Deoxyribonuclease A, a novel deoxyribonuclease highly active toward Polydeoxyadenylic acid and polythymidylic acid from Achatina fulica. Two novel deoxyribonucleases, termed DNases A and A', have been purified from the hepatopancreas of Achatina fulica (agate snail). DNases A and A' were obtained in 3.6 % and 7.7% yields by acetate buffer extraction and successive chromatography on hydroxyapatite, phosphocellulose and poly(A)-Sepharose. The two DNases are highly active toward poly(dA) and at a salt concentration of 0.15 m and pH 5.0 exhibit 10-fold higher hydrolyzing activities toward poly(dA) than toward calf thymus denature DNA. The enzymes also act considerably on poly(dT) at pH 4.0, but do not appreciably digest other synthetic homopolymers such as poly(dC), poly(dG) and poly(A). The limit products obtained by exhaustive digestion of poly(dA) with DNases A and A' are 98% and 99% acid-soluble and consist of oligonucleotides with an average chain length of about 5 nucleotides containing barely detectable dimers and trimers, respectively. These fragments have terminal 5'-hydroxyl and 3'-phosphate groups. The mode of action appears to be endonucleolytic. Both enzymes have the same pH optimum of 5.0 for poly(dA-hydrolyzing activity. Ionic strength is critical for the maximum activity.


Acetyl-CoA-dependent elongation of fatty acids in Mycobacterium smegmatis. An enzyme system of Mycobacterium smegmatis catalyzing the elongation of medium-chain fatty acids with acetyl-CoA was obtained free from de novo fatty acid synthetase by ammonium sulfate fractionation. The system was resolved by gel filtration and DEAE-cellulose chromatography into three fractions, all of which were required for reconstitution of the elongation activity. The three fractions were highly purified enoyl-CoA hydratase, highly purified 3-hydroxyacyl-CoA dehydrogenase, and a fraction containing both enoyl-CoA reductase and thiolase. The reconstituted system was avidin-insenstive, required NADH as a sole hydrogen donor, and was sensitive to pCMB, but not to N-ethylmaleimide or monoiodoacetate. Decanoyl-CoA and octanoyl-CoA were the best primers for the elongation system. When decanoyl-CoA was used as the primer, the major product was found to be a lauroyl derivative (probably lauroyl-CoA). Evidence was obtained suggesting that acyl-CoA dehydrogenase, catalyzing the first step of beta-oxidation, was not functional in the elongation system.


Isolation and characterization of malic enzyme from the pupa of Bombyx mori. Malic enzyme, which requires NADP+ as a coenzyme, was isolated and purified  from pupae of the silkworm, Bombyx mori. The purified enzyme appeared homogeneous and had a molecular weight of 195,000 on polyacrylamide gel electrophoresis. The optimum pH for the oxidative decarboxylation of malate, measured in terms of the increase of NADPH (MH activity) and CO2 (MC activity), was pH 7.5, while that for the decarboxylation of oxaloacetate measured in terms of the increase of CO2 (OC activity) was pH 4.6. Several differences between MH  and OC activity were investigated.


The structure and function of ribonuclease T1 XXIV. Preparation and properties of a stable water-insoluble polyacrylamide derivative of ribonuclease T1. Ribonuclease T1 [EC 3.1.4.8] was coupled to a water-insoluble cross-linked polyacrylamide (Enzacryl AH) by the acid azide method. The immobilized enzyme exhibited about 45% and 77% of the original activity toward yeast RNA and 2', 3-cyclic GMP, respectively, as substrates. Although the specific activity was lowered by the coupling, the immobilized enzyme was found to be far more stable to heat and extremes of PH than the native enzyme. The immobilized enzyme was active toward RNA even above pH 9 (at 37 degree C) or above 60 degree C (at pH 7.5), where the native enzyme was inactive. The immobilized enzyme retained much of its activity as assayed at 37 degree C after incubation in the range of pH 1 to 10 at 37 degree C, or after heating at 100 degree C (at pH 7.5) under conditions where the native enzyme was inactivated to a considerable extent. The enzyme derivative could be repeatedly recovered and reused without much loss of activity. The active site glutamic acid-58 in the immobilized enzyme appeared to be nearly as reactive with iodoacetate as that in the native enzyme.


Reactivity of ferrous myoglobin at low pH. The rates of reaction of myoglobin with carbon monoxide at low pH are reported. The pH versus rate profile of these kinetics resembles that found for heme model compounds, revealing an increase in combination rate at low pH. These facts suggest that CO binding by myoglobin changes from a mechanism of "direct ligant association" at pH 5 to a mechanism, similar to that proposed for heme model compounds, which assumes a tetracoordinated intermediate as a result of the protonation of the proximal imidazole.


Endogenous activating factor for guanylate cyclase in synaptosomal-soluble fraction of rat brain. When the crude mitochondrial fraction of rat brain was homogenized with distilled water and centrifuged, most of guanylate cyclase activity was detected in the soluble fraction. The total guanylate cyclase activity recovered in the soluble fraction was 5- to 8-fold higher than that of the crude mitochondrial fraction. The greater recovery of guanylate cyclase activity was found to be due to a release of an endogenous activating factor for guanylate cyclase. The activating factor was partially purified by acid extraction followed by a gel filtration and ion exchange resin columns. The factor was a dialyzable small molecule. The molecular weight was estimated to be between 300 and 600 by a Sephadex G-15 column and Diaflo ultrafilter membranes. It was stable in dilute acids, but labile in alkaline solution. It was readily soluble in water, but insoluble in organic solvents. Treatment with various enzymes, so far as tested, failed to abolish the activity. The activating factor stimulated the initial velocity of the reaction. It altered neither the Km value for GTP nor the dependency of the enzyme on divalent metals. The activation by the factor was due to an increase in the Vmax of the reaction. The activation was prevented by lysolecithin, Lubrol PX, hydroxylamine, methylhydroxylamine, or hemoglobin.


Studies of the beta-galactoside transporter in inverted membrane vesicles of Escherichia coli. I. Symmetrical facilitated diffusion and proton gradient-coupled transport. Facilitated diffusion of [14C]lactose into inverted membrane vesicles of Escherichia coli was measured using HgCl2 as a stopping reagent and polylysine to flocculate the vesicles for filtration. Equilibration of lactose between the internal and external volumes required expression of the y gene of the lac operon and was inhibited by thiodigalactoside or by prior incubation with N-ethylmaleimde or HgCl2. The initial rate of uptake was saturable, with a Kt of 0.95 mM. Counterflow of [14C]lactose was demonstrated in either direction. ATP hydrolysis or respiration drove the efflux of internal lactose. The effect of ATP required addition of F1 coupling factor (ATPase) from E. coli when lactose transport was studied in F1-deficient inverted vesicles. Accumulation of lactose against a concentration gradient was achieved by forming an artificial electrochemical proton gradient consisting of a membrane potential negative inside or a pH gradient basic inside. Addition of ATP inhibited this proton driven uptake showing that it occurred in inverted vesicles. It was concluded that the lactose-proton co-transport protein (M protein) is qualitatively symmetrical with respect to the facilitated diffusion of lactose and the coupling of proton and lactose transport.


Nucleus-dependent regulation of tyrosine aminotransferase degradation in hepatoma tissue culture cells. Upon removal of the  nucleus from rat hepatoma tissue culture cells, levels of the enzyme tyrosine aminotransferase no longer change in response to withdrawal of glucocorticoids. The rate of tyrosine aminotransferase degradation is drastically reduced in rat hepatoma tissue culture cytoplasts leading to stabilization of pre-existing levels of tyrosine aminotransferase. Moreover, the rate of synthesis of the enzyme in cytoplasts is very low near that observed in uninduced whole cells. These effects of enucleation occur very rapidly and appear to be specific for tyrosine aminotransferase and a small number of other unstable hepatoma proteins. A nuclear effect is thus directly involved in the control of tyrosine aminotransferase degradation and synthesis.


Stabilization of adenylate energy charge and its relation to human sperm motility. The adenylate energy charge of human ejaculated spermatozoa was studied when the sperm motility was perturbed by varying pH, prolonged incubation, and caffeine. Between pH 8 and 9, which was optimal for the sperm motility, the energy charge was in the physiological range of 0.8 to 0.9. Above pH 9, the mobility, ATP content, and adenine nucleotide pool declined rapidly but the energy charge was maintained slightly below 0.8. Below pH 8, the motility also dropped drastically, but the ATP, nucleotide pool, and energy charge fell only slightly. Prolonged incubations of the spermatozoa decreased the motility, ATP, and nucleotide pool. However, the energy charge would remain above 0.6. Caffeine stimulation of the motility caused a rapid fall of ATP and the reduction of the physiological energy charge by 0.2 unit, unless glucose was added. Imidazole which reduced the caffeine-stimulated motility did not alter the physiological energy charge of the spermatozoa. The study showed that the spermatozoa could maintain the energy charge above 0.6 under stress.


Hydrogen exchange study of membrane-bound rhodopsin. I. Protein structure. Structural parameters of rhodopsin in disc membrane preparations from frog and cattle were studied by hydrogen exchange methods. The method measures the exchange of protein amide hydrogens with water and can distinguish protons which are internally bonded from those which are hydrogen-bonded to water. The results show that about 70% of rhodopsin's peptide group protons are exposed to water. The identification of these groups as free peptides was made initially on the usual basis of the identity of their exchange rate with the well characterized free peptide rate; other experiments specifically excluded contributions from lipids, protein side chains, adventitious mucopolysaccharides, and intradisc water. In contrast to rhodopsin, other proteins generally have only 20 to 40% free peptide groups. Apparently rhodopsin has some unusual structural feature. Our results together with available information on rhodopsin suggest that a considerable length of its polypeptide chain is arranged at the surface of a channel of water penetrating into the membrane. Physicochemical considerations indicate that such a channel would have to be quite wide, 10 to 12 A or more, to explain the hydrogen exchange results.


Hydrogen exchange study of membrane-bound rhodopsin. II. Light-induced protein structure change. Hydrogen exchange studies of rhodopsin in disc membranes demonstrated that photolysis induces changes in the protein itself. Two different altered forms were detected. A late photointermediate in the bleaching sequence, which can be identified with metarhodopsin II, displays accelerated exchange. Subsequently, at the stage of fully bleached opsin, exchange becomes even slower than in rhodopsin. These changes involve only a small fraction of the protein's internally hydrogen-bonded peptide groups. The unusually large fraction of exposed peptide hydrogens observed previously for rhodopsin is unaltered in the photolyzed forms.


