with 0.16 M KCl, whereas urate oxidase, NADH cytochrome c reductase and D-amino acid oxidase were not solubilized. After the solubilization of peroxisomal membrane by 0.2% Triton X-100, the remaining core contained about 50% amino acid oxidase activity, and had 1.28--1.30 g/cm3 density. These results suggest that D-amino acid oxidase associates with urate oxidase in the peroxisomal core.


[Acid-base state disorders and their correction in acute mesenteric vascular obstruction]. The results of the investigation of main indices of acid-base equilibrium in 28 patients with acute obstruction of the mesenteric vessels are set forth. 12 patients out of 28 developed subcompensated or decompensated metabolic acidosis. The authors believe that the correction of acid-base equilibrium disorders is an efficacious factor of intense therapy and recommend to use it at all the stages of the treatment.


Absorption of the beta-adrenergic-blocking agent, nadolol, by mice, rats, hamsters, rabbits, dogs, monkeys, and man: an unusual species difference. 1. Dogs absorbed [14C]nadolol almost completely (88--104%) after oral administration of 10--75 mg/kg. In contrast, mice, rats, hamsters, and rabbits, given oral doses of 20 mg/kg, absorbed, at most, 25%. After administration of oral doses of 2--75 mg/kg, monkeys absorbed no more than an average of 30%. Six monkeys given oral doses of 10 mg/kg absorbed 12--44%. 2. In rats, [14C]nadolol was orally absorbed to an average of 13.7--17.5%, whether given in suspension or in solution, indicating that absorption was not limited by rate of dissolution nor by solubility. 3. Mildly hypertensive humans given single doses of [14C]nadolol (2 or 80 mg, orally) and normal subjects given single doses (10 mg, orally) absorbed an average of 20.4--33.3%.


Glutamate metabolism in a monokaryon of Coprinus cinereus. The possible involvement of the glutamine synthetase/glutamate synthase pathway in ammonia assimilation is reported in a monokaryon of Coprinus cinereus.