Nicotinamide adenine dinucleotide-specific glutamate dehydrogenase of Neurospora. IV. The COOH-terminal 669 residues of the peptide chain; comparison with other glutamate dehydrogenases. A sequence is presented for the COOH-terminal 669 residues of the NAD-specific glutamate dehydrogenase of Neurospora crassa. Comparison of this sequence with those of the vertebrate glutamate dehydrogenases of chicken and bovine liver and with the NADP-specific enzyme of Neurospora shows some similarities in sequences around residues previously identified as important for the function of these enzymes. These are: (a) the reactive lysine residue of low pK in the NADP and the vertebrate enzymes; (b) the tyrosine residue of the NADP enzyme that is readily nitrated by tetranitromethane with inactivation, a residue protected by NADP or by NMN; and (c) the arginine residue of the NADP-enzyme that is reactive with 1,2-cyclohexanedione with inactivation. Despite these similarities, comparison of the sequence of the NAD-enzyme with those of the other glutamate dehydrogenases of known sequences revealed relatively little overall homology as determined by computer analysis.


Membrane isolation on polylysine-coated beads. Plasma membrane from HeLa cells. HeLa cell plasma membranes have been purified after binding cells to polylysine-coated polyacrylamide beads. Cell attachment to beads and membrane recovery were maximal in a sucrose-acetate buffer, pH 5.0, at 25 degrees C. Measurements of ouabain-sensitive NaK-adenosine triphosphatase, membrane-bound 125I-wheat germ agglutinin, and chemical analyses showed that membranes on beads were of comparable or greater purity than membranes isolated by conventional methods. Because the isolation procedure is rapid (approximately 2.5 h), and produces membranes whose protoplasmic surfaces are fully exposed, it should be a useful supplement to standard isolation techniques.


Patterns of plasminogen activator production in cultured normal embryonic cells. Cultured normal low-passage embryo fibroblasts, from a number of species, and two untransformed clones of a Balb/3T3 line elaborate increasing amounts of plasminogen activator (PA) as they approach confluence; the low-passage cells then lose this PA activity after reaching confluence, while the 3T3 cells retain it indefinitely. Even at their peaks, however, the PA activities of the low-passage cells remain well below those of the corresponding virally or spontaneously transformed cells. The PA increases in normal cells are probably a result of PA production rather than of adsorption of secreted PA to the cell surface, or of changes in cell-associated protease inhibitors. The elaboration of PA by normal cells is dependent upon their metabolic activity, such that the level of serum supplementation and the growth phase of the culture directly influence the level of cell-associated PA observed. In addition, there may be a component of serum which exerts a negative control on PA production and which is not an acid-labile protease inhibitor.


Complementary use of the reversed-phase and anion-exchange modes of high-pressure liquid chromatography for studies of reduced nicotinamide adenine dinucleotide. The reversed-phase and anion-exchange modes of high-pressure liquid chromatography were used to separate breakdown products and impurities in solutions of the reduced form of nicotinamide adenine dinucleotide (NADH). The two chromatographic modes are compared for studies of NADH. Their use in following the course of acidic breakdown of NADH is described.


Fractionation and characterization of acidic ogligosaccharides and glycopeptides from normal and pathological urines. A general procedure is described for the isolation of urinary acidic oligosaccharides and glycopeptides resulting from catabolism of glycoproteins. This procedure has been applied to normal urine and to urine from patients with diseases of the metabolism, including mucolipidosis and fucosidosis.


Bromothymol blue and carbohydrate-sensitive plating media. A new plating medium using bromothymol blue (BTB) indicator is described and compared with eosin-methylene blue (EMB), MacConkey, and Endo media. These media were tested with L-arabinose by plating fermenting and nonfermenting mutant strains of Escherichia coli. The minimum concentrations of L-arabinose that permitted differentiation of these strains were determined. Different concentrations were required for differentiating confluent patches of cells, isolated colonies, and closely spaced or adjacent colonies. L-Arabinose, L-rhamnose, D-lactose, and D-galactose were tested with modified enteric media and with BTB medium, again to determine minimum usable concentrations. BTB media and reformulated conventional media allowed detection of acidification, aerobically, at one-fifth to one-hundredth the (1%, wt/vol) concentration of carbohydrate used in standard indicator plates.


Effect of aldosterone on the coupling between H+ transport and glucose oxidation. The mode of action of aldosterone on the energetics of H+ transport in the turtle bladder was examined with the rate of glucose oxidation as an index of the metabolic activity of the epithelium (we show that H+ transport is not coupled to fatty acid oxidation). Within 6 h of addition of aldosterone H+, transport increased; so did glucose oxidation. The amount of H+ transport per mole of 14CO2 produced from glucose oxidation was 15.6 eq-mol-1 in the control hemi-bladder, while in the aldosterone-treated bladder it was 13.6, delta = 2.0+/-4.0 (n = 6). However, in bladders exposed to aldosterone for 20 h, the relation of transport to glucose oxidation was significantly altered: control 10.8, aldosterone 16.4, delta = 4.5+/-2.5, P less than 0.02, n = 7. The slope of H+ transport on the applied electrochemical gradient was steeper during both short- and long-term incubations. However, the maximum gradient necessary to nullify the net rate of secretion was unaltered in both experiments. Evidence is presented that aldosterone does not alter the passive backflux into the cell. In five additional experiments where aldosterone produced no significant stimulation of H+ transport, no change was noted in any of the metabolic or transport characteristics measured, suggesting that the alterations discussed above are dependent on the stimulation of H+ transport by the hormone. These results, along with some thermodynamic considerations, suggest that the effect of aldosterone is primarily exerted on the transport process rather than on metabolism. Further, it appears that prolonged stimulation of transport work leads to secondary alterations in the metabolic pathways reminiscent of the changes that occur in skeletal muscles of athletes undergoing physical conditioning.


The maladaptive renal response to secondary hypocapnia during chronic HCl acidosis in the dog. It has generally been thought that homeostatic mechanisms of renal origin are responsible for minimizing the alkalemia produced by chronic hypocapnia. Recent observations from this laboratory have demonstrated, however, that the decrement in [HCO(-) (3)], which "protects" extracellular pH in normal dogs, is simply the by-product of a nonspecific effect of Paco(2) on renal hydrogen ion secretion; chronic primary hypocapnia produces virtually the same decrement in plasma [HCO(-) (3)] in dogs with chronic HCl acidosis as in normal dogs (Delta[HCO(-) (3)]/DeltaPaco(2) = 0.5), with the result that plasma [H(+)] in animals with severe acidosis rises rather than falls during superimposed forced hyperventilation. This observation raised the possibility that the secondary hypocapnia which normally accompanies metabolic acidosis, if persistent, might induce an analogous renal response and thereby contribute to the steady-state decrement in plasma [HCO(-) (3)] observed during HCl feeding. We reasoned that if sustained secondary hypocapnia provoked the kidney to depress renal bicarbonate reabsorption, the acute salutary effect of hypocapnia on plasma acidity might be seriously undermined. To isolate the possible effects of secondary hypocapnia from those of the hydrogen ion load, per se, animals were maintained in an atmosphere of 2.6% CO(2) during an initial 8-day period of acid feeding (7 mmol/kg per day); this maneuver allowed Paco(2) to be held constant at the control level of 36 mm Hg despite the hyperventilation induced by the acidemia. Steady-state bicarbonate concentration during the period of eucapnia fell from 20.8 to 16.0 meq/liter, while [H(+)] rose from 42 to 55 neq/liter. During the second phase of the study, acid feeding was continued but CO(2) was removed from the inspired air, permitting Paco(2) to fall by 6 mm Hg. In response to this secondary hypocapnia, bicarbonate concentration fell by an additional 3.0 meq/liter to a new steady-state level of 13.0 meq/liter. This reduction in bicarbonate was of sufficient magnitude to more than offset the acute salutary effect of the hypocapnia on plasma hydrogen ion concentration; in fact, steady-state [H(+)] rose as a function of the adaptive fall in Paco(2), Delta[H(+)]/Delta Paco(2) = -0.44. That the fall in bicarbonate observed in response to chronic secondary hypocapnia was the result of the change in Paco(2) was confirmed by the observation that plasma bicarbonate returned to its eucapnic level in a subgroup of animals re-exposed to 2.6% CO(2). These data indicate that the decrement in plasma [HCO(-) (3)] seen in chronic HCl acidosis is a composite function of (a) the acid load itself and (b) the renal response to the associated hyperventilation. We conclude that this renal response is maladaptive because it clearly diminishes the degree to which plasma acidity is protected by secondary hypocapnia acutely. Moreover, under some circumstances, this maladaptation actually results in more severe acidemia than would occur in the complete absence of secondary hypocapnia.


Solubility and heat stability of whey protein concentrates. Rennet whey protein concentrates have excellent nutritional properties, but their use in fluid food systems is impaired by the poor heat stability of the protein. Heating whey protein concentrated solutions at neutral pH caused up to 70% losses in solubility. In the absence of added calcium, protein coagulation occurred. near the iso-electric zone whereas in the presence of .03 M calcium chloride, similar protein coagulation occurred in the whole pH range (pH 2 to pH 12). Tryptic hydrolysis of the protein increased the heat stability of whey protein concentrates considerably.


Lymphocyte subpopulations of atopic children and the effect of therapy upon them. Recent reports note decreased T cell function in association with certain atopic conditions in man. This study was performed to determine whether numbers of circulating T cells are decreased in atopic children and adolescents in comparison with nonatopic age-matched control subjects. The subjects were not selected on the basis of a particular atopic diagnosis, but relatively more had allergic rhinitis and/or asthma (52) than had atopic eczema (7). Numbers of circulating T cells were not found to be significantly different in allergic children aged 2 to 10 yr than in control subjects. Atopic children and adolescents over age 10 yr had significantly fewer T cells in relative percentages (p less than 0.05), but when absolute numbers were considered significance was lost. Atopic children aged 2 to 10 years had significantly more B cells in both relative percentages and absolute numbers than did control subjects (p less than 0.02 and p less than 0.05, respectively). When those subjects treated with corticosteroids were separated from the total atopic group, there were no significant differences between the atopic and control subjects. The effects of corticosteroids, bronchodilators, antihistamines, and immunotherapy were considered and could be shown to produce no consistent effect on lymphocyte subpopulations.


[Cervical mucus. III. Physiological roles]. The first part of this large review on the bibliography of cervical mucus was concerned both with the mechanism and the determinism ensuring its secretion. A second paper reviewed the present knowledge on chemical, physical ultrastructural characteristics of cervical mucus. This paper, the last part of the review, will be concerned with the different functions of the mucus ensuring synchronisation and encounter of the germ cells. From a physiological point of view, the following properties may be ascribed to the cervical secretion: - protection of sperm from the hostile environment of the vagina, and from being phagocytized along the cervical canal ; - cyclic receptivity to sperm penetration at or near ovulation and interference with entry at other periods ; - filtering effect on sperm cells; - possible intracervical sperm storage ; - supplementing the energy requirements of sperm ; - protection of the uterine cavity against bacteria.


[Neuroleptic analgesia in the management of labor]. Neuroleptic analgesia, which is used for the major part of the conduct of labour, has the advantage of abolishing the pain of uterine contractions without altering the mother's state of consciousness. A study of the effects of droperidol together with phenoperidine on the mother and the child has been carried out. Clinical results, the parameters of the intra-uterine pressure curves and the fetal heart rates as well as of the acid base balance and the pO2 of the mother and infant during dilatation of the cervix and the two first hours of life have been monitored. The conclusion is that neuroleptic analgesia does not cause neonatal depression and can be used as a method for conducting labour so long as very strict monitoring conditions are applied.


The supply of physicians and physicians' incomes: some projections. This study identifies trends that will lead to a dramatic increase in the number of active physicians in the United States during the next decade. The supply of active medical doctors (MDs) and doctors of osteopathy (DOs) as well as active post-graduate MDs and DOs in the U.S. is projected to increase by approximately 50 percent in the decade ending in 1985. The number of active physicians per 100,000 population is similarly expected to increase by approximately one-third. The production of surgical specialists, in particular, appears to be excessive. In response, the average length of physician graduate training programs is anticipated to be shortened as more MD and DO graduates enter shorter, general practice residencies. The authors expect that the effects of this projected increase in the supply of physicians may relieve geographic disparities in physician distribution, rationalize the organization of medical practice, and reduce physicians' incomes relative to other professional groups and possibly in absolute terms. The projected increases in the supply of physicians will give the federal government much more flexibility and bargaining power should it choose to implement a national health insurance program with salaried physicians.


Adsorption of horseradish peroxidase, ovomucoid and anti-immunoglobulin to colloidal gold for the indirect detection of concanavalin A, wheat germ agglutinin and goat anti-human immunoglobulin G on cell surfaces at the electron microscopic level: a new method, theory and application. A method is described for the adsorption of selected macromolecules to colloidal gold which is then used as an electron dense marker for the indirect detection of specific cell surface molecules. Membrane bound concanavalin A, which binds specific sugars on horseradish peroxidase, and wheat germ agglutinin, which binds specific sugars on ovomucoid are detected indirectly with gold labeled horseradish peroxidase and ovomucoid, respectively. Goat anti-human IgM on blood lymphocytes is detected with gold labeled rabbit anti-goat IgG. In the preparation of colloidal gold labeled proteins, the problems of flocculation of colloidal gold by proteins and nonadsorption of proteins to colloidal gold, are solved through a combination of concentration of protein and pH variable adsorption isotherms, which allows one to determine the conditions for adsorption of proteins to colloidal gold. Adsorption is pH dependent, the pH conditions correlating with the isoelectric point(s) of the major protein fraction(s); adsorption is influenced by interfacial tension, solubility and by the electrical charge on the molecules. Colloidal gold is inexpensive and preparation of a useful label is rapid, reproducible and the results easily quantitated from electron micrographs.


Enumeration of t rosette-forming cell population in rabbit peripheral blood using human, sheep, guinea-pig, rat, mouse and chicken red blood cells under different experimental conditions. Attempts were made to estimate peripheral T-lymphocytes in rabbits by their ability to form rosettes with sheep, human, guinea-pig, rat, mouse and chicken RBC. The reaction was studied under different conditions of pH and temperature. The test was also carried out in the presence or absence of glutaraldehyde. With sheep RBC, lymphocytes from 2 of the 5 rabbits studied formed rosettes to the extent of 1.5% in the system containing glutaraldehyde. These results however were not reproducible when studied after a week. The RBCs from other species, under similar experimental conditions, did not give rise to rosettes. The test carried out at pH 6.8 and pH 7.2 showed up to 4 and 8% rosettes respectively with rat RBCs, while RBCs of no other species showed rosettes. When rabbit lymphocytes preincubated at 37 degrees C for 1 h were used for rosette test with guinea-pig RBC, a maximum of 14.6% rosettes were obtained. Here, again the percentage of rosettes formed in the 5 animals, over a period of one month, varied widely. The reasons for the variations of the percentage of rosettes formed are discussed.


[Multiple forms of NADPH-dependent glutathione reductase in serum. Studies on the NADPH-dependent glutathione-reductase in serum II. (author's transl)]. Sera with elevated activities of glutathione reductase were investigated by gel electrophoresis in agar or polyacrylamide, and by gel filtration. In both separation methods the glutathione reductase activity in individual samples was resolved, showing up to three fractions differing in rate of migration and molecular size. The fractions with the lowest and highest molecular weight, corresponded respectively to the slowest and fastest migrating bands in agar gel electrophoresis. Preincubation of the serum samples with FAD or neuraminidase had no effect on the rate of migration of the three fractions. After the addition of beta-mercaptoethanol to the serum, gel electrophoresis and gel filtration showed only the enzyme fraction with the slowest rate of migration and the lowest molecular weight (140,000). The other two fractions reappeared after removal of the thiol from the serum. Further studies on the isolated (agar gel electrophoresis) fractions showed the existence of oligomeric forms of the enzyme, which are reversibly interconvertible.


Liquid and solid-state Cl- -sensitive microelectrodes. Characteristics and application to intracellular Cl- activity in Balanus photoreceptor. When intracellular chloride activity (aiCl) was monitored with chloride-sensitive liquid ion exchanges (CLIX) microelectrodes in Balanus photoreceptors, replacement of extracellular chloride (Cl0) by methanesulfonate or glutamate was followed by a rapid but incomplete loss of aiCl. When propionate was used as the extracellular anion substitute, CLIX electrodes detected an apparent gain in aiCl, while a newly designed Ag-AgCl wire-in glass microelectrode showed a loss of aiCl under the same conditions. This discrepancy in Cl- washout when propionate replaced Cl0 is explained by the differences in selectivity of CLIX and Ag-AgCl electrodes for native intracellular anions and for the extracellular anion substitute which also replaces Cli and interferes in the determination of aiCl. Both electrodes indicate that ECl approximately Em when the cells are bathed in normal barnacle saline, and both electrodes showed the rate of Cl washout (tau approximately 5 min) to be independent of Cli when Cl0 was replaced by glutamate. Details of Ag-AgCl microelectrode construction are presented. These electrodes were tested and found to be insensitive to the organic anion substitutes used in this study. Selectivity data of CLIX electrodes for several anions of biological interest are described.


Exopolysaccharide production by Pseudomonas NCIB11264 grown in batch culture. Fermentation studies using batch culture indicated that exopolysaccharide production by Pseudomonas NCIBI1264 in a chemically defined medium increased under conditions of nitrogen limitation and excess carbon substrate at pH values above 6. The polysaccharide was formed from a variety of carbon substrates and its composition was not affected by the nature of the carbohydrate source. Polysacharide formation did not increase in media containing small amounts of phosphate, and, as in secondary metabolite production, it started late in the exponential growth phase continuing maximally after growth had ceased. The efficiency of glucose conversion into exopolysaccharide was low. Colorimetric, viscometric, and total carbon estimation techniques are described for determining exopolysaccharide levels in cell-free culture supernatants.


Ribonucleic acid polymerase activity associated with purified calf rotavirus. The presence of an RNA-dependent RNA polymerase is demonstrated in purified rotavirus particles. Optimum polymerase activity was found between 45 to 50 degrees C, at pH 8, and in the presence of 10 mM-magnesium ions. The polymerase product was highly sensitive to pancreatic RNase (97%) in low or high salt concentration. The enzyme was activated by EDTA treatment of intact particles or heat shock. The similarities between reovirus, blue-tongue virus and rotavirus polymerases are discussed.


Varicella-zoster virus-induced DNA polymerase. Nuclear extracts of varicella-zoster virus (VZV)-infected human embryo lung (HEL) cells were found to contain DNA polymerase activity not present in uninfected HEL cells. This enzyme was designated the VZV-induced DNA polymerase. The VZV-induced polymerase was partially separated from the cellular alpha- and beta-polymerases by fractionation of the cells and by phosphocellulose chromatography. The separated enzymes were examined for the effect of added (NH4)2SO4, activity with synthetic templates, optimal pH, and the effect of phosphonoacetic acid. The VZV-induced DNA polymerase was distinct from cellular enzymes and had the properties of a typical herpesvirus-induced DNA polymerase.


[A new complex polymalformative syndrome (author's transl)]. The authors discuss the nosological position and the aetiology of a complex of malformations affecting a boy, born at the end of a 40-week pregnancy with a weight of 1420 g and height of 42.5 cm, who died at the age of 3 1/2 months. The case presented an association of an acrocephalosyndactyly, phalangeal hypoplasias with anonychia, aplasia of the abdominal muscles, a genital hypoplasia with cryptorchidism and hypospadias, patches of lipodystrophy localised on the outside of the arms, at the folds of the elbow and the popliteal fossae. The autopsy revealed a large interventricular communication, absence of common mesentery and especially important cerebral anomalies (smooth brain, absence of interhemispherical suture, disorganization of the basal ganglia). The genetic study suggested a spontaneous abortion 2 1/2 months before the birth of the patient and revealed, in the mother, the presence of bilateral clinodactylies of the fingers and a unilateral syndactyly of the big toes.


Brain tyrosine level controls striatal dopamine synthesis in haloperidol-treated rats. Animals received either haloperidol (2 mg/kg) or probenecid (200 mg/kg) in conjunction with tyrosine (100 mg/kg) or its diluent. Striatal homovanillic acid levels increased in probenecid-treated animals to the same range whether they were given tyrosine or not. In haloperidol-treated animals the levels of homovanillic acid were significantly elevated in animals receiving tyrosine. Tyrosine and homovanillic acid levels were highly correlated as determined by linear regression analysis.


The interactions of bromocriptine and lergotrile with dopamine and alpha-adrenergic receptors. Bromocriptine and lergotrile, which are clinically used as antiparkinsonian (AP) agents, compete for the binding of H3-dopamine, H3-apomorphine, and H3-haloperidol to striatal membrane sites. Lergotrile has a higher affinity for the H3-dopamine binding to bovine striatal membranes than bromocriptine. Lergotrile and bromocriptine are almost equipotent in competing for the binding of H3-apomorphine to rat striatal membranes, but bromocriptine is more potent in competing for the binding of H3-haloperidol than lergotrile. These results indicate that lergotrile and bromocriptine are mixed putative agonist-antagonist with respect to the postsynaptic dopamine receptors. Lergotrile and bromocriptine at higher concentrations inhibit synaptosomal tyrosine hydroxylase activity, and reverse the apomorphine elicited enzyme inhibition. Thus, these ergot alkaloids behave as mixed agonist-antagonist also with respect to the presynaptic dopamine receptors. Bromocriptine and lergotrile, as well as other tested DH-ergot alkaloids and neuroleptics, compete for the binding of the alpha-antagonist H3-WB-4101 to rat cerebral cortical membranes. The displacing potencies of the tested DH-ergot alkaloids and of the neuroleptics indicate that they have a high affinity for the alpha-adrenoreceptors in the CNS.


Comparison of gamma-glutamyl transpeptidase activity in tissues of normal and dystrophic hamsters and mice. The activity of gamma-glutamyl transpeptidase (gamma-GT), a membrane-bound enzyme, was assayed by a sensitive fluorometric method in the brain, heart, kidney, liver, skeletal muscle and serum of normal and dystrophic hamsters and mice. Normal and dystrophic hamsters were of the golden brown and BIO 14.6 line respectively. Both normal and dystrophic mice were of the C57 BL/6J -dy2j strain. Kidneys of both hamsters and mice contained the highest enzyme activity followed by the brain, skeletal muscle, liver and heart. Gamma-GT activity was elevated in dystrophic skeletal muscles but decreased in the dystrophic kidneys and sera. In the brain, heart and liver of dystrophic animals from both species enzyme activity was similar to that in normals.


8alpha-hydroxyflavinmononucleotide and related compounds. 2', 3', 4'-Triacetyl-FMN has been transformed by selective radical bromination into 2', 3', 4'-triacetyl-8alpha-bromo-FMN, and the following hydrolysis of the latter has afforded 8alpha-hydroxy-FMN. The presence of the hydroxy group in the 8alpha position of 8alpha-hydroxy-FMN is confirmed by its acetylation into 2', 3'-diacetyl-8alpha-acetoxyriboflavin-4', 5'-cyclophosphate. The absorption spectra of the synthesized compounds have shown the reduction of the extinction ratios of the first and second absorption maxima in comparison with the extinction of the same maxima for 8alpha-hydroxyriboflavin. Unlike FMN, fluorescence quenching for 8alpha-hydroxy-FMN has been found.


The presence and possible function of methylmalonyl CoA mutase and propionyl CoA carboxylase in Spirometra mansonoides. Both spargana and adult forms of Spirometra mansonoides were shown to accumulate lactate, succinate, acetate, and propionate upon in vitro incubation. Adults differed markedly from the spargana in that quantitatively the most significant products of the former were acetate and propionate while the latter formed primarily acetate and lactate. The adults accumulated approximately 32 times more propionate than the spargana per gram of tissue. In accord with this propionate formation, propionyl CoA carboxylase and methylmalonyl CoA mutase have been found to be present in both stages of the parasite. As might be predicted, however, the activities of the carboxylase and mutase were 100-fold and 10-fold higher, respectively, in the adults as compared to the larvae. A possible physiological relationship between propionate formation and energy generation is suggested. Accordingly, inorganic 32P was incorporated into ATP upon incubation of methylmalonyl CoA with a homogenate obtained from adult S. mansonoides. Since methylmalonyl CoA mutase requires vitamin B12 coenzyme, a relationship between vitamin B12 content and propionate formation in helminths is suggested.


A comparison of the dissolution rates of caffeine tablets in a rotating-basket with those in a Sartorius dissolution tester. Uncoated caffeine tablets of four different hardnesses were tested for dissolution rate by the Sartorius (S.S. method) and by the rotating basket method of the U.S.P. XVIII. In both methods the dissolution rate decreased with increasing hardness, and the rate obtained with the S.S. method was always less than that by the U.S.P. method. This result cannot be explained as being due only to the difference in the volume of dissolution medium. Also it was difficult to ensure that the characteristic changes in the process of dissolution paralleled the curves obtained from a plot of % caffeine dissolved vs time. Accordingly, the dissolution rate constants were calculated from the slope of each straight line in a plot of ln W infinity/(W infinity --W) vs time.


Plasma concentrations and excretion of zipeprol in man under acidic urine conditions. Plasma concentrations of zipeprol, and the urinary excretion of it and two of its basic metabolites, compound II and compound III were examined after oral administration of zipeprol hydrochloride to man. The drug was rapidly and extensively metabolized, and the amount of unchanged drug excreted in the acidic urine varied from 1-5% of the dose; the two basic metabolites accounted for 19-38% of the dose.


The effect of age and thyroid hormones upon the ability of the chick heart to deaminate monoamines. The effect of age and thyroid hormones upon the ability of chick heart homogenates to metabolize monoamines has been investigated. 5-Hydroxytryptamine is entirely metabolized by a monoamine oxidase (MAO) with the characteristics of MAO-A, whereas some of the tyramine and all of the benzylamine are oxidatively deaminated by a clorgyline-resistant, but semicarbazide-sensitive enzyme, with a similar subcellular distribution to that of MAO. The remainder of the tyramine deamination is brought about by MAO-A and MAO-B. The specific activities of both clorgyline-sensitive and resistant enzymes are increased by the same proportion by increase in age or by treatment with (--)-thyroxine, and decreased by 2-thiouracil. The significance of these results is discussed.


Evidence from agonist and antagonist studies to suggest that the beta1-adrenoceptors subserving the positive inotropic and chronotropic responses of the heart do not belong to two separate subgroups. The positive inotropic and chronotropic responses of guinea-pig isolated left and right atria to 17 sympathomimetic amines were examined under conditions selected to control the pharmacological environment. Each agonist was compared with (--)-isoprenaline as the reference by constructing dose-response curves. Equiactive molar concentration ratios relative to (--)-isoprenaline were calculated at the EC50 for rate and tension responses. Statistical analysis revealed that (--)-noradrenaline and (+/-)-alpha-methylnoradrenaline were tension selective whereas (+/-)-alpha-ethylisoprenaline and N-methyldopamine were rate selective relative to (--)-isoprenaline. However, no structural trends emerged. The rank order of potency varied slightly between rate and tension, but an analysis of the regression and correlation coefficients indicated respectively that the equiactive molar concentration ratios and the rank orders could be considered identical on rate and tension. Antagonism of the (--)-isoprenaline-induced rate and tension responses by acebutolol, atenolol, practolol and propranolol was assessed from the pA2 values which were almost identical for both parameters with each antagnosti. It is concluded that the beta1-adrenoceptors mediating the positive inotropic and chronotropic responses do not warrant subdivision into two separate groups.


The role of dopaminergic systems in gamma-hydroxybutyrate-induced electrocorticogram hypersynchronization in the rat. The effects of dopaminergic agonists and antagonists on the duration of hypersynchronization induced in the electrocorticogram (ecog) by gamma-hydroxybutyrate (gamma-HB) were tested in rats. Apomorphine (0-2-8 mg kg-1), piribedil (2-5-10 mg kt-1) and haloperidol (0-5-1 mg kg-1) had no influence on the duration of the hypersynchrony. Amphetamine (1-5-6 mg kg-1) inhibited the hypersynchrony, while (3,4-dihydroxyphenylamino)-2-imidazoline (DPI; 5, but not 1, mg kg-1) prolonged its duration. The lack of effect of the dopamine receptor agonists apomorphine and piribedil, and the dopamine receptor blocker haloperidol, on the gamma-HB-induced hypersynchrony might indicate that the inhibition of the impulse flow in the nigrostriatal dopamine system by gamma-HB is not involved in the generation of the hypersynchrony. DPI is thought to be an agonist at a dopamine receptor not sensitive to apomorphine, and its facilitatory effect on gamam-HB-hypersynchrony can be interpreted in terms of a possible involvement of another dopamine system in the ecog hypersynchrony induced by gamma-HB. The antagonism of gamma-HB by amphetamine is possibly due to an indirect stimulatory effect on noradrenergic receptors.


The central and peripheral effectiveness of two oxotremorine-antagonists determined using oxotremorine-induced tremor and salivation. Two oxotremorine antagonists have been examined using oxotremorine as the tremor salivation inducer and the ratios between the central and peripheral nervous systems determined. Under these conditions these compounds are still more effective antagonists centrally than peripherally. Differences in distribution to the various muscarinic receptors involved is believed to be the major factor in determining this separation of central and peripheral activities.


A sex difference in the interaction between promethazine and morphine in the mouse. The effects of promethazine on the antinociceptive and respiratory actions of morphine have been examined in the mouse. Moderate doses of promethazine (5 and 10 mg kg-1) potentiated morphine's action in male mice but inhibited it in female mice. Gonadectomy abolished the interaction between promethazine and morphine in both sexes, although the intensity and duration of morphine's activity was greatly enhanced in these mice. Replacement of oestradiol in ovariectomized mice restored morphine's activity to intact female control values. However, interactions between promethazine and morphine required progesterone, as well as oestradiol, replacement to obtain results approaching those obtained in intact female mice.


Some antagonists of dantrolene sodium on the isolated diaphragm muscle of the rat. The effects of a number of potential antagonists of dantrolene sodium have been studied on twitches of the isolated hemidiaphragm preparation of the rat stimulated directly at a frequency of 0-1 Hz, after complete neuromuscular block produced by tubocurarine or erabutoxin a. The substances selected as possible dantrolene antagonists were uranyl ions, thiocyanate ions, adrenaline, caffeine, quazodine, quinine, 4-aminopyridine and the calcium ionophore a23187, all of which facilitate excitation-contraction coupling in one way or another. Contracture was the main feature of the response to A23187, the increase in the tension of the dantrolene-depressed twitches being very slight. All the remaining compounds increased the amplitude of the twitches, but only 4-aminopyridine, quinine, quazodine and caffeine were capable of restoring to the control amplitude twitches that had been maximally depressed by dantrolene. OF these, 4-aminopyridine and quinine were the most potent on a molar basis.


The dissolution mechanism in a system undergoing complexation: salicylamide in caffeine solution. The dissolution rate of compressed salicylamide discs has been measured in water and in caffeine solutions of increasing concentration at 15, 25, 37 and 45 degrees in an apparatus rotating at 48 rev min-1 or more. Dissolution rate profiles showed breaks indicative of a shift in the mechanism of dissolution from interfacial towards transport control. The shifts occurred at higher caffeine concentrations on increasing the agitation rate or temperature. The dependencies of dissolution rates on agitation rates typified the intermediate type of dissolution and Arrhenius plots indicated that interfacial and transport processes participated in salicylamide dissolution.


Dilute solution properties of clindamycin 2-palmitate hydrochloride. Clindamycin 2-palmitate hydrochloride forms micelles in aqueous solution at concentrations above 2-2 X 10(-3) M. Dilution of micellar solutions of the drug to concentrations less than 2-0 X 10(-3) M produces turbid systems. Turbidity is caused by unionized drug, produced by hydrolysis of the salt, phasing out of solution at concentrations too low for micelles to be formed. Changes in specific conductivity and turbidity associated with the phasing-out process have been measured and explained in terms of the rates of the various processes involved, including demicellization and emulsion stability. The phenomenon of phasing-out, below a critical concentration in aqueous solution, is predicted to be a general one for weakly ionized surfactants where the unionized species possesses low water solubility.


Relaxations mediated by adrenergic and non-adrenergic nerves in human isolated taenia coli. In human taenia coli electrical field stimulation after cholinergic blockade excited mainly non-adrenergic inhibitory nerves. Adrenergic relaxations during stimulation were demonstrated only with short electrical pulses at high frequencies or with ganglion stimulants. Reduction by tetrodotoxin of relaxations to indirectly acting sympathomimetics supports the histochemical finding that colonic adrenergic nerves are mainly preganglionic, and possibly synapse with non-adrenergic inhibitory nerves. When the latter undergo direct maximal stimulation, responses to adrenergic nerve excitation are masked.


Evidence for more than one type of 5-hydroxytryptamine receptor in the human colon. The actions of methysergide, a 5-hydroxytryptamine (5-HT) antagonist, have been examined on muscle strips taken from the circular and longitudinal layers of the human colon. Relaxations of the longitudinal muscle to 5-HT were antagonized by concentrations of methysergide known to be selective. Relaxations of the circular muscle to 5-HT were unaffected by similar concentrations of methysergide. Responses of both types of muscle to 5-HT were partially reduced by tetrodotoxin. Furthermore there was evidence for 5-HT receptors in circular colonic muscle which were unaffected either by selective concentrations of methysergide or tetrodotoxin.


Influence of pyridylisatogen tosylate on contractions produced by ATP and by purinergic stimulation in the terminal ileum of the guinea-pig. 2-2'Pyridylisatogen tosylate (PIT), which antagonizes the inhibitory action of ATP in the taenia caeci, did not antagonize the excitatory effects of exogenous ATP and of purinergic stimulation of the terminal guinea-pig ileum. PIT (0-5--2-5 muM) potentiated the ATP-induced contractions and also the contractions produced by potassium chloride, though the potentiation could not be related to the dose in every experiment. The responses to noradrenaline, adrenaline and histamine were slightly inhibited. PIT also potentiated the contractions produced by electrical stimulation of intramural purinergic nerves when either an alternate or uniform stimulation pattern was used. The present results in which the preparation is contracted by ATP are opposite to those obtained with PIT on the taenia caeci, which is relaxed by ATP. This raises a question of duality or plurality of receptors for ATP.


Effect of two non tricyclic antidepressant drugs on [14C]5-hydroxytryptamine uptake by rat platelets. The uptake of 14C-5-HT by rat blood platelets was examined in vitro in experimental conditions which allowed measurement of the initial velocity and excluded other passive processes across the cell membrane. In these conditions, the effect of two non tricyclic antidepressant drugs (Lilly 110140 and trazodone) was investigated. Lilly 110140 was as active as chlorimipramine and several times more active than imipramine as an inhibitor of 14C-5-HT uptake. Like chlorimipramine, Lilly 110140 appeared to be either a non-competitive or an uncompetitive inhibitor, according to the concentration of drug used. Trazodone also inhibited 14C-5-HT uptake by platelets but to a lesser extent than chlorimipramine, imipramine or Lilly 110140. m-Chlorophenylpiperazine, a possible metabolite of trazodone, was about 3 times more potent an inhibitor than the parent molecule. Both compounds acted non-competitively. Compared with published data on the effect of Lilly 110140 and trazodone on brain 5-HT, the present results support the suggestion that rat platelets are a useful pharmacological model of serotoninergic nerve endings.


Analysis of the beta-receptor mediated effect on slow-contracting skeletal muscle in vitro. Subtetanic contractions of the guinea-pig isolated soleus, a slow-contracting skeletal muscle, were evoked by transmural field-stimulation. Isoprenaline caused a dose-dependent depression of the contractions. This effect was inhibited by propranolol and H 35/25 (1-(p-tolyl-2-isopropylamino-1-propanol) but not by practolol. Similar results were obtained for terbutaline. Tazolol and H 80/62 (1-isopropylamino-3-(p-hydroxyphenoxy)-2-propanol (HCl), selective beta1-agonists, had no effect per se but inhibited the effect of terbutaline. Adrenaline, noradrenaline, and dopamine all caused a dose-dependent decrease in the force of the soleus contractions, their potencies being in that order. Tyramine did not appreciably affect the contractions nor did it inhibit the effect of terbutaline. Pretreatment with reserpine, if anything, increased the response to terbutaline. It is concluded, in conformity with previous in vivo studies, that the adrenergic receptor mediating the effect on the soleus muscle contractions is of the beta2-type. Indirect sympathomimetic effects do not contribute to the responses observed on the isolated soleus muscle.


Biochemical and pharmacological studies on amineptine (S 1694) and (+)-amphetamine in the rat. The pharmacological activities of amineptine (S 1694) and (+)-amphetamine and their interaction with biogenic amines have been examined in rats. The locomotor activity, stereotyped behaviour and hypothermia induced by amineptine were similar to but not as marked as those produced by (+)-amphetamine, and there was little or no anorectic action. Amineptine does not modify the concentrations of brain noradrenaline or acetylcholine which are respectively reduced and increased by (+)-amphetamine. Moreover, amineptine does not affect significantly the decrease of brain noradrenaline induced by an intraventricular injection of 6-hydroxydopamine, an effect significantly antagonized by (+)-amphetamine. On the other hand, like amphetamine, amineptine significantly reduces the effect of 6-hydroxy-dopamine on brain dopamine. Both drugs increase the striatal concentrations of homovanillic acid and show a cross tolerance in this action. Therefore they could act similarly on the striatal dopaminergine system. Amineptine thus appears to be a new type of antidepressant with a brain biochemical profile differing from that of other drugs used in depressive disorders.


pH-stat titration of aluminum hydroxide gel. The pH-stat titration of aluminum hydroxide gel was evaluated and was affected by pH, temperature, concentration, and ionic strength. Control of these parameters resulted in a highly sensitive and reproducible in vitro antacid test. The utility of the pH-stat test was illustrated by monitoring the aging of several carbonate-containing aluminum hydroxide gels and by comparing the antacid properties as measured by the pH-stat titration, the acid-consuming capacity, the Rossett-Rice test, and the test proposed by the food and Drug Administration Drug Evaluation Panel. The pH-stat titration  also was useful for relatively nonreactive aluminum hydroxide gels. The use of sodium fluoride as the reaction medium extended the capability of the pH-stat titration to monitor the aging of chloride-containing gels. The pH-stat titrigram was interpreted in terms of a previously published polymer model of the structure of a chloride-containing aluminum hydroxide gel. The acid reactivity of relatively nonreactive gel is believed to be due totally to the chemical neutralization of acid, because the milliequivalents of aluminum  ion appearing in solution is the same as the milliequivalents of acid neutralized throughout the ph-stat titration.


Relationship between zero point of charge and solubility product for hydroxides of polyvalent cations. The zero point of charge (ZPC) of slightly soluble compounds is the pH at which their particles suspended in water have zero charge. The ZPC values of slightly soluble hydroxides were compared with their solubility product in the form of its negative logarithm, pKSP, and with th pH of their suspensions in pure water, pHSP, which is a function of pKSP. The ZPC-pKSP relation was nonlinear while the ZPC-pHSP relation was linear. Either equation can used to estimate the ZPC value of a hydroxide from its solubility product. The ZPC of a given hydroxide was higher than its pHSP because polyvalent cations are more extensively adsorbed and less extensively desorbed from the particle surface than the monovalent hydroxide ion. At the pHSP, there are equivalent amounts of the cation and of the  hydroxide anion in solution, but the surface layer of the hydroxide particle contains an excess cation on an equivalent basis. This imbalance confers a positive charge to the particle. The solubility product of aluminum hydroxide, redetermined at 25 degrees by means of pH measurements, was 8 X 10(-33). Its ZPC, redetermined by microelectrophoresis, was 8.5 +/- 0.1.


In vitro photodecomposition of uric acid in presence of riboflavin II. In vitro studies on the photodecomposition of uric acid in the presence of the monosodium salt of riboflavin 5'-phosphate in buffers at various pH values, in methanol, and in human plasma are reported. The decomposition rate increased with increasing pH and was independent of solvent or buffer species. The mechanism appears to be an energy transfer process involving triplet riboflavin and single oxygen. Riboflavin-enhanced photodecomposition of uric acid occurred in vitro in hyperuricemic human plasma.


Effects of adenine nucleotides on oxidation of phenothiazine tranquilizers. The effects of adenosine diphosphate and triphosphate on the periodic  acid oxidation of the phenothiazine tranquilizing drugs were studied. The principal effect was a marked reduction in the rate of formation and decay of the drug free radical. The oxidation rates of the nucleotide free drugs seemed to be most strongly influenced by the inductive effects of substituents at the 2-position of the phenothiazine nucleus. However, the oxidation rates of the drugs in the presence of nucleotide were most strongly influenced by the substituents at the 10-position. Variations of the structure of substituents at the 10-position have only a modest effect on the electronic state of the phenothiazine nucleus. Therefore, the marked effect of structural variation in the 10-substituents in the presence of nucleotide on the periodate oxidation rate most likely is an expression of steric effects related to an interaction between drug and nucleotide.


Stability of prostaglandin E1 and dinoprostone (prostaglandin E2) under strongly acidic and basic conditions. The stability of prostaglandin E1 and dinoprostone was investigated at the extremes of the pH range (less than or equal to 3 and greater than or equal to 10) in the sequence prostaglandin E leads to prostaglandin A leads to prostaglandin B. The degradation rate is first order with hydrogen-ion and hydroxide-ion concentrations. Separation and analysis of the E prostaglandins were accomplished by TLC and UV spectrophotometry. At the lowest pH values and at elevated or low temperatures, significant amounts of 15-epiprostaglandin E were present. Apparent activation energies for the total dinoprostone loss, calculated from elevated temperature data, were 21 kcal/mole in the strongly acidic region and about 18 kcal/mole at pH 3. Corresponding studies in the alkaline  region led to a derived arrhenius activation energy of 15 kcal/mole with the appearance of significant amounts of 8-isoprostaglandin E. This difference in activation energies may reflect the different mechanisms operant at high and low pH values.


Tetrazole analogs of phenylalanine derivatives as tyrosine hydroxylase inhibitors. Eight new tetrazole analogs of phenylalanine derivatives in which the carboxyl group was replaced by a tetrazole ring were synthesized. At a concentration of 2.5 time 10(-5) M, each compound exhibited greater than 50% inhibition of tyrosine hydroxylase activity.


Kinetics of concomitant degradation of tetracycline to epitetracycline, anhydrotetracycline, and epianhydrotetracycline in acid phosphate solution. The concentrations of tetracycline, epitetracycline, anhydrotetracycline, and epianhydrotetracycline in pH 1.5 phosphate solution were followed as a function of  time at four temperatures. Separation and quantification of all four species were accomplished using high-pressure liquid chromatography. Through nonlinear regression analysis, rate constants for the reversible first-order epimerization of tetracycline and anhydrotetracycline and for the first-order dehydration of tetracycline and epitetracycline were obtained. Solutions to the differential equations obtained through Laplace transforms successfully predict concentrations found experimentally. The energy of activation for each reaction  step was calculated and ranged from 15 to 27 kcal/mole. The rate constants for tetracycline and epitetracycline dehydration conform with those of earlier studies that used different experimental methods. The study shows that epimerization of tetracycline and anhydrotetracycline can take place at a low pH.


Evidence for antral inhibition of pentagastrin from experiments using mucosal cooling. 1. The acid secretion of the fundic mucosa in Heidenhain pouches in response to pentagastrin became progressively less as the pouch mucosa was cooled. 2. When a cooled Heidenhain pouch in an animal receiving pentagastrin was warmed, acid and pepsin secretion from the main stomach was depressed. Change from warm to cool produced no obvious effect. 3. In animals receiving pentagastrin continuously, but not in those receiving histamine, lowering the temperature in an antral pouch, or the application of local anaesthetics to its mucosa, increased acid and pepsin secretion from the main stomach when the antral pouch was fully innervated. 4. This effect could readily be abolished by ganglionic and beta-adrenergic blockade, but not by bilateral vagal block in the neck, thus suggesting a sympathetically mediated inhibitory mechanism of pyloric origin. 5. The effect of indirect vagal stimulation, using 2-deoxy-D-glucose on secretion from the main stomach, was augmented by pyloric antral local anaesthesia and depressed by antral cooling.


Effects of neurochemicals upon a dinoflagellate photoresponse. Photoresponsiveness by Gymnodinium splendens Lebour was monitored quantitatively by a microscope-television system. Exposure to the catecholamines DOPA and Dopamine caused a decrease in light sensitivity, while 0.01 mM norepinephine, or isoproterenol did not affect photoresponsiveness. Classical catecholamine blocking agents, dichloroisoproterenol, propranolol, and dibenzyline, and an inhibitor of DOPA synthesis, alpha-methyl-rho-tyrosine, caused an increase in sensitivity. In addition, acetylcholine and an inhibitor of acetylcholinesterase activity, eserine, caused an increase in sensitivity, while an inhibitor of acetylcholine action atropine, had the opposite effect. These experiments suggest that G. splendens may have an antagonistic catecholamine-cholinergic system which participates in regulating photosensitivity.


Toxoplasma gondii-vertebrate cell interactions. I. The influence of bicarbonate ion, CO2, pH and host cell culture age on the invasion of vertebrate cells in vitro. A controlled-environment culture system was used to show that both physical and biologic parameters can influence the penetration of vertebrate cells by Toxoplasma gondii. The optimum bicarbonate ion concentration for the penetration of bovine embryo skeletal muscle (BESM) cells is 36.25 mM. Higher or lower bicarbonate ion concentrations are increasingly inhibitory to penetration. As CO2 increases in the range from 0.5-3.7 mM, penetration is progressively inhibited. No relationship was found between penetration and pH in the pH range of 6.949-7.765. The culture age of the BESM cells directly influenced the ability of the parasites to penetrate the cells. Older BESM cells were more refractory to penetration than younger cells.


Specific labeling of intracellular Toxoplasma gondii with uracil. Radioactive uracil was not significantly incorporated into the nucleic acids of human fibroblast cells. Infection of these cells with Toxoplasma gondii resulted in an exponential increase in the rate of uracil incorporation that paralleled the exponential growth of the parasite. One day after infection the rate of uracil incorporation was increased 100-fold. It was established by autoradiography that all of the [3H] uracil was incorporated into the intracellular parasites. A possible explanation for this difference in ability to use uracil is our observation that the specific activity of uridine phosphorylase was 100-fold greater in partially purified parasites than in the host cell.


Monoamine oxidase and catechol-O-methyl transferase activity in Tetrahymena. Tetrahymena pyriformis strain HSM was found to have monomine oxidase (MAO) and a catechol-3-methyl transferase-like (COMT) activity. As in mammalian tissues, the MAO activity is predominantly localized in the mitochondrial pellet and COMT in the cytosol. The COMT-like activity was present in amounts comparable to several mouse tissues and was inhibited by tropolone. MAO activity was much lower than in any of the mouse tissues tested, and its activity varied greatly from preparation to preparation. The substrate preference of Tetrahymena MAO was tryptamine greater than serotonin greater than dopamine, and activity increased with increasing pH from pH 6.5 to pH 7.8, as does that of mouse liver MAO. Teh Km of Tetrahymena MAO for tryptamine was approximately 4 micrometer, an order of magnitude lower than that of mouse liver MAO. Sensitivity of inhibition by MAO inhibitors was variable. In some preparations, no inhibition was observed. In others clear inhibition was obtained, harmine and clorgyline being among the most potent inhibitors.


Poly(ethylene oxide), a new slowing agent for protozoa. The water-soluble, viscoelastic resin Polyox WSR 301), a poly(ethylene oxide) of high molecular weight (approximately 4 million) is introduces as a new slowing agent for protozoa. Generally, as the kinetic viscosity of the resin increased from 0.25% to 1% (w/v), the swimming velocity of Euglena gracilis, Didnium nasutum, Paramecium aurelia, Blepharisma undulans, and Prorodon platyodon decreased. The 1.0% solution had the highest viscosity and decreased velocity more effectively than 1.0% methyl cellulose and Protoslo solutions. The Polyox solutions differed from those of methyl cellulose and Protoslo by having, in addition to viscous drag, an elastic recoil that pulled the protozoa backwards when their swimming efforts stopped. The toxicity of these slowing agents was determined using 10 P. aurelia/test slide preparation. Paramecium numbers decreased in 1.0% methyl cellulose and Protoslo to nearly zero by 24 hr; in Polyox, not only were most these ciliates alive after 24 hr, but many survived for 96 hr and divisions occurred in 0.25% and 0.50% solutions.


Synthesis and pharmacological activity of some N-alkyl-substituted 9alpha-ethyl-2'-hydroxy-5-methyl-6,7-benzomorphans. A series of N-substituted 9alpha-ethyl-2'-hydroxy-5-methyl-6,7-benzomorphans was synthesized and evaluated for their narcotic analgesic and antagonistic activities. Compounds 2a and 22 were as potent as morphine in the writhing (PPQ) and hot-plate tests, while a number of compounds demonstrated antagonistic activities greater than nalorphine. Generally, the compounds in this series show activities somewhat greater than the comparable compounds in the 5,9alpha-dimethyl-6,7-benzomorphan series for analgesic effect and similar or slightly less antagonistic potency.


Application of the Free-Wilson technique to structurally related series of homologues. Quantitative structure-activity relationship studies of narcotic analgetics. A series of benzomorphans with ED50 values determined in vivo by the hot-plate method in mice is analyzed by the modified Free-Wilson method. The QSAR yields 36 substituent constants (ai) with contributions to the overall activity in agreement with experimental data. Substituent constant values obtained for benzomorphans are used in calculating log (1/C) values for six morphinans. An excellent correlation is obtained (r = 0.95) between the six calculated and observed activities. The possibility of extending the Free-Wilson approach from one series of homologues to another is demonstrated.


Synthesis of 1-substituted analogues of trimetoquinol possessing differential and selective beta-adrenergic properties. The synthesis of the 1,1-disubstituted tetrahydroisoquinoline analogues, 1-methyl-1-(3,4,5-trimethoxybenzyl)-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline hydrochloride (2) and 1-benzyl-1-(3,4,5-trimethoxybenzyl)-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline hydrochloride (3), is described. The profile of beta-adrenergic activity for these analogues was determined and compared to that of trimetoquinol (1) in isolated guinea pig atrial, tracheal, and rat adipocyte preparations. Unexpected selective beta1-blocking activity in guinea pig trachea was noted with analogue 3. With the exception of 2 in guinea pig atria, 2 and 3 did not possess any beta-stimulant activity. Substitution at the 1 position of trimetoquinol (1) has revealed qualitative differences in beta-adrenergic activity.


A systematic and comprehensive approach to teaching and evaluating interpersonal skills. This study addressed one problem with current methods for teaching and evaluating interpersonal skills: the failure to include a wide range of behaviors reported in the literature as contributing to patient dissatisfaction and noncompliance. To address this concern, the authors developed a comprehensive interpersonal skills training program and a pretest-posttest evaluation. The tests were administered to two student groups one of which received the interpersonal skills instruction. The student group exposed to the training exhibited a significant positive change from pretest to posttest. Additionally the change for this group was significantly greater than the change for the group not exposed to interpersonal skills instruction.


The effect of certain rater roles on confidence in physician's assistant ratings. Previous research on the psychology of confidence suggests that the more confident a rater is in his judgment the more accurate is his rating. The purpose of the present study was to investigate possible differences among raters in their confidence in competency ratings which they had provided. Results indicated significant differences due to the rater's interpersonal role with the ratee and the particular aspect of competence rated. Greater simple structure of competence ratings when adjusted for rater confidence is also shown. Rater confidence is discussed as an index for rater selection and as a moderator variable for competence ratings.


Normal acid-base status of arterial blood from the conscious, chair-restrained squirrel monkey. Normal acid-base status of arterial blood (pHa =7.40, PaCO2=38.1 Torr) was demonstrated for conscious, restrained squirrel monkeys when environmental stimuli were minimized and monkeys are habituated to experimental procedures. These results indicate the potential for using squirrel monkeys in experiments in which normal acid-base status is a significant factor.


Multifocal candidiasis in a capuchin monkey (Cebus apella). Candidiasis involving nasal, pharyngeal, and intestinal mucosal surfaces and a pharyngeal lymph node was demonstrated microscopically in a young adult female capuchin monkey (Cebus apella) experimentally infected with Schistosoma haematobium (Iran strain). Persistent nasal exudation and weight loss characterized the clinical disease preceding the animal's death.


Management of dental trauma in children and adolescents. The emergency treatment of dental trauma has traditionally been handled in the emergency rooms of medical centers, or in the dental office. With increasing emphasis on comprehensive medical care, and the tendency of more recent graduates to locate in rural areas, there is a growing trend for physicians to be involved in providing emergency care for traumatic injuries to the dental and oral tissues. The authors discuss the etiology of dental trauma and the highly psychologic impact of such injuries upon the patient and parents. Techniques for prevention and a comprehensive plan are outlined for systematic history taking and examination. A classification of eight levels of dental injuries (5), techniques for the emergency, and subsequent restorative care are given. Emphasis is placed upon immediate care which can, if necessary, be provided by the physician pending referral to a dentist.


Nonstructural proteins of herpes simplex virus. I. Purification of the induced DNA polymerase. Herpes simplex virus-induced DNA polymerase purified by published methods was found to be contaminated with many others proteins, including virus structural proteins. Thus, DEAE-cellulose and phosphocellulose chromatography were used in combination with affinity chromatography to purify DNA polymerase from herpes simplex virus type 1- and type 2-infected cells. The purified enzyme retained unique features of the herpesvirus-induced DNA polymerase, including a requirement for high salt concentrations for maximal activity, a sensitivity to low phosphonoacetate concentrations, and the capacity to be neutralized by rabbit antiserum to herpesvirus-infected cells. By polyacrylamide gel electrophoresis, the purified DNA polymerase was associated with a virus-induced polypeptide of about 150,000 molecular weight.


Properties of deoxynucleoside 5'-monophosphatase induced by bacteriophage T5 after infection of Escherichia coli. Bacteriophage T5 induced a deoxynucleoside 5'-monophosphatase during its infection of Escherichia coli. The enzyme was purified about 100-fold. It was clearly distinct from the host 5'-nucleotidase activity in its physical characteristics and substrate specificity. The enzyme was active on deoxynucleoside 5'-monophosphates but was not active as a phosphatase on ribonucleotides, deoxynucleoside 5'-triphosphates, deoxynucleoside 3'-monophosphates, or deoxyoligonucleotides. Furthermore, it did not have oligonucleotidase or exonuclease activity. The enzyme could exist in multimeric form but had a monomer molecular weight of about 25,000.


Renal acidifying ability in subjects with recurrent stone formation. To determine the incidence of an acidification defect in men in whom calcium stones form and its relationship to parathyroid function 120 such patients were given an acute dosage of 0.1 gm. per kg. oral ammonium chloride and circulating immunoreactive parathyroid hormone was determined. The subjects were divided into 2 groups, according to normal or high parathormone levels. Group 1 consisted of 46 men in whom immunoreactive parathyroid hormone was less than or equal to 60 mulEq. per ml. and group 2 consisted of 74 men with immunoreactive parathyroid hormone greater than 60 mulEq. per ml. Of 8 men in whom the urine failed to acidify to less than a pH of 5.3, 3 were from group 1 and 5 were from group 2. None of the patients had an active urinary tract infection. There was no difference in minimal urine pH among the patients in whom the urine acidified normally regardless of immunoreactive parathyroid hormone. The incidence of abnormal acidification in our population was 6% and all of these patients had the incomplete form of renal tubular acidosis. These findings have important therapeutic implications.


The effect of bicarbonate and distilled water on sickle cell trait hematuria and in vitro studies on the interaction of osmolality and pH on erythrocyte sickling in sickle cell trait. The effect of intravenously administered distilled water was examined alone and during alkalization in a patient with gross hematuria associated with the sickle cell trait. On each of 4 occasions hematuria ceased promptly after the infusion of distilled water. Bicarbonate therapy also consistently decreased hematuria. In vitro studies on erythrocytes from another patient with sickle cell trait and hematuria demonstrated that slight increases in urinary pH similar to those that occur in the urine during alkalization can reverse or prevent erythrocyte sickling in the sicle cell trait. If patients with the sickle cell trait are hydrated adequately and have a good rate of urine flow distilled water can be given intravenously with virtually no danger of acute tubular necrosis secondary to erythrocyte hemolysis.


Bilateral seminoma 34 years after orchiopexy. A case of bilateral seminoma in a 59-year-old patient who had undergone bilateral orchiopexy 34 years previously is presented. To our knowledge this is the longest interval between an orchiopexy and diagnosis of bilateral malignancy.


Occurrence of rumenitis in a supplementary fed white-tailed deer herd. Rumenitis was observed with increased frequency in a herd of white-tailed deer (Odocoileus virginianus) maintained on a high carbohydrate supplemental ration. Healing rumen scars were found in 4.4% (n=225) of animals examined in 1973; 24.1% (n=278) in 1974; and 42.5% (n=308) in 1975. The lesions often involved nearly the entire ventral blind sac of the rumen. Histopathologic studies did not define the etiologic agent and invasion by either fungi or Fusiformis necrophorus was not a prominent feature in the cases examined. Recovery appeared to be complete and the disease was not considered to be an important herd mortality factor.


Demonstration of vaccinia virus antigen in brains of postvaccinal encephalitis cases. Five lethal cases of postovaccinal encephalitis (PVE) were examined in attempts to detect the viral antigen by the fluorescent antibody techniques and to isolate the infectious vaccinia virus. Histologically, the brain tissues were characterized by meningoencephalitis, with perivascular infiltrations and inflammatous reactions mainly in leptomininx, chroid plexus and ependyma. By direct immunofluorescence staining, vaccinia virus antigen was found in the brain specimens of one case, but not in the other four cases. The existence of antigen-antibody complexes was suggested by the deposition of immunoglobulins in the brain specimens. After treatment with 3 M NaSCN solution to dissociate the immunoglobulins from the complex, the viral antigen was demonstrated in specimens of two cases. Localization of viral antigen(s) thus detected coincided with that of meningoencephalitic reactions in the histological examination. Virus isolation on chick embryo chorioallantoic membrane was negative in all cases. Participation of viral pathogenicity in the etiology of PVE is discussed.


Prostaglandin synthetase systems in rat and rabbit renal medulla and inhibition by non-steroidal anti-inflammatory drugs. The properties of prostaglandin synthetase systems (PSSs) in the renal medulla of the rat and rabbit, and inhibition by ketoprofen, indomethacin, ibuprofen, phenylbutazone and aspirin were investigated in relation to their anti-inflammatory, analgesic, antipyretic and ulcerogenic activities. Rat and rabbit PSSs produced prostaglandin (PG)E and PGF from arachidonic and dihomo-gamma-linolenic acids and had an optimal pH of 8.5 and 7.5 for PGE formation, respectively. Only a slight loss of activity occurred with lyophilization. In the rat PSS, all drugs tested were inhibitory in the order of ketoprofen, ibuprofen, indomethacin and aspirin, respectively. In the rabbit PSS, the same potency relationship was also found. Drug sensitivity of the rat PSS was remarkably lower than that of the rabbit PSS. Significant correlations were noted between the inhibitory potencies of the drugs against both PSSs and other in vivo pharmacological activities within the same species.


[Evaluation of antireflux effect of fundic patch operation by withdrawal pH curve (author's transl)]. With the use of gastroesophageal withdrawal pH curve, pathophysiology after various surgical procedures for achalasia of the esophagus was investigated experimentally from the view point of postoperative reflux. A total of 68 dogs were divided into four groups and were prepared with proximal gastrectomy with end-to-end esophagogastrostomy, Wendel procedure, Heller procedure and Fundic patch operation, respectively. In the group with Fundic patch operation, efficacy of the flap valve, mucosal valve and fundoplication was also evaluated preparing them in various sizes. The results obtained may be summarized as follows: 1) Withdrawal pH measurement is a sentive mean to detect the gastroesophageal reflux. 2) Proximal gastrectomy with end-to-end esophagogastrostomy and Wendel procedure yielded worst results with severe gastroesophageal reflux. On the contrary, Fundic patch operation best controlled the gastroesophageal reflux. 3) Gastroesophageal reflux could be prevented by the Fundic patch operation with the flap valve 6 cm in length and fundoplication enclosing around 1/2 to 2/3 circumference of the distal esophagus.


[Prostaglandin E1: effect on blood gases in infants with cyanotic congenital heart disease (author's transl)]. 15 infants with cyanotic congenital heart disease (pulmonary atresia, Ebstein-Syndrom) were treated with Prostaglandin E1. During treatment there was a significant rise of PO2 and SO2 in the arterial, capillary and central venous blood. There was a minor rise of pH and HCO3; the arterial pCO2 fell slightly. We used PGE1-therapy as pretreatment before the heart operation, so that the critical babies go to the operation well oxygenated and in a better state. The side effects we observed (tachypnoe, tachycardia, hyperpyrexia, augmented urine output, erythemas, apnoic spells) seemed less important than the great advantage of a better oxygenation of these hypoxic babies.


Detection of circulating immune complexes with a modified Raji cell technique. A modification of the Raji cell technique for the detection of circulating immune complexes is described. Raji lymphoid cells bind immune complexes via the C 3 receptor. Prior to incubation with the test serum it is necessary to block the IgG Fc receptor which would interfere with the assay. In the present modification the IgG Fc receptor is blocked with monoclonal IgG myeloma proteins thereby facilitating the detection of C 3 bound immune complexes with fluoresecein-conjugated anti-light chain serum. This modification is presently used in a survey of immune complexes in a number of autoimmune diseases.


Polyanion and complement receptor of the glomerular epithelium. Relationship to pH. Two markers on the surface of glomerular epithelial cells were studied on human and rat tissue. The glomerular polyanion is known to be a sialoglycoprotein present profusely on glomerular epithelial cell surfaces, and it is detected by binding with cationic dyes. This staining is lost after incubation in the pH range 3.8 to 4.4, yet retained at a more acidic or physiologic pH. A receptor for complement is also present on human glomerular epithelial cells. This receptor is also lost as a function of varying pH incubation and this loss parallels the pattern observed for glomerular polyanion. Neuraminidase treatment of renal tissue removes the sialic acid-dependent glomerular polyanion staining but preserves and stabilizes the complement receptor. Both functions are markers of the cell surface of glomerular epithelial cells and behave as peripheral membrane proteins.


On the inactivation of hepatic tyrosine aminotransferase. Tyrosine aminotransferase from frog liver requires no exogenous pyridoxal-5'-phosphate for maximum activity. The cofactor cannot be removed from the enzyme by dialysis as in the case of the rat enzyme. Pyridoxal phosphate also fails to elevate frog liver tyrosine aminotransferase activity in vivo. The enzyme activity decreases rapidly after administration of cycloheximide, which indicates that its turnover is rapid. These results strongly contradict the cofactor-dependent model of enzyme degradation. Rat and frog liver tyrosine aminotransferases are stable in neutral homogenates at 37 degrees C but are rapidly inactivated after addition of cysteine in millimolar concentrations. This effect is probably due to cystine formed during the incubation. The rates of inactivation of the different subforms of the enzyme in this system were identical. No membrane-bound system is needed for the inactivation by cystine. It is possible that the denaturation occurs by sulfide exchange. Fructose and glucose lower the enzyme activity in both rat and frog liver to an equal extent. This effect is not due to instability of the enzyme activity in both rat and frog liver to an equal extent. This effect is not due to instability of the enzyme in the presence of sugars of their metabolites. Theories on the inactivation of enzymes will be discussed in the light of the present results.


Model membranes for the study of active transport phenomena. Models for active transport are prepared by the covalent bonding of molecules containing a catalytic site to the membrane asymmetrically with respect to its thickness. The membrane separates two compartments of unequal volume, both initially containing reactant at the same concentration. We observed the time dependence of the reactant and product concentrations, and of the osmotic pressure, and these results are compared with theoretical predictions. The two reactions studied are the hydrolysis of ethyl N-acetyl-L-tyrosinate catalyzed by alpha-chymotrypsin and the acid-catalyzed hydrolysis of triethoxymethane. The asymmetric membrane consumes reactant, and discharges product, at different rates through its two faces. The concentration of products, and of non-rate determining reactants, may exhibit maxima or minima with time, and computer simulation indicates that oscillatory behavior could be expected under appropriate conditions. Due to the different exit path lengths, the osmotic response (determined by the solute concentrations at the two membrane surfaces) may be opposite in sign to that expected from the concentrations in the external compartments.


Use of prazosin at the dunedin hypertension clinic controlled and open studies and pharmacokinetic observations. Prazosin has been used as an antihypertensive drug in the treatment of 93 patients in Dunedin since 1971-1972, mainly those whose previous control was unsatisfactory. It has led to very significant improvement of control in many cases; 48 patients now receiving the drug have taken it for an average of 30 months. Forty-five patients have stopped taking the drug for various reasons, such as failure to respond (11-8%), side effects (20-4%), myocardial infarction (8-6%) or unrelated reasons (7-5%). This reflects to some extent the use ofa new drug in patients who are difficult to control, and its use initially without concomitant diuretic and beta-blocker therapy. We believe that the place of praxosin is primarily as a third drug for patients whose hypertension does not respond satisfactorirly to a diuretic and a beta-blocker. The first dose must be kept small, preferably 0-25 mg, and even with this dose a small proportion of patients receiving diuretic and beta-blocker therapy seem to become hypotensive; patients should be warned about this. The plasma half-life in normal volunteers was 3-8 hours but in some patients it seems to be considerably longer.


[On the action of antihypotensive agents in sympathicotonic orthostatic hypotension in geriartric patients: comparison between placebo and etilefrin (author's transl)]. The action of etilefrin (Effortil Depot Perlongets) was compared with placebo in a double-blind study in 24 female geriatric patients with sympathicotonic orthostatic hypotension. After determining the basic values, all patients were treated with the active substance in a titration phase lasting one week (dosage 2-3 Perlongets/day). The decrease in systolic blood pressure and the increase in pulse rate while standing were then statistically significant lower (p less than (p less than 0.001) when compared with the basic values. In 17 out of 21 cases of collapse which occured while standing before therapy was commenced no further collapse occurred (p less than 0.005). Following the titration phase treatment with etilefrin was continued in 12 patients in a double-blind study while the remaining 12 patients were changed over to placebo as a withdrawal study. The effect continued in those patients treated with the active ingredient and the average values and incidence of effects improved further. On the other hand, in the placebo group there was one again a marked decrease in systolic blood pressure and increase in pulse rate as observed in the basic values. The symptomatology, particulary episodes of collapse increased again. There was a statistically significant difference between the action of etilefrin and placebo for all criteria (both objective and subjective parameters). Side-effects were not observed.


Mutagenesis by simian virus 40. II. Changes in substrate affinities in mutant hypoxanthine-guanine phosphoribosyl transferase enzymes at different pH values. A number of 8-azaguanine-resistant clones selected from Chinese hamster cells infected with SV 40, and supposed to originate by virus infection was investigated to demonstrate and analyze genetic alterations occurring in the cells after infection. All resistant clones tested showed reduced but detectable activity levels of the enzyme hypoxanthine-guanine phosphoribosyltransferase. The extent of reduction in the activity was not identical for different substrates. In all the clones tested, spontaneous mutants included, the pH optimum for the enzymic reaction with guanine was shifted to lower values. The reduced enzymic activities of resistant clones correlated with their colony-forming ability in corresponding selective media. The results support the suggestion that SV 40 is able to induce gene mutations.


Bacterial stimulation and granulocyte inhibition of granulopoietic factor production. We attempted to determine the effect of live bacteria (Staphylococcus epidermidis) on granulocyte colony-stimulating-factor production by human peripheral blood mononuclear cells (monocytes and lymphocytes) in vitro. Addition of bacteria to mononuclear-cell cultures enhanced colony-stimulating-factor production by these cells, as assayed on both human and mouse bone marrow. Addition of peripheral blood granulocytes to parallel cultures eliminated this enhancement effect, presumably by bacterial removal or inactivation. These data suggest that micro-organisms may have a pivotal role in granulocyte production and maturation by serving as a stimulus to increase colony-stimulating-factor production and also as negative control through their removal by the newly formed granulocytes.


Reconstitution in severe combined immunodeficiency by transplantation of marrow from an unrelated donor. A patient with severe combined immunodeficiency received seven transplants of bone marrow from an HLA-B-compatible and HLA-D-compatible unrelated donor in an attempt to provide immunologic reconstitution. The first four transplants achieved restricted engraftment with evidence of rudimentary immunologic function. A fifth transplant, given after low-dose cyclophosphamide, produced reconstituion of cell-mediated immunity. Marrow aplasia developed after recontamination with a nonpathogenic microflora. Transplantation of marrow previously stored in liquid nitrogen was ineffective. A subsequent transplant, administered after high-dose cyclophosphamide, achieved durable engraftment, with complete hematopoietic and immunologic reconstitution. Seventeen months after transplantation, full functional engraftment persists. Graft-versus-host disease has been chronic and moderately severe, but limited to the skin and oral mucosa. Transplantation of marrow from unrelated histocompatible donors may provide a useful treatment for patients with severe combined immunodeficiency or aplastic anemia who lack a matched sibling or related donor.


[Information transmission by means of neurosecretory peptides as mediators]. Morphological indications and new experimental results demonstrate that neurosecretory peptides or low-molecular proteins, respectively, appear to effect as mediators in information transmission. They are acting as releasers triggering specific behavior programs in the nervous system or as modulators in the regulation of different functions. This can be observed in different groups of the animal kingdom including mammals and man as well as different levels of organization or systems. Some of such facts at behavior level, concerning regulation of organ function and finally at cellular level of distinct single neurons are discussed.


[Possibilities of treatment of maldescensus testis in the dog (author's transl)]. The diagnosis of inguinal retention of the testis can be made in the dog from the 10th week of life. Like in man, therapy of this condition can be done with gonadotropins or releasing hormones. In conditions with retention of the testicles in the upper inguinal region LH-RH proved to be more successful than gonadotropin. 10 out of 16 patients showed a complete descent of the testes. In addition the effect of plasmagonadotropin as well as urinary chorionic gonadotropin, and of LH-RH on the plasma testosterone levels of juvenile and adult dogs was studied. It could be seen that in the young dogs all 3 hormones caused an increase in plasma testosterone, while in the adult animal only LH-RH was able to produce a significant rise.


Comparative study of beta andreoceptor antagonists on the isolated  atria of control and of the reserpine treated rabbits. The effect of four adrenoceptor antagonists, chosen because of reported differences in their selectivities, were compared using the positive chronotropic response to isoprenaline on isolated rabbit atria. Oxprenolol was the most potent antagonist of the isoprenaline induced cardiac arrhythmias. Alprenolol, practolol and propranolol also blocked the positive chronotropic responses to isoprenaline effectively. All the antagonists lowered the resting heart rate. The effects of all four selected antagonists on heart rate probably result from cardiac beta adrenoceptor blockade. Atria isolated from reserpine treated rabbits, showed a reduced percentage of increase in the heart rate in response to addition of isoprenaline. This decreased response has been related in some cases to direct myocardial inhibition.


Arginine esterase and lysosomal hydrolases in liver from cystic fibrosis subjects. The total activity and isoelectric focusing patterns of arginine esterase, cathepsin B1, and several lysosomal hydrolases were normal in liver from two patients with cystic fibrosis. No abnormalities were observed in values for pH optimum, Km, and Vmax for arginine esterase and cathepsin B1 in liver from cystic fibrosis patients compared to those values for liver from the control subject. Soybean trypsin inhibitor at concentrations up to 100 microgram/ml had no effect on liver arginine esterase or cathepsin B1.


Problems in assessment of vividness and control of imagery. Difficulties in assessing individual differences in vividness and control of mental imagery hinder understanding of the role of of imagery in various phenomena. This study examined the relationships among the Gordon test of imagery control, the Betts questionnaire on imagery vividness, and a research questionnaire on control of imagery developed for this study. The research questionnaire demonstrated satisfactory internal consistency and scores were more widely distributed than were those on the Gordon test. Correlations among the instruments ranged from .47 to .57, suggesting that vividness and control of imagery are not clearly differentiated in practice. Discussion of these results suggests that further empirical work is needed to establish the differential validity of the two constructs.


Heat stable inhibitors produced in raw milk. Twenty raw farm milk samples were tested for inhibitory substances before and after being kept at room temperature until the pH fell to less than 6.3 but greater than 6.2. The tests were done using the IDF-method and the commercial Thermocult method. In both methods B. stearothermophilus var. chalidolactis is used as the test organism. In two instances a thermostable inhibitor was produced with an inhibitory potential greater than 0.01 IU/ml penicillin standard. The IDF-method was found unsuitable for the study of relatively small concentrations of antibiotics or other chemotherapeutics in milk.


[Septicemia in the elderly (author's transl)]. Out of 1,251 patients above 65 years of age staying at the Charles Foix Hospital (prolonged hospitalization) and the St. Joseph Hospital (acute cases), 168 had one or more positive blood cultures. Urinary tract infection is a major source of septicemia due to gram negative bacilli. It is important to stress cases of septicemia due to pneumococcal pneumoniae, eschars, and other skin lesions. Mortality varies between 33 and 36%, depending upon the hospital. Collapse, although infrequent, still portends a grave prognosis (61% of cases of collapse led to death at Charles Foix Hospital). The combination of more than two risk factors considerably worsens the prognosis. Hypoproteinemia and dementia are every bit as grave as diabetes and cancer. A better isolation of the microorganisms involved in cases of septicemia in the elderly will lead to a more judicious choice of antibiotics. The administration of chemotherapy immediately after the samples were obtained remains the main guarantee of successful therapy